نشریه تحقیقات ژنتیک و اصلاح گیاهان مرتعی و جنگلی ایران
سال هجدهم شماره 1 (پیاپی 35، بهار 1389)

Alfalfa is one of the most important forage crops in the world. The west and northwest parts of Iran are the important regions for Alfalfa planting. Because few studies are available on the genetic diversity of this crop in the regions, therefore, the objective of this study was to determine the genetic diversity in 34 populations of alfalfa from west and northwest of Iran.These were analyzed using 10 microsatellite markers. Totally, 51 bands were obtained from which 45 bands were polymorphic corresponding to an average of 5 alleles per locus. The percentage of polymorphism with an average of 0.55 varied from 0.29 for MAL369471 to \0.8 for ENOD20 and also for MI with an average of 2.1 from 0.29 for MAL369471 to 4.01 for ENOD20. Chaharmahal va Bakhtiari province had maximum of both gene diversity and Shannon's Information index (He=0.19, I=0.29), while the minimum amounts (He=0.13, I=0.13) were obtained in Tehran province. Dendrogram constructed based on dissimilarity matrix grouped populations in two main groups; the first group belongs to populations from Karaj, while in the other populations were located in second group. The result showed that microsatellite markers are effective tools for evaluation genetic diversity in Iranian alfalfa populations. The results of this study provided useful information for the future collection and for the of genetic resource management in the regions.

Montpellier Maple (Acer monspessulanum) is one of the broad leaf species grows in semiarid and cold climate of I.R. Iran. Preservation of this wild woody species has a greatimportance in natural resource point of view. Cryopreservation technique is used to preserve seed and vegetative organs under -196 ºC conditions. In the mentioned temperature, all of the metabolic and physiological activities of the seed or organ almost is stopped. As the result, seed and organ can survive for a long period of time. In order to preserve A. monspessulanum seed under -196 ºC condition, three pre-cryopreservation treatments, PVS2 solution, Desiccation and 30% Glycerol were applied. The treated seeds were transferred into Liquid Nitrogen (LN) and stored for 7 and 30 days respectively. After finishing storage period, the seeds were removed from the LN, imposed to heat shock and transferred between wet-papers in germination boxes.The cryogenic seeds together with control seeds were incubated at +4 ºC for 105 days to germinate. Mean germination percentage of the cryopreserved seeds was 19.18% and, there were no significant differences between the control and cryopreserved seeds. No adverse effects or abnormalities were observed on seedlings developed from cryopreserved seeds. The results showed that, the long-term preservation of the species, seed in -196 ºC using suitable precryopreservation treatment is possible.

Inter-specific hybrids of Lolium with various capabilities provide possibilities to integratesuitable characteristics into a cultivated variety of the genus. Baring this objective in mind,inter-specific hybrids had been produced which are investigated in this research. Four genotypes of the inter-specific hybrids between Lolium perenne and L. rigidum, and their parents were investigated for their protein profiles using seed storage protein electrophoresis. They were compared to make sure their hybrid originality as well as investigating their similarity to their parents. SDS-PAGE method of electrophoresis was used to provide the profile of the seed storage proteins. Presence or absence of the protein bands in the profile column of the genotypes were scored as one and zero respectively. The provided data were analyzed by cluster analysis. A dendrogram produced by the analysis, was used along with the proteins profile for classifying and estimating the hybrids and their parents similarities. Twenty protein bands were observed in the profile of the studied genotypes in which there were differences between thehybrids and their parents based on limited number of the bands. There were also differences on thickness and stain intensity of the protein bands of the studied genotypes, which might be due to differences in gene expression of parents and progenies. On the whole the profile revealed remarkable differences between the parents and progenies as well as approving the hybrid origins of the progeny genotypes.

