فهرست مطالب

علوم باغبانی - سال سی و سوم شماره 3 (پاییز 1398)
  • سال سی و سوم شماره 3 (پاییز 1398)
  • تاریخ انتشار: 1398/10/10
  • تعداد عناوین: 15
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  • خدایار همتی، بختیار اردوان پور، وحید اکبرپور* صفحات 349-361

    مرزه (Satureja hortensis L.) گیاهی است از خانواده نعناعیان که اسانس آن در صنایع دارویی و آرایشی و غذایی کاربرد زیادی دارد. هدف از این تحقیق مطالعه اثرات چند نوع بستر کشت آلی شامل: 1- خاک مزرعه (شاهد)، 2-کود دامی،3-کمپوست زباله شهری،4- خاکبرگ، 5- مخلوط بسترها به نسبت مساوی بر روی خصوصیات رویشی و زایشی و نیز کمیت و کیفیت اسانس در سه اکوتیپ مرزه تابستانه (کردستان، شیراز و یزد) بود. آزمایش به صورت فاکتوریل بر پایه طرح بلوک‌های کامل تصادفی در چهار تکرار در مزرعه تحقیقاتی شماره یک دانشگاه علوم کشاورزی و منابع طبیعی گرگان انجام گرفت. اکوتیپ شیراز دارای بیشترین درصد اسانس (2/1 درصد) در بین اکوتیپ‌های مختلف بود، ولی بین تیمارهای مختلف و بین اکوتیپ ها تفاوت معنی داری نشان نداد. اثر تیمارها بر روی اجزای اسانس شامل کارواکرول، گاماترپینن و پاراسیمن معنی‌دار بود. اسانس گیاهان در بستر مخلوط بیشترین میزان کارواکرول (69/74 درصد) را داشت. اکوتیپ یزد در بین اکوتیپ‌ها بیشترین میزان کارواکرول را نشان داد. در میان بسترهای کشت، تیمار شاهد بیشترین درصد گاماترپینن (48/5درصد) و بیشترین درصد پاراسیمن (76/1) را داشت و همه بسترهای آلی باعث کاهش معنی‌دار این ترکیب ها در اسانس شدند. همچنین در مقایسه اکوتیپ‌های مختلف، اکوتیپ شیراز دارای بیشترین میزان گاماترپینن و اکوتیپ کردستان بیشترین درصد پاراسیمن (69/1) بودند. با توجه به نتایج حاصله و اینکه اختلافی بین مصرف انواع کودهای آلی از لحاظ اجزای عملکرد و درصد اسانس مرزه مشاهده نشد، بنابراین در صورتی که هر یک از انواع کودهای آلی مورد استفاده در این آزمایش در دسترس بوده و از لحاظ اقتصادی مقرون به صرفه‌تر باشد، قابل توصیه برای کشاورزان و تولیدکنندگان گیاه دارویی مرزه می‌باشد.

    کلیدواژگان: اکوتیپ، کیفیت اسانس، کود دامی، کارواکرول
  • محمد حسین شیخ محمدی، نعمت الله اعتمادی*، علی نیکبخت، مصطفی عرب، محمد مهدی مجیدی صفحات 363-375

    خشکی و شوری از جمله تنش‌های غیرزنده مضر برای رشد چمن در طیف گسترده‌ای از جهان می‌باشند. هدف از اجرای این آزمایش، انتخاب توده‌های مقاوم به خشکی و شوری چمن جهت استفاده در برنامه مدیریت چمن در شرایط تنش خشکی و شوری بود. آزمایش به‌صورت کرت‌های خردشده بر پایه طرح کاملا تصادفی با سه تکرار انجام شد. عوامل آزمایش شامل سه تیمار آبیاری بود که به مدت 45 روز اعمال گردید: 1) گیاهان شاهد (بدون تنش)، 2) تنش خشکی (قطع آبیاری به مدت 45 روز) 3) تنش شوری (روزانه 100 میلی‌لیتر با شوری 9 دسی‌زیمنس بر متر). تیمارهای آبیاری به‌عنوان فاکتور اصلی در سه سطح (شاهد، تنش خشکی و تنش شوری) و توده‌های علف گندمی در شش سطح به‌عنوان کرت فرعی در نظر گرفته شدند. نتایج حاصل از این تحقیق نشان داد تنش خشکی و شوری باعث کاهش کیفیت در توده‌های علف گندمی تاجدار شدند و سطح کاهش کیفیت بین توده‌ها متفاوت بود. نتایج ما نشان داد، توده سبزوار تحت تنش خشکی و شوری به ترتیب، بالاترین ارتفاع (52/5 و 86/5 سانتی‌متر) و بیشترین محتوی کلروفیل (85/2 و 75/2 میلی‌گرم برگرم وزن‌تر) را نشان می‌دهد. نتایج به‌دست‌آمده از بررسی کربوهیدرات‌های محلول اندام هوایی، محتوی آب نسبی و میزان نشت الکترولیت نشان داد که در گیاهان تحت تنش خشکی توده‌های دماوند و سبزوار بیشترین مقدار کربوهیدرات‌های محلول (16/332 و 14/345 میلی‌گرم برگرم وزن خشک)، بیشترین محتوی آب نسبی (18/59 و 18/62 درصد) و کمترین میزان نشت الکترولیت (16/48 و 14/59 درصد) را به خود اختصاص دادند. در شرایط تنش شوری، توده دماوند (15/322 میلی‌گرم برگرم وزن خشک) بیشترین کربوهیدرات محلول، توده سبزوار (42/68 درصد) و اراک (14/65 درصد) بیشترین محتوی آب نسبی و توده سبزوار (17/54 درصد) کمترین میزان نشت الکترولیت را نشان دادند. مطابق نتایج این آزمایش رتبه‌بندی مقاومت به تنش خشکی به صورت سبزوار <دماوند<  اراک < ارومیه = تاکستان < هشتگرد و رتبه‌بندی مقاومت به شوری به‌صورت سبزوار <اراک< دماوند < هشتگرد = تاکستان< ارومیه می‌باشد. توده‌های سبزوار و دماوند مقاومت خوبی نسبت به خشکی، و توده‌های سبزوار و اراک مقاومت خوبی نسبت به تنش شوری در مقایسه با سایر توده‌های علف گندمی تاجدار از خود نشان دادند.

    کلیدواژگان: تنش، چمن، خشکی، شوری، علف گندمی
  • مهدیه خرازی، علی تهرانی فر*، احمد شریفی صفحات 377-386

    موفقیت در تکنیک کشت بافت، به خصوص در رابطه با گیاهان سوخ‌دار، به میزان کنترل آلودگی میکروبی در طی دوره کشت وابسته می‌باشد. اعمال تیمارهای مختلف نظیر تیمار گرمایی و کاربرد قارچ‌کش‌های مختلف می‌تواند به کنترل آلودگی میکروبی و به دنبال آن، افزایش درصد زنده‌مانی ریزنمونه‌ها کمک نماید. لذا پژوهش حاضر با هدف بررسی اثر تیمار گرمایی و کاربرد قارچ‌کش بنومیل بر کاهش آلودگی کشت درون شیشه‌ای گیاه نرگس، به صورت آزمایش فاکتوریل در قالب طرح کاملا ‌تصادفی با 2 عامل شامل غلظت قارچ‌کش بنومیل در محیط کشت (یک و دو در هزار)، اعمال و یا عدم اعمال تیمار گرمایی (2 سطح) با 10 تکرار انجام شد. به منظور اعمال تیمار گرمایی از آب گرم 54 درجه سانتی‌گراد به مدت یک ساعت استفاده گردید. از محیط کشت پایه MS تکمیل شده با تنظیم‌کننده‌های رشد BA (1 میلی‌گرم در لیتر) و NAA (2/0 میلی‌گرم در لیتر) برای کشت ریزنمونه‌های فلس جفتی استفاده شد. نتایج پژوهش حاضر نشان داد با وجود آنکه کمترین درصد آلودگی در شرایط اعمال تیمار گرمایی و کاربرد دو گرم در لیتر بنومیل حاصل شد، ولی با این حال بیشترین درصد قهوه‌ای شدن ریزنمونه‌ها نیز در این تیمار مشاهده شد که منجر به کاهش پتانسیل باززایی ریزنمونه‌ها شد. لذا با در نظر گرفتن درصد قهوه‌ای شدن، درصد باززایی و درصد آلودگی ریزنمونه‌ها، عدم اعمال تیمار گرمایی و کاربرد محیط کشت حاوی یک گرم در لیتر قارچ‌کش بنومیل جهت کشت ریزنمونه های فلس جفتی گیاه نرگس توصیه می‌گردد.

    کلیدواژگان: بنومیل، درصد زنده مانی، درصد قهوه ای شدن، کشت بافت
  • محمد بهزاد امیری*، پرویز رضوانی مقدم، محسن جهان صفحات 387-403

    به منظور بررسی اثر کودهای آلی و شیمیایی بر خصوصیات کمی گاوزبان ایرانی (Echium amoenum) در تراکم های مختلف گیاهی، آزمایشی در سال های زراعی 92-1390 در دانشگاه فردوسی مشهد به صورت آزمایش فاکتوریل در قالب طرح پایه ی بلوک‌های کامل تصادفی با سه تکرار انجام شد. فاکتورهای آزمایشی شامل 3 تراکم گیاهی (3، 5 و 10 بوته در متر مربع) و 5 نوع کود مختلف (کمپوست زباله شهری، ورمی‌کمپوست، گاوی، شیمیایی (اوره) و شاهد) بودند. نتایج آزمایش نشان داد که با کاهش تراکم گیاهی، تاثیر کودهای آلی در افزایش عملکرد گل تشدید شد و در کمترین تراکم گیاهی، کودهای کمپوست زباله شهری، ورمی‌کمپوست و گاوی، عملکرد گل را به ترتیب 112، 79 و 223 درصد نسبت به شاهد بطور معنی‎داری افزایش دادند. در تمامی تراکم های مورد مطالعه، طول ساقه فرعی در شرایط استفاده از کودهای آلی نسبت به شاهد بطور معنی‎داری بیشتر بود. کودهای کمپوست، ورمی کمپوست، گاوی و شیمیایی (اوره) در تراکم 10 بوته در متر مربع به ترتیب افزایش 106، 54، 66 و 173 درصدی میزان فنول کل را در مقایسه با شاهد سبب شدند. در تراکم متوسط گیاهی (5 بوته در متر مربع) استفاده از کودهای کمپوست، ورمی‌کمپوست و گاوی به ترتیب منجر به افزایش 83، 74 و 57 درصدی میزان آنتوسیانین کل نسبت به شاهد شدند. به طور کلی نتایج این پژوهش نشان داد که استفاده از کودهای آلی مختلف و کود شیمیایی اوره در تراکم های مطلوب گیاهی می‌تواند منجر به بهبود خصوصیات مورفولوژیکی و عملکرد کمی و کیفی گاوزبان ایرانی شود.

    کلیدواژگان: آنتوسیانین کل، فنول کل، کمپوست، گیاه دارویی، نهاده بوم سازگار
  • مهناز صیادی نژاد، سید مصطفی صادقی* صفحات 405-415

    امروزه پرورش و تکثیر گیاهان زینتی به خصوص گیاهانی که تکثیر آنها به سختی صورت می گیرد، از طریق کشت بافت از ارزش و اهمیت زیادی برخوردار است. تکثیر گیاه زینتی زامیفولیا با ارزش اقتصادی بالا از طریق قلمه برگ و تقسیم ریزوم به سختی صورت می‌گیرد و به مدت زمانی حدود 7 ماه نیاز دارد لذا جهت تسریع در تکثیر آن کشت بافت مفید واقع می‌گردد. جهت بهینه سازی تولید کالوس و باززایی، ریزنمونه‌های دمبرگ، ریزوم، برگ و شاخه، در محیط کشت های با سطوح متفاوت تنظیم کننده‌های رشد در دو آزمایش جداگانه به صورت فاکتوریل در قالب طرح کاملا تصادفی در سه تکرار مورد بررسی قرار گرفتند. در آزمایش اول ریزنمونه‌های دمبرگ و ریزوم در سه مرحله کالوس، شاخه و ریشه‌زایی به ترتیب در سطوح هورمونی] BA (4، 2، 0 میلی‌گرم بر لیتر)، 2,4-D (2، 1، 0 میلی‌گرم بر لیتر)[،] BA (2، 1، 0 میلی‌گرم بر لیتر) و 2,4-D (1، 5/0، 0 میلی‌گرم بر لیتر)[و] BA (2، 1، 0 میلی‌گرم بر لیتر) و NAA (1، 5/0، 0 میلی‌گرم بر لیتر)[قرار گرفتند و در آزمایش دوم ریزنمونه‌های برگ و شاخه نیز در سه مرحله کالوس، شاخه و ریشه‌زایی به ترتیب در سطوح هورمونی] BA (4، 2، 0 میلی‌گرم بر لیتر) و NAA (1، 5/0، 0 میلی‌گرم بر لیتر)[، BA] (2، 1، 0 میلی‌گرم بر لیتر) و NAA (1، 5/0، 0 میلی‌گرم بر لیتر)[و] BA (2، 1، 0 میلی‌گرم بر لیتر) و NAA (2، 1، 0 میلی‌گرم بر لیتر)[قرار گرفتند. در مرحله کالوس‌زایی، صفات مدت زمان تا شروع کالوس‌زایی و پس از گذشت 5 هفته درصد کالوس‌زایی و وزن ترکالوس اندازه گیری گردید. در مرحله باززایی مدت زمان تا شروع ریشه‌زایی، مدت زمان تا شروع شاخه‌زایی و پس از گذشت 5 هفته طول شاخه و تعداد برگ اندازه‌گیری گردید. بالاترین درصد کالوس‌زایی، کوتاهترین زمان برای رسیدن به کالوس و بیشترین وزن تر کالوس در محیط کشت با هورمون های 2 میلی‌گرم بر لیتر BA و 1 میلی‌گرم بر لیتر NAA در ریز نمونه برگ به ترتیب با 5/94 درصددرصد، 14 روز و 1/1 گرم از آزمایش دوم مشاهده گردید. بهترین تیمار در مرحله شاخه‌زایی با کوتاهترین زمان تا شروع شاخه‌زایی، بلندترین طول شاخه و بیشترین تعداد برگ به ترتیب با 5/10 روز ، 10/4 سانتی متر و 8 برگ مربوط به ریزنمونه برگ با غلظت هورمونی 2 میلی‌گرم بر لیتر BA و 5/0 میلی‌گرم بر لیتر NAA از آزمایش دوم بود و در مرحله ریشه‌زایی بهترین تیمار برای ریزنمونه دمبرگ با غلظت هورمونی 1 میلی‌گرم بر لیتر BA و 5/0 میلی‌گرم بر لیتر NAA با 14 روز زمان تا ریشه‌زایی از آزمایش اول بود. از نتایج آزمایش‌ها در مرحله باززایی چنین بر آمد که نوع ریزنمونه در شاخه‌زایی و ریشه‌زایی همانند مرحله کالوس‌زایی موثر بوده‌است به طوری که در مرحله شاخه‌زایی نتایج به دست آمده از ریزنمونه ریزوم نسبت به ریزنمونه دمبرگ بهتر بود اما در مرحله ریشه‌زایی ریزنمونه دمبرگ بهتر از ریزنمونه ریزوم عمل کرده ‌است.

