نشریه تحقیقات ژنتیک و اصلاح گیاهان مرتعی و جنگلی ایران
سال بیست و هشتم شماره 2 (پیاپی 56، پاییز و زمستان 1399)

Phylogenetic study of savory (Satureja spp.), which has different species and valuablecompounds such as thymol and carvacrol, is of special importance among medicinal plants. In this study, 21 accessions belonged to nine Satureja species were identified and collected from main habitats in eight provinces of Iran, including East Azarbaijan, Kurdistan, Gilan, North Khorasan, Yazd, Lorestan, Ilam, and Kermanshah. Genomic DNA extraction was performed using the modified CTAB method. In the phylogenetic study, in addition to the nine collected species, the sequences of ITS and matK genes of Satureja species registered with the NCBI were also added to the phylogeny tree as part of the group. In addition to Satureja genus, the sequences of Thymus, Ziziphora, Clinopodium genera were also used NCBI as an outgroup to root the phylogenetic tree. The results showed that in the study of Satureja. spp kinship relationships, the ITS nuclear gene was more appropriate than the matK chloroplast gene. In the native species of Iran, the maximum genetic distance was between S.mutica and S.atropatana and the minimum genetic distance was between S.rechingeri and S.khuzistanica. In the phylogenetic tree, five species of S.macrantha, S.atropatana, S.sahendica, S.edmondi, and S.bachtiarica with %100 bootstrap value, two species of S.mutica and S.spicigera with %96 bootstrap value, and two species of S.khuzistanica and S.rechingeri with %100 bootstrap value were close relatives. The results showed a significant consistency with Flora Iran and Flora Iranica in terms of morphological traits.

Fagus orientalis is one of the most important broadleaf species in Hyrcanian forests. The phenotypic and genetic adaptation potential of this species to climate change is very important. Quantifying and evaluating genetic diversity between and within oriental beech populations makes it possible to best preserve the diversity of populations. This study was conducted on mature trees of four different populations (regions) of this species in two drought index gradients from West to East and lowland to highland in Hyrcanian forests (Shafarood forest of Gilan, Khairudkenar forest of lowland and highland of Mazandaran and Shast Kalateh forest of Golestan). Genetic diversity was studied in native populations using seven microsatellite markers and one biochemical marker. The results showed high diversity in all populations, especially in the population of Shast Kalateh, Golestan. The results of molecular analysis of variance with SSR microsatellite marker showed that 77% of the total diversity was within populations and 23% was among the populations. The results of this study showed that the presence of more genetic diversity within the beech populations of Hyrcanian forests indicates their strong genetic basis for adaptation to environmental conditions and may facilitate short-term adaptation to climate changes. On the other hand, the presence of higher genetic diversity in Shast Kalateh than in other regions indicates stronger evolutionary forces for adapting trees to environmental conditions. Additionally, higher gene flow among allogamous populations of oriental beech has led to low genetic differentiation between populations in the drought index gradient of Hyrcanian forests.

To evaluate the genetic variation in an in vitro culture of two samples of Hyoscyamus niger callus including new (one-month-old) and old (one-year-old) calli versus fresh leaves of the plant, the Inter Simple Sequence Repeat (ISSR) marker was used. Using 10 primers, 6.49% polymorphism was observed in the three mentioned samples. Analysis of Molecular Variance (AMOVA) revealed 68% genetic variation among samples. Jaccard and Dice coefficients (similarity matrix) were also used by UPGMA and NJ methods. The best dendrogram to show genetic diversity (and distance or proximity of individuals) was known as the Jaccard similarity coefficient. The results of cluster analysis grouped the samples into two different groups. This classification was also confirmed by principal component analysis (PCoA). The first major group consisted of old calli and the second involved new calli and plant leaves. The result showed that the frequency of somaclonal variation in H. niger calli tended to increase with increasing the number of subculture times.

