Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research
Year:2018 | Volume:26 | Issue:1 (51) |Papers Count:18

Papaver somniferum is one of the important commercial sources for several pharmaceutical alkaloids including the narcotic analgesics codeine and morphinane and the semi-synthetic drugs oxycodone, buprenorphine and naltrexone. One of the downstream genes in alkaloid biosynthesis pathway in the species is thebaine-6-O demetylase (T6ODM) that inverts thebaine and oripavine to codeinone and morphinone, respectively. In this study, a transient silencing construct based on virus-induced gene silencing (VIGS) technique was used to knockdown the T6ODM expression. In this regard, cDNA of T6ODM was prepared using specific primers and the results of reaction were sequenced for confirmation. Then, a part of resulted cDNA (silencing fragment) was selected for cloning in pTRV vector. The silencing construct was infiltrated to plant leaves via Agrobacterium tumefaciens. The presence of cp (coat proteine) gene of TRV vector was assayed by PCR and the positive TRV plants were screened for analysis of gene expression by semi quantitative and real-time PCR. Results of gene expression analysis showed that transgenic plants had a significant decrease in expression (with average amount of 83. 71%), comparing to control plants.

An anti-cancer drug called Taxol, or paclitaxel is one of the secondary metabolites of yew tree (Taxus bacatta). Taxol is one of the most important and most useful anticancer drugs in the treatment of lung cancer. Taxol prevents the depolymerization of microtubules and inhibits cell division in tumor tissues. JAMYC is a transcription factor from MYC family with a DNA-binding motif called bHLH. The gene involved in Taxol biosynthesis has been identified in other yew species. The aim of this study was isolation and cloning of TbJAMYC gene and bioinformatics analysis of its structure. Initially to obtain yew seedling, embryos were removed from seeds of the species and cultured on MS medium containing activated charcoal and ascorbic acid. Total RNA was extracted from the seedlings and cDNA was synthesized from extracted RNA. The gene of TbJAMYC was amplified by PCR from cDNA. Amplified fragment cloned in pTG 19 vector using TA cloning method and sequenced. Bioinformatics analysis was carried out by MODELLER and Pfam softwares. The cloning was confirmed by endonuclease digestion and sequencing. TbJAMYC bioinformatics analysis was carried out by Pfam and MODELLER softwares that revealed three-dimensional structure of the DNA binding domain. These analyzes showed that TbJAMYC protein is a member of MYC transcription factors family that binds to DNA and regulates gene expression by bHLH domain. TbJAMYC transcription factor involved in the biosynthesis of valuable anticancer Taxol drug. Hence Taxol biosynthesis pathway manipulation via genetic engineering can be a new way to increase production of the metabolite in yew plant.

Soapwort (Saponaria officinalis L. ) is a native plant to central and southern Europe. It is an important and rare plant species in Iran. The species has numerous applications in medicinal, agricultural and health fields. Saponins is one of the bioactive compounds in the species that is used in chemotherapy. Tissue culture is one of the ways of proliferation of the species that is used for production of active ingredients. Saponin has ten isoforms that are different to inhibition of translating ribosomes and cytotoxicity. SO6 is one of the isoforms of saponins. Presence of SO6 protein in soapwort callus was studied through plant tissue culture and different concentrations of growth regulators and environmental factors. After achieving success in tissue culture, protein was extracted from its callus and presence of SO6 protein was confirmed by western blot analysis and measured by ELISA. Results showed that seed germination was possible through using appropriate treatment and soapwort root callus had the highest amount of SO6 protein.