    کلیدواژگان: کشت بافت، ریزازدیادی، واکشت، BA
  • محترم عباسپور اسفدن، سپیده کلاته جاری*، فواد فاتحی صفحات 417-432

    پریوش (Catharanthus roseus) گیاهی زیبا و رونده از خانواده خرزهره ئیان و دارای کاربردهای زینتی و دارویی است. این گیاه همانند سایر گونه‌های فضای سبز تحت تاثیر آثار سوء ناشی از تنش خشکی قرار می گیرد، اما در این راستا استفاده از محرک های رشد گیاهی می تواند برای گیاه تحت تنش مفید باشد. بنابراین تحقیق حاضر به منظور تاثیر اسید سالیسیلیک و ال-آرژنین بر صفات مورفو-فیزیولوژیکی گیاه پریوش تحت تنش خشکی انجام شد. این آزمایش به صورت فاکتوریل با دو عامل شامل محلول پاشی در 5 سطح (شاهد، ال-آرژنین 5/1 و 3 میلی مولار، اسید سالیسیلیک 100 و 200 میلی گرم در لیتر) و تنش خشکی در سه سطح (100، 70 و 40 درصد ظرفیت زراعی) در قالب طرح پایه کاملا تصادفی با 3 تکرار انجام شد. صفات مورد بررسی شامل کارایی مصرف آب، نسبت وزن ریشه به شاخساره، تعداد گل، قطر گل، محتوای کلروفیل برگ، فعالیت آنزیم های پراکسیداز و سوپر اکسید دیسموتاز برگ و عناصر پرمصرف (نیتروژن، فسفر و پتاسیم) برگ بود. کارایی مصرف آب در تیمارهای تحت تاثیر تنش 70 درصد ظرفیت زراعی بیشتر از سایر تیمارها بدست آمد. نتایج تجزیه واریانس نشان داد که برهمکنش محلول پاشی و تنش خشکی بر نسبت وزن ریشه به شاخساره، فعالیت آنزیم پراکسیداز، مقدار فسفر و نیتروژن برگ معنی دار شد. نسبت وزن ریشه به شاخساره در تیمارهای عدم محلول پاشی (شاهد) × 40 درصد ظرفیت زراعی و ال-آرژنین 5/1 میلی مولار × 40 درصد ظرفیت زراعی بیشتر از سایر تیمارها گزارش شد. فعالیت آنزیم پراکسیداز در تیمار برهمکنش عدم محلول پاشی و 100 درصد ظرفیت زراعی کمتر از سایر تیمارهای آزمایشی بود. بیشترین مقدار فسفر و نیتروژن در تمامی سطوح محلول پاشی در 100 درصد ظرفیت زراعی مشاهده شد. حداقل مقدار عناصر پرمصرف در تیمارهای حاوی 40 درصد ظرفیت زراعی مشاهده شد. قطر و تعداد گل در تیمارهای 100 درصد و 70 درصد ظرفیت زراعی بیشتر از 40 درصد ظرفیت زراعی به‌دست آمد و در بین تیمارهای محلول پاشی میانگین قطر گل و کلروفیل کل در تیمار ال-آرژنین 3 میلی مولار بیشتر بود. بیشترین و کمترین فعالیت آنریم سوپر اکسید دیسموتاز به ترتیب در تیمار 40 درصد ظرفیت زراعی و 100 درصد ظرفیت زراعی مشاهده شد و در بین تیمارهای محلول پاشی، فعالیت این آنزیم در تیمارهای ال-آرژنین 3 میلی مولار و اسید سالیسیلیک 100 میلی گرم در لیتر بیشتر از سایر تیمارها بود. بنابراین با هدف مصرف بهینه آب و محلول پاشی، در تیمار 70 درصد ظرفیت زراعی و استفاده از ال-آرژنین 3 میلی مولار یا اسید سالیسیلیک 100 میلی گرم در لیتر موجب افزایش عناصر فسفر و ازت گردید. همچنین در مجموع تیمارهای ال-آرژنین 3 میلی مولار یا اسید سالیسیلیک 100 میلی گرم در لیتر موجب افزایش تعداد و قطر گل پریوش شدند.

    کلیدواژگان: عناصر پرمصرف، کارایی مصرف آب، کلروفیل کل، محرک های رشد
  • محمد نورانی یزن آباد، محمدتقی عبادی، مهدی عیاری نوش آبادی* صفحات 433-449

    گیاه دارویی پای خر با نام علمی Tussilago farfara L.، چندساله و از تیره کاسنی (Asteraceae) می‏باشد. پای خر مصارف دارویی دارد و از زمان‏های بسیار دور برای درمان سرفه، مشکلات ریوی و خلط استفاده می‏شود. در این تحقیق برای کشت پای خر، از ریزوم‏ این گیاه استفاده شد و آزمایش به صورت فاکتوریل در قالب طرح بلوک های کامل تصادفی در سه تکرار انجام گرفت که تیمارها عبارت از هفت جمعیت از شهرهای فیروزکوه، نور، دیلمان، کلیبر، نمین، دماوند و پل زنگوله گیاه پای‏خر کشت شده در تهران بودند. صفات مورفولوژیکی و فیزیولوژیکی مانند سطح برگ، تعداد گل در بوته، قطر گل، طول ساقه گل‏دهنده، ارتفاع گل، وزن خشک گل، طول ریشه و میزان فتوسنتز اندازه‏گیری شد. برگ و گل گیاهان کشت شده برای بررسی فیتوشیمیایی مانند فنول و فلاونویید کل آماده گردید. نتایج مربوط به گیاهان کشت شده نشان داد که اختلاف معنی‏داری در سطح احتمال یک درصد بین جمعیت‏ها وجود داشته و جمعیت پل زنگوله از لحاظ برخی خصوصیات مورفولوژیکی مانند تعداد گل در بوته با میانگین 15، بیشترین قطر و ارتفاع گل به ترتیب با میانگین 12 و 16 میلی‏متر، وزن خشک گل با میانگین 2/5 گرم و همچنین بالاترین میزان فتوسنتز، به عنوان جمعیت برتر در بین گیاهان کشت شده مشخص گردید. نتایج تجزیه واریانس نشان داد که بین فنول و فلاونویید کل جمعیت‏های مورد بررسی اختلاف معنی‏داری در سطح احتمال یک درصد وجود داشته و جمعیت نور با میانگین 242 میلی‏گرم گالیک اسید بر گرم عصاره خشک بیشترین مقدار فنول کل را داشت و عصاره برگ جمعیت نور با IC50  برابر با 271، بیشترین فعالیت آنتی‏اکسیدانی را نشان داد.

    کلیدواژگان: بررسی فیتوشیمیایی، فعالیت آنتی اکسیدانی، فنول کل، Tussilago farfara L
  • علی ناصری مقدم، حسن بیات*، محمد حسین امینی فرد، فرید مرادی نژاد صفحات 451-466

    در بین تنش‌های محیطی، خشکی و شوری جزء مهم‌ترین تنش‌ها هستند که رشد و نمو گیاهان را در مناطق خشک و نیمه خشک محدود می‌کنند. هدف از تحقیق حاضر بررسی اثر تنش‌های خشکی و شوری بر صفت‌های رویشی، زایشی و بیوشیمیایی گل نرگس شهلا (Narcissus tazetta L.) بود. این آزمایش گلدانی، به صورت فاکتوریل در قالب طرح کاملا تصادفی با سه تکرار در دانشکده کشاورزی دانشگاه بیرجند در سال 1396 انجام شد. تیمارها شامل کلرید‌سدیم در چهار سطح صفر (شاهد)، 20، 40 و 60 میلی‌مولار و تنش خشکی در چهار سطح 70،50،30 و 90 درصد ظرفیت زراعی بودند. نتایج اثرهای ساده نشان داد که تنش‌های خشکی و شوری باعث کاهش مقدار قطر ساقه گل‌دهنده، قطر گل، طول ریشه، حجم ریشه، وزن تر ریشه، طول و وزن تر سوخ، وزن تر اندام هوایی، وزن خشک کل و تعداد روز از گلدهی تا پیری گل شدند. در مقابل صفت‌های تعداد روز از کاشت تا گلدهی، میزان قندهای محلول کل برگ و ریشه، فعالیت آنتی اکسیدانی و محتوای فنول کل تحت تاثیر این دو تنش افزایش یافتند. نتایج اثرهای متقابل نشان داد که اثر مخرب تنش‌های شوری و خشکی در شرایط کاربرد توام دو تنش، تشدید شد به طوری که کمترین مقدار صفت‌های رشدی و زایشی در شدیدترین سطح تنش (30 درصد ظرفیت زراعی × شوری 60 میلی‌مولار) مشاهده شد. به طور کلی، نتایج نشان داد که در محدوده تیمارهای اعمال شده، اثر مخرب تنش شوری بر صفت‌های رشدی، زینتی و فیزیولوژیکی گل نرگس بیشتر از تنش خشکی بود.

    کلیدواژگان: حجم ریشه، فعالیت آنتی اکسیدانی، قطر گل، قندهای محلول کل، محتوای فنول کل
  • مائده عقدایی، سید حسین نعمتی*، لیلا سمیعی، احمد شریفی شریف آباد صفحات 467-479

    پپینو یک سبزی‌ میوه‌ای متعلق به خانواده سولاناسه بوده که مشکل اساسی در توسعه‌ی کشت این گیاه تکثیر آن می‌باشد که از طریق بذر موفقیت اندکی دارد. از این‌رو این پژوهش با هدف تکثیر انبوه از طریق کشت بافت انجام گردید. به‌منظور دستیابی به بهترین نوع محیط کشت و ترکیب تنظیم کننده‌های رشد گیاهی از ریزنمونه تک گره پپینو استفاده گردید. این تحقیق در قالب سه آزمایش جداگانه با استفاده از چهار نوع محیط کشت (MS، ½ MS، SH و B5)، دو نوع سیتوکنین (بنزیل آدنین (BA) و کینتین (Kin)) و دو نوع اکسین (ایندول بوتریک اسید (IBA) و نفتالین استیک اسید (NAA)) برای تعیین بهترین محیط کشت و دستیابی به ترکیب مناسبی از تنظیم کننده‌های رشد گیاهی برای پرآوری شاخساره و ریشه‌زایی انجام شد. نتایج نشان داد که بهترین محیط کشت برای ریزازدیادی ریزنمونه تک گره پپینو با توجه به تعداد شاخساره، طول شاخساره، تعداد ریشه، طول ریشه، تعداد برگ و طول برگ محیط کشت پایه MS بود. در آزمایش پرآوری شاخه، بیش‌ترین تعداد شاخساره، تعداد برگ، رنگ شاخساره و کیفیت شاخساره القا شده در اثر کاربرد محیط کشت MS غنی شده با دو میلی‌گرم در لیتر BA به‌همراه یک میلی‌گرم در لیتر Kin به‌دست آمد. همچنین بیش‌ترین میزان طول شاخساره و طول برگ به‌ترتیب با کاربرد محیط کشت MS غنی شده با یک میلی‌گرم در لیتر BA همراه با دو میلی‌گرم در لیتر Kin و تیمار یک میلی‌گرم در لیتر BA همراه با یک میلی‌گرم در لیتر Kin حاصل شد. در آزمایش ریشه‌زایی، بیش‌ترین میانگین تعداد ریشه و کیفیت ریشه با استفاده از IBA با غلظت 6/0 میلی‌گرم در لیتر به‌دست آمد، در حالی که بیش‌ترین میانگین طول ریشه با کاربرد IBA با غلظت 3/0 میلی‌گرم در لیتر مشاهده شد. در مجموع بهترین نتایج با محیط کشت MS، غلظت دو میلی‌گرم در لیتر BA همراه با یک میلی‌گرم در لیتر Kin جهت پرآوری و همچنین غلظت 6/0 میلی‌گرم در لیتر IBA جهت ریشه‌زایی ریزنمونه پپینو به‌دست آمد.

