نشریه تحقیقات ژنتیک و اصلاح گیاهان مرتعی و جنگلی ایران
سال بیست و یکم شماره 2 (پیاپی 42، تابستان 1392)

Nowadays production of secondary metabolites from plant cell suspension cultures has become an attractive subject. In this study, callus induction of black zira, an important medicinal plant, was investigated in two MS and B5 solid media, supplemented with 2 mg/l NAA + 0.5 mg/l BA, and 2 mg/l 2,4-D + 0.5 mg/l Kin. Another experiment was also carried out for callus culture in MS liquid medium with four different combinations of plant growth regulators, 2,4-D (2 and 0.5 mg/l) + 0.5 mg/l Kin, and NAA (2 and 0.5 mg/l) + 0.5 mg/l BA. For the cell suspension cultures, the hormonal combinations, as above mentioned in callus induction, were used in liquid MS medium. Results showed that the best response for the callus induction percentage and callus dry weight in solid medium as well as callus fresh weight in liquid medium was obtained from MS medium supplemented with 2 mg/l NAA + 0.5 mg/l BA. It was also found that liquid MS medium with 2 mg/l NAA + 0.5 mg/l BA was suitable for the cell suspension cultures of black zira due to the higher cell dry weight in this stage compared to other treatments. The stationary phase in growth course was observed in MS medium supplemented with 2 mg/l 2,4-D + 0.5 mg/l Kin and 2 mg/l NAA + 0.5 mg/l BA after three and four weeks, respectively.

Moringa peregrina is one of the important species which is growing in a vast area of the Southeast part of Iran. Despite of its importance in medicinal industry and living environment, the species did not receive enough attention by now. In other words, there was not enough research on its genetic potentials. Due to various reasons, the species is under genetic erosion; therefore, in order to optimize its remarkable characteristics exploitation, its conservation through various methods is considerable. Current research, multiplication of the species through lateral buds by tissue culture, was employed in order to find a suitable culture media for the species multiplication as well as investigating its plant population's different responses to micropropagation and several culturing media. Immature seeds of three plant populations were collected from three locations of its habitats, Chanf, Benet and Kenshki, and cultured on aseptic conditions. The seedlings containing one or two buds were transferred to the multiplication culturing media. MS and Half-MS culturing media along with four hormone combinations (0.5 and 1 mg/l BAP, combination of BAP and 2ip, 0.5 mg/l) were used in the study. Several characteristics such as number of branches, active buds, and branches average length were recorded. There were significant differences between the studied plant populations based on the number of branches. Whereas, there wasnt significant differences between the populations based on branches length. Chanf population had the highest value in branch production between the studied plant populations. The longest branch was also observed on the Chanf plant population.

Calligonum species are distributed in centeral Asia desert and are effective to stabilize sand dunes. RAPD is a useful genetical marker to determine polymorphism and relationship among plant species. In order to evaluate Genetic diversity within and between species of Calligonum was evaluated using 15 RAPD primers in 26 samples of 11 species collected from different parts of Iran. According to diversity indices, Dice similarity, cluster analysis and band information a total of 191 bands was amplified and 94% of the bands were polymorphic. Cluster analysis revealed a high amount of diversity among the samples. The most similar samples with 0.81 similarity was found within C. bungei species, while the least similarity (0.45) obtained in samples from other different species. All samples clustered in four groups, samples of C. polygonoides and C. leucocladum located in two groups separately. However, other species distributed in two other groups. There was low relationship between genetic divergence and geographical origins that could be because of seed migration between the locations by wind and human.

Genetic diversity is a basic element in plant breeding which could be used to improve new lines with favorable traits. Because of importance of high molecular weight seed storage proteins in commercial wheat cultivars, investigation of diversity of the proteins in wheat lines and its wild relatives is an essential factor in breeding programs. Genetic diversity of seed storage proteins among 105 accessions of the 9 Aegilops species including Ae. columnaris, Ae. umbellulataa, Ae. triuncialis, Ae. neglecta, Ae. kotschyi, Ae. cylindrica, Ae.crassa, Ae. tauschii, and Ae. speltoides were evaluated by SDS-PAGE method. Twenty two protein bands were observed among the accessions, which grouped in 24 different patterns. The most diversity was observed on Ae. neglecta which its diversity index was 0.12 and the least was observed on Ae. cylindria with diversity index of 0.03. Among the origin provinces of the accessions, Tehran and East Azerbaijan with index of 0.13 showed the most diversity and Lorestan with index of 0.06 showed the least diversity. Cluster analysis using between group linkage method, clustered both the species and origin province of accessions into three groups. The most genetic distance was observed between Ae. triuncialis and Ae. crassa, and the least genetic distance was observed between Ae. neglecta and Ae. umbellulata. Moreover, the most genetic distance was observed among the samples of Lorestan and Semnan while the least was observed among the samples of Kermanshah and Ilam provinces.

