نشریه تحقیقات ژنتیک و اصلاح گیاهان مرتعی و جنگلی ایران
سال سی‌ام شماره 2 (پیاپی 60، پاییز و زمستان 1401)

Juniperus is one of the few conifers in Iran, which has six species and forms mostly sparse forests with scattered trees. Among them, Juniperus polycarpos and J.excela are more widespread than the other species. Due to the great similarity in their phenotypes, it is difficult to distinguish the two species from each other. In this study, pollinological, anatomical, and molecular indicators were evaluated to identify and differentiate these two Juniper species in the Chahar-Bagh region of Gorgan province, Iran. Using nrDNA ITS nuclear marker, molecular relationships between the two species were investigated. Phylogeny analyzes showed that two species are monophyletic and the populations were subdivided into unresolved subclades I and II. Pollen grains of the two species, J. polycarpos and J. excelsa, were examined using light (LM) and scanning electron (SEM) microscopes. The results showed that all pollen grains were monad, ulcurate, asymmetric, and small in size. In terms of shape, both species were spheroidal (prolate spheroidal) with granular exine ornamentations. The general shape of the pollens in the LM was circular with a smooth surface and no fovea, but the pollen grains examined under SEM were irregular and had fovea. The results of the anatomical studies of the stems also showed little difference, and it was not possible to distinguish and identify the two species. The results indicate that one species is probably distributed in the Chahar-Bagh region and these two taxons are synonymous.

miRNAs genes are regulatory elements that their main role is to downregulate gene expression at the mRNAs level. miRNAs also play important roles in several plant pathways related to important cellular activities such as growth, reproduction, differentiation, morphogenesis, apoptosis and, response to abiotic and biotic stresses. This research was conducted to identify conserved miRNAs and their target genes using next generation sequencing data in the damask rose (Rosa damascena Mill.). Although bioinformatics methods have been developed as the most efficient strategy for target miRNA identification, high-throughput experimental strategies are still in high demand. Bioinformatics tools (Cmii, psRNATarget, WEGO, Blast2GO) and laboratory method (real-time PCR) were used to identify new miRNAs, predict and determine the ontology of the target genes involved in the production of scent and color in the damask rose, respectively. Using EST and RNA-seq data based on Rosa lucieae homology, four miRNA families, including: miR5021, miR2673, miR156, and miR838, were selected as candidate miRNAs. In the next step, for the quantitative evaluation of the real-time PCR to validate the expression level of the selected miR5021 in two samples (white and hot-pink) and the biological stage (young and bud) of the damask rose, foliar spraying with the concentrations of zero (control) and 300 μM of methyl Jasmonate for 48 hours was made. Result showed the highest relative expression of miR5021 belonging to the pink sample on the young development stage. These findings accelerate future prospective studies on the regulatory mechanisms of miRNAs in Rosa damascena.

In order to identify the ecological, micro-morphological and molecular characteristics of six alfalfa species (Medicago L.), this research was conducted by collecting data of habitat information, vegetation characteristics and the soil characteristic in sampling depth (0 to 30 cm) of the species in their habitat. The pollen grain morphology of the species was investigated using scanning electron microscopy (SEM). Investigation of the molecular relationships between six species was done using nuclear nrDNA ITS and rpl32-trnL(UAG) chloroplast sequences. The results of the ecological study showed that the most species in the study area grew in arid to semi-arid climate and in silty, silty-clay and silty-clay-loam soils. Based on the results, most of the pollen grains were medium-sized and trizoncolporate and rarely had four to six colpus. In addition, molecular studies showed that all the tested species formed a monophyletic group with high support. The perennial Medicago species was located in a separate clade in the vicinity of the annual species, which indicates that the annual species are monophyletic. The results of this study indicate the efficiency of molecular markers in determining the relationship of taxa at the species level, and it can be used as a complementary tool in the taxonomic identification of Medicago species.