Persian oak (Quercus brantii Lindl. ) with a vast habitat in Zagros mountain range, has a specific importance in Iran, based on economic, sociologic and environmental aspects. A complex of human and non-human factors caused to show genetic erosion in several plant populations of the species. Therefore, evaluation of genetic variation and architecture of plant population could be informative for forest managers and breeders. The research was performed to evaluate genetic potentials of a plant population of the species located in Lorestan provinces, west of Iran. Seed was collected randomly on twelve single plants to be used in assessment of the parent plants by a progeny test experiment. The seeds were evaluated as twelve half-sib and Full-sib families planted in a completely randomized block design with three replications. Biometric parameters were converted to genetic parameters using nested mating design model, by which genetic variance components and heritability values were estimated. The studied families showed a range of 6 to 92 percent establishment. Analysis of variance revealed significant difference between the parents based on all of the recorded traits. Majority of the variance components was observed between the studied families, for which heritability of the characters was also strong, varying between 57% on collar diameter, to 95% on leaf length. Therefore, the studied oak plant population has the capability to be used for selecting suitable materials for seed orchard establishment as well as performing other breeding projects.

Urtica dioica L. is dioecious plant and cross pollination with active components and special medicinal properties with economic values. Due to high level of heterozygosity of the species, homogeneity in fiber production and harvesting of selected plants is not guaranteed by seed propagation. For this reason micropropagation and asexual proliferation is suggested for production of selected plants. For micropropagation of the species, apical buds of Nettle shoots were used as explant. After sterilization, the explants were transferred into MS medium containing BAP, 2iP and IBA. Then, to evaluate the culture conditions, the explants were located on culturing mediums (MS, DKW, WPM) with different concentrations of growth regulators (IBA, 2, 4-D, BAP, KIN, 2iP, TDZ) in a factorial experimental model based on a completely randomized design with three replications. The best medium for shooting initiation was WPM culture. Between rooting treatments, the best root number and secondary roots belonged to KIN (0. 5 mgl-1) + IBA (0. 5 and 1 mgl-1) and BAP (0. 5 mgl-1) + IBA (0. 5 mgl-1) treatments, respectively. Then rooted plantlets transferred to the greenhouse and successfully were acclimatized (55. 5%).

Satureja khuzistanica Jamzad is an endemic species of Iran. The species is an endangered and valuable medicinal plant. In order to propagate through micropropagation, lateral buds were collected from Lorestan province, region of Pol of Dokhtar. The best treatment of sterilization of the explants was 0. 1% HgCl2 for 6 minutes. The best medium for shoot regeneration was MS with BA, 2iP and IBA 0. 3, 0. 3 and 1 mg/l, respectively. Applying 12 different treatments, the mentioned medium was better than other treatments in term of the coefficient of elongation, leaf elongation and leaf greenness. The best medium for root regeneration was MS with 1 mg/l IBA and 0. 1 mg/l NAA. It initiated the highest number of primary and secondary roots. Various combinations of different soils were evaluated for survival during the acclimatization, and sand: soil: peat: perlite at a ratio of 1: 1: 1: 1 showed the highest percentage of survival and growth. The best propagation method of the species was developed and the propagated plants could be transplanted into the habitat of the species.

Zhumeria majdae is one of the most important aromatic plant species from Lamiaceae family. The valuable species has medicinal and industrial applications. It is an endemic and endangered species of Iran. Therefore, tissue culture techniques are proposed for propagation and conservation purposes of the species. Usual regeneration of the species is by seed. Apical buds of seedlings from Hormozgan genotypes were sterilized by HgCl2 0. 1% solution for 5 minutes during spring season. The best medium for shoot proliferation was WPM containing BA (0. 5 mg/l), 2iP (0. 3) and IBA (0. 1 mg/l). The highest rooting of the shoots was obtained by WPM supplemented with 1 mg/l of IBA. The plants were acclimatized in greenhouse after transferring in soil. The protocol of this research could be used for culture and developmental programs of the species in its original habitat.

