نشریه تحقیقات ژنتیک و اصلاح گیاهان مرتعی و جنگلی ایران
سال بیست و هشتم شماره 1 (پیاپی 55، بهار و تابستان 1399)

The genus Tulipa belongs to the Liliaceae family and is one of the most important ornamental bulbous plants, which has been cultivated all over the world to produce cut flowers, potted plants and use for landscaping. In this study, the genetic diversity of 46 accessions belonging to 9 species of Tulipa was investigated using 19 CDDP primers. A total of 265 bands were amplified, all of which were polymorphic. The average number of amplified bands per primer was 13.9. Primer 5 (ABP1-1) with the best marker parameters was introduced as the best primer for investigating genetic diversity in this study. Calculation of genetic diversity parameters showed the greatest diversity among T. systola species. Results of analysis of molecular variance (AMOVA) showed that within species diversity (77%) was higher than between species diversity (23%). Evaluation of distance matrix based on the Dice similarity coefficient of the studied Tulipa species showed that the highest and the lowest distance were related to T. armena and T. clusiana (0.684) and T. systola and T. schrenkii (0.226), respectively. Due to the higher contribution of within species variance than to between species, grouping of the species based on cluster analysis did not fully correspond to those in the principal coordinate analysis. However, different accessions belonging to T. micheliana, T. lehmanniana, and T. clusiana were well separated. The present study showed that the remarkable efficacy of the CDDP marker  in relation to the genus Tulipa requires a comprehensive study of the genetic diversity of this genus in different distribution points of Iran.

Tall fescue (Festuca arundinacea) is one of the most important forage plants in the Gramineae family. Due to its resistance to salinity, drought, heavy metals, excessive grazing, as well as preventing soil erosion, tall fescue can be an appropriate choice for large-scale cultivation in Iran because of recent droughts and declined rangelands area. In the present study, the genetic diversity of 90 tall fescue populations was evaluated using 10 morphological and yield related traits including; canopy diameter, days to flowering, days to pollination, plant height, panicle length, collar diameter, number of stems, leaf length, dry forage yield and seed yield in Basmanj region, Tabriz (East Azerbaijan), Iran, during 2015 to 2017. According to the mean of the populations,there were significant differences (p < 0.01) among populations for all of studied traits except for canopy diameter. The results of correlation coefficients revealed that the number of stems, peduncle length, collar diameter and canopy area had a positive and significant correlation with dry forage and seed weight. The heritability of all studied traits was moderate to high. The highest heritability was related to seed yield, dry forage yield and flowering date, respectively. Based on the results of cluster and principal component analyses, studied tall fescue populations were divided into two main groups of ornamental and forage and four clusters (clusters I and II including early- and late-maturing forage populations and clusters III and IV including ornamental populations). The cluster IV included ornamental populations showed the highest difference and genetic distance with cluster I including forage populations and then more heterosis is expected by realizing cross between them. The results showed that the populations investigated in this study had the necessary diversity to running tall fescue breeding programs and can be potentially used in future breeding programs of this plant.

Haloxylon species are adaptable plants that have had satisfactorily results in sand fixation programs. To investigate the genetic potential of progeny of the superior Haloxylon aphyllum genotypes, an experiment with 19 genotypes was performed using a randomized complete block design with three replications at Hossein Abad sand dune stabilization station, Qom, Iran in 2013,. During five years, various morphological traits were evaluated. Combined analysis of variance over years for all traits showed that there was a significant difference between many of  traits among genotypes in all years. Tree height, main trunk diameter and crown diameter showed a positive and significant correlation with each other in all years. Height in the fifth year showed a positive and significant correlation with the height and diameter of the main trunk in the preceding years. The first five factors resulting from principal component analysis with specific values higher than one were entered into factor analysis as the main factors. The first to fifth factors were identified as "crown diameter", "height", "survival and seedling survival", "pest and disease damage " and "powdery mildew infection", respectively. Based on the mean comparison of the four groups obtained from cluster analysis, the groups showed significant differences in all the traits at p < 0.01 or p£0.05. The genotypes in cluster2 were superior to other groups in terms of height, main trunk diameter and crown diameter in all years. This group included genotypes of primary origins of Yazd 4 and Semnan 3, 5 and 6.