    کلیدواژگان: ایندول بوتریک اسید، بنزیل آدنین، پپینو، پرآوری، ریشه زایی، کشت بافت
  • محمد حسین عالم خومرام، امیر حسین کشتکار، اصغر میرزایی اصل صفحات 481-497

    سیر ایران از کیفیت عطر و طعم خوبی برخوردار است، ولی تا به امروز به جز رقم مازند که مناسب مازندران است، رقم بومی دیگری از سیر اصلاح و معرفی نشده است. به منظور شناسایی پربازده ترین توده های محلی برای اصلاح و معرفی رقم مناسب شرایط آب و هوایی همدان، در سال زراعی 95-1394 آزمایشی بر روی 18 توده محلی مختلف در قالب طرح بلوک های کامل تصادفی با چهار تکرار در مرکز تحقیقات کشاورزی همدان اجرا شد. نه توده از مناطق کشت سیر استان همدان و نه توده از سایر استان های سیرکار کشور انتخاب گردیدند. توده های محلی از نظر 33 صفت مهم زراعی- مورفولوژیکی از جمله عملکرد سوخ؛ و میزان اسیدپیروویک کل (مجموع اسید پیروویک حاصل از فعالیت های آنزیمی و غیرآنزیمی)، میزان اسید پیروویک غیرآنزیمی، میزان اسید پیروویک آنزیمی و میزان آلیسین مورد بررسی قرار گرفتند. نتایج این بررسی نشان داد که صفات مورد بررسی توده های محلی کشور از تنوع بالایی برخوردارند که لازمه برنامه های اصلاحی است و همچنین معرفی توده ها به مناطق دیگر می تواند با نتایج مطلوب همراه باشد. چنانکه از نه توده محلی غیر بومی همدان سه توده محلی ارومیه، اراک و بردسیر کرمان در شرایط آب و هوایی همدان تولید قابل قبول داشتند. توده های محلی ارومیه، سولان و حیدره بر اساس عملکرد سوخ، اسید پیروویک آنزیمی و رابطه مولی دو به یک آن با آلیسین بیشترین ماده عطر و طعم دهنده آلیسین را در هکتار تولید کردند. صفات وزن سوخ به طور مستقیم و قطر یقه به طور غیرمستقیم از طریق وزن سوخ، با عملکرد سوخ و صفات شاخص سطح برگ، شاخص برداشت، وزن خشک برگ بوته و قطر سوخ با میزان آلیسین تولیدی ارتباط معنی دار بالایی نشان دادند. بر اساس تجزیه خوشه ای توده ها به دو گروه اصلی مناسب و نامناسب کشت در همدان تقسیم شده که گروه مناسب کشت در همدان خود به چهار زیرگروه سه تایی تقسیم شد. خوشه بندی امکان کاهش هزینه های حفظ منابع ژنتیکی سیر را با کم کردن میزان مواد نگهداری شده با حداقل افت تنوع صفات، فراهم می سازد.

    کلیدواژگان: اسید پیروویک، تجزیه خوشه ای، تنوع ژنتیکی، رگرسیون گام به گام، سیر
  • نوشین کاظمی*، علی اکبر حبشی، وهب اسدی صفحات 499-599

    این پژوهش با هدف تولید نهال عاری از سه ویروس ACLSV، ASPV، ASGV سیب "گوشت سرخ" (Malus pumila Mill.)، در پژوهشگاه بیوتکنولوژی کشاورزی ایران (ABRII) در سال 1394 انجام شد. آزمایش ها با ارزیابی اثربخشی تیمار‌ گرمادرمانی (0، 7، 14 و 21 روز در دمای 38 درجه سانتی‌گراد) و کشت مریستم انتهایی (در سه اندازه کوچک‌تر از 2/0 میلی‌متر، بین 2/0 تا 7/0 میلی‌متر و بزرگ‌تر از 7/0 میلی‌متر) بر نرخ حذف ویروس از ریزشاخه ها انجام شد. در ابتدا حضور ویروس هایACLSV، ASGV و ASPV در نمونه‌های مادری با روش های آزمون الایزای ساندویچ دو طرفه آنتی بادی و RT-PCR مورد ارزیابی قرار گرفت. سپس تیمارهای گرمادرمانی و کشت مریستم در شرایط درون شیشه‌ای انجام شد. ریزشاخه های رشد یافته از مریستم توسط الایزا و RT-PCR برای هر سه ویروس بررسی شدند. به طورکلی نرخ حذف سه ویروس ACLSV (9/25 درصد)، ASGV (4/7 درصد) و ASPV (4/44 درصد) با یکدیگر متفاوت بود. نتایج نشان داد افزایش طول مدت گرمادرمانی و کاهش اندازه مریستم با اثر گذاری متفاوت بر ویروس های مورد مطالعه باعث افزایش نرخ عاری شدن ریزنمونه ها از ویروس شده است. افزایش دوره زمانی گرمادرمانی در 21 روز باعث کاهش رشد و تکثیر و حتی از بین رفتن ریزنمونه ها شد. بنابراین 14 روز گرمادرمانی موثرترین تیمار جهت حذف آلودگی ویروس هایASGV  (11/11درصد)، ASPV (89/88 درصد) و ACLSV (44/44 درصد) از ریزنمونه های مورد مطالعه بود. در پایان آزمایش نمونه‌هایی که توسط هر دو روش الایزا و RT-PCR سالم تشخیص داده شدند، تکثیر و ریشه‌دار شدند و در شرایط گلخانه ای سازگار شدند.

    کلیدواژگان: الایزا، Apple chlorotic leaf spot virus، Apple stem grooving virus، Apple stem pitting virus، RT-PCR
  • محبوبه زمانی پور* صفحات 511-524

    این مطالعه با هدف بهبود ویژگی‌های رویشی و زایشی بر روی گیاهچه‌های گل جعفری آفریقایی در قالب دو آزمایش جداگانه در گلخانه تحقیقاتی دانشگاه ولایت ایرانشهر در سال 1396 اجرا شد. تیمارهای آزمایش اول شامل غنچه‌گیری در دو سطح (عدم غنچه‌گیری و غنچه‌گیری در 20 روز بعد از کاشت) و نیتروژن در چهار سطح (0 درصد، 2 درصد، 4 درصد و 6 درصد) و تیمارهای آزمایش دوم شامل سه سطح ایندول استیک اسید (50، 100 و 150 میلی گرم بر لیتر) و سه سطح اسید جیبرلیک (100، 200 و 300 میلی گرم بر لیتر) و آب مقطر به عنوان شاهد بودند. کیفیت گیاهچه‌ها بر اساس ارتفاع و قطر گیاهچه، تعداد برگ، تعداد و طول شاخه فرعی، تعداد و قطر گل، وزن تر و خشک گل و تعداد روز تا گلدهی اندازه‌گیری شدند. نتایج نشان دادند که بیشترین ارتفاع گیاه (5/47 سانتی متر)، طول شاخه (50/21 سانتی متر)، قطر گل (25/5 سانتی متر)، وزن خشک گل (60/3 گرم) و وزن تر گل (8/0 گرم) در تیمار عدم غنچه‌گیری به همراه 6 درصد نیتروژن بدست آمد. بیشترین تعداد برگ (91)، تعداد شاخه (10) و تعداد گل (7) با غنچه‌گیری همراه با تیمار 6 درصد نیتروژن بدست آمد. کیفیت گیاهچه‌ها با محلول‌پاشی اسید جیبرلیک نسبت به ایندول استیک اسید افزایش چشمگیری داشت و با افزایش غلظت، کیفیت گیاهچه‌ها نیز افزایش یافت، به‌طوری‌که بیشترین ارتفاع گیاه (25/34 سانتی متر)، طول شاخه (23 سانتی متر)، تعداد برگ (42)، تعداد شاخه (14)، قطر گل (3 سانتی متر)، وزن خشک گل (65/0 گرم)، وزن تر گل (13/3 گرم)، تعداد گل (6) و کمترین تعداد روز تا گلدهی (31 روز) در محلول پاشی با اسید جیبرلیک در غلظت 300 میلی گرم بر لیتر ایجاد گردید.

    کلیدواژگان: اسید جیبرلیک، ایندول استیک اسید، اوره، کیفیت گیاهچه
  • سید مهران علوی، اسد معصومی اصل*، ناصر زارع، رسول اصغری زکریا، پریسا شیخ زاده مصدق صفحات 525-536

    گیاه چویل دارای خواص داروئی مختلف بوده و تاکنون گزارشی از کشت تعلیقی سلولی این گیاه بومی ایران ارائه نشده است. به ‌منظور ارزیابی تاثیر عوامل مختلف بر پینه‌زایی چویل، آزمایشی به صورت فاکتوریل بر پایه طرح کاملا تصادفی اجرا شد. فاکتورها شامل اکوتیپ (آب نهر، گایونه، وزگ و سی‌سخت)، ریز نمونه (ریشه، برگ و ساقه) به همراه ترکیبات هورمونی مختلف NAA و BAP (هر کدام در غلظت‌های 0، 5/0، 1 و 2 میلی‌گرم در لیتر) در محیط کشت پایه موراشیگ و اسکوگ بودند. نتایج نشان دادند که بهترین اکوتیپ در واکنش به پینه‌زایی، اکوتیپ آب‌نهر و بهترین ریز‌نمونه، ریزنمونه برگی و بهترین ترکیب هورمونی شامل 1 میلی‌گرم در لیتر NAA و 5/0 میلی‌گرم در لیتر BAP بود. به‌ منظور بررسی تاثیر عوامل مختلف بر کشت تعلیقی سلولی چویل، علاوه بر هورمون NAA، از سطوح مختلف (غلظت‌های 0، 5/0، 1 و 2 میلی‌گرم در لیتر) هورمون 2,4-D نیز در ترکیب با هورمون BAP استفاده شد. بهترین ترکیب هورمونی برای کشت تعلیق سلولی، 2 میلی‌گرم بر لیتر NAA یا 2,4-D به همراه 5/0 میلی‌گرم بر لیتر BAP بود و بهترین ریزنمونه و اکوتیپ به ترتیب، ریزنمونه‌ برگی و اکوتیپ آب‌نهر بود. در مجموع، اکوتیپ آب نهر و ریزنمونه برگی هم در کشت پینه و هم در کشت تعلیقی سلولی نسبت به بقیه اکوتیپ ها و ریزنمونه ها برتری نشان دادند و بهترین ترکیب هورمونی در هر دو نوع کشت 2-1 میلی‌گرم در لیتر NAA و 5/0 میلی‌گرم در لیتر BAP بود.

    کلیدواژگان: پینه، تنظیم کننده رشد، ژنوتیپ، SCV، PCV
  • پریا دهخدایی، سعید ریزی، مسعود قاسمی قهساره* صفحات 537-548

    یکی از روش های بهبود ویژگی های مورفولوژیکی گیاهان، تغییر کیفیت نور است. به همین منظور آزمایشی جهت بررسی اثر کیفیت نور ال ای دی بر ویژگی های مورفولوژیک و فیزیولوژیک دانهال های حسن یوسف و اطلسی انجام شد. تیمارها شامل نور طبیعی گلخانه (شاهد) و نور ال ای دی (نور ترکیبی 50 درصد آبی + 50 درصد و دیگری نور 100 درصد سفید) بود. در پایان، تعدادی از صفات مورفولوژیک و فیزیولوژیک اندازه گیری شد. نتایج نشان داد که سطح برگ، ارتفاع، طول میانگره، قطر ساقه، وزن تر شاخساره و فلورسانس کلروفیل برگ دانهال حسن یوسف در نور 100 درصد سفید بیشتر از نور ترکیبی 50 درصد آبی + 50 درصد سرخ ال ای دی و نور طبیعی گلخانه بود. همچنین، بیشترین دمای سطح برگ (4/27 درجه سلسیوس)، میزان کلروفیل کل (8/0 میلی‌گرم بر گرم) و کاروتنوئید (30/2 میلی‌گرم بر گرم) در تیمار شاهد و بیشترین تعداد برگ (21)، وزن خشک شاخساره (17/0 گرم)، وزن تر (65/1 گرم) و خشک ریشه (11/0 گرم) در ترکیب نوری 50 درصد آبی + 50 درصد سرخ مشاهده شد. نتایج در نشاء اطلسی نشان داد که نور سفید باعث افزایش سطح برگ، وزن تر شاخساره و وزن خشک ریشه شد. دمای سطح برگ و میزان کلروفیل کل در شاهد نسبت به سایر تیمارها بیشتر بود و بیشترین میزان کاروتنوئید (12/3 میلی‌گرم بر گرم) اطلسی در نور ترکیبی 50 درصد آبی + 50 درصد سرخ حاصل شد. بر اساس نتایج بدست آمده، با جایگزینی شرایط گلخانه با نور ال ای دی می توان، نشاء حسن یوسف و اطلسی را با کیفیت بالاتری تولید کرد.

    کلیدواژگان: فلورسانس کلروفیل برگ، کیفیت نور، نشاء، مورفولوژی
  • عابدین مشعشعی، محمدمهدی جوکار* صفحات 549-565

    قرنفل (Dianthus barbatus L.) یکی از مهمترین گیا‌هان فضای‌ سبز در مناطق معتدل است که به‌عنوان گیاه باغچه‌ای، گلدانی و همچنین گل‌بریدنی کشت می‌گردد. شوری خاک و آب در بخش‌های وسیعی از کشور مهمترین مشکل پیش‌رو در گسترش فضای ‌سبز شهری آن می‌باشد. باتوجه به اهمیت قرنفل در فضای ‌سبز شهری و درک بهتر مبانی فیزیولوژیکی پاسخ گیاه به تنش در محیط درون شیشه‌ای، در این پژوهش تاثیر تنش شوری بر گیاه قرنفل در شرایط درون شیشه‌ای مورد بررسی قرار گرفت. بدین منظور پینه‌های حاصل از ریزنمونه‌های برگ که در محیط‌کشت MS حاوی 2 میکرومولار BAP + 6 میکرومولار NAA پرآوری گردیده بودند، تحت تاثیر سطوح مختلف کلریدسدیم (0، 1، 3، 6، 9 و 12 گرم‌درلیتر) در محیط کشت MS قرار گرفتند. سپس شاخص‌های مختلف فیزیولوژیکی (همچون رشد، نرخ رشد نهایی، وزن‌تر و خشک) و بیوشیمیایی (همچون میزان تغییرات پروتئین‌کل، اسمولیت گلاسین-بتائین، فعالیت آنزیم بتائین آلدهایددهیدروژناز و همچنین میزان فعالیت آنزیم‌های کاتالاز، پراکسیداز، سوپراکسیددسموتاز و میزان بیومارکرتخریبی مالون‌دی‌آلدهاید) در پینه‌های تنش‌یافته مورد بررسی قرار گرفت. نتایج نشان داد با افزایش سطح تنش، رشد پینه‌ها کاهش و درنتیجه از وزن‌تر و خشک آن‌ها نیز کاسته شد. از سوی دیگر میزان پروتئین کل، فعالیت آنزیم بتائین آلدهایددهیدروژناز و درنتیجه اسمولیت گلاسین-بتائین با افزایش سطح تنش، افزایش یافت. فعالیت آنزیم‌های شکارکننده رادیکال آزاد همچون کاتالاز، پراکسیداز و سوپراکسیددسموتاز نیز با افزایش میزان تنش به‌طور‌معنی‌داری افزایش یافت. در بین سه آنزیم شکارکننده رادیکال‌آزاد، پراکسیداز کمترین میزان فعالیت و کاتالاز بیشترین میزان فعالیت را درنتیجه تنش داشتند. پراکسیداز همچنین کمترین میزان افزایش فعالیت در نتیجه تنش را به خود اختصاص داد. این درحالی بود که افزایش میزان فعالیت در دو آنزیم کاتالاز و سوپراکسیددسموتاز تقریبا به یک اندازه مشاهده گردید. بیومارکر تخریبی مالون‌دی‌آلدهاید نیز علیرغم افزایش فعالیت آنزیم‌های شکارکننده رادیکال‌آزاد با افزایش تنش شوری، افزایش یافت. به طور کلی نتایج این تحقیق نشان داد که گیاه قرنفل در محیط درون شیشه‌ای به تنش شوری حساس بوده و آستانه تحمل آن به شوری محیط غلظت 1 گرم در لیتر کلریدسدیم می‌باشد. این درحالی است که، پینه‌های این گیاه تا شوری 12 گرم در لیتر کلریدسدیم مقاوم بوده و ضمن رشد بطئی به حیات خود ادامه دادند.