Fritillaria raddeana is one of the most fascinating wild flowers, belonging to Lilliaceae family, native to Iran. It is an important ornamental-medical bulbous plant, tolerant to arid conditions and stony slopes but facing extinction. There have been very few reports exist, so any studies on the species would be of great importance. Regardless of mutation occurrence, successful regeneration will be a prerequisite for further researches that must be done on the endangered plant species. This research presents an in vitro callus induction and indirect regeneration of F. raddeana via culturing petal and leaf. The leaf (before flowering) and petal (green to yellow flower buds) explants excited from plant and surface sterilized and were cultured on MS media containing different concentrations of Auxin and Cytokinin. The explants were first kept at darkness and 18º C for callus induction, then were transferred to light condition with the same temperature. The temperature was raised to 20º C, when the calli sub cultured into regeneration media. Although due to the low amount of bacterial and fungal contamination, conservation of parent plants, adequate and ready available explants, both petals and leaves are acceptable explants, among them petal showed better and quicker response to variable plant growth regulators. The best medium for callus formation (82. 66%) and indirect regeneration (36. 66%) via petal explants was MS medium supplemented with 2 mgl-1 TDZ and 0. 5 mgl-1 IBA. Therefore, the mentioned plant growth regulators were more suitable than the others for indirect regeneration of F. raddeana.

Amygdalus scoparia spach. a multi-purpose species which is growing in most of vegetation regions of Iran. Knowledge of genetic variability and heritability is essential prerequisite for conservation, forest development and sustainable use of forest plants. Genetic parameters on early growth were estimated in open-pollinated combined provenance-progeny tests. The experiment was conducted based on a nested model using randomized complete block design with 7 population, 10 mother trees per population 7 individuals per subplot and 3 replications in nursery of Kohmareh-Sorkhi of Fars (1660 m a. s. l. ). Significant differences among the populations and mother trees within populations were observed for all of the studied traits, except for the taproot length among mother trees within populations. Evaluation of components of variance showed that the population effects (ranging from %5. 9 to % 34. 2) contributed to total variance more than mother trees within populations effects (ranging from%1. 8 to %11. 1). Additive genetic variation coefficient in populations varied from 0 to 58. 6%. In general, Khonj and Kavar populations performed better growth. Heritability was low and not yet stable for some traits in some populations. Therefore, selection should be postponed until the heritability values reach to relatively high and stable values.

Assessment of genetic diversity and germplasm classification of plant species are important activities in plant improvement and management of genetic resources. Regarding observed variation on various traits of Zataria multiflora Bosis. random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to assess genetic diversity of the species in Iran. Leaves were collected on thyme plants of 15 different regions of Kerman, Sistan and Balochistan, Boushehr, Fars, and Khuzestan provinces. Leaf DNA extraction was carried out by cetyltrimethyl ammonium bromide (CTAB) method with small modifications. Ten RAPD markers and 10 ISSR markers with sharper bands in polymerase chain reaction (PCR) were used for data analysis. NTSYS-pc software and dice similarity coefficient, as well as Unweighted Pair Group Method with Arithmetic Averages (UPGMA) were used to draw the related dendrogram. Cophenetic correlation coefficient was calculated and a two-dimensional graph was created based on principle component analysis (PCA) results. Range of the bands produced by the primers varied from 120 to 3100bp. The 15 selected regions were classified into three separate groups. The established clusters were in a good harmony with geographical conditions. Entirely the markers produced 207 bands with polymorphism percentage of 86. 9. The results of the dice similarity coefficient produced by NTYSIS suggested that genetic similarity of thyme varied between 0. 4390– 0. 8460. The most genetic similarity was observed between Andimeshk and Eizeh samples and the least similarity was observed between Iranshahr and Jiroft samples. Results of principle components analysis and cluster analysis were similar. Cophenetic correlation coefficient was 74. 60%. The results suggested that RAPD & ISSR markers are useful for investigating genetic variability of Zatraia multiflora germplasm.