To investigate the broad-sense heritability of some Haloxylon traits, an experimental design was performed using native populations collected from different provinces of Iran. 32 native populations were planted based on a randomized complete block design with three replications in Shaheed Zendeh Ruh's research station in Kerman. During three years, seven traits including main stem length, collar diameter, number of tillers, sub-stem length1, sub-stem length2, the small and large diameter of the canopy on shrubs were examined. The data were assessed by simple and combined analysis of variance. For all traits, the difference between the mean of genotypes and years was significant (p < %1). The genotype× year interaction effect was significant for all other traits except the number of tillers, which indicates different effects of genotypes in different years. Heritability of traits was estimated based on simple and combined analysis of variances. The amount of heritability was ranged from 75 to 87% for simple analysis and 51 to 64% for combined analysis, which indicates a decrease in heritability in a combined analysis. This is due to the genotypes by the year interactions effects. In both methods, the main stem length and number of tillers had the highest and lowest broad-sense heritability and coefficients of genetic and phenotypic variation, respectively. Due to the high genetic diversity among the studied Haloxylon genotypes, it will be possible to select the best rootstocks for making windbreaks in desertification projects.

The objective of this study was to evaluate the genetic variation of 26 selected genotypes of Phalaris aquatica L. based on important agronomic traits. Each selected genotype was divided into four clones and they were space planted according to a randomized complete block design with four replications under irrigation conditions in the Khalatposhan research station of Tabriz University, Iran. Data collection was started in March 2017 for two years. The results of analysis of variance showed significant differences among the studied genotypes for most traits (P<0.01). Accordingly, the mean comparisons of the traits among genotypes were conducted. Using principal component analysis (PCA), the first two components were accounted 66% of the total variation. The traits like forage yield in both cuts, crown diameter, plant height, and seed yield had the most important role in determining the first component. Based on cluster analysis, the 26 genotypes were grouped into four distinct groups. Based on the mean comparison results, principal component, and cluster analyses, 12 genotypes with the highest forage and seed yields in both years were selected as superior genotypes. Therefore, this population provided a suitable genetic source for breeding improved varieties using polycross progeny tests and breeding improved synthetic variety.

To evaluate the genetic variation of Rosa damascena as an important aromatic and medicinal species, 38 genotypes of this species were studied for the diversity of phenological and morphological traits (flowering period, plant height, canopy diameter, number of leaves at the plant, total green area of the plant, number of flowers per plant, fresh weight and percentage of dry matter content of flowers and flower yield per plant) in a randomized complete block design with three replications in Zalleh Research Station of Sanandaj, Iran during 2004-2008. The results showed significant differences (p < /em>≤ 0.01) among the genotypes, in terms of the studied traits. The average total yield of fresh flowers was 2427.4 g/plant. The genotypes belonging to temperate and cool areas had lower flower yields than those originated from hot and dry areas. Flower yield showed a significant  and positive correlation with the crown height and diameter, the number of leaves, total green surface, and the number of flowers per plant (p ≤ 0.01). The results of stepwise regression showed that the flower weight and number positively, while the total green surface and flowering period negatively correlated with flower yield. The results of cluster analysis (CA) grouping were relatively consistent with segregation based on the flower yield and characteristics of genotypes origin areas. The results of principal component analysis (PCA) showed high consistency with the CA results, but the genotypic separation in PCA method was clearer. The overall results of this study along with CA and PCA analyzes showed considerable rich genetic variation among the genotypes in terms of phenological and morphological traits and the possibility of using this diversity in Iranian Damask rose breeding such as improving flower yield through direct and indirect selections (through flower yield components) or hybridization of genotypes with a greater genetic distance for the use of higher heterosis.

This study aimed to estimate the genetic parameters of Haloxylon ammodendron located in the seed orchard of Yazd province, Iran, using REML method and repeatability model. For this purpose, among the stands of black saxaul in Yazd province, several rootstocks with suitable appearance, desirable quality, the relative balance between crown and trunk, resistance to pests and diseases selected and their seeds were sown in nursery, and the seedling were planted in the field based on a completely randomized design with unequal replications, 6 to 20 seedlings. The genotypes of each rootstock were planted on rows at 6-m intervals. Data collected and genetic parameters estimated for seedling traits as tree height, canopy expansion, trunk diameter, number of sub-branches, and freshness. Data were analyzed of variance using REML method, two different repeatability, and non-repeatability models. The results based on both models showed that the highest heritability was related to tree height. But these estimates between two methods were different so that, the genetic correlations and heritability values in the repeatability model were significantly lower than that for non-repeatability model. Repeatability was estimated based on the intragroup correlation between two independent variables on individuals and expressed as the ratio of differences between individuals to the existing phenotypic differences. Based on the results, the repeatability of the most traits was high. This result enables breeders to make good decisions about keeping rootstocks in the seed orchard during the growth stage in the early years. Finally, it was concluded that the repeatability model, could estimate moderate heritability for all of traits, which is more in line with the reality and quantitative nature of the studied traits and they could be mare affected by the environment factors.

