نشریه تحقیقات ژنتیک و اصلاح گیاهان مرتعی و جنگلی ایران
سال بیست و هفتم شماره 1 (پیاپی 53، بهار و تابستان 1398)

MicroRNAs are small non-coding molecules that regulate the expression of genes through mRNA digestion and/or translation inhibition. By accurately regulation the expression of the genes, these molecules allow the plant to respond appropriately to the changes in growth stages or environmental conditions. In this study, the miRNA159, miRNA160, miRNA398, and the miRNA398 targeted gene (CSD) were investigated in six species belonging to the Triticum and Aegilops species, along with the susceptible (DARYA) and tolerant (SIRVAN) check cultivars under drought (FC=25%) and non-stress (FC = 100%) conditions using real-time PCR. The qRT-PCR analysis showed that all three miRNAs had a significant (P≤0.01) increase in expression under drought stress. The highest expression of miRNA159, miRNA160, miRNA398, and CSD gene was observed in the species Ae.tauschii, T.urartu, T.durum, and T.durum, respectively. By the mean comparing of species × stress interaction, the highest expressions were observed in the three species T.durum, T.urartu and Ae.tauschii for all three miRNAs under drought stress. Also, the highest increase in CSD expression under drought stress was observed in T.urartu, T.durum and Ae.tauschii equal to 6.62, 6. 21 and 0.63 fold in compared to the non-stress condition, respectively. It was concluded that the species T.urartu, T.durum and Ae.tauschii due to their superiority in terms of containing miRNAs causer tolerance, could be suitable candidates for bread wheat germplasm enrichment and breeding to drought stress tolerance.

 In order to evaluate genetic variation and relationship among 20 genotypes of sainfoin, an experiment was conducted as a randomized complete block design with 3 replications in Agricultural and Natural Resource Research Center of Isfahan, during 2015-2016. Results of variance analysis showed significant differences among the genotypes for all traits, in exception of number of nodes, lateral branches, leaflets and leaf width. PLC population had the highest forage yield (3179 kg/h). 9147 population was the second population for forage yield (2965 kg/h). Correlation coefficients showed that forage yield had a positive correlation with total stems, number of lateral branches, nodes, leaves and leaf and stem dry weight. Using principal component analysis, the first three components accounted for 72% of the total variation. Number of lateral branches, nodes, leaf, leaf and stem dry weight and wet and dry forage yield were the most important traits in the first component. Stem height, leaf width, leaf and stem percentages and leaf/stem in the second component and leaflets, total stems, lateral stem height and dry matter percentage in the third component were important. Genotypes were classified into 4 groups with distinct variations for forage yield.

In this research a total of 49 accessions in oat collection of National Plant Gene Bank of Iran belonging to four species of Avena barbata, A. wiestii, A. fatua and A. sativa were studied. The experiment was performed in an observatory designin research field of Seed and Plant Improvement Institute located in Karaj, Iran. The agromorphological traits were measured according to IBPGR descriptor. Based on Shannon index, lemma color had the largest variety (1.42). In principal component analysis, the first seven components comprised 68.43% of total variation. Two accessions of 439 (A. barbata) and 449 (A. barbata) were the nearest and two accessions of 447 (A. wiestii) and 115059 (A. wiestii) and were the farthest based on genetic distance. The studied accessions were placed  in five groups by K means clustering method. Three discriminant functions were developed justifying total variance in data. The first and second discriminant functionsmaintained the most distinction between two species of A. barbata and A.sativa and between two species of A. fatua and A. wiestii, respectively. Total results indicated presence of suitable diversity in the studied population based on the measured traits which could be exploited in oat breeding in future. In addition, the discriminant functions developed in this research could be used in distinguishing the relating oat species.