Propagation of Pronus avium L. through conventional micropropagation is essential for forest development and production of scion for grafting of commercial Pronus varieties. For micropropagation and production of juvenile tissue as an initial source of proliferation, mature embryo explants were used. For in vitro shoots preparation, isolated embryos of mature trees were transferred into MS medium containing 30 and 60 g/l sucrose. There were no significant differences between MS media containing 30 and 60g/l for percentage of embryo germination using t-Student test. Factorial analysis of collected data for further shoot proliferation indicated that there were highly significant differences between the used growth regulators for in vitro shoot length and number of produced explants at α =0. 01 level. The highest average shoot length (5. 86 cm) and the highest mean number of explants production were observed on MS medium supplemented with 4 mg/l BA and 0. 2 mg/l IBA. Plantlet at 4 to 5 cm in height were rooted in MS hormone free medium and after successful acclimatization transferred to a green-house.

Berberis vulgaris var. asperma, is a medicinal plant of Iran and lack of healthy vigorous seedlings is the main limitation of barberry cultivation. This research was carried out to develop a suitable technique for mass micropropagation of the species. Effects of different plant growth regulators, different gelling agents and different carbon sources on in vitro growth of barberry were studied in several experiments. Based on the results of the experiments, nodal segments containing 1-2 axillary buds of semi-hardwood shoots were the best explants for establishment of clean in vitro cultures. Surface disinfection of plant materials originated from adult trees was performed with immersion in 2 gl-1 of Benomyl solution for one hour, then 1% solution of commercial bleach for 15 minutes and rinsed three times with sterile distilled water. Explants cultured on media solidified with Phytagel were significantly better than the media with DifcoBacto agar and Agargel for shoot production. Addition of 3 gl-1 activated charcoal resulted in control of nutrient medium and explants browning, improvement of survival and in vitro growth of the explants. DKW medium containing 2 mgl-1 BA, 0. 5 mgl-1 IBA, 2 mgl-1 GA3, 3 gl-1 activated charcoal and solidified with 2. 5 gl-1 Phytagel was optimum for in vitro growth of barberry and recommended for shoot multiplication of the species. Rooting experiment was carried out using a two phase procedure, root induction in media with high concentration of IBA, followed by transferring to growth regulators free medium for root development. No rooting was observed and further research is needed.

Agropyron pectiniforme having moderate to high yield and quality traits is cultivated for pasture regeneration and forage production in rangelands of Iran. In order to study genetic variability in Agropyron pectiniforme based on morphological traits, 16 populations were examined in the field trial using a complete block design with three replications during 2004-2005 in Karaj, Iran. Data were collected on heading date, plant height, tiller number, spike length, forage yield, seed yield and thousand grain weights. Results showed significant differences between the populations for all of the recorded traits (P≤ 0. 01). Populations Zangan-487, Tabriz-707 and Gorgan-7565 with averaged values of 96 to 119 g/plant and 25 to 33 g/plant had higher forage and seed production, respectively. Among them, Zangan-487 had early maturity date and Tabriz-707 had higher seed thousand grain weights. Phenotypic correlation between forage and seed yield was positively significant (P<0. 01). Both traits positively correlated to plant height, tiller number and spike length. The populations were classified into three groups based on cluster analysis. Cluster number 1 contained populations with late maturity and moderate yield. The populations in cluster 2 had higher forage and seed production and populations in cluster 3 were early maturity and low productivity. Distance between the clusters were assessed using principal components analysis (PCA) and Mahalanobis Distance. Result showed higher genetic distance between clusters 2 and 3. It was suggested to use populations located in the separate clusters for improving composite varieties. The population Zangan-487 having higher both forage and seed yield was introduced as promising population for future research. Result of mantel correlation test between population’ s geographical origin and matrix genetic distance of morphological traits was not significant. This finding indicated a lack of association between genetic variation and their geographic distribution. It was concluded that morphological traits provide important information for the genetic conservation and utilization of future breeding programs in A. pectiniforme.

In order to determine relationship among forage dry matter yield and soeveral associated agronomic traits in Agropyron genus, 31 populations of three Agropyron species (A. desertorum, A. cristatum and A. elongatum) were evaluated in a randomized complete block design (RCBD) with three replications in research farm of Agricultural Biotechnology