Drought stress is one of the most important factors limiting the survival and growth of plant in different parts of Iran. In this study, the morphological characteristics of four perennial chamomiles species, Anthemis tinctoria, Matricaria recutita, Tripleurospermum sevanense and Tanacetum parthenium, were evaluated in rainfed and greenhouse conditions in Alborz research field, Research Institute of Forests and Rangelands, Iran. The results showed that M. recutita had the highest plant crown surface, height, flower number,plant dry weight; and early flowering date in rainfed conditions compared to the other three species. In the greenhouse experiment in order to study the physiological mechanisms of response to water deficit, the four species were subjected to four drought treatments including well-watered (control), mild drought stress (75% of field capacity (FC)), moderate drought stress (55% of FC), and severe drought stress (35% of FC). Changes in compatible solutes (proline and soluble sugars), relative water content (RWC), antioxidant enzymes (peroxidase and polyphenol oxidase) and pigments of stress plants were investigated. The result of the physiological response of the chamomile species in greenhouse condition showed that M. recutita was the most tolerant species to severe drought stress conditions due to its the higher osmotic regulation capacity, while the other three species A. tinctoria, T. servanes and Tanacetum parthenium showed good tolerant to moderate stress conditions.

Tecomella undulata is one of endemic and endangered plant species in Iran. It is belong to  Bignoniaceae family grown in southern of Iran. Due to the specific characteristics of this plant in various aspects of medicine, ornamentation and natural resources, its mass production through tissue culture and creation of identical plants seems necessary. Effects of growth regulators on plant growth indices such as number of buds and branches, shoot length, branch greenness and rooting were studied by bud explants under in vitro conditions. The optimal aseptic technique was obtained by immersing the buds in 0.1% mercuric chloride solution for 4 minutes in spring. The best shooting was observed in MS basal medium containing BA 1 treatment and K0.5 mg/L. This treatment was the best selection for this phase with the average shoot growth of 4.9 cm and shoot length of 7.78 cm. At the rooting stage, despite the numerous treatments of the medium and hormones (various concentrations of auxin hormone), the onset of rooting was only observed in 1/2 MS IBA2.

Senna (Cassia angustifolia Vahl.), known as an important medicinal plant, belongs to the Caesalpiniaceae family used in the treatment of some diseases such as skin infections, constipation, and intestinal worms. The aim of this study was to optimize the micropropagation of this plant. For this purpose, senna seeds were first disinfected and cultured in MS medium and the nodular stem parts were used as explants. The treatments were used to optimize the propagation medium supplemented with different concentrations of cytokinins, BA and Kin (0, 1, 2, and 4 mgL-1) in combination with NAA (0 and 0.25 mgL-1). After finding the best type and concentration of cytokinin for propagation (cytokinin BA), this growth regulator was tested in combination with various auxins to optimize the proliferation of the explants. In this experiment, BA was evaluated at three concentrations (0.5, 1, and 2 mgL-1), in combination with NAA, IAA, and IBA at two concentrations (0 and 0.25 mgL-1).  The propagated shoots (at least 3 cm long) were then exposed to two types of auxin NAA and IBA at different concentrations (0, 0.3, 0.6, and 0.9 mgL-1) to induce rooting. The results indicated that 1 mgL-1 BA treatment in combination with 0.25 mgL-1 NAA induced the maximum shoot proliferation (2.93 shoots per explant) and leaf number (21.6 leaves per shoot). Finally, NAA with 0.25 mg / l BA treatment at the concentration of 1 mgL-1 in combination with 0.25 mgL-1 NAA, was selected as the best treatment for the propagation of the explants. Maximum rooting induction obtained in 0.3 mgL-1 IBA treatment which was not significantly different from 0.3 mgL-1 NAA. Moreover, application of IBA at the concentration of 0.9 mgL-1 caused the higher mean values of root number and root length, so that it increased by 172.22 and 73.41% of the mentioned traits compared to the control, respectively. The results of this study can be used in micropropagation and extensive proliferation of this plant.