In order to evaluate root yield and morphological traits of 15 endemic and imported chicory Cichorium intybus L.) accessions and cultivars, an experiment was conducted in a randomized complete block design with five replications in Khoshkedaran Research Station of Mazandaran province, Iran, during 2016-2018 years. In this study, the number of leaves per plant, leaf length and width, plant height, fresh and dry root yield, and some qualitative traits such as root shape and diameter, hairy leaf and stems, bolting time, lignified top root, longevity, and reproductive cycle of chicory accessions were measured. In the first year, the root was not harvested and in the second year, at the end of the rosette stage (rootstock start), the roots were harvested and the relevant measurements were made. Data for each morphological and functional trait were analyzed separately. The results showed that the Selenite cultivar had the highest fresh (35.458 t/ha) and dry (9.574 t/ha) root yield and ranked in the first group of means. The results of principal component analysis (PCA) based on yield and morphological traits showed that the first two components accounted for 72% of the total variation. Based on the results of cluster analysis (Ward method),   the genotypes were classified into four groups. The highest genetic distance was obtained between cluster two (Behshahr accession) and cluster four (foreign cultivars). The results of cluster analysis and PCA were in agreement with each other. According to the results, It was concluded that Tonekabon accession from the first cluster can be cross with the high-yielding cultivar Selenite to breeding improve new varieties of endemic chicory.

In order to study the effect of foliar application of humic acid on the yield, yield components, and some physiological and phytochemical properties of Stevia rebaudiana under different irrigation regimes, a factorial experiment was conducted based on a randomized complete block design with three replications in the research campus of University of Agricultural Sciences and Natural Resources, Gorgan, Iran. The treatments included four levels of irrigation regimes (70, 85, 100 (control), and 115% of field capacity) and humic acid foliar application at four concentrations (zero, 250, 500, and 750 mg/L). Irrigation regimes and humic acid treatments started from the 10-leaf stage and continued until the flowering stage. At full flowering stage, data collected for shoot and root length, root diameter, shoot and biomass fresh and dry weights, number of sub branches, flowers number, internodes number, leaves number, leaf area, root antioxidant activities, total phenol, total flavonoid, proline, ion linkage, and leaf relative water content. According to the results, the effect of different irrigation regimes on shoot height, shoot and biomass fresh and dry weights, sub-branches number, internodes number, was significant (p<0.01). By flooding, the plant height, shoot and biomass fresh and dry weights were increased. The interaction effect of drought stress by humic acid was significant for plant height, root antioxidant activity, and proline. Low irrigation stress had negative effect on the growth traits but humic acid foliar application had moderated the negative effects caused by low irrigation stress. Therefore, based on the findings, it was concluded that stevia is a resistant plant to both flooding and drought stress and its resistance mechanisms to drought stress for maintaining vegetative yield components, are to increase osmolytes and phenolic compounds.