Eucalyptus grandis is a fast growing species with economic and industrial value. It iscultivated for its wood fibers which is widely used in the paper industry, domestic fuel andcharchoal production. For clonal propagation of superior genotypes of the species, apical and axillary buds from adult elite trees in forests of northern Iran (Chamestan- vase research station) were collected in different seasons. Application of 0.1% mercuric chloride solution for 7 minutes was the best treatment for surface sterilization of the samples in summer. Different compositions of hormones were evaluated for regeneration. The best shoot multiplication was obtained using a modified MS medium containing half strength Nitrate containing BA, IBA, Kin and GA3 with concentration of 0.1, 0.01, 0.2 and 0.1 mgl-1, respectively. Shoots from the multiplication medium were transferred in MS medium (half strength Nitrate) containing 1 mgl- 1 Zeatin and 0.2 mgl-1 IAA for shoot elongation. Rooting of shoots were achieved in MS with 1/4 strength of macroelemants and 0.5 mgl-1 IBA plus 0.5 mgl-1 NAA. The plantlets were successfully established in greenhouse and field conditions.

Understanding genotype * environment interaction structure for investigating genotypesstability and adaptation is very important in plant breeding programs. Study was carried out on 18 genotypes of Agropyron elongatum performance based on randomized complete block design with three replications during three years (2004, 2005 and 2007) at two environments, dry(stress) and irrigated environments (non-stress). The experiment was conducted at Islam Abade Gharb Research station, Kermanshah, Iran. Results of analysis of variance showed significant differences (p<0.05) between the studied genotypes. Genotype × environment interaction was not significant but the first and second components of interaction (IPCA1 & IPCA2) were highly significant (p<0.01) and justified 59.92% and 29.38% of the G×E interaction SS, respectively. Mean comparison for forage yield revealed that the genotypes 10 and 15 had the highest (4.49 and 4.50 t ha-1, respectively) and genotypes 1 and 16 had the least (3.14 and 3.49 t ha-1, respectively) forage yields. AMMI1 model showed that genotypes 2, 3, 4, 5, 6, 7, 8, 9, 11 and 17 had the highest general stability and AMMI2 model showed genotypes 3,6, 11 and 17 had the highest specific adaptation to the stress environment.

In this study, we investigated karyological characteristics variation of four Anthemis speciesselected from various locations of Iran. We used of α-bromonaphthalene for pretreatment,Lewitsky solution for fixation, 1N NaOH for hydrolysis and finally, aceto iron-hematoxylin forstaining stage. Detailed karyotype features, i.e. total chromosome length (TL), total formpercent (TF %), asymmetry index (AI), intrachromosomal asymmetry (A1) andinterchromosomal asymmetry (A2) were also estimated. Somatic chromosome number were 2n=18 for all species. Karyotypic analysis showed a predominance of metacentric chromosomes and satellite was observed in the shape of microsatellite in almost all of them. The largest chromosome and genome length belong to A. altissima. In addition, the four studied species showed varying interachromosomal (A1; range = 0.28-0.40), and interchromosomal (A2; range = 0.12-0.15) asymmetry indices. According to Stebbins categories, they were separated to 2A (A. altissima, A. cotula & A. hyalina) and 1A (A. pseudocotula) karyotypic symmetry classes. A. pseudocotula had more symmetric karyotype than other species. Clustering of the Anthemis species and accesions based on cytogenetic data showed close relationship between A. cotula and A. hyaline. Chromosome count of A. hyaline and A. pseudocotula are reported here for thefirst time. Other two counts are agreement with the previous reports.

