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بیوتکنولوژی کشاورزی - سال چهارم شماره 2 (پیاپی 8، پاییز و زمستان 1391)

مجله بیوتکنولوژی کشاورزی
سال چهارم شماره 2 (پیاپی 8، پاییز و زمستان 1391)

  • تاریخ انتشار: 1391/10/10
  • تعداد عناوین: 9
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  • Asghari F., Hossieni B., Hassani A., Asghari M.R., Farrokhi J Page 1
    Basil (Ocimum basilicum) is an annual, herbaceous plants belonging to the family of lamiaceae. Basil is used to as a herb, a spice as well as fresh vegetables. The present study aimed to investigate the effects of different concentrations of growth regulators such as auxin and cytokinin to identify the best hormonal concentration to obtain the highest yield of in vitro produced plantlets using nodal explants. Experiment was performed in factorial completely randomized design, using the basic MS. In this experiment the effects of different concentrations of BAP(0, 2/5, 5 and 7/5 mg/l) and indol acetic acid (0, 0/1 and 0/5 mg/l) was studied alone or together. Regenetaion percent, number of regeneration, vitrification and rooting evaluated. The results showed that the highest percentage and number of regeneration obtained in the medium included 2/5 mg/l BAP concentration and 0 mg/l IAA, respectively.Vitrification was increased with increasing BAP concentration, maximum rooting was observed in the medium containing 0/5 mg/l IAA.
    Keywords: Basil, Nodal, Regeneration, Vitrification, In vitro
  • Taghizadeh N., Farsi M., Pakdin Parizy A Page 17
    Actinomyces bacteria are the best parser microrganisms in environment and agriculture.These bacteria are frequent in compost and produce antibiotics & extracellular enzymes. In this study, some strains of actinomycetes were separated from different steps in composting process. After that, morphological, biochemical and molecular tests were performed for identification of them. Carboxy Methyl Cellulase assay was applied for endoglucanase activity. For optimizing conditions different treatments including pHs: 5.7, 6, 6.5, 7, 7/5, temperatures 45-65°C and times 30, 60 and 90 minutes were used. Optimum temperature for high endoglucanase activity was 60°C and isolate No.19 with 0.289 unit/ml showed the highest endoglucanase activity in this temperature. According to molecular result obtained from RFLP of 16SrRNA, most of actinomyces isolates were from streptomyces genus.
    Keywords: Actinomyces, Carboxy methyl cellulase, Enzyme activity
  • Mehdi Shirazi, Javad Mozafari, Farshad Rakhshandehroo, Masood Shams, Bakhsh Page 29
    Tomato yellow leaf curl virus (TYLCV) is considered as one of the most important tomato pathogens in Iran. During the years 2007 to 2009, a total number of 284 symptomatic samples were collected from tomato fields and greenhouses from eight provinces of the country. The PCR test showed that 153 tomato samples from six provinces were infected by TYLCV. This is the first report of TYLCV in Rezvan (Hormozgan), Dashtestan (Bushehr),Dezful (Khuzestan), Pakdasht and Islamshahr (Tehran) regions. Based on geographical region and symptoms, nine isolates from six provinces were selected for further studies.Comparison of an amplified DNA fragment of 670 bp, including a part of coat protein and movement protein genes of the virus, with those of previously reported isolates from Iran and closely related isolates available in GenBank, suggested that Iranian isolates examined in this study were grouped into two major groups. Isolates collected from Hormozgan, Khuzestan and Kerman provinces were grouped with other Iranian isolates including TYLCV-Ir2 and TYLCV-Kahnooj and an isolate from Oman. Whereas, isolates collected from Bushehr, Fars, and Tehran were placed close to the Iranian isolate of TYLCV-Abadeh and isolates from occupied Palestine and Egypt. Based on our study, TYLCV has a high genetic diversity in Iran.