    کلیدواژگان: بتائین آلدهایددهیدروژناز، سوپراکسیددسموتاز، کاتالاز، گلاسین-بتائین، مالون دی آلدهاید
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  • Kh. Hemmati, B. Ardavanpour, M. Ghazaeian, V. Akbarpour* Pages 349-361
    Introduction

    Essential oils make up a large part of aromatic substances in plants. Generally, essential oils are considered as remnants of main processes of plants metabolism, especially in stressful conditions. Summer savory (Satureja hortensis L.) is one of the most important plants in Lamiaceae family, which it’s more than 30 species grown in the East Mediterranean. It is one of the oldest plants that have been used as vegetables and medicinal and aromatic plants. Dried summer savory has been introduced as one of the most pleasant spices that are planting in large areas of farmland in many countries. Generally, the aerial parts of it, that is usually harvested at flowering stage, has therapeutic effects such as facilitating digestion, stomach tonic, diuretic, astringent, carminative, anti-diarrhea and anti-worm. Summer savory essential oil is used in food (conserves and beverages) and pharmaceutical industries. Various studies on S. hortensis essential oil has been showed that it contains high amounts of phenolic compounds such as Carvacrol, γ-Terpinene, Thymol, p-Cymene, β-Caryophyllene, Linalool and other terpenoids. Organic fertilizers have been known to improve the biodiversity and may prove a large depository for excess carbon dioxide. These fertilizers increase the number of soil organisms by providing organic matter and micronutrients for organisms such as fungal mycorrhizae, (which aid plants in absorbing nutrients), and can drastically reduce external inputs of pesticides, energy and fertilizer, at the cost of decreased yield. Nevertheless they are as effective as chemical fertilizers over longer periods of use. The aim of this project was to study the effects of different organic matters on vegetative and reproductive growth and percentage and components of essential oil in three Satureja ecotypes in Gorgan, Golestan, Iran climate.

    Materials and Methods

    This study was designed as a factorial experiment based on Randomized Complete Block design with four replications and done in research farm of Agriculture and Natural Resource, University of Gorgan in 2010. Treatments were cattle manure, municipal solid waste, leaves compost, the mixture of them and control. Three ecotypes of summer savory studied in this experiment were Kordestan, Shiraz and Yazd. The plants were harvested at full flowering stage, when they have the highest essential oil and were dried in oven at 45 °C. The essential oils of dried samples were isolated by hydro distillation for 3 h, using a Clevenger-type apparatus. Statistical analysis was performed by SAS software and the mean comparison were measures by using Duncan tests at 5% level of probability.

    Results and Discussion

    The analysis of variance showed that media had significant effect on yield components in summer savory. There were no significant differences between ecotypes and media*ecotype effect on some parameters of yield components. The cattle manure showed positive effects on bush height in this experiment. Hendawy et al (2010) showed that the application of cattle manure and sheep manure had reached significantly the 5% level of plant height and essential oils percentages. They concluded that sheep manure have macro and micro nutrients which provide thyme plants with their requirements from these elements. Essential oil percentage also, tended to increase with application of organic fertilizers either cattle manure, sheep manure or compost. Khalil (2002) showed the effect of two kind of organic fertilizers on Rosmarinus officinalis and Jasminum spp growth parameters and yield components. In this experiment organic manure treatments showed increasing in plant fresh and dry weight. This could be related to positive effect of organic manure on soil texture and fertility. Ahmadian et al. (2006) showed that the animal manure significantly enhanced the number of umbers per plant, the number of seed per plant, the biological and seed yield in Cuminum cyminum. Animal manure significantly enhanced cumin aldehyde and r-cymene and decreased b-pinene, g-terpinene and a-pinene in cumin oil. Due to the results, treatments had no significant effects on essential oil percentages, but were effective on essential oil components. Hussein et al. (2006) showed that compost levels combined with different distances had a pronounced effect on the various essential oil constituents in Dracocephalum moldavica. In this experiment, Although Shiraz ecotype showed the maximum essential oil percentage (1.2 percent) but there was no significant differences between ecotypes. Among essential oil components, the maximum percentages of Carvacrol achieved in mixture media and Yazd ecotype. Control treatment showed the highest amount of γ- terpinen and paracymene in compare to other media. Shiraz and Kordestan showed also, the highest percentages of γ- terpinen and paracymene, respectively. Plants in mixture media had maximum levels of Carvacrol (74.69 percent). Between ecotypes, Yazd had maximum Carvacrol percentages. Between media, Control treatment was maximum contents of γ –terpinen (5.48) and paracymene (1.76) and other treatments had reduction effects on essential oil components. Between ecotypes, the highest γ-terpinen percentage (5/79) was in Shiraz ecotype and the highest p-cymene percentage (1/69) was in kordistan ecotype.

    Conclusion

    Therefore, by considering the fertilization of savory plants with different organic manure fertilizers in the form of compost that are economic in each region is recommended.

    Keywords: Carvacrol, Cattle manure, Ecotypes, Essential oil quality, γ-terpinen
  • M.H. Sheikh Mohammadi, Nematollah Etemadi*, A. Nikbakht, M. Arab, M.M. Majidi Pages 363-375
    Introduction

    Drought and salinity are the most detrimental abiotic stresses for turfgrass growth across a wide range of geographic locations. Most cool season grass species are not well adapted to extended periods of drought and salinity stress. The decline in turf quality caused by drought and salinity stresses is a major concern in turfgrass cultivation and management. Therefore, developing management practices for improving drought and salinity resistance of turfgrasses has become imperative in arid and semiarid regions. Grass genotypes differ in their responses to drought and salinity stresses, which involve changes in morphological and physiological aspects. Understanding of relative involvement of each morphological and physiological characteristic in drought and salinity tolerance is important in selecting grass genotypes to facilitate breeding of drought and salinity-tolerant genotypes. The purposes of this research were to make selections of genotypes tolerant to drought and salinity stress for turfgrass management program.

    Materials and Methods

    To study some morphological and physiological responses of six Iranian crested wheatgrasses (Agropyron cristatum L.) under drought and salinity, an experiment was conducted in the greenhouse of College of Abureyhan, University of Tehran, Iran. Six Iranian Agropyron cristatum genotypes were collected from six locations in Iran. Agropyron cristatum genotypes were planted in polyvinyl chloride tubes and kept in the greenhouse. Pots were filled with sandy loam soil which had been sterilized in an oven at 160ºC for 6 h. Irrigation was applied as needed to prevent any visible stress during grass establishment. Grasses were watered three times weekly to maintain plants under well-watered conditions and soil moisture at field capacity. The experiment consisted of three treatments: 1) well-watered plants were irrigated three times per week with distilled water (control), 2) Drought stress was imposed by withholding irrigation for 45 days (drought stress), and 3) plants were irrigated daily with 100 mL of 9 dS.m–1 NaCl solution (salinity stress). To avoid primary salinity shock, the soil in each pot was drenched with 100 mL NaCl solution at incremental electrical conductivity (EC) by 3 dS.m–1 per day until the final EC reached 9 dS.m–1. Data were subjected to analysis based on a split-plot design with water treatments as main-plots and genotypes as sub-plots. Irrigation treatment as the main factor in three levels (control, drought, and salinity) and crested wheatgrass at six levels were considered as sub-plots. Studied characteristics such as height, turf quality, chlorophyll content, soluble carbohydrates, relative water content, electrolyte leakage, root penetration, and effective root depth were recorded. Statistical significance was tested using the analysis of variance procedure in SAS 9.1 (SAS Institute Inc., Cary, NC). Differences between the means were determined using the Fisher’s protected LSD test at the 5% probability level.

    Results and Discussion

    The results of this study showed that drought and salinity stress decreased the quality of crested wheatgrass masses, and reduced the level of quality varied among the masses. Throughout the experiment, ‘Sabzevar’ and ‘Damavand’ under drought conditions and ‘Sabzevar’, ‘Arak’ and ‘Damavand’ under salinity conditions maintained higher Turf quality compared with other genotypes. Total chlorophyll content of ‘Sabzevar’ and ‘Damavand’ were higher than other genotypes under drought and salinity conditions. The maintenance of higher chlorophyll content has been associated with better drought and salinity tolerance in plant. The soluble sugar content of ‘Sabzevar’ and ‘Damavand’ under drought conditions and ‘Sabzevar’, ‘Arak’ and ‘Damavand’ under salinity conditions were higher than other genotypes during the experiment. Soluble sugar content is an important compatible osmolyte in plants. Increased accumulation of soluble sugar content in stressed plants may be an adaptation process and resistance strategy to abiotic stresses in plants. Throughout the experiment, ‘AEKQI’, ‘Sabzevar’ and ‘Damavand’ genotypes under drought conditions and ‘Sabzevar’ genotypes under salinity conditions maintained higher relative water content in compared with other genotypes. Higher RWC indicates the ability of the leaf to maintain its higher water content under stress conditions with the simultaneous capability of the root system to take up adequate water. Based on morphological and physiological analysis for drought and salinity tolerance in investigated genotypes, the tolerance ranking would appear to be ‘Sabzevar’ > ‘Damavand’ > ‘Arak’ > ‘Urmia’ = Takestan > ‘Hashtgerd’ under drought stress and ‘Sabzevar’> ‘Arak’ > ‘Damavand’ > ‘Takestan’ = ‘Hashtgerd’> ‘Urmia’ under salinity stress. The results of this study showed that ‘Sabzevar’ and ‘Damavand’ genotypes had good tolerance to drought stress, and ‘Sabzevar’ and ‘Arak’ genotypes had good tolerance to salt stress than other Iranian crested wheatgrass genotypes.

    Keywords: Agropyron cristatum, Drought, Grass, Salinity
  • M. Kharrazi, A. Tehranifar*, A. Sharifi Pages 377-386
    Introduction

    Success in tissue culture technique, especially in bulbous plants, depends on the microbial contamination control during in vitro culture. Applying different treatments, such as heat treatment and usage of fungicides, can control the microbial contamination and consequently increase the percentage of explant survival.

    Materials and Methods

    This study aimed to investigate the effect of heat treatment and fungicide on reducing the contamination during in vitro culture of narcissus. So, an experiment was done as a factorial experiment in a completely randomized design with two factors, including benomyl concentration in the medium (1 and 2 g/l) and heat treatments (two levels, with and without heat treatment), with 10 replications. In order to sterilizing the plant materials, damaged and infected scales were removed firstly and then bulbs were washed for 30 minutes with running tab water and a few drops of dishwashing liquid. For applying heat treatment, bulbs were divided into two groups. In the first group, heat treatment was not applied and in the second group heat treatment 54 °C was applied for one hour using water bath. After this step, bulbs surface were sterilized by dipping in 70% ethanol for one minute and rinsed with sterile distilled water, followed by immersing in 1.5% sodium hypochlorite solution for 30 min. After sterilization with sodium hypochlorite solution, bulbs were washed three times with sterile distilled water under laminar air flow hood. After sterilization step, bulbs were cut into 32 twin scales explants and cultured in MS medium supplemented with 1 mg/l BA and 0.2 mg/l NAA + benomyl (1 or 2 g/l). After 30 days, the response of explants (number of produced bulbs, percentage of explant survival, percentage of bacterial contamination, percentage of fungal contamination, percentage of browning) was evaluated. Data preparation was done in the Excel program and data analysis was done using JMP-8 software. Mean comparison of the treatments was done by LSD test and finally the charts were drawn using the Excel program.

    Results and Discussion

    The results showed that increasing the concentration of benomyl in the medium and applying heat treatment had negative effect on regeneration potential of explants, so that the maximum regeneration mean were observed when heat treatment was not applied for explants and medium contains 1 g/l benomyl. Using the heat treatment and application of 2 g/l benomyl in the medium leads to the lowest regeneration amount. On the other hand, evaluating the browning percentage of explants showed that the effect of treatments was significant in this trait. Applying the heat treatment and using 2 g/l benomyl in the medium had severe effect on the increasing of explant browning and the maximum mean was observed in this treatment. But reducing the benomyl concentration in the medium and none application of heat treatment caused the lowest amount of explant browning. Contamination percentage that includes bacterial and fungal contamination is an important parameter in this study. Explants that cultured in the medium containing 1 g/l benomyl and applying heat treatment showed the highest contamination percentage, which contains 21% fungal and 14% bacterial contamination. The lowest percentage of contamination was observed when heat treatment applied and medium contains 2 g/l benomyl. However, this treatment caused the highest percentage of explant browning that lead to reduction of explants regeneration potential. Researches showed that the use of fungicides can help to control tissue culture contamination and according to previous studies, benomyl is the most effective treatment against fungal infection. As benomyl is considered as a systemic fungicide, so it is useful to eliminate the internal fungus. On the other hand, there are some reports about the positive effect of heat treatment on the control of tissue culture contamination. As regards this investigations were done in 1914 to 1997 and then stopped, so it seems that application of this treatment had no sufficient efficiency for contamination control during in vitro condition.