Alfalfa (Medicago sativa L.) is one of the most important forage crops in the world that have a significant role in forage supplying, nitrogen fixation and reduction of soil erosion. The species is self-incompatible and cross pollinate, so maintenance and propagation of desired genotypes are difficult and production of pure and hemogeneous seeds is faced with difficulties.Somatic embryogenesis is one of the most useful techniques that can help us to solve theproblems. Therefore, recognition of the affecting factors on somatic embryogenesis isimportant. This study was aimed to evaluate the effects of two growth regulators, 2,4-D (0.5 and 1 mgL-1) and Kinetin (0 and 1 mgL-1), carbon source (sucrose and maltose) and Glutamine and Serine as amino acids, on alfalfa somatic embryogensis in an embryogenic cell suspension culture using 4 explants (hypocotyl, cotyledon, petiole and leaf). The treatments showed significant differences. There was a correlation between 2,4-D concentration and the number of embryos in maturity stage, so that increasing the concentration of 2,4-D resulted in lower number of embryos. Presence of Kinetin in the culture media increased number of somatic embryos, significantly. Despite significant effects of Glutamine and Serine on embryo development, application of these amino acids in suspension culture media decreased the regeneration of somatic embryos in the liquid cultures. Maltose in liquid culture was the most effective agent on the maturity of somatic embryos and increased the number of embryos in embryogenic cell suspension culture. Four types of the examined explants, showed significant differences of embryogenic ability; hypocotyl and petiole explants produced the most and the least number of embryos, respectively

Smirnovia turkestana is an endemic plant species of sandy deserts of Iran. This shrub andpsamophyte plant belongs to the Papilionaceae family. It is an important species for protecting sandy soil, forage production and medicinal value. The aim of micropropagation of S. turkestana is for using it for desertification, overcome on its propagation, cryopreservation and …. Seeds were collected from sandy dunes of Kashan, Isfahan province. Seeds were scarified and sterilized by NaOCl (1.25% v/v) for 15 min; washed in distilled water and cultured on MS (half sucrose) medium. Shoots were isolated after 3 weeks and cultured on 8 medium treatments. The best treatment for shooting was MS basal medium supplemented with BA (0.5 mg/l), 2ip (0.5 mg/l) and IBA (0.01 mg/l). The best treatment for rooting achieved in modified MS medium without hormone. Rooted plantlets were transferred to pots containing field soil and grew well under greenhouse conditions.

In order to investigate genetic diversity in Zelkova carpinifolia population, sampling ofleaves of 30 trees in three lowland habitats were done. For evaluating enzyme activities in theleaves, the samples extracted immediately and peroxidase studied quantitatively andqualitatively. Quantitative studies accomplished by spectrophotometer (in 530 NM wavelengths) and qualitative studies performed by polyacrilamide gel electrophoresis (PAGE).Quantitative activity analyzed with ANOVA method and showed no significant differences in the three habitats, but Mazandaran habitat showed the most average quantitative activity. Grouping of isoenzyme bands showed three ecotypes in three habitats. All trees of the habitats were classified into 6 groups using Ward method of cluster analysis on quantitative data.Assessing genetic distance showed the most distance between Golestan habitat individuals and the least genetic distance in Guilan habitat. Mazandaran and Guilan habitats exhibited low level of isoenzyme pattern relative to Golestan habitat. Golestan habitat had the most within group diversity and is suitable for Zelkova carpinifolia development. Result emphasized on utilizing of the most effective approaches for conservation of genetic biodiversity of the species.

Effects of subculture media on somatic embryogenesis and rhizogenesis in different parts of mature embryo of Olea europaea L. (cv. Kroneiki) was studied. Radicle and proximal part of cotyledon were cultured in OMc medium containing or IBA (5 mg/l) and 2ip (0.5 mg/l). twenty one days old calli were subcultured on OMc or OM media without any hormones. Cultures were maintained in darkness. Differentiation in the calli depended on kind of explants and subculture media. After the third subculture calli of radicle and proximal part of cotyledon had the potential of somatic embryogenesis. In both media, the potentiality of embryogenesis in calli from radicle was more than calli from proximal. In addition, somatic embryogenesis in OM medium showed significant increase. After the second subculture rhizogenesis occurred in both explants and different subculture media. Differences between rhizogenesis were significant. In both subculture media the potentiality of rhizogenesis in calli from proximal part of cotyledon was much more than calli of radicle and in the both explants rhizogenesis in OM medium were more than OMc medium.