Leonurus cardiaca is a medicinal plant of the family Lamiaceae, due to the effects of blood dilutioning, the species is used for centuries. The species has a high diversity and widely distributed in Iran, so that from 4 sub species in the world, 3 sub-species are grown in Iran. In this study AFLP markers were used to investigate genetic diversity among 47 samples of 6 motherwort populations and the kinship ties between them. The results showed that 6 selected AFLP primer combinations (MseI /EcoRI) amplified 608 scorable DNA bands, of which 355 were polymorphic (polymorphic 58%). The average genetic similarity of samples was estimated based on the Nei coefficient, 0.26 (0.21-0.30). Analysis of molecular variance (AMOVA) showed that the genetic variation was mainly within populations (57%), but variance among the populations was only 43%. A UPGMA dendrogram based on genetic distance also revealed five major branches in accordance to geographical regions. Maximum and minimum values of genetic similarity of the populations were recorded between “Taleghan 6” and “Taleghan 7” (0.7) and “Dargaz 1” and “Taleghan 7” (0.06) respectively. Data showed a completely different banding pattern of subspecies L. Cardiaca subsp Turkestanicus. of Dargaz region. Coefficient of genetic differentiation among the populations (Gst) and gene flow (Nm), average number of emigration exchanged per generation estimated among the populations, were estimated 0.37 and 0.85, respectively.

Genetic diversity of 39 genotypes from 25 populations of Achillea biebersteinii Afan., collected from different parts of Iran, were evaluated using random amplified polymorphic DNA (RAPD) data. From a total of 18 primers used, 322 recognizable RAPD bands were detected of which 279 were polymorphic (86.6%). The average number of bands per primer was 17.9. Total resolving power was 147 with the mean of 8.17. Among the primers, maximum resolving power was recorded for the primer BA03 (16.67). Jaccard’s similarity index was used for measuring genetic similarities among the genotypes. The highest similarity (0.95) was found between Mohammad Shahr-2 and Mohammad Shar-5 genotypes, whereas the lowest was observed between Meshkin Shahr-2 and Piran Shahr-1, and also Meshkin Shahr-6 and Piran Shahr-1 (0.24). UPGMA algorithm was used for cluster analysis. Cluster analysis separated the 39 genotypes into 15 main groups. The obtained clusters based on RAPD markers to some extent matched with the geographical origin of the studied populations of A. biebersteinii. Genotypes of Piran Shahr-1 and Tangehgol-2 showed the highest difference to the other genotypes and classified in separate groups, and maximum similarity was observed among the genotypes of Mohammad Shahr population with over 0.90 similarity. Results showed a high level of genetic diversity among the investigated genotypes. The results also indicated that RAPD technique was a suitable approach for assessing genetic diversity and germplasm management in populations of the species.

Chamomile is one of the most important medicinal plants in commerce that has many applications in drug and sanitary industries. In order to evaluate the genetic diversity of different scentless chamomile landraces (Matricaria inodora L.) based on morphological traits and molecular markers, 17 landraces were collected from different areas of Iran. In order to evaluate the morphological traits, the landraces were planted in an augmented design in Fozveh station of Agriculture Research Center of Isfahan. Forty-four arbitrary 10, 15 and 18-mer primers were used for PCR amplification of total genomic DNA. Twenty nine primers were selected based on the quality and reliability of their amplification. Genetic similarity between the genotypes was estimated using Jaccard's similarity coefficient and UPGMA method of cluster analysis was conducted on the collected data. Results showed that yield of flower per plant, number of flower per plant, and 1000 grain weight had maximum variance coefficient. Days to end of flowering and Days to flowering had minimum variance coefficient. Genetic similarity coefficient was estimated in a range of 0.20–0.72, with a mean of 0.39. According to the cluster analysis on both morphological and molecular markers, the studied landraces were classified into 5 clusters, but intra-groups landraces were different. Results of cluster analysis showed that genetic diversity was not coordinated to the geographical distribution.