    Conclusion

    Therefore, by considering the browning, regeneration and contamination percentage, non-application of heat treatment and usage of 1g/l benomyl fungicide in the medium for in vitro culture of Narcissus twin scales explants is recommended.

    Keywords: Benomyl, Percentage of survival, Percentage of browning, Tissue culture
  • M.B. Amiri*, P. Rezvani Moghaddam, M. Jahan Pages 387-403
    Introduction

    Maintenance of soil fertility as a permanent bed for continuous production of agricultural products is one of the most important issues affecting the sustainability of food production. In order to achieve healthy food production, application of ecological inputs such as organic fertilizers are inevitable. Organic fertilizers are fertilizer compounds that contain one or more kinds of organic matter. They can improve the soil’s ability to hold water and nutrients. They create a beneficial environment for earthworms and microbial organisms that break the soil down into rich, fine humus. Compost is organic matter that has been decomposed and recycled as a fertilizer and soil amendment. Compost can greatly enhance the physical structure of soil. The addition of compost may provide greater drought resistance and more efficient water utilization. Vermicompost is the final product of composting organic material using different types of worms, such as red wigglers or earthworms, to create a homogenized blend of decomposed vegetable and food waste, bedding materials and manure. Vermicompost helps store nutrients and keeps them safe from leeching and irrigation, functioning to balance hormones within plant physiology, and adding beneficial biology to soil. Vermicompost improves soil aeration, enriches soil with micro-organisms (adding enzymes such as phosphatase and cellulose), attracts deep-burrowing earthworms already present in the soil and improves water holding capacity, enhances germination, plant growth and crop yield, improves root growth and structure. Cow manure is an excellent fertilizer containing nitrogen, phosphorus, potassium and other nutrients. It also adds organic matter to the soil which may improve soil structure, aeration, soil moisture-holding capacity, and water infiltration. Plant density is the number of individuals of a given plants that occurs within a given sample unit or study area. Planting density can impact the overall health of plants. Plantings that are too sparse (the density is too low) may be more susceptible to weeds, while planting that are too dense might force plants to compete over scarce nutrients and water and cause stunted growth. Despite of many researches on the effect of organic fertilizers and plant density on different crops, information on the effects of these factors for many medicinal plants is scarce, therefore, in this study effect of organic fertilizers and plant density on morphological characteristics and yield of Echium amoenum in a low input cropping system was studied.

    Materials and Methods

    In order to evaluate the effect of different fertilizers on quantitative characteristics of Echium amoenum at different plant densities, a factorial experiment based on RCBD design with three replications was conducted in 2011-2013 growing seasons, in Ferdowsi University of Mashhad, Iran. Experimental factors included three plant densities (10, 5 and 3 plant per m2) and five different types of organic and chemical fertilizers (municipal waste compost, vermicompost, cow manure, chemical (urea) fertilizer and control).

    Results and Discussion

    The results showed that with reduction of plant density, effect of organic fertilizers on increasing the flower yield was exacerbated and at the lowest density, municipal waste compost, vermicompost and cow manures increased flower yield 112, 79 and 223% compared to control, respectively. In all studied densities, length of lateral branches in conditions of organic fertilizers application was more than control. Municipal waste compost, vermicompost, cow manure and chemical fertilizer increased total phenol 106, 54, 66 and 173% compared to control, respectively. In plant mean density (5 plant per m2), application of municipal waste compost, vermicompost and cow manure increased total anthocyanin 83, 74 and 57% compared to control, respectively. In general, result of this study showed that application of organic and chemical fertilizers at optimal plant densities, can improve growth characteristics and quantitative and qualitative yield of Echium amoenum.Organic fertilizers are the most significant resources for development of agricultural soil quality and increase in the yield of different medicinal plants. It has been reported that this ecological inputs provide favorable conditions for plant growth and development through improvement of physical, chemical and biological properties of the soil, therefore, it can be concluded that improvement of the most studied traits in the present study were due to use of organic fertilizers. Fallahi et al., (20) have reported the positive effects of organic fertilizers on the improvement of quantitative and qualitative characteristics in chamomile (Matricaria chamomilla L.). It seems that plants compete with each other over scarce nutrients and water in high plant density and cause stunted growth. Some other studies have reported that suitable plant density can increase the growth and yield of some medicinal plants such as coriander (Coriandrum sativum L.), Ajwain (Carum copticum L.), lemon balm (Melissa officinalis L.) and anise (Pimpinella anisum L.).

    Conclusion

    In general, results of this study showed that application of organic and chemical fertilizers at optimal plant densities, while improving growth and yield characteristics of Echium amoenum flower can be ensured health of production of this important medicinal plant.

    Keywords: Evaluation of Morphological Characteristics, Quantitative, Qualitative Yield of Echium amoenum as Affected by Plant Density, Application of Chemical (Urea) Fertilizer, Different Organic Fertilizers
  • M. Sayadi Nejad, S.M. Sadeghi* Pages 405-415
    Introduction

    Zamiifolia is a perennial ornamental plant and is one of the most important medicinal plants of the Aracea family. The origin of this evergreen, low-anticipated plant is East Africa. Zamiifolia spreads through the leaves and split rhizomes, which is very time-consuming. The traditional Zamiifolia proliferation method have been done by dividing rhizomes and leaf cuttings, but the production efficiency is low due to the slow growth of the plant, tubers and roots. In addition, due to the warm and humid environment, reproduction is limited to summer season. Due to the traditional reproductive problems in this plant, tissue culture or microbial culture is the best way to replicate rapidly and to achieve a large number of plants with the same genetic structure, as well as the elimination of diseases in the short term and reducing the costs. The aim of this study was to compare different microorganisms in terms of calogenesis and regeneration, as well as to determine the optimum culture medium for Zamiifolia tissue culture.

    Materials and Methods

    In this study, the explants prepared for the first experiment, including rhizome and petiole and the explants for the second experiment were the leaeaves and shoots. In the first experiment, rhizome and petiole were cultured in three replications in ½ MS medium containing BA (0, 2, 4 mg / L) and 2,4-D (0,1,2 mg / L) in combination with vitamins, 30 g/l sucrose, 5 g / L agar and adjusted to 5.8 PH. The cultivars were cultured for the callus induction under temperature of 27-25 ° C and light conditions of 16 hours light and 8 hours darkness. After 5 weeks, the percentage of callus and fresh callus weight were measured. The callus generated from rhizome and petiole in three replicates on ½ MS medium containing BA (0, 1, 2 mg / L) and 2,4-D (0, 0.5, 1 mg / l) for shoots and after the observation of branches and leaf buds were grafted on to ½ MS medium containing BA (0, 1, 2 mg / L) and NAA (0, 0.5, 1 mg / L) for rooting. Traits such as time to shoot elongation were recorded at regeneration stage, and after 5 weeks, shoot length and the number of leaves were measured. The time to rootstock was also recorded. In the second experiment leaf and shoot explants were cultured in ½ MS medium containing BA (0, 2, 4 mg / l) and NAA (0, 0.5, 1 mg / L) in combination with vitamins, 30 g/l sucrose, 5 g / L agar and PH adjusted to 5.8. The cultivars were cultured for the callus induction under temperature 27-25 °C and light conditions of 16 hours light and 8 hours darkness. The time to reach the callus was recorded and after 5 weeks, the percentage of callus and fresh callus weight were measured. The calli generated from the leaves and shoots were cultures on ½ MS medium containing BA (0, 1, 2 m g / L) and NAA (0, 0.5, 1 mg / L) for shoots and after observation of branch and leaves were transplanted to the ½ MS medium containing BA (0, 1, 2 mg / L) and NAA (0, 1, 2 mg /L) for rooting. The traits such as time to shoot elongation were recorded at the regeneration stage, After 5 weeks, the shoot length and the number of leaves were measured.The time to rootstock was also recorded.

    Results and Discussion

    The results of the first experiment showed that the effect of the rhizome and petiole type on the callus formation was significant at 1% level. So that the rhizome showed greater ability to callogenesis. The results of the second experiment showed that the effect of the type leaf and shoot on the callus formation was significant at 1% level. So that the leaf showed greater ability to callogenesis. The highest percentage of callosing (94.5%), the shortest time to reach the callus (14 days) and the highest callus weight (1.1 g) in culture medium with 2 mg / l BA and 1 mg / l hormones NAA was observed in leaf samples from the second experiment. The best treatment in the shoot elongation stage, which included the shortest time to shoot formation (10.5 days), the longest shoot length (4.10 cm), and the highest leaf number (8 leaves) in the leaf extract with hormonal concentrations of 2 mg / 0 mg / L NAA was observed from the second experiment. In the rooting stage, the best treatment for petiole extracts with hormonal concentrations was 1 mg / l BA and 0.5 mg / l NAA with 14 days to rooting from the first experiment.

    Conclusion

    In this study, explants and various concentrations of growth regulators had significant effect on the response to callus induction in Zamiifolia. In the first experiment, the rhizome and in the second experiment the leaf showed a better reaction to callus induction. According to this research, it can be suggested that the treatments applied in both experiments should be applied on all four leaves, petiole, rhizome and shoot samples, and the best culture type and the best culture medium for the cultivation of Zamiifolia plant tissue should be determined in subsequent studies.

    Keywords: BA, Micro propagation, Sub culture, Tissue culture
  • M. Abaspour Esfaden, S. Kallaterjari*, F. Fatehi Pages 417-432
    Introduction

    Catharanthus roseus is a beautiful herbaceous plant, belonging to Apocynaceae family, which is widely cultivated due to its unique and beautiful appearance. It is native to the Caribbean Basin and has historically been used to treat a wide assortment of diseases. European herbalists used this plant for different conditions such as headache and a folk remedy for diabetes. The effects of drought range from morphological to molecular levels and are evident at all phonological stages of plant growth at whatever stage the water deficit takes place.The limitation of water resources in our country is one of the important issues in landscape. However, we can alleviate the adverse effects of water stress with the use of some biological and non-biological stimulators. Under severe stress conditions, the antioxidant capacity may not be sufficient to minimize the harmful effects of oxidative damage. Therefore, synthesis of signal molecules in plants is an important step in our better understanding of how plants respond to environmental stresses. Several such signal molecules have been identified in plants such as jasmonic acid, ethylene and salicylic acid (SA). SA is considered as a hormone-like substance, which plays an important role in regulating a number of plants’ physiological processes including photosynthesis. Besides, L-arginine is an amino acid, which can alleviate the adverse effect of drought. Among the 21 proteinogenic amino acids, arginine has the highest nitrogen to carbon ratio, which makes it especially suitable as a storage form of organic nitrogen. Synthesis in chloroplasts via ornithine is apparently the only operational pathway to provide arginine in plants. Therefore, the present study was conducted to evaluate the effect of salicylic acid (SA) and L-arginine on morpho-physiological properties of C. roseus.

    Materials and Methods

    This experiment was carried out as a factorial with two factors including the foliar application at five levels (control, L-arginine 1.5 and 3 mM, SA 100 and 200 mg /L) and water stress at three levels (100, 70 and 40% field capacity (FC)) in a completely randomized design with three replications. Water use efficiency (WUE), the root/shoot weight, flower number, flower diameter, leaf chlorophyll content, peroxidase and superoxide dismutase activity of leaf, and leaf macronutrients (nitrogen, phosphorus and potassium) were measured.

    Results and Discussion

    The results showed that WUE in plants under moderate stress (70% FC) was higher than the plants in control and severe drought condition (40% FC). Root/shoot weight at no foliar application (control) × 40% FC and L-arginine 1.5 mM × 40% FC was higher than other treatments. The number of flowers in the treatments of 100% FC and 70% FC was more than 40% FC. The highest flower diameter was obtained from the L-arginine 3 mM. Total chlorophyll of L-arginine 3 mM was higher relative to other treatments. Drought stress significantly increased the activity of antioxidant enzymes. The lowest amount of phosphorus and nitrogen was observed at no foliar application ×40% FC. Therefore, according to the optimum application of water and amendment substrates, at 70% FC stress, L-arginine 3 mM and SA 100 mg /l increased the nitrogen and phosphorus content. Overall, L-arginine 3 mM and SA 100 mg /l increased the flower number and flower diameter significantly. Accumulation of these organic solutes either actively or passively helps the plants to retain water within cells and protect cellular compartments from injury caused by dehydration or maintain turgor pressure during water stress. Turgor maintenance plays an important role in drought tolerance of plants which may be due to its involvement in stomatal regulation and hence photosynthesis. Foliar application with arginine resulted in elevated proline levels and radiotracer experiments demonstrated that both 3H and 14C from arginine can be recovered as proline. The physiological relevance and the biochemical pathway of the conversion of arginine to proline in plants remain unclear. The most prominent hypothesis is that ornithine, derived from arginine catabolism, is converted by δOAT to GSA/P5C, which then serves as substrate for proline synthesis by P5CR. This model has been doubted, since Arabidopsis δOAT was found to be exclusively localized in mitochondria, while P5CR is localized in the cytosol.

    Conclusion

    The findings of our study showed that water stress can morphologically and physiological change C. roseus. There was no significant difference between 70% and 100% FC for root/shoot, flower number and flower diameter traits. So, we can reduce the use of water to 70% FC increased the flower number and flower diameter and can be used to alleviate the adverse impact of water stress.