Recognition of plant genetic resources, investigation of ecological and morphologicalconditions, and determination of geographical position of the plant species as well asimplementation of research projects on conservation and protection of these resources with the use of different conservation techniques seem to have a special importance. In this research, geographical position and distribution of the trees and shrubs of Markazi province were determined with GPS and recorded in digital map as a segment with 1/50000 scale and then polygonized. Geographic information system (GIS) and ilwis academic software were applied on mapping the trees and shrubs species distribution with two types of polygon and point maps. All trees and shrubs of the province were listed. Several characteristics of the trees and shrubs species such as seed production term, multiplication, utilization, health, pest and disease were determined and registered. Finally, geographical distribution of 11 tree and shrub species wer determined and mapped based on their priorities, as: Pistacia atlantica, Pistacia khinjuke, Nitraria shobiri, Rus cororaria, Quercus persica, Amygdalus scoparia, Lonicer Nummulariifolia, Berberris Integrrima, Cerataegus Zsovitsii, Cerataegus Pseudoheterophylla,Creatagous Atrosanguinea. The results showed that trees and shrubs are crowded in 40geographical areas and distributed individually in various and many parts of the Lorestan forest regions and among them most of the area is covered by Amygdalus scoparia species.

Genus of Juniperus is one of the widely distributed plants from coniferales in the worldwhich is native to moderate regions of central and south Asia. In this study, RAPD markerswere used to evaluate the genetic variation among 21 samples of Juniperus (collected fram 6 habitat of Tandure National Park of Iran). The 7 RAPD primers produced 58 scorable bands of which 33 (57%) were polymorphic. The pair-wise genetic distance was from 0.129 to 0.788. The average Heterozygosity within accessions (Hs) was 0.213 and the total heterozygosity was 0.326. The Fst Index was 0.346 and the gene flow (Nm) was 0.944. The dendrogram constructed using UPGMA method, distinguished 2 main groups among 6 accessions of Juniperus that had good compatibility with geographical map of park and was also confirmed by PCoA.

Morphological characteristics of Arabidopsis thaliana, A. pumila and A. wallichii wereevaluated in Iran. Shape of the trichomes, shape of the basal and cualine leaves length and shape of the fruits, presence or absence of trichome in fruits flowers, color of the petals and shape of silique in cross section are the differentiating characteristics of the three species.Anatomical features of stems and leaves of the species were observed and analyzed by light microscope. Shape of the stem in transverse sectios thickness of derm presence ofcollanchyme and scloranchyme around vascular bundle the number of vascular bundle andwavy shape of the stele are the distinctive features in stems. The anatomical features of the leaves such as shape of leaves in cross section The number of palisade tissue layers, uniformity or non-uniformity of mesophyll tissue were distinctively different in the three species. Floral meristem of the two species were observed by light microscope. No difference was observed in tissue structure.

Effects of pre-chilling temperature was studied on seed characteristics including,percentage and speed of germination, seedling length, seedling dry weight, ratio of dry weight to fresh weight and vigor index, on five ecotypes of Festuca arundinacea under laboratory, greenhouse and growth chamber conditions. Tiller number and leaf area were also studied in greenhouse and growth chamber conditions. In laboratory condition, pre- chilling germination temperature (4°C) was applied by two weeks on seeds of the five ecotypes before standard germination test. In greenhouse, seeds of the five ecotypes were sown on pots under day temperature of 20±5°C and night temperature of 5-12°C and then they were subjected to cold treatment, on 4°C at stages of 15 and 35 days of seedling ages for two weeks compared to control. In growth chamber condition, the 15 days old seedlings were transferred into a cold room in 4°C for two weeks. Ecotype like Esfahan, Brojen and Kamyaran had high values for seed characteristics in growth chamber. Also, the leaf area of the ecotypes increased with cold treatment. Tiller numbers of the ecotypes, particularly the two ecotypes of Kamyaran and Mashhad were affected by cold treatment. In greenhouse condition, the ecotypes of Esfahan, Brojen and Kamyaran had higher values for leaf area and tiller numbers in cold treatment comparing to non-cold treatment. These ecotypes also had higher values for seed characteristics in response to cold treatment on the two stages of seedling growth.