    Keywords: Geminivirus, TYLCV, PCR, Solanum lycopersicom
  • Ghasemkhani M., Mohammadi, Nejad G Page 43
    In order to identify the responsible QTLs attributed to salinity tolerance at seedling and reproductively stages in rice and detection of the ratio of each QTL in phenotypic variation, eighty RILs derived from IR29/Pokkali cross were used. The Sodium (Na) and Potassium (K) concentration, Na/K ratio at seedling stage and height, number and grain weight, number of panicle, number of tillers, fertility and biomass of rice in salinity stress at reproductive stage were assessed as phenotypic traits. Using 81 polymorphic SSR markers, a major QTL related to seedling stage tolerance was detected on the opposite arm of Saltol QTL in chromosome 1.Moreover, a major effected QTL for Na, K and their ratio was found on chromosomes 1 and 10 which totally controlled the tolerance at seedling stage. For tolerance to salinity at reproductive stage; major QTLs detected on chromosome 7, 9 and 10 for Reduction of Filled Grain Weight and Reduction of Biomass Weight. Multi trait QTL which were located on chromosome 10 had the same location with the QTL mapped for seedling stage tolerance, so it can be a good candidate for marker assisted breeding. It can be expected to achieve to the high tolerant genotypes through QTL pyramiding of saltol and chromosomal region of 10 together.
    Keywords: QTL mapping, Recombination inbred lines (RILs), Rice, Salinity tolerance
  • Kabirnataj S., Zolala J., Nematzadeh G. A., Shokri E Page 61
    Agrobacterium rhizogenes induced hairy roots gain of the potential of production and accumulation of plant secondary metabolites. In recent decades, many researchers have focused on the biosynthesis of valuable secondary metabolites in hairy roots, because genetically modified roots produce such compounds with more genetic and biosynthetic stability than cell suspension cultures. Chicory (Cichorium intybus L.) is a biennial or perennial medicinal plant from Asteraceae. The plant contains many important metabolites including inulin, scoline, coumarin, vitamins, flavonoids and aromatics. In this study, with the aim of optimizing hairy root culture establishment, cotyledons of Chicory plants inoculated with different strains of Agrobacterium rhizogenes were cultured on different plant tissue culture media. Numbers of hairy roots produced in different experiments were recorded and molecular confirmation of hairy root clones was performed through specific amplification of rolB and rolC genes in PCR reactions. Based on the efficiency of hairy root induction in different experiments, the best combination of bacterial strain-culture medium for establishing hairy root cultures of Chicory were determined. Agrobacterium rhizogenes strains A4, A13 and 15834 induced hairy roots in Chicory and MS medium was considered as the best culture medium for developing and establishment of hairy root cultures.
    Keywords: Agrobacterium rhizogenes, hairy root, Chicory, secondary metabolites
  • Mehrabani B., Nazeri S., Piri K Page 77
    Many reports have demonstrated that many medicinal valuable secondary metabolites such as Phenolic compounds can be produced and accumulated by in plants in vitro culture. Polyphenols or phenolic compounds have potentially antioxidant activity. In past few years, pharmacological studies have confirmed that components and extracts of plants belong to the genus Stachys exert significant antibacterial, antitoxidant, antinephritic, antihepatitis, hypotencive and anxiety effects. Stachys lavandulifulia Vahi, popularly named "Chai Koohi" is widely distributed in different regions of Iran. In this study, effect of different concentrations of 2,4-D, on callus induction and effect of two elicitors (metyl jasmonate and salisylic acid) in different time points(2, 10 and 20 days after culture) on the level of total phenol were evaluated in callus. The results showed that, the best concentration of callus induction and growth was at the 1 and 2 mg/l 2,4-D. Also the effects of elicitors were significant in total phenol production.Cultivation time indicate significant difference on total phenol production, whereas interaction between time and elisitore was not significant. The highest total phenol concentration was found in the treatment with 10 μm of metyl jasmonate, after 10 days of culture.