This study was aimed to assess genetic variation and characterize 50 selected genotypes of Agropyron desertorum Fisch. ex Link. using agro-morphological traits and forage yield. Each genotype was clonally propagated into four clones and they were space planted in a randomized complete block design with four replications on the research farm of the Agricultural Biotechnology Research Institute of the Northwest and West regions, Tabriz, Iran. Data collected from March 2017 for two years. The results of the variance analysis showed significant differences among the genotypes for most of the traits (P<0.01), which indicated high variation among the studied genotypes for all the traits. Based on the mean comparison of annual forage dry matter yield over two years, the 14 genotypes with higher annual forage production (more than 350 g/plant equal to 9.8 t.h-1) and also higher seed yield (more than 40 g/plant equal to 1.12 t.h-1) were selected as the superior genotypes. Based on the cluster analysis, the 50 genotypes were divided into four clusters. The results indicated that there were significant variation and considerable distance for forage, seed yield and other traits among the studied genotypes. Thus, these genotypes could be suitable genetic sources for selecting superior parental genotypes to improve breeding composite varieties.

Black poplar (Populus nigra L.) is one of the dioecious species that is planted in a large area for forestry and wood production in different regions of Iran, but there is less information about its sexual response to water scarcity. In the present study, this plant was used as a model species against water deficit stress, and some morphological and physiological responses of a male clone (62/127) and a female clone (62/154) of black poplar were investigated. A factorial experiment was conducted based on completely random design with four replications. There were four irrigation levels (100, 75, 50, and 25% of field capacity) and two gender or sexes (male and female). Seedlings were irrigated for 120 days in a pot experiment. The results showed that the effect of drought and gender (sex) was significant on many of investigated traits. The sex × irrigation regime interaction effect was significant for height growth, total biomass, stomatal conductance, and transpiration. In all the drought treatments, males plants had higher values of many traits than that for females. It was concluded that that, in the studied black poplar clones, males black poplar were less sensitive to water deficit stress than that for females.

Cannabis (Cannabis sativa L.) is an important dicotyledonous medicinal plants that used in many developed countries of the world. The present study was aimed to evaluate the effect of salinity stress on morphological, physiological, and biochemical traits of male and female plants of cannabis (Cannabis sativa L.). A factorial experiment was conducted using salinity stress treatments at four levels (0, 50, 75, 100 mM) and plant sex in two levels (male and female genotypes) based on a completely randomized design with three replications under greenhouse condition. The results showed that the salinity stress by plant sex interaction effects were significant for plant yield components, total phenol, antioxidant activity, total flavonoid, proline and carbohydrate (P<0.01). Increasing the salinity level significantly decreased plant height, number of leaves, plant fresh and dry weights in both sexes. But, the root weight, root length, and number of flowers increased in female plants up to 75 mM salinity. Also, increasing salinity level significantly increased secondary metabolites such as phenol, flavonoid, proline, and soluble carbohydrates in both males and female plants. However, by increasing salinity level, the amount of antioxidants increased in the females and decreased in the male plants than that for control, respectively. The sodium to potassium ratio (N/K) was considered as an index of salinity tolerance. The highest N/K ratios were observed at 75 and 100 mM salinity levels in shoot and root of the female plants, respectively. The results indicated that cannabis female plants can tolerate salinity up to 75 mM salinity level, and the female biomass was increased, but above the 75 mM level, the female plants were more sensitive than that for male plants in terms of salinity.

The rapid and mass production of secondary metabolites through chemical methods is mostly expensive, difficult, or impossible. Considering the economic importance of these metabolites and their small production in medicinal plants, some methods such as the cultivation of hairy roots and the use of biotic and abiotic stimuli have been suggested to increase these metabolites. The accumulation of the tropane alkaloid atropine in Atropa belladonna L. is of great importance in the pharmaceutical industry. In this research, transgenic hairy roots of A. belladonna was produced using an A7 strain of Agrobacterium rhizogenes. The transformation of hairy roots was confirmed by PCR test. The results of the PCR test showed the correctness of the amplification of the bp780 fragment for the rolB gene in hairy roots. In addition, the effect of different concentrations (0, 125, 250, and 500 μM) of salicylic acid in two times of 24 and 48 hours on the amount of atropine content, the plant growth rate and the activity of antioxidant enzymes of A. belladonna hairy roots was investigated. The data statistically analyzed using factorial experiment based on a completely random design with three replications. The results of the analysis of variance showed significant effect of salicylic acid on all the traits (P<0.01). The effect of time was significant for the amount of atropine and ascorbate peroxidase (APX). The salicylic acid by time interaction effects were significant for the amount of atropine, catalase (CAT), and guaiacol peroxidase (GPX) (P<0.01, 0.05). The results showed that by increasing salicylic acid concentrations the growth of hairy roots significantly decrease compared to the control and the lowest values of fresh and dry weight of the hairy roots (2.79 and 0.148 g) were obtained in 500 μM of salicylic acid compared to the control (6.76 and 0.273 g). However, based on GC/MS analysis, the highest amount of atropine (11 times more than the control) was obtained in the 250 μM salicylic acid in a period of 24 hours. The highest catalase and guaiacol peroxidase activities were obtained at concentrations of 250 and 125µM salicylic acid for 24 hours, respectively. The highest level of ascorbate peroxidase enzyme activity was obtained at the concentration of 250 µM salicylic acid compared to the control. It was concluded that the salicylic acid stimulant at low concentrations is useful for cell signaling and stimulating the production of secondary metabolites, but at high concentrations, it disrupts the growth of hairy roots.