 Whortleberry is one of the important medicinal plants in traditional medicine that has been used in Iran for decades in reducing blood sugar and blood pressure adjustment. In the present study, the effect of auxin, gibberellin (GA3), casein hydrolysate (CA), putrescine (PU), and salicylic acid (SA) on in vitro establishment and growth of Whortleberry explants and the amount of anthocyanin, flavonoid and total phenolic compounds in in vitro condition were evaluated. The apical bud and nodal explants were collected from natural habitat, surface sterilized and cultured on the MS medium supplemented with 2 mg/L BAP and different concentrations of NAA or IBA (0.01- 1 mg/l) and MS medium supplemented with 2 mg/L BAP and 0.1 mg/L NAA or IBA and different concentrations of gibberellin (0- 0.5 mg/l), CA (0- 150 mg/l), PU and SA (0- 10 mg/l). The results indicated that the percentage of explant viability and leaf producing explants were significantly influenced by auxin type and concentration. So, the percentage of explant viability and explants leaf production on MS media supplemented with low concentration of NAA were significantly higher than those of the higher concentration of NAA and all levels of IBA. Addition of GA3 to the culture medium was significantly increased the number of leaves per explant. Furthermore, CA, PU and SA led to an increase in the percentage of explants leaf production and number of leaves per explant, but this increase was not statistically significant in some of these treatments. Moreover, the amount of secondary metabolites was significantly affected by medium compositions. So, the highest amount of anthocyanin was observed in the MS + 2mg/l BAP+0.1mg/l IBA and the highest amount of flavonoids and total phenolic compounds were observed in the same medium 15 mg

 Seed priming is an effective method for enhancement of seed germination and seedling establishment in range plant species. In order to study of priming effects on seed germination of deteriorated seeds of Astragalus Spp., a factorial experiment was conducted based on completely randomized design with three replications in glasshouse condition in research institute of forests and rangeland, Tehran, Iran in 2016. The factor A was Astragalus species in three levels (Astragalus cyclophyllon, A. siliqusus andA. homosus), Factor B was seed deterioration in two levels (seed preserved in cold store (+4°C for 12 years), and accelerated aged seeds (by incubation in 100% relative humidity and temperature 40°C for 48 h). Factor C was priming treatments in four levels as: osmoprimings using PEG (-0.3Mpa and -0.6Mpa), hydropriming (soaking seeds in distilled water for 24 h) and control. Data were collected for seed germination, rate of germination, seed vigor, shoot, root and seedling length, root/shoot length ratio and seedling fresh weight. Result of analysis of variance showed significant effects of main factors and their interaction for most of traits. The A. siliqusus had higher mean values for all of traits except root length followed by A. cyclophyllon and A. homosus, respectively. The priming by deterioration interaction effect, showed that, for natural deteriorated seeds, both hydropriming and osmopriming -0.3Mpa had increased 25 to 96% means of traits except root length, whereas for artificial aged seed only hydropriming had improved 37% of seed vigor. It was concluded that for regeneration of aged seeds preserved for a long time in gene banks both hydropriming and osmopriming are effective methods to recover of deteriorated seeds of Astraglus spp.

 Persian Silk tree (Albizia julibrissin Durazz.) is one of the native leguminous species in plain and lowland hyrcanian forests, which has been severely damaged by fungal disease. In this research, feasibility of preservation of A. julibrissin seeds in cryo condition was studied. Seeds of A. julibrissin were collected from 10 trees in lowland forests of western part of Haraz watershed, Mazandaran province, Iran. Seed germination characteristic of this species were analyzed in cryopreservation with four pretreatments including: vitrification solutions, 30% glycerol, desiccation and without cryoprotectants as well as control treatment, storage in dry condition at 15°C temperature. Seeds after one week and one month storage in liquid nitrogen condition were removed from the liquid nitrogen and the seeds were soaked into water at +42°C (heat shock). Then, seed were placed in petri dishes and so on were transferred into germinator at +22 ºC. Seed germination was daily checked as long as no seed has been germinated. Results of two way ANOVA followed by Duncan as a post hoc multiple comparisons showed that there were no significant differences between germination traits of A. julibrissin seeds in four pretreatments of cryopreservation with control treatment. So we concluded that cryopreservation of A. julibrissin seeds is feasible. Our result indicated that having hard seed coat and as well as very low moisture content enable A. julibrissin seeds to be preserved in cryo condition without any cryoprotectants. Therefore, cryopreservation of A. julibrissin seeds could be done without any pre-treatment and it could be proposed as an effective strategy to protect the genetic recourse of this valuable species of the Hyrcanian forests in Iran.