    Keywords: Stachys lavandulifolia Vahi., callus culture, total phenol, Elicitors
  • Hashemi F., Shobbar Z.S., Majidi M.M Page 89
    ABA-Insensitive3/Viviparous1 (ABI3/VP1) genes encode transcription factors involved in ABA signal transduction. The objective of this research is the functional analysis of OsVP1 gene - ortholog of the maize VP1 and Arabidopsis ABI3 in rice- through studying related mutant lines. Therefore, characteristics of the mutants (T-DNA inserted lines in the promoter or coding region of OsVP1 gene) and wild type rice plants were compared in the vegetative and reproductive stages to clarify the role of this gene. As well, the comparative analysis of the wild and mutant lines (T-DNA inserted in the promoter of OsVP1 gene) response to abscisic acid treatment and abiotic stresses were done, in addition to the expression analysis of OsVP1 gene and the candidate downstream genes in 12 day seedlings using real time PCR.Mutant plants in OsVP1 coding region were pale, subtle and short compared to the wild type,in both vegetative and reproductive growth stages with no yield. Plant height of mutants in OsVP1 promoter region were less than the wild type. Panicle emergence and grain filling were also delayed compared to the wild type. Abiotic stress induced growth reduction in seedlings of both wild type and mutant plants. Gene expression analysis was revealed that OsVP1 transcripts levels have been increased under drought, salinity, cold and ABA treatments compared to the control condition. Also, OsVP1 has up regulated considerably in mutant plant at OsVP1 promoter region, which were expected because the T-DNA insertion carries several enhancers from 35S promoter of cauliflower mosaic virus. Therefore, it seems OsVP1 encodes a transcription factor which can play a role in ABA signal transduction pathway and stress induced growth arrest.
    Keywords: Rice (Oryza sativa), OsVP1 (OsViviparous1), abiotic stress, ABA (Abscisic acid)
  • Honarvar M., Moradi, Shahrebabak H., Sadeghi M., Behzadi Sh, Mohamadabadi M.R Page 103
    The Insulin-Like Growth Factor I (IGF-I) Gene has a similar structure with Insulin hormone. That has an important functional in growth and development different tissue and transcribe. The IGF-I gene coded this hormone which located on Chromosome five which has six exons and five introns. The single nucleotide polymorphism was occurred in IGF-I gene and that associated with carcass traits in Zel (tailed) and Lori-Bakhtiari (fat-tailed) breed sheep with using PCR-SSCP marker. For this purpose, 4 mL of total blood was collected from the left jugular vein using vacuum tubes from Zel and Lori-Bakhtiari sheep breeds. Genomic DNA was purified from 1000-μl of blood samples that using the salting-out procedure and amplified region is located in the 5’ flanking region of the ovine IGF-I Gene. The IGF-I gene 5’flanking fragment was amplified which investigated with Single-Strand Conformational Polymorphism (SSCP) method for genotyping. Electrophoresis of PCR products were done on acrylamide gel that observed the polymorphic in this region. The frequency of patterns in Zel and Lori-Bakhtiari breeds were A (37.40%), B (41.98%), C (2.29%), D (18.32%) and A (58%), B (32%), C (4%), D (6%) respectively. Finally, for analysis of some traits was used SAS program. No significant associations of the SNPs in the 5’ flanking region of the ovine IGF-I Gene were observed for any trait.
    Keywords: 5 flanking region of the ovine IGF, I Gene, SNP, PCR, SSCP, Ovine
  • Hassanzadeh Davarani F., Rezaee S., Mahmoudi S.B., Norouzi P., Safarnejad M.R., Safarpour H Page 117
    In order to design a precise diagnostic and quantitative system to detect of P. betae in sugar beet roots, three procedures including microscopical, serological and molecular methods were compared on a sugar beet cultivar susceptible to causal agent and vector of rhizomania disease.At first, susceptible cultivar Regina was cultivated in infested soil in greenhouse condition. After 5 weeks, roots of plants were stained with fuchsin- acid- lactophenol solution. Quantitative ELISA test was optimized with specific antiserum against recombinant protein glutathione stransferase (GST) of P. betae. Running ELISA test, extracts of health and infected plants were prepared and used in DAS- ELISA test. Presence of P. betae in roots of plants was detected with amplification of rDNA of Plasmodiophoromycetes and P. betae species, simultaneously. Existence of cystosori of P. betae was observed in all of the plants planting in infested soil.Results of ELISA test were distinguished healthy plants from infected plants. Average of optical density at 405 nanometer for healthy plants, infected plants, protein PBS (negative control) and GST (positive control) was 0.126, 0.75, 0.11 and 2.45 respectively. Duplex PCR method was amplified two fragments of 454bp and 170bp in infected samples relating to rDNA region and specific region of the P. betae, respectively. Based on the results, it seems that, rapid detectionand identification of P. betae in sugar beet and other hosts of the vector could be done on the basis of PCR method. Evaluating of resistance germplasm of sugar beet to P. betae could be recognized using quantitative ELISA tests.
    Keywords: Rhizomania, Glutathione, S, transferase, ELISA, PCR, ITS