For the past 30 years or so, Iranian foresters have attempted to improve management of broad-leaved stands of Hyrcanian forests, of which beeches are among the most important species. Silviculture of beech requires skill, and the slightest error can cause reduced quality, often as a result of the formation of forks on the stem. This phenomenon can be related to species suggesting that there may be genetic determination in some species; and linked to external elements, which is improvable by appropriate management. Aim of this work is the investigation of a possible relation between the variability of the trunk morphology and the genetic variation. Based on two polymorphic chloroplast microsatellites that had been previously identified and sequence characterized, genetic variation was studied in a total of 176 individuals from the forked and monopodial beech (Fagus orientalis Lipsky) tree groups in 13 oriental beech (Fagus orientalis Lipsky) populations distributed all over the natural range in Hyrcanian forests. We found six and two different length variants at each locus, respectively, which combined into ten different haplotypes. Genetic distances between most of the populations were high and significant, whereas not between the forked and monopodial beech tree groups in each population. There is also evidence for spatial organization of the distribution of haplotypes, as shown by permutation tests, which demonstrate that genetic distances increase with spatial distances. Besides, the correlation between morphology and geographical distance matrices was significance (R2=0.116, P=0.01), indicating existence of a relatively clinal trends in variation of microsatellite loci. A large heterogeneity in levels of diversity across populations was observed, but not between the forked and monopodial beech tree groups in each population. Furthermore, there is good congruence in the levels of allelic richness of the two loci across populations. The present organization of levels of allelic richness across the range of the species did not show any significant differentiation between the forked and monopodial beech tree groups.

One of the approaches to agricultural sustainability is using diversity of plant species, including forage crops. A total of 64 accessions of forage grass genus: Agrostis, Bromus, Dactylis, Festuca and Phalaris were sown in a field experiment, and their agro-morphological traits were characterized according to standard descriptors. A high diversity was observed for most of the studied traits on the genus. F test was significant for most of the traits based on one-way ANOVA. Duncan mean comparison method was performed to grouping genus based on the recorded traits. Phalaris located in the highest rank for herbage summer yield followed by Bromus, Festuca and Dactylis in the next group and Agrostis and Poa located in the last group. Results showed forage grasses can be seemed as candidate to be used in agronomy systems due to the existence of high potential for forge yield in the studied materials.

In consideration of nutrition deficiency in grasslands, utilizing of biological fertilizer and cultivation of resistant plants such as pigweed may help to amend grasslands. Thus, in order to evaluation nitroxin effects on pigweed, a factorial experiments was performed based on a completely randomized design at Damavand during 2010. The first factor consisted of three levels of nitroxin (0, 1, 2 Kilogram per hectares) and the second factor consisted of eight species of Amaranthus from different regions of Iran. Evaluated traits consisted of fresh and dry weights, stem height, dry matter percent, leaf number and area, chlorophyll a, b and total chlorophyll contents, chlorophyll a to b ratio, carotenoids contents, carotenoids to total chlorophyll ratio, anthocyanin and flavonoid contents. Analysis of variance and mean comparison showed that the first and second factors and their interaction were significant on most of the studied traits. A. retroflexus from Zarand Kerman was the best based on the most of the traits. The best answer were achieved in 1 kg/ha nitroxin for the most of traits. The best results for stem fresh and dry matters were achieved in 1 and 2 kg/ha nitroxin for A. retroflexus from Zarand Kerman. The best results for plant pigments were achieved in 2 kg/ha nitroxin in A. retroflexus from Tehran. Based on principal components analysis, the first three components could explain 88% of the total variance.