    Keywords: Growth stimulators, Macronutrients, Total chlorophyll, Water use efficiency
  • M. Norani, M.T. Ebadi, M. Ayyari Noushabadi* Pages 433-449
    Introduction

    Coltsfoot (Tussilago farfara L.) from Asteraceae family is a perennial plant. T. farfara is native and widespread from Europe to Western and Northern Asia and North Africa. Coltsfoot distributed in wet mountainous regions of Iran, such as Azerbaijan, Tehran and Northern provinces. Its flowers and leaves have been used traditionally for the treatment of cough, bronchitis and phlegm disorders. T. farfara leaves and flowers have expectorant activity and are used for chronic dry cough and various pulmonary diseases. The extracts of T. farfara were shown to exhibit various activities, such as antioxidant and antimicrobial activity. Biologically active agents of T. farfara have been studied due to their antimicrobial and antioxidant characteristics.

    Materials and Methods

    For morphological study, the rhizomes of seven Iranian coltsfoot populations were collected in August 2016 from different regions of Iran including Pol-e zangholeh, Damavand, Firoozkooh, Nur, Deylaman, Kaleybar and Namin. The collected samples were planted in the same condition during 2016-2017 in research field of Tarbiat Modares University in Tehran (51º10ʹ23ʺ N, 35º44ʹ17ʺ E), with a randomized complete block design experiment and three replications. The average annual rainfall and temperature of cultivating place is about 220 mm and 16.4 ºC. In order to study the morphological characteristics between different samples, traits such as the number of flowers per plant, stem height, flower length, flower diameter, flower dry weight, root length, root dry weight, leaf area and leaf dry weight had been measured. Morphological traits were measured under the same conditions and for this research, were used ruler, caliper and balance. The leaves and flowers were prepared for phytochemical studies. DPPH method has been used to evaluate the antioxidant activity, and the IC50 was used to compare the antioxidant properties. The absorbance of the samples was measured at 517 nm with ELISA reader. The radical scavenging capacity (RSC) was calculated by the following formula: % In=[(Ab-As)/Ab]×100, where In is DPPH inhibition, Ab is the absorbance of the blank, As is the absorbance of samples including extracts and BHT as a positive control. The phenolic contents of different extracts were determined by Folin-Ciocalteu method and the aluminum chloride method was used to measure total flavonoid.

    Results and Discussion

    Analysis of variance showed that there is a significant difference between all evaluated traits in studied populations (p ≤0.01). Pol-e zangholeh population of T. farfara, has shown the best performance for all morphological traits. The results of the flowers per plant showed that the Pol-e zangholeh population was the highest and the Deylaman population had the lowest number of flowers. The maximum length of the stem was related to the Pol-e zangholeh population while the population of Namin had the smallest amount. Comparison of flower diameter showed that the Pol-e zangholeh population had the largest diameter and Nur population of all the smaller. The results of the flower length showed that the maximum flower length was related to the Pol-e zangholeh population and Deylaman population had the smallest amount of flower length. Comparison of mean of flower dry weight showed that the Pol-e zangholeh population of T. farfara had the highest value while the population of Nur had the smallest amount. Pol-e zangholeh population of T. farfara, considered as superior populations and its flowering time were earlier than the others. It was found that there is a significant difference (p ≤0.01) between different populations. This variation may be due to Population differences of People, either due to differences in environmental conditions or sometimes due to the interaction of the population and the environment. Our results also indicate the presence of comparable genetic potentials of T. farfara in these population for any further cultivar development. Analysis variance showed that there was significant difference between populations for total phenol, flavonoid and antioxidant activity (p ≤0.01) and the Nur population had the highest total phenolic content and highest antioxidant activity. Comparison of antioxidant activity of different extracts from leaves and flowers of T. farfara showed that the most antioxidant activity was related to leaf extract of Nur population with IC50 271 µg/ml closer to BHT (33 µg/ml) as a synthetic and industrial antioxidant. The least amount of this activity was related to Kaleybar leaf extract with IC50 888 µg/ml. Nur population showed the highest total phenol content with 242 mg GAE/g dried weight. Damavand population showed the highest total flavonoid content with 40 mg QE/g DW extract.

    Conclusion

    Our results indicate the presence of comparable potentials of T. farfara in these populations for any further cultivar development. Study of antioxidant activity in different habitats shows that Nur habitat is a suitable place for the cultivation of T. farfara to increase the amount of phenolic compounds and antioxidant activity.

    Keywords: Antioxidant activity, Coltsfoot, Morphological study, Phytochemical studies, Total flavonoid
  • A. Naseri Moghadam, H. Bayat*, M.H. Aminifard, Farid Moradinezhad Pages 451-466
    Introduction

    Salinity and drought have adverse impacts on crop production throughout the world, especially in arid and semiarid areas. Salinity decreases crop growth and yield through modifications of ion balance, water status, stomatal behavior, photosynthetic efficiency, salinity-induced nutrient deficiency and changes to the soil structure. Drought is another one of the main factors limiting the growth and development through the anatomical, morphological, and physiological and biochemical changes that the severity of drought damage varies depending on the duration of the stress and plant growth stage. Water stress reduces relative water content, photosynthesis pigments, stomatal conductance, biomass, growth and ultimately plant performance. Narcissus (Narcissus tazetta L. cv. ‘Shahla’) belongs to the Amaryllidaceae family is grown as a cut flower, landscape and medicinal plant that grows throughout the world except tropical regions. It is necessary to know the tolerance of N. tazzeta to drought and salinity stress in order to produce optimal product. According to previous studies, no complete research has been done on the effects of drought and salinity stress on N. tazzeta. Therefore, the present study was carried out with the aim of investigating the combined effects of drought and salinity stress on growth, flowering and biochemical characteristics of N. tazzeta.

    Materials and Methods

    This research was carried out in Faculty of Agriculture, University of Birjand, in 2017. A pot experiment was conducted in completely randomized deign with factorial arrangement and three replications. The treatments included sodium chloride (NaCl) in four levels 0 (control), 20, 40 and 60 mM and drought stress in four levels 30, 50, 70 and 90% of field capacity. The plants were harvested four months after the start of salt and drought treatments. The investigation traits were included vegetative, reproductive and biochemical characterizes. Measured traits were included root length, volume of root, root fresh weight, bulb length, bulb fresh weight, shoot fresh weight, total dry weight, flower diameter, flower crown diameter, stem diameter, days from planting to flowering, days from flowering to senescence, antioxidant activity, total phenolic content and total soluble sugar of leaf and root. The data were analyzed by SAS version 9.4 and the means separated by Duncan's multiple range test at p < 0.05. Excel was used to draw graphs.

    Results and Discussion

    The results of simple effects showed that drought and salinity stresses decreased the values of flowering stem diameter, flower diameter, root length, root volume, root fresh weight, bulb fresh weight, bulb length, shoot fresh weight, total dry weight and days from flowering to senescence. In contrast, the values of the number of days from sowing to flowering, total soluble sugars of leaf and root, antioxidant activity and total phenolic content increased under the influence of these two stresses. The results of interaction effects showed that the destructive effects of salinity and drought stress were intensified in co-application conditions, so that the lowest values of growth and reproductive traits were observed in the most severe stress (30% field capacity × 60 mM salinity). Usually, root and shoot length in sodium chloride solution is reduced due to the toxicity of ions and their negative effects on cell membranes. Drought stress and salinity reduce cell division and also reduce the size of cells and consequently the length of the plant decreases. Delay in flowering is due to multiple stresses (osmotic imbalance, nutrient insufficiency and cellular toxicity) that is caused by salinity and drought stresses. These stresses produce ROS compounds that damage the proteins, lipids, carbohydrates, and nucleic acids. Plants for scavenging and detoxifying these compounds from the cell surface use enzymatic (catalase, superoxide dismutase, etc.) and non-enzymatic (phenolic compounds and carotenoids) defense systems that increase the antioxidant activity of the plant.

    Conclusion

    The results of this study showed that drought and salinity stresses had negative effects on growth and flowering traits, which was exacerbated by the combined application of these two stresses. On the other hand, the highest levels of antioxidant activity, total phenolic content and total soluble sugars were obtained under severe stress conditions (drought or salinity). Salinity and drought stress reduced the flowering rate, quality of flowers and the flower life on the plant, but all the levels of stress reached to flowering stage. Also, the growth of narcissus plant was not affected by the highest levels of drought (30% crop capacity) and salinity (60 mM) stress. In general, the results showed that both drought and salinity stress reduced the growth and yield of narcissus flower, but the destructive effects of salinity stress on the growth, ornamental and physiological traits of narcissus flower were more than drought stress.

    Keywords: Antioxidant activity, Flower diameter, Root volume, Total phenolic content, Total soluble sugar
  • maedeh aghdaei, S.H. Nemati*, L. Samiei, A. Sharifi Pages 467-479
    Introduction

    Pepino (Solanum muricatum Aiton) is a diploid herbaceous plant belongs to the Solanaceae family, which is growing in subtropical zone, originates from Andes in South America. It is commercially grown for its fruit, which is appreciated not only for food but also for its appearance, in South American countries, including Bolivia, Colombia, Ecuador and Peru, as well as in countries such as New Zealand and Australia. Pepino is propagated by seed, cutting, and tissue culture methods. Most pepino cultivars are sexually fertile and produce viable seeds, but their seeds have poor germination and high level of heterozygosis causing to highly variable plants. Both mentioned negative aspects have limited the mass production of this plant through seed. In this case, stem cutting is used as the most common way of propagating pepino led to transmission of viral diseases and increasing propagation costs as two main limiting factors of pepino propagation. So, micropropagation systems are a promising tool to produce disease-free clonal plant material with low costs. Therefore, the present study was aimed to assess the effect of different media and plant growth regulators on micropropagation traits of pepino.

    Materials and Methods

    Three separate experiments were carried out in institute of plant sciences of Ferdowsi University of Mashhad in 2016. Pepino seeds were bought from company of Plant World Seed, UK, were cultivated on MS medium. Grown plants were used as source of providing explants. Four mediums, including MS, ½ MS, SH and B5 were used to determine the best culture medium for shoot regeneration of pepino using single node explant. A factorial experiment was conducted based on a completely randomized design. Some growth properties such as number of shoots, shoot length, number of roots, root length, leaf number and leaf length were evaluated after two and four weeks. In proliferation experiment, MS medium was compared with MS supplemented with different concentrations of BA (0.5, 1 and 2 mg L-1) and Kin (0.5, 1 and 2 mg L-1) applied as combined treatments, and also BA used alone at concentrations of 2, 4 and 6 mg L-1 that was conducted based on a completely randomized design. For rooting of explants, an experiment was conducted based on a completely randomized design containing of two concentrations of IBA (at 0.3 and 0.6 mg L-1) and three concentrations of NAA (at 0.3, 0.6 and 0.9 mg L-1) in MS medium. Some growth properties including root number and length, root density and root quality were evaluated after four weeks

    Results and Discussion

    Results indicated that micropropagation rate of pepino was affected by culture medium type. The highest shoot length, number of root, root length and leaf number were obtained in MS medium, although statistically there was no significant difference between MS and ½ MS media. The highest number of shoots and leaf length were observed in MS medium, which led to a significant difference with other media (½ MS, SH and B5). Overall, Based on obtained results MS medium was the best culture medium for micropropagation of pepino using single node. In the proliferation experiment, the highest shoot and leaf number and plant color were obtained with using 2 mg L-1 BA + 1 mg L-1 Kin, whereas the highest shoot length and leaf length were observed in the 1 mg L-1 BA + 2 mg L-1 Kin and 1 mg L-1 BA+1 mg L-1 Kin treatments, respectively. Increasing in concentration of BA up to 2 mg L-1 in combination with Kin had a positive effect on shoot proliferation, while applying BA at concentration 2, 4 and 6 mg L-1 alone led to decrease in proliferation. Results obtained from rooting experiment showed that the highest root number, root density and root quality were obtained using IBA at the concentration of 0.6 mg L-1, whereas the highest root length was observed by applying IBA at concentration of 0.3 mg L-1, which led to a significant difference with other treatments. Furthermore, results indicated that the effect of IBA on rooting of pepino microshoots was more than NAA.

    Conclusion

    Generally, the best results were obtained by MS medium, 2 mg L-1 BA with 1 mg L-1 Kin for shoot proliferation, and IBA at concentration of 0.6 mg L-1 for the rooting of pepino nodal segments.

    Keywords: BA, IBA, Proliferation, Rooting, Tissue culture
  • M.H. Alemkhoumaram, A.H. Keshtkar*, A. Mirzaie Asl Pages 481-497
    Introduction

    Garlic is native to central Asia, with a three thousand years history of human consumption and use. Global consumption has increased fourfold during the 24-years period from 1989 to 2013. Area under garlic cultivation, average yield/ha and total production of the world were 1,468,811 ha, 18,092 kg/ha and 26,573,001 tons, respectively in 2016; and China produces about 80% of world garlic production. The maximum global record has been registered at 2012 with an average bulb yield of 45,270 tons per hectare belongs to Uzbekistan. The area under cultivation of this plant in Iran is about 9000 hectares with an average yield of 10 tons per hectare. Garlic has been used both as a food flavoring and as a traditional medicine, and these characteristics are affected by sulfur compounds. Alliin (C6H11NO3S) with 177.22 molecule weight is a sulfoxide that is a natural constituent of fresh garlic. It is a derivative of the amino acid cysteine. When fresh garlic is chopped or crushed, the alliinase enzyme, which contains pyridoxal phosphate (PLP), cleaves alliin, generating allysulfenic acid, pyruvate (C3H4O3), and ammonium (NH3). At room temperature two moles of allysulfenic acid as an unstable and highly reactive compound that through a dehydration reaction form one mole of allicin (C6H10OS2) with 162.26 molecule weight, which is responsible for the aroma of fresh garlic. In general, alliinase needs few minutes time to effect on every two molecules of alliin to form one molecule of allicin, two molecules of pyruvate and two molecules of ammonium. The generated allicin is unstable and quickly breaks down, for example during sixteen hours at 23°C it converts to a series of other sulfur-containing compounds such as diallyl disulfide. Allicin is part of a common defense mechanism in garlic plants against pest attacks. It is produced and activated after causing physical injuries. Iranian garlic has a good flavor, while with the exception of Mazand cultivar so far there is no other introduced bred cultivar. In this effort, diversity of agromorphological traits, bulb yield and allicin content of some Iranian landraces were studied to breed promising cultivar/s by single-bulb selection method for Hamedan climatic condition.