The medicinal plant Lavender (Lavandula angustifolia) is cultivated all over the world due to its economic and ornamental potential and its wide application in the cosmetic and pharmaceutical industries. To investigate the effect of abscisic acid foliar application on increasing drought stress tolerance in Munstead Organic cultivar, a factorial experiment was conducted in a randomized complete block with three replications in the Faculty of Plant Production of Gorgan University of Agricultural Sciences and Natural Resources, Iran in year 2016-2017. Experimental treatments were considered at four levels of irrigation regimes (including 90-100, 70-80, 50-60, and 30-40% of field capacity(FC)) and three levels of abscisic acid foliar application (including zero, 15, and 30 mM/l). Data analysis showed that the highest percentage of the essential oil (1.18%) was obtained in 15 μM abscisic acid and 30-40%FC. While the highest yield of the essential oil (0.055 g/plant) was related to the application of 30 μM abscisic acid in 70-80%FC. The category of the essential oil components also showed that out of 26 identified components, six compounds were hydrocarbon monoterpene, 10 compounds were oxygen monoterpene, six compounds were hydrogen peroxide and four components were oxygen sedimentation. With increasing drought, stress, the amount of hydrocarbon and oxygen monoterpene compounds decreased, but hydrocarbon and oxygen sesquiterpene compounds increased. The results also showed that the highest amount of limonene (10%) was obtained from adding 15 μM abscisic acid in irrigation of 70-80%FC. The highest amounts of camphor (10.47%) and bernoulli (51.58%) were achieved in complete irrigation and non-use of abscisic acid. However, the highest amounts of cariofillenoxide (7.80%) and α-Muurolene (24.90%) was observed in 30 μM abscisic acid and 60-50%FC and in 30 μM abscisic acid and 30-40%FC, respectively. Overall, it was found that foliar application of abscisic acid under drought stress 30-40%FC, could increase the percentage of the essential oils and sesquiterpene compounds to cope with drought conditions in lavender.

Pot marigold (Calendula officinalis L.) is an important ornamental-medicinal plant in Iran, that is widely used alone or in combination with other medicinal to produce low-risk drugs for human. In order to mass-production of this plant by tissue culture, four separate factorial experiments were conducted using completely randomized design with three replications (10 samples) at four stages of callus induction, indirect organogenesis, rooting, and acclimatization The results showed that the highest callus weight was observed in the interaction of leaf explant and medium supplemented with 2,4-D (2.0 mg/l). The tallest and highest branching number was related to the callus obtained from leaf explant in medium containing thidiazuron (1.0 mg/l TDZ) and putrescine (100 mg/l PUT), in indirect organogenesis. Tissue culture cuttings had the best rooting in 2.0 mg/l NAA medium. The obtained seedlings planted in pots containing perlite showed better acclimatization. By optimizing callus production from Calendula officinalis explants and propagation by tissue culture method can be an important step towards producing plants with the same quality and free of pathogens.

The genus Ferulago includes multi-purpose plants with valuable pharmaceutical, industrial, and soil protective functions which are native to Iran and recently have been classified as endangered plants. Ferulago subvelutina Rech.F. is an endemic species to Iran and there is little information about its ecological, physiological, and morphological characteristics. Due to the importance of this species, the present study was conducted to investigate the mentioned characteristics and evaluate the antioxidant activity of three ecotypes in the natural distribution areas (including Izman-e Olya, Izman-e Sofla, and Darkesh) in North Khorasan province, Iran. Investigation of the plant growth stages (Izman-e Sofla ecotype) showed that its vegetative growth began with increasing temperature in the second decade of April. Full flowering begins in the second decade of July and continues until the first decade of August with decreasing rainfall and increasing temperature. In the third decade of July, the plants produce seeds and the seed dispersal begins in the third decade of August. It grows in altitude ranged 1360-1137 m above sea level, and it grows in soil with a pH of 7.7-7.81, organic carbon of 0.31-1.73%, loamy to sandy loamy texture, with average annual temperature of 13.6°C and annual rainfall 354.56 mm. Study of morphological traits showed that ecotype 3 (Darkesh) had higher values of plant height (48.80 cm), crown large and small diameter (98.60 and 93.40 cm), number of leaves per plant (4.67 pcs), and leaf length and width (58.1 and 52.5 cm), than that for other ecotypes (Izman-e Olya and Izman-e Sofla). The studied ecotypes had lower antioxidant activity than ascorbic acid as a control (13.97 μg ml-1), of which ecotype 2 (Izman-e Sofla) with an IC50 level of 566.72 μg ml-1 is noticeable due to its antioxidant activity compared to other ecotypes. Therefore, due to the importance of medicinal-rangeland species of F. subvelutina, the results of the current study could be effective in maintaining plant survival, breeding, and domestication, and also because of the high antioxidant activity of this species, could be used in the food industry.