Teucrium stocksianum Boiss. is one of the important medicinal plant species that is naturally grown in the Southern highlands of Hormozgan and Sistan & Baluchestan provinces, Iran. The aim of this study was to investigate the diversity of some morpho-physiological traits of different populations of T. stocksianum collected from natural habitats. The results showed significant differences among the populations for all the morpho-physiological traits. Plant height was positively correlated with plant height and width. Shoot dry weight was also positively correlated with root dry weight. Principal component analysis (PCA) showed that the first five components accounted for 84.15% of the total variation. In the first component, the inflorescence length, plant height, root dry weight, plant width, and stem dry weight were important traits (%31). Cluster analysis divided the populations into three groups and showed that the cluster analysis using morpho-physiological did not correspond to the geographical diversity. Based on the results, the populations of the second group due to having higher values for desirable traits such as large diameter, small diameter and plant length, stem and root dry weight, main stem diameter and the weight of 1,000 seeds, were introduced to breeding improved varieties.

Kelus (Kelussia odoratissima Mozaff.) is one of the medicinal, spicy, aromatic and endemic herb that grows in restricted regions of Iran,. Due to the importance of its pharmaceutical compounds in traditional medicine, the excessive harvesting of kelus has put it at the risk of extinction. To protect kelus from extinction, plant tissue culture can be used to produce useful compounds and secondary metabolites. The hairy root culture is an efficient system for the production of medicinal compounds and valuable secondary metabolites in plants. For this purpose, two pathogenic strains of Agrobacterium rhizogenes (A4 and ATCC15834), two types of explants (cotyledon and hypocotyl), three levels of acetosringon (a1= 0 µmol, a2=50 µmol, and a3= 100 µmol), and three levels of glucose (b1=0, b2=0.5%, b3=1%) were applied using a factorial experiment based on a completely randomized design with three replications. The effect of such different factors on the induction and production of hairy roots in kelus was investigated. The results showed that both Agrobacterium strains were able to induce hairy roots in kelus. Also, ATCC15834 strain in MS medium with 100 µmol of acetosringon in the presence of 0.5% glucose had the highest efficiency in the production of hairy roots in cotyledon explants. The lowest amount of hairy roots was observed for A4 strain in MS medium without both acetosringon and glucose in both explants. PCR analysis was performed with rolC specific primers to ensure hairy root induction has occurred by A. rizhogenes. The results of this research showed that for the first time in Iran, it was possible to induce hairy roots in kelus explants.

Lavender (Lavandula officinalis L.) is one of the rangeland species in the many parts of the world, which is cultivated as a medicinal and ornamental plant. Plant tissue culture techniques are an alternative method for the mass production of lavender colons and can overcome the problems caused by reproductive propagation. To determine the effect of different combinations and concentrations of growth regulators, and also explant type in the process of callogenesis, organogenesis, and rooting of lavender, a factorial experiment was conducted based on a completely randomized design with three replications. In this research, different responses to growth regulatory compounds were assessed at different stages. The best treatment for callogenesis was cotyledon explants in culture medium containing 0.5 mg/l 2.4-D along with 0.25 mg/l BAP. In indirect organogenesis, the highest number of branches and the rate of organogenesis were obtained in the medium containing 2 mg/l of KIN. Also, the highest number of branches was obtained from using 0.5 mg/l BAP treatment with 0.5 mg/l of KIN along with 0.1 mg/l IBA. The results showed that the best culture medium for rooting was the medium containing 1 mg/l IBA. The highest explants survival rate was related to using perlite and coco peat as a bed of cultivation. The finding of this research introduced an efficient regeneration system for this plant that can be used in rapid vegetative propagation, germplasm maintenance, and molecular modification programs through genetic engineering.