    Materials and Methods

    The experiment was carried out on the basis of a randomized complete block design (RCBD) with four replications in 2015-16 agronomy season at the Agricultural Research, Education and Natural Resources Center of Hamedan Province Located about 6 km north of Hamedan at the altitude of 1740 meters above sea level, longitude 48°, 52̍ E, latitude 34°, 52̍ N. The soil texture of the test site was loam with 24.4, 40.6 and 35 percent of clay, silt and sand, respectively. The soil tillage operation included plowing with the moldboard plow, disc and leveler. The furrow and stack were created with a width of 50 cm. Nine landraces from Hamedan province (Maryanaj, Bahar, Sheverin, Soolan, Toyserkan, Heidareh, Toeen, Hossein Abad and Ali Abad, and other nine ones from Gorgan (Golestan), Oromyeh (West Azarbaijan), Arak (Markazi), Bardsir (Kerman), Amol (Mazandaran), Ramhormoz (Khuzestan), Isfahan (Isfahan), Tarom (Zanjan) and Zabol (Sistan) cultivated on plots having 2 ridges with 4 meters long. Two rows were planted on the ridge with 25 and 10 cm spacing between rows and plants on the row, respectively (40 plant m-2). Bulb planting was carried out on November 17th, and first irrigation was rain-fed on 18th and 19th of November, with 19 and 9.9 mm rain, respectively, and during the fall and winter seasons, all water requirements were met through precipitation. Different landraces germinated from March 12th to March 16th. Pressure irrigation system was installed by re-growing in spring and plants were irrigated every seven to 10 days, depend on air temperature severity, until June 5th. Harvesting of different landraces was carried out by hands from June 26th to July 6th. Evaluated traits for each plot included weight of 30 seed clove, date of germination and maturity, number of plant per plot, chlorophyll a and b and carotenoids content of leaf, chlorophyll index by SPAD, relative water content of leaf, bulb yield, and weight of 30 produced clove, total pyruvic acid, and non-enzymatic pyruvic acid. The evaluated traits for five normal random plants (healthy plants on both sides, and on the opposite side of cultivation lines adjacent to the healthy plant) of each plot were including height, crown diameter, number of leaf, number of fallen leaf, length and width and leaf area, dry weight of plant leaf, dry weight of plant root (to a radius of five centimeters around the bulb) height and width and length of bulb, dry matter of bulb, number of skin layers, number and weight of clove, dry matter percentage. Assessment were also considered based on traits such as, moisture percentage of bulb, leaf area index, sum of chlorophyll a and b, loss weight of cuddle, harvest index, sphericity coefficient of bulb, total number of plant leaf, days number of germination to maturity, enzymatic pyruvic acid and allicin. Traits measurements were done based on "Descriptors for Allium". To measure pyruvic acid, 25 g of cloves with 25 ml of water were completely homogeneous in a mixer and passed through a filter paper after ten minutes. Ten ml of filtrated extract was increased into 100 ml volume with water and a half-milliliter of it was taken and added to 1.5 ml of 5% trichloroacetic acid solution, and after one hour, 18 ml of water was added to sample. Then one ml of resulting solution and each standard solution were transferred to separate test tubes. One ml of water and dinitrophenyl hydrazine 0.0125%, were added to each sample, and were heated in bain-marie bath at 37°C for ten minutes. At the end, by adding five ml of 0.6% normal caustic soda to the test tubes, the concentration of samples were determined in the presence of standard solutions (0.01, 0.225, 0.05, 0.1 and 0.2 μmol / ml sodium pyruvate) at 420 nm wavelength using spectrophotometer Pharmacia-Biotech model Novaspec II. To assess non-enzymatic pyruvate activity, alinase enzyme must become inactivated, so 40 to 60 grams of garlic clove is initially heated in microwave at 900 watts for three seconds for every gram of sample (120 to 180 seconds). To find relationship between variables, the correlation coefficients between them were calculated. The study of the relationship between traits with bulb yield and alicin yield was done by stepwise regression method. Causal relationships and direct and indirect effects of traits on bulb and allicin were determined by path analysis method. Analysis of variance, mean comparison of traits by LSD method at 1% probability level, correlation determination, stepwise regression analysis and cluster analysis were done by Minitab software version 17.3.1, while Microsoft Word and Excel 2003 softwares were used to draw the graphs and path analysis, respectively.

    Results and Discussion

    The results shown that Iranian garlic landraces have noticeable diversity of agro-morphological traits, such as weight of 30 seed clove from 21 (Ramhormoz landrace) to 177 g (Hossein Abad landrace), bulb yield from 2059 (Ramhormaoz landrace) to 12090 kg ha-1 (Soolan landrace), Allicin yield from 4.9 (Ramhormoz landrace) to 53 kg ha-1 (Ali Abad landrace), bulb weight from 12.8 (Ramhormoz landrace) to 48 g (Bardsir landrace) and bulb diameter from 28 (Isfahan landrace) to 35 mm (Hossein Abad landrace), which are necessary for breeding programs. It is also confirmed that the adapted landraces from one region may have acceptable production in other regions. According to the present results, landraces from Oromyeh (10866 kg/ha), Arak (9941 kg/ha) and Bardsir (11444kg/ha) produced high yield in Hamedan climatic condition, so that the maximum allicin content were produced by landraces from Ali Abad (53 kg/ha), Maryanaj (51 kg/ha) and Oromyeh (51 kg/ha). Allicin content is calculated based on the bulb yield, and enzymatic pyruvic acid content and its molar relationship with allicin. In the recommended equilibrium of chemical reaction, the alliin coefficient on the one side is two and the coefficients of allicin and pyruvic acid on the other side are two and one, respectively. Bulb weight directly and crown diameter indirectly had highly correlation with bulb yield; and leaf area index, harvest index, bulb diameter and dry weight of plant leaf showed a high correlation with allicin content. The cluster analysis used standardized traits to divide landraces into two main groups as suitable and unsuitable sets to plant under Hamadan climatic conditions.

    Conclusion

    According to the results of the present study, it is recommended that Oromyeh, Soolan and Heidareh landraces can be used as valuable populations for breeding and introducing the suitable cultivars for Hamedan climatic condition through single-bulb selection method. Characteristics with easy and low cost assessment, such as crown diameter, leaf area index, bulb diameter under field condition; and after harvest measurements, such as bulb weight, harvest index, dry weight of plant leaf traits can also be considered as suitable selection indicators in breeding programs. Thirdly, the current study indicated that the cost of preserving garlic germplasm resources can be decreased by reducing the amount of stored materials, without the significant decline in genetic diversity.

    Keywords: Cluster analysis, Garlic, Genetic diversity, Pyruvic acid, Stepwise regression
  • N. Kazemi*, A.A. Habashi, W. Asadi Pages 499-599
    Introduction

    Apple (Malus spp.) is one of the most economically important fruit crop worldwide. This crop is highly affected by various virus infections, leading its considerable devastation and eventually results in yield loss in the whole world. Among effective viruses, Apple stem grooving virus (ASGV), Apple stem pitting virus (ASPV), and Apple chlorotic leaf spot virus (ACLSV) which have been firstly characterized in apple (Malus domestica), play important roles in altering the plant defense mechanism, leading a low performance. According to the references, heat treatments can reduce the movement of virus particles into the apical meristem through inhibiting viral RNA synthesis, so high temperature over a long period is an efficient method for virus elimination. In additions, meristem culture is also a common method to eradicate viruses from horticultural plants. This study was done to produce virus-free (ACLSV, ASPV, ASGV) apple (Malus pumila Mill.) plantlets.

    Materials and Methods

    The effect of different thermotherapy duration (0, 7, 14 and 21 days at 38 °C) and the sizes of apical meristems for meristem culture(less than 0.2 mm, between 0.2 and 0.7 mm and larger than 0.7 mm) was assessed on virus eradication. Our plant material was a genotype of red flesh apple belongs to Budagovsky Bud.9 (Malus+pumila) "Niedzwetzkyana" of Rosaceae family from Shahroud, that has high levels of important phytochemicals like antioxidants, flavonoids and anthocyanins in its cortex, which in addition to attractiveness, creating special healing properties in some disease. At first the presence of ACLSV, ASGV and ASPV were checked in mother samples by ELISA and RT-PCR methods. Then we performed the thermotherapy treatments and later meristems were cultivated and grown in vitro. Regenerated shoots from meristem were tested by ELISA and RT-PCR methods for all three viruses. Samples that were diagnosed virus-free by both techniques, were proliferated, rooted and transferred into the pots to be used for later propagation and establishment of the mother orchard. Results and Discussion ELISA results for the presence or absence of ACLSV, ASGV and ASPV were indeterminate; they were neither negative nor positive for each virus, indicating ELISA was not an accurate method to study virus infections in our samples. Therefore, careful examination of initial infection of samples was performed by RT-PCR. Examining mother samples by RT-PCR showed that all samples were infected by ASG, ASP and ACLS viruses. Results of RT-PCR testing suggested that the number of days in thermotherapy and the size of meristem had a significant effect (P<0.01) on the ACLSV, ASGV and ASPV elimination. Increasing in the duration of the thermotherapy decreased the survival rate of the explants and it was difficult to acquire virus-free explants after 21 days treatments, because all the explants were destroyed in this treatment. The relation between the size of meristem and its survival has been examined in different studies, whose results are consistent with the present research and confirms that increasing the size of the meristem and the number of leaf primordium, will increase the vitality. However, it should be noted that larger meristems are more susceptible to viral contamination. In the previous study, the percentage of ASGV and ACLSV elimination was significantly influenced by the size of the meristem and the duration of the thermotherapy. In the same study, ACLSV, ASGV and ASPV could not be detected in plants grown from meristems smaller than 1 mm after 35 days of thermotherapy. They also showed that virus eradication was happened after shorter period of thermotherapy if smaller meristems were cultured (0.7 mm). The results of our study were consistent with these observations; ACLSV, ASGV and ASPV appeared to be eliminated more frequently from the smallest meristem treatment (smaller than 0.2 mm) compared to the other treatments (0.2-0.7 mm and larger than 0.7 mm). Elimination rates of ACLSV, ASGV and ASPV were different in each case and ASPV-free frequency obtained in regenerated shoots treatment was higher than other viruses (ASGV and ACLSV). This observation was similar to the results of some previous studies. It could be due to differences in the morphology of the viruses and transmission factors in plant cells that affect viruses, which has been investigated and confirmed in previous studies. Meristem culture alone, without other treatments, had very low efficiency in virus elimination from explants. The best results of virus eradication from apple explants was obtained after 14 days of thermotherapy, that we had 44.44%, 11.11% and 88.89% of virus-free samples from ACLSV, ASGV and ASPV, respectively. Generally, our results showed that by increasing the thermotherapy duration, the elimination of all three types of viruses was increased. Many researchers also found that thermotherapy is a very effective treatment in virus eradication.

    Conclusion

    Results showed that increasing the duration of thermotherapy (14 days) and reducing the size of cultivated meristem (smaller than 0.2mm) increased the elimination rate of all three viruses (ACLSV، ASPV، ASGV) from apple explants.

    Keywords: Apple chlorotic leaf spot virus, Apple stem grooving virus, Apple stem pitting virus, ELISA, RT-PCR
  • M. Zamanipour* Pages 511-524
    Introduction

    African marigold is one of the most popular annual flowers in almost of countries. In the most ornamental plants, the flowering and yield depends on the flowering branches that can be increased with pinching. Nitrogen spraying is beneficial on the growth, production and quality improvement of marigold flowers. Among plant growth regulators, auxin and gibberellin play a crucial role in regulating the development processes of African marigold.

    Material and Methods

    This study was carried out in the research greenhouse of Velayat University of Iranshahr in two separate experiments in December 2017. The beds were prepared after mixing the soil with FYM (Farm Yard Manure) and the seeds of African marigold var. "African Double Orange" were sown on the bed at a distance of 10 cm between the row and 2 to 3 cm within the row. Transplanting of plantlets were performed in the 3 to 4 leaves stages in distances of 45 cm×35 cm. Treatments of the first experiment were contains of two stages of pinching (no pinching and pinching in 20 days after planting) and nitrogen spraying in four stages (0%, 2%, 4% and 6%). A factorial experiment was laid out in a complete randomized block design with three replications where each replication was comprised of six plantlets. Treatments of the second experiment consisted of three levels of indole acetic acid (50, 100 and 150 mgL-1) and three levels of gibberellic acid (100, 200 and 300 mgL-1) and distilled water as control. The first stage of auxin and gibberellin spraying, 20 days after planting and the second stage of spraying, was performed 50 days after planting. On the harvest time (when the last flower harvested), measurement factors were contains of vegetative parameters (plant height and diameter, branch number, leaf number and branch length) and reproductive parameters (flower number, flower diameter, flower fresh weight and flower dry weight). Days taken to flowering were counted from transplanting till the date of flower bud opening.