Purple coneflower (Echinacea purpurea (L.) Moench) is a plant species of high medical value. Somatic embryogenesis is considered to be the most efficient micropropagation technique. Considering the importance of this plant, the current study has been performed with the aim of optimizing somatic embryogenic of that. Embryogenic callus induction and embryogenesis were investigated using different concentrations of plant growth regulations including Benzyl adenine (BA) in three levels (3, 4 and 5 mg/l), Naphtalene Acetic Acid (NAA) at three levels (0.1, 0.2 and 0.5 mg/l) with or without 1 g/l active charchoal, 50 ml/l coconut milkand 50 mg/l casein hydrolysate in leaf explants in MS culture medium in a completely randomized design with three replications. Induction of embryogenesis occurred indirectly with callus formation. Embryogenic calli were dark green, coherent with faster growth. The highest percentage of embryogenic callus (100 %) and also regeneration of callus (plantlents) were obtained in combination of BA and NAA with active charcoal, coconut milk and casein hydrolysate treatmens. Whereas the combination treatments of plant growth regulators without coconut milk and casein hydrolysate failed to produce somatic embryos. The maximum number of shoots (43.3 shoots) was observed in 3 mg/l BA + 0.1 mg/l NAA + 1 mg/l activate charcoal + 50 ml/l coconut milk + 50 mg/l casein hydrolysate treatment.

The genus Astragalus of the Leguminosaefamily is one of the largest genera among flowering plants. Astragalus verus is one of the suitable species for tragacanth production. The purpose of this study was to investigate the effect of different hormonal compounds on callus induction of A. verus different explants . Callogenesis from root, hypocotyl and cotyledon explants was studied in MS basal medium supplemented with different concentrations of BAP, 2,4-D and NAA. The highest percentage of callogenesis was obtained in 2,4-D (0.5 mgl-1) treatment with a mean of 56% and the lowest percentage of callogenesis was in 0.5 mg -1L NAA treatment with a mean of 4%. The highest callogenesis percentage was achieved from root explant with an average of 67%. The highest and lowest callus dimensions were obtained in treatment of 9 mgl-1NAA and 9 mgl-12,4-D with average values of 62 and 0.1 mm3 , respectively. The highest fresh and dry weights of calli were obtained with an average of 0.148 and 0.022 g of root explant in 2,4-D treatment, respectively. The separability of calli in medium containing 2,4-D was better than NAA. The results of this study can be used in indirect regeneration studies, gene transfer, and cell suspension culture to produce secondary metabolites such as tragacanth under in vitro conditions.

Tamarix aphylla is used as a windbreaker, shade tree and ornamental plant in gardens and urban landscape. The browning phenomenon is an important inhibitory factor in various stages such as disinfection and regeneration of Tamarix aphylla. To optimize the disinfection conditions and control the browning process of the explants, experiments were carried out with disinfectants and eugenol compound to disinfect the current and annual stem explants. Then, callus induction was studied in a medium complemented by two types of auxins (2,4-D and NAA). In the third experiment, the optimization of shoot induction medium was investigated on current and one year old shoots explants. The best disinfection treatment to control fungal and bacterial contamination and phenol secretion from the current explants was obtained using a 60-second ethanol application combined with 10 minutes of 20% sodium hypochlorite disinfection. Spraying of donor plants with thiophanate methyl fungicide was also able to reduce fungal contamination by almost 50% compared to the control. Application of 0.01% eugenol in the medium by injection reduced the contamination and phenol secretion from the explants. The highest survival and callus induction was acquired at the concentration of 1 mgL-1 2, 4-D, in the shortest time. Shoot induction in the explants of current and one year old shoots was obtained in the culture medium containing 2 mgL-1 BA and 0.5 mgL-1 IBA In conclusion, the application of younger plant materials, spraying of donor plant with fungicides and the use of 0.01% eugenol in the medium can reduce the browning phenomenon and proper control of contamination and finally could provide micropropagation of Tamarix aphylla shrubs using appropriate growth regulators

Jashir (Prangos spp.) is forage, medicinal and indigenous herb from some parts of Iran that is used in traditional medicine in the treatment of many diseases. This study was conducted to investigate the effect of altitude on morphological traits and essential oil compounds of Prangos ferulacea (L.) Lindl. at the flowering stage in Kohgiluyeh and Boyer-Ahmad province, Iran. For this purpose, 10 plants were selected from each of three altitudes (2000-1600, 2400-2000 and 2800-2400 m above sea level). Measurements were made for 30 morphological traits and essential oil content and compositions of the plants in each altitude. Result of one-way ANOVA showed a significant difference among altitudes classes (p < /em><0.01) for most of traits. The results showed that with increasing altitude, the number of branches, the number of leaves per plant and the number of primary leaflets decreased and the number of umbrellas per plant, leaf length, internode length, fresh and dry weight of plant increased. Significant correlations were observed between some important morphological traits such as (leaf number and umbrellas number, leaf length, internode length, fresh and dry weights) with the essential oil content. Path coefficients analysis of morphological traits showed that altitude had the greatest effect on the canopy diameter (P = 0.733) and leaf width (P= 0.857). In total, 22 compounds of essential oil were identified in three different altitude classes. The major compounds were Δ-3-carene, α-pinene, (Z)-β-ocimene, myrcene, β-phellandrene, and α-terpinolene. Altitude had a negative effect on the essential oil yield. Overall, the results of this study showed that the many of the studied traits in P. ferulacea were strongly affected by altitude and there were significant differences among the three altitude classes. Finally, the observed diversity can be related to the role of altitude and the genetic structure of the populations, which can also be considered as a kind of adaptation and physiological response of Jashir to environmental conditions.

Ferulago angulata as a medicinal plant belongs to the Apiaceae family, and propagated by seed. But, seed dormancy is a main limiting factor in its propagation. To evaluate the efficiency of in vitro culture as well as the combination of culture medium and explant type on breaking seeds dormancy of Ferulago angulata, a factorial experiment was conducted based on a completely randomized design with five replications. The treatments consisted of four ecotypes (Abenahr, Gauonhe, Vezg, Sisakht) from Kohgiluyeh and Boyer-Ahmad province, Iran and three concentrations of Murashige and Skoog basal medium (1/4, 1/2 and 1MS) and explant type (seed or embryo). Collected seeds were surface sterilized and then were planted as intact seeds, and isolated embryos in the above mentioned media. In this experiment, isolated embryos germinated after 3 to 6 days, depending on the ecotype. However, no germination was observed in the intact seeds in the used media. The results of embryo culture showed that culture medium and ecotype significantly affected all measured seedling traits. Mean comparison indicated that there were significant differences between different media and 1/4MS medium was superior to the other two culture media in terms of embryo germination percentage and seedling growth. There also were significant differences between different ecotypes, and Gauonhe ecotype showed higher embryo germination percentage and seedling growth than other ecotypes. So, in vitro culture of Ferulago angulata seed embryos on 1/4MS medium provides an effective method to overcome seed dormancy. Seedlings resulting from germination and embryo growth were successfully transferred to pots and placed in natural environment. According to the results of this study, germination inhibitors in F. angulata are located in the tissues surrounding the embryo and the embryo would not be able to germinate until the inhibitory factors are removed. Therefore, isolation and in vitro culture of the embryo provides a suitable approach to overcome seed dormancy in this plant, especially for laboratory studies.

Seed dormancy is one of the most complex processes in response to adverse environmental conditions that has been reported in wild pear seeds. This research was conducted to investigate stratification and gibberellic acid treatments on wild pear seed germination from two intermediate and high altitude origins (2150 and 2450 m above sea level, respectively) located in the southeastern forests of Yasuj, Iran. For this purpose, seeds of wild pear were collected from two different provenances and then treated with gibberellic acid (0 and 1500 mg/l) for 48 hours followed by stratification treatment (at zero, 30, 60 and 90 days) at 4°C. The germination test was performed through a factorial experiment with two factors of seed origin at two levels and dormancy treatments at eight levels in completely randomized design with four replications in the seed laboratory of Yasouj University. The highest germination percentage (100%), germination rate (49.75 seed/day) and seed vigor index (17.9) were observed in seeds collected from high altitude and treated with cold-stratification (90 days) + gibberellic acid (1500 ppm). The results showed that the germination percentage of intermediate seeds with gibberellic acid + cold stratification treatment for 90 days (88%) was lower than the germination percentage of high altitude seeds treated with gibberellic acid + stratification for 60 days (93%) and the high altitude seeds were reached the maximum germination percentage in shorter period of time. According to the obtained results, seed dormancy of wild pear is of physiological type. In addition, for the production of seedlings in the nursery, as well as for sowing to reforestation and enrich the degraded stands in Zagros forests; the use of high altitude seeds is suggested.