The study was carried out in order to investigate the effects of water deficit on forage yield and agronomic traits on 12 accessions of the species. The treatments included 3 irrigation regimes, without stress, moderate stress and severe stress. At harvesting date, data were collected on survival rate, fertile tiller percent, flag leaf length, peduncle length, plant height, spike length, number of spikelet, spike yield and straw yield. Results showed that the drought stress had no significant effect on survival rate, flag leaf length, spike length and number of spikelet. Whereas, low and moderate drought stress showed significant differences on fertile tiller percent, peduncle length, plant height, spike length and yield without spike. For straw yield, moderate and sever drought had significant different effects. Comparison of the means of survival rates indicated that the accessions of Parvar with average values of 85% and Arasbaran and Semirom with 45% had the higher and lower survival rate, respectively. Regarding fertile tiller percent, Mianeh with average values of 50% showed the lowest value. Regarding spike yield, Semirom and Mianeh with average values of 39.6 and 7.9 kg/ha had the higher and lower spike yield, respectively. Parvar and Mianeh with average values of 67.1 and 21.9 kg/ha had the higher and lower straw yield, respectively.

In order to evaluate genetic variability, 36 populations of Dactylis glomerata were investigated under irrigated conditions, using a randomized complete block design with three replications during 2009. Morphological traits were recorded. According to analysis of variance, a significant variation observed for forage yield, number of stems per square meter, plant hieght, flag leaf length and internod length. The highest value of genetic coefficient of variation and broadsense heritability were obtained for forage yield and number of stems per square meter, whereas the lowest value belonged to flag leaf width. A significant phenotypic correlation (P<0.01) were observed between forage yield with flag leaf length, days to pollination, plant hieght, internod length and number of stems per square meter. Comparing the means of cluster groups showed that populations number 1, 12, 3, 5, 15 and 8 had a desirable forage yield and vegetative growth. Factor analysis indicated that, 34.87 and 19.21 percent of total variance were explaned by the first and second factors respectively. Results of mean classification and cluster analysis of the triats confirmed by populations grouping based on first and second factors. Populations 1, 12, 15, 22 and 3 showed the highest vegetative and generative growth, based on the first two factors. Consequently, populations 1, 3, 12 and 15 showed superiority for vegetative and generative triats and could be selected for synthetic cultivar development in breeding programs under Kermanshah province climate conditions.

Thyme is an herb of the Lamiaceae family. The plant is very important in the world of medicinal properties. In order to study morphologic variation among a number of ecotypes of thyme, an experiment was performed based on a randomized complete block design with three replications and 24 treatments at Research Center of Agriculture and Natural Resources of Zanjan. Plant height, length, width, canopy, yield, branch length, leaf length, leaf width, days to first flowering, days to 50% flowering, days between first flowering and 50% flowering dates were recorded. Correlation between the studied agronomic traits was significant. Stepwise regression for plants yield showed that plant length and number of days between first flowering and 50% flowering had a positive effect on yield and plant height. Seed weight had a negative impact on yield. Cluster analysis based on the studied agronomic traits divided the ecotypes into three major clusters. Ecotypes number 1, 3, 5, 7, 9, 11, 15, 17, 20, and 24 located in cluster number 1. Ecotypes number 8, 14, 18 located in cluster number 2 and the rest of the ecotypes were classified in cluster 3.

Relationships between different morphological traits were assessed on 75 tall fescue genotypes under moisture stress and non-stress conditions, using several multivariate statistical methods including regression analysis, path analysis and factor analysis during 2011 and 2012. Results showed that drought stress significantly affected all the studied traits except for days to ear emergence and plant height. There were significant differences among the genotypes for all the studied traits indicating a broad genetic diversity in the germplasm. Correlation coefficients of dry forage yield of the first and second cuts with most of the recorded traits under moisture stress and non-stress conditions were significant. The results of stepwise regression showed that crown diameter, days to pollination and the number of fertile spikes contributed in 63% of observed variation for forage production under non stress condition. Under stress condition, the number of fertile spikes, crown diameter and plant height contributed in 73% of the observed variation for forage production. Path analysis indicated that crown diameter and number of fertile spikes had the strongest direct effects on forage yield. Factor analysis for all of the traits revealed four factors under non-stress and stress conditions which justified 80 and 77 percent of the total variation, respectively. In factor number 1 traits such as forage yield, number of fertile spikes and crown diameter and in factor 2, days to ear emergence and pollination had the highest factor loads.