    Result and Discussion

    Analysis of variance showed that pinching and nitrogen spraying had significant effects on plant height and diameter, branch number, leaf number and branch length per plant (P< 0.01). The highest plant height (47.5 cm), plant diameter (2.5 cm) and branch length per plant (21.5 cm) obtained in no pinching with 6% N treatment. This result showed that spraying of nitrogen could increase apical dominance and growth of longitudinal growth of plant. The maximum leaf number (91) and branch number (10) obtained in pinching 20 days after harvesting with 6% N treatment. When the apical dominance is removed, the lowering of IAA (Indol acetic acid) concentration increased the new branches. Increasing in new branches and positive effect of nitrogen spraying created new leaves. Analysis of variance showed that pinching and nitrogen spraying were not effective on flower diameter but had significant effects on flower number, flower fresh weight and flower dry weight. The highest flower number (7) were created in pinching supplemented with 6% N treatment. The highest flower fresh weight (3.60 g) and flower dry weight (0.8 g) were obtained in no pinching and 6% N treatment. Sing et al. (2017) reported that no pinching increased availability of more food material and better allocation of energy pertaining to less number of flowers. Therefore, this action increased the fresh and dry weight of flower. Analysis of variance showed that pinching and nitrogen spraying had significant effects on the number of days to bud flower initiation. The highest number of days to flowering (75 day) were obtained in pinching supplemented with 6% N treatment. Pinching of apical bud prevents from cell division in the lateral meristem. Nitrogen spraying also increased vegetative growth of plant and therefore, resulted in delaying the initiation of flower bud. The quality of seedlings significantly increased with gibberellic acid spraying compared to indole acetic acid. As the concentration increased, seedling quality increased, so that the highest plant height (34.25 cm), the length of each branch (23 cm), Leaf number (42), the number of branches (14), flower diameter (3 cm), dry weight of flower (0.65 g), fresh weight of flower (3.13 g), flower number (6) and the minimum number of days to flowering (31 days) were sprayed with gibberellic acid at a concentration of 300 mgL-1. Increasing the number of flowers by application of gibberellic acid treatments resulted from an increasing in plant height and the number of leaves per plant. Increasing the number of leaves increased the production and accumulation of photosynthetic assimilates, and these assimilates were directed to the flowers and resulted in the production of more flowers.

    Conclusion

    Pinching and Spraying of nitrogen had significant effects on the vegetative and reproductive traits of African marigold. Pinching reduced the apical dominance and induced more lateral branches that ultimately increased the flower number of marigold., Nitrogen spraying was also effective on the growth and quality improvement of marigold flowers and the highest number of flower and fresh and dry weight of flowers induced with application of 6% N. In addition, the application of spraying of gibberellic acid had a direct correlation with height, the number of branches and the diameter of flowers and inversed relationship with the number of days to flowering. The concentration of 300 mgL-1 gibberellic acid had the greatest effect on the growth and reproductive characteristics of African marigold. Since, the African marigold is widely used in the garden and park landscapes, the application of 6% nitrogen or 300 mgL-1 of gibberellic acid is recommended to improve the reproductive characteristics of the flower.

    Keywords: IAA, GA3, Plantlet quality, Urea
  • S.M. Alavi, A. Masoumiasl*, N. Zare, R. Asghari Zakaria, P. Sheikhzade Mosaddegh Pages 525-536
    Introduction

    The main habitat of Chavil, Ferulago angulata, in Iran is Zagros area. This plant has a rejuvenating effect and is used to treat digestive diseases and intestinal worms. Because the different explants show different amounts of callogenesis under the effect of different growth regulators, selection of an optimal explant and suitable plant growth regulators combination has a significant effect on the production of callus and their suspension culture. There is no reports on Ferulago angulata callogenesis and its cell suspension culture. Therefore, this study was designed and implemented to optimize callus production and cell suspension culture in this important medicinal plant.

    Materials and Methods

    Seeds of Chavil were collected from four different habitats in Kohgilooyeh and BoyerAhmad Province in Southwest of Iran include Abenahr, Guayoune, Vezg and Sisakhat. Seedlings obtained from embryo culture were used to prepare the explants. Various explants (leaf, root and stem) were cultured on MS medium supplemented with different concentrations (0, 0.5, 1 and 2 mgl-1) of NAA and BAP. Callus traits were evaluated and from the best culture medium, the best explants and the best PGRs composition for callogenesis of each ecotype were used to cell suspension culture. In order to study the growth rate of cells in suspension culture and plotting the curve of cells growth, two cell density indices and packed cell volume index were evaluated. To determine the cell density index, every 3 days, 10 ml of cell suspension were transferred to the graded falcon and centrifuged at 5000 g for 5 minutes, and the percentage of sediment cells was calculated as the total volume. To determine the packed cell volume index, also every 3 days, 10 ml of culture medium containing cells were transferred to the graded falcon and stored for 30 minutes to precipitate cells and cell masses. Finally, the cell volume was recorded and was calculated as percentage of the total collected medium.

    Results and Discussion

    According to the callogenesis percentage, the best ecotype is Abenahr and best explant is leaf explant. The highest level of NAA is 2 mgl-1, and the best level of BAP is 2 mgl-1, which causes 100 callogenesis percentage. The best medium for cell suspension culture is MS medium containing 2 mgl-1 NAA and 0.5 mgl-1 BAP for callus was obtained from leaf explant of Abenahr ecotype. Along with these plant growth regulators, 2,4-D was used in combination with BAP to form suspension culture. The results also showed that 2 mgl-1 2,4-D plus 0.5 mgl-1 BAP were useful in producing suspensions. The difference between 2,4-D +BAP and NAA + BAP combinations more cell volumes were observed, and cell suspension was created at a faster rate and in less time, which is an advantage in research work. Growth rate of cell suspension originated from the leaf explant was higher than root explant. In terms of culturing cell suspension, the Abenahr ecotype was favorable compared to other ecotypes. During cell suppression culture of Cyperus aromaticus by applying different levels of NAA, cell growth was increased up to 3 weeks after application, and then decreased. By applying 2,4-D, cell growth also increased until the third week, and after the third week, cell growth declined, which was very low growth rate compared with the NAA. In cell suspension culture of sugar beet, using 2,4-D was much more effective than NAA on all explants. In the present study, 2,4-D was also more effective than NAA for cell suspension culture of Chavil.

    Conclusion

    In general, the Abenahr was the best ecotype among of investigated. The explants in both callus culture and the suspension culture, and the best combination of plant growth regulator in both culture was 2 mgl-1 NAA plus 0.5 mgl-1 BAP.

    Keywords: Callus, Genotype, PCV, Plant growth regulator, SCV
  • P. Dehkhodaei, S. Reezi, M. Ghasemei Ghehsareh* Pages 537-548
    Introduction

    The light is not only a photosynthesis energy source but also is a plant growth and development stimulation from germination to flower initiation process. The light quality plays an important role in all steps of growth process particularly in photosynthesis and morphological properties of plant species. Studies have reported that, LEDs present the maximum PAR efficiency among artificial lighting systems. LED lamps contribute to energy saving by 75 and 30 percent as compared to incandescent light bulbs and fluorescent lamps, respectively. LEDs emit blue, red, orange, yellow, green, red, and infrared light and can be used as a hybrid spectrum. For these reasons, an experiment was conducted to investigate the effect of quality of LED light on morphological and physiological characteristics of Solenostemon and petunia.

    Materials and Methods

    Research experiments were conducted on Solenostemon escutellariodes ‘Wizard Scarlet’ and Petunia × hybrida ‘Scarlet Eye’ in a completely randomized design with three treatments of different light quality and 10 replications. The seeds were planted in 105-cell seedling trays and in a mixture of 70% peat moss and 30% perlite. Seedlings were grown in natural greenhouse (control) and LED (50% blue + 50% red and 100% white light). The light intensity was 60 ± 5 µmol.m-2.s-1 for LEDs and the daily greenhouse cavity was 5 ± 14 µmol.m-2.d-1. Since the main goal of the study was to compare the effect of LED light quality with sunlight in conventional greenhouse conditions. The LED treatments were applied from the germination until seedling production stage in a growth chamber with the light/dark regime of 18/6 hours, 23±1°C temperature, and 50±5% relative humidity. While, their tray in the greenhouse with 55±5 mol.m-2.d-1 DLI, 21±5°C average daily temperature and 50±5% relative humidity (Data logger 8808 temp. + RH) were regarded as the control treatment. After eight weeks, the morphological and physiological traits were recorded. Finally, the data were statistically analyzed with SAS (9.4) software package, and the means were compared by LSD test at p < 0.05 level.

    Results and Discussion

    At the end of study, some morphological and physiological traits were evaluated. The results showed that the leaf area average (3.63 cm2), height (5.04 cm), internode length (18.62 mm), stem diameter (2.52 mm), shoot fresh weight (1.74 g) and chlorophyll fluorescence (0.83) of Solenostemon seedlings in 100% white light were more than 50% blue + 50% light green and greenhouse light. Also, the highest leaf area temperature (27.4 ° C), total chlorophyll (0.8 mg g-1 fw) and carotenoids (2.229 mg g-1 fw) were related to control treatment and the highest number of leaves (21), shoot dry weight (0.165 g), fresh (1.65 g) and root dry weight (0.114 g) were observed in 50% blue + 50% red light composition. According to the results of petunia, white LED light increased leaf area (1.74 cm2), shoot fresh weight (0.51 g) and root dry weight (0.03 g). The leaf surface temperature (26 oC) and total chlorophyll content (1.44 mg g-1 fw) in the control group were higher than the other treatments, and the carotenoids content (3.12 mg g-1 fw) was obtained in 50% red + 50% blue LED. The leaf surface temperature and total chlorophyll content in greenhouse light were higher than other treatments and the highest carotenoids (3.119 mg / g) were obtained from 50% red + 50% blue LED.

    Conclusion

    Light quality has a great impact on the growth and development of plants and is a powerful tool for controlling various processes .The results showed that different light sources with different qualities had different effects on different plant species and that LEDs could replace natural light in growing chambers and in areas where light was insufficient. Due to the increased quality and reduced transplant losses in the transplant phase and their low utilization, they are justified in terms of production costs. It should be noted that in addition to light; temperature, humidity and other environmental factors are also effective in transplant production. Based on the measured indices in Solenostemon, it can be concluded that the red + blue LED light transplanting stage is superior to other treatments. In petunia, white LED treatment caused the highest shoot and fresh and dry weights of roots and leaves respectively. Like Solenostemon, the highest chlorophyll and leaf surface temperatures were related to the control treatment and was superior to the petunia transplant in terms of the sum of measured indices of white light treatment.

    Keywords: Leaf chlorophyll fluorescence, Light quality, Morphology, Transplant
  • A. Moshashaei, M.M. Jowkar* Pages 549-565
    Introduction

    Dianthus barbatus L. from Caryophyllaceae is an important herbaceous landscape ornamental plant in the temperate regions throughout the world. Although this ornamental plant is used as bedding plant, it also has other extensive applications such as cut flower and pot plant. Besides, in some countries the leaves of Dianthus barbatus are used for treatment of digestive disorders in their traditional medicine. Landscape and its quality, plays an essential role in human life. Nowadays, maintenance and development of urban landscape in many regions such as Iran is facing major challenging issues. Soil and water salinity are the most important challenges of urban landscape. Considering the challenges which landscape development faces in many parts of the country and the importance of this plant in landscape, the effect of salinity on D. barbatus during in vitro condition was studied in order to obtain some basic physiological information for future stress studies on this plant.

    Materials and Methods

    In vitro sterile plants were initially obtained from seeds by culturing them on basal MS medium. Calli were obtained by culturing leaf, stem and root explants on previously reported callogenesis optimization medium which was MS medium supplemented with 2µM BAP + 6µM NAA. Calli proliferation was then conducted on the same media. Proliferated calli with same size were subjected to MS medium supplemented with the mentioned plant growth regulators and various NaCl concentrations (0, 1, 3, 6, 9, 12 gl-1) in order to induce salinity stress. Various physiological aspects and stress related mechanisms such as growth, osmo-protection and oxidative stress mechanisms were consequently studied in the stressed Dianthus barbatus calli. Studied growth features were calli growth, final growth rate, fresh and dry weight. Total protein content, glycine-betaine content, and the activity of betaine aldehyde dehydrogenase (BAD) were studied as a part of osmo-protection mechanism. On the other hand, the activity of free radical scavenger enzymes such as sccccuperoxide dismutase (SOD), catalase (CAT), peroxidase (POX) and malondialdehyde damage biomarker content were studied in stressed calli as a part of oxidative stress mechanism. Total protein was measured using Bradford method. Glycine-betaine content was measured using spectroscopy method by KI-I2 reaction agent. The activity of betaine aldehyde dehydrogenase was evaluated using spectroscopy method by reduction with NAD+. SOD activity was measured using spectroscopy method and formation as an indicator. CAT activity was measured using spectroscopy method by measuring hydrogen peroxide breakdown. POX activity was measured using spectroscopy method by nitro-blue tetrazolium reduction activity. Malondialdehyde content was also measured by using spectroscopy method.

    Results and Discussion

    Results indicated that by increasing salinity level, calli growth and consequently fresh and dry weight reduced. Calli growth was dramatically reduced as salinity level reached above 1 gl-1. This slow growth was also observed at high stress levels such as 12 gl-1. By salinity increment, total protein content of the stressed calli significantly reduced. On the other hand, BAD activity increased as a result of salinity stress and consequently glycine-betaine content also increased significantly. The highest level of glycine-betaine content and also the activity of BAD activity were observed in 12gl-1 NaCl treated calli. A significant correlation between glycine-betaine content and BAD activity was observed. The activity of free radical scavenger enzymes such as SOD, CAT, POX also significantly increased by salinity stress increment. The highest activity of the mentioned free radical scavenger enzymes were observed in 12 gl-1 NaCl treated calli. CAT had the most activity among the mentioned studied free radical scavenger enzymes. The highest increase in POX activity was half of the control and also half of the activity increment of other studied free radical scavenger enzymes. Although the activity of free radical scavenger enzymes increased, the level of malondialdehyde damage biomarker also increased as the result of salinity stress increment. The highest amount of malondialdehyde content was seen in the 12gl-1 NaCl treated calli. A significant correlation between malondialdehyde content and the activity of free radical scavenger enzymes was observed.

    Conclusion

    In general, the calli of Dianthus barbatus cultured in vitro are sensitive to salinity stress induced by more than 1 gl-1 NaCl; indicating the stress tolerance threshold. The tolerance and resistance mechanisms performed well under stress condition and the stress calli resisted to salinity up to 12 gl-1 NaCl and survived without chlorosis.

    Keywords: Betaine aldehyde dehydrogenase, Catalase, Glycine-betaine, Malondialdehyde, Superoxide dismutase