Among species of hardwood forest trees, Eucalyptus citriodora is a fast growing plant that is cultivated for aromatic essential oils in perfumes, pharmaceuticals, and its soft and light wood in the paper and furniture industry. The micropropagation of E. citriodora Lemon-scented Gum was carried out in two medium culture of MS (1/2N) and WPM through bud culture. Sterilization was done using solutions of mercuric chloride 0.1% and sodium hypochlorite 20%. Shoot multiplication was done with cytokinin (BAP, Kin), auxin (IBA) and gibberellin (GA3) hormones and rooting with auxin hormones IBA, NAA and IAA in only MS (1/2N) medium. The shoot multiplication was assessed in two medium using factorial experiment based on the completely randomized design (CRD) and rooting multiplication in one medium using CRD. Propagation coefficient, shoot length, greenity, total root and rooted seedlings were measured. The best sterilization was immersion seeds in 20 % sodium hypoclorite solution in 18 min. The best propagation and shoot elongation was obtained in MS (1/2N) medium coupled with BAP, Kin, GA3 cytokinin hormones and auxin hormones IBA in 0.3, 0.2, 0.1, 0.01 mgl-1 and 0.3, 0, 0.1, 0.01 mgl-1concentrations respectively by 200 mgl-1 P.V.P. The best rooting treatments were a modified MS (1/2N) medium, auxin hormone NAA and IBA+IAA in 1 mgl-1, 0.5mgl-1. These plantlets transferred to soil and were kept in the greenhouse conditions. The results of this study show that tissue culture is a suitable method for the propagation of this valuable species in a short time.

Sweet pea (Lathyrus odoratus L.), belongs to Fabaceae family, is an ornamental herbaceous plant with fragrant flowers and climber. This plant is as a source of genetic for important traits such as resistance to edaphic stresses. In order to keep genetic sources, plant propagation through tissue culture can be effective. To establish an efficient protocol of shoot regeneration from callus, effects of explant type, culture media and plant growth regulators on callus induction and shoot regeneration of sweet pea were evaluated. The results showed that internode was the best explant for callus induction (0.77 g callus per each explant) and MS was the best medium to induce callus formation with 0.78 g callus per each explant. The highest callus induction (1.9 g callus per each explant) was achieved planting internode on MS medium supplemented with 0.5 mg.l-1 6-benzylaminopurine (BAP) and 2.0 mg.l-1 1-Naphthaleneacetic acid (NAA) after 25 days of culture. Shoots regenerated at the highest frequency with 5.33 shoots when calli were cultured on MS medium with 1.0 mg.l-1 Thidiazuron (TDZ). This protocol provides a basis for future studies on genetic improvement and could be applied to large-scale multiplication systems for commercial nurseries of Lathyrus odoratus L.

Stevia plant(Stevia rebaudiana Bertoni.) is an important plant species due to its high content of recoverable natural sweeteners. Considering the industrial and medical importance of plant, the current study was performed with the aim of optimizing somatic embryogenic of this plant. Embryogenic callus induction and embryogenesis were investigated using different concentrations of Benzyl Adenine (BA), Naphthalene Acetic Acid (NAA) and 2,4-Dihydrophenoxy acetic acid (2,4-D) in leaf and bud explants in MS culture medium in a factorial experiment based on completely randomized design (CRD) with five replications. The highest callus induction was obtained in culture media contained 0.01 mg/l (BA + 0.5 mg/l 2, 4-D and the highest embryo induction was obtained in 0.01 mg/l BA + 1 mgl-1 2, 4-D treatment. Also the effect of Casein hydrolysate on the embryogenesis of the Stevia plant were investigated in five concentrations (0, 50, 100, 150 and 200 mgl-1) and coconut milk in two levels (0 and 55 ml) by both explants of leaf and bud in a factorial experiment based on CRD with six replications.  In concentrations of 0 and 50 mg/l of Casein hydrolysate respectively obtained the highest (20.2%) and lowest (7.84%) embryos. The percentages of embryogenesis in MS medium without coconut milk was 78.82% for bud explants and 25.09% for leaf explants, and finally, embryos were transferred to medium containing 0.1 mgl-1 of GA3 for organogenesis, and 18.03% of the embryos were grown to complete seedling.

 The roots and stolons of Glycyrrhiza species constitute one of the most important crude drugs in the world and contain a large amount of glycyrrhizin, an oleanane-type triterpenoid saponin. Tissue culture and callus production in licorice species are prerequisites for studies on secondary metabolites and cell transformation. In order to effect of antioxidant to reduce the browning and growth of callus, was used Ascorbic Acid and PVP in different levels. Also in order to influence these antioxidants in the callus growth index, were cultured both species as cell suspension. Callus was produced from hypocotyl related to two weeks’ seedlings. Callus was induced from NAA (0.5 mg/L) and BA hormones (2 mg/L) as well as ascorbic acid and PVP treatments at different concentrations as two separated factorial experiments based on CRD design. The results showed that in consecutive sub culture 1200 mg/L of PVP had the least generation, necrosis and the most callus production in both species. Whereas, in cell suspension, with increasing PVP concentration, was decreased callus growth index and 1200 mg/L PVP has shown the lowest callus growth index in both species. It can be said that PVP absorb NAA and BA hormones in cell suspension and therefore it was reduced concentration of hormones in medium. Ascorbic acid treatment with 80 and 100 mg/L had the highest callus growth index in both species respectively and it is recommended for cell suspension culture in licorice.

 Hymenocrater yazdianus is one of the important medicinal and range plant. This plant belongs to Lamiaceae family that grows in arid land regions. In this research mass production of Hymenocrater plant via tissue culture technique was studied. Lateral and apical buds were used as a primary explant for cultivation. For sterilization, Mercuric chloride was used at 0.1% concentration for 2 minutes. After sterilization, explants were washed 3 times with sterilized water containing 0.1% Citric acid for browning prevention of theme. Then all explants transfer to MS medium without any plant growth regulators as an establishment stage. After 4 weeks, explant transported in MS medium containing 0, 0.5, 1, 1.5 mgl-1, Benzyl adenine (BA) and Indole-butyric acid (IBA) concentrations at 0, 0.01 and 0.1 mgl-1 in 12 treatments for proliferation. The results showed, highest shoot number were obtained when we used MS medium at a concentration (1 mgl-1BA and IBA 0.1 mg l-1) In this culture medium we obtained 5.5 shoots in each tube. For rooting stage, MS and LS culture media were studied. In these culture media IBA was used in 0.0, 0.05, 0.1, 1 and 2 mgl-1. The results indicated using MS medium supplemented to 2 mgl-1 95% of explants were rooted.  Finally rooted plantlets were adapted successfully in peat and perlite complex to ratios of 2:1 and in vitro culture plant successfully transferd to the field.

 Medicinal and aromatic medicinal plant, Ziziphora clinopodioides L., belonging to the genus Ziziphora and Lamiaceae family. The aim of this study was to optimize asexual regeneration in two genotypes of this species as code 5 and 6. For this purpose, the first experiment was conducted for contamination control using sodium hypochlorite 1% and alcohol 96%. In the next step, the control of produced phenolic compounds was investigated by combining three polyvinylpyrrolidone, active charcoal and ascorbic acid. Multiple shoots induction were performed using lateral bud explant in a MS medium containing the BAP at five levels in combination with NAA (0.01 mg/L). Rooting of branches was performed in MS medium containing NAA at two levels (0. 1 and 0.3 mg/L). Result showed 1% sodium hypochlorite treatment coupled with 96% alcohol eliminated contamination up to 74%. The treatment of combination of polyvinylpyrrolidine, ascorbic acid and activated charcoal with average value of 8.25% had the lowest browning in both Genotype of 6 and 5. The highest branching values of (88% and 100%) were obtained in the treatment of 2 and 2.5 mg/L BAP combined with 0.01 mg/L NAA in genotype 5 and 6, respectively. The highest value of rooting (33.3%) was observed in genotype 6 using 0.3 mg/l NAA. The results of this research could be  used in the micropropagation and proliferation of this plant.

 In this experiment morphological diversity of seven wild populations of Bunium persicum from Razavi and South Khorasan provinces of Iran were studied. Results showed significant differences among the populations for some morphological traits such as umbellate per umbel, leaf length, number of seeds per plant and inflorescence width. Cluster analysis of morphological traits divided populations into two groups and showed that the geographical and genetic diversity does match. Plant height with number of umbels, leaf length, inflorescence length and width and inflorescence length with inflorescence width had a high positive correlation. principle component analysis showed that four major components accounted 93.81 percent of the total variation, which the first components  including plant height, number of umbels, number of umbellate per umbel, leaf length, inflorescence length and width and number of seeds per plant traits had the most proportion (%52.08). According to obtaining results, populations 1, 2 and 3 due to idealistic traits such as higher values for plant height, leaf length, umbellate per umbel and seeds per plant were the best populations for breeding improved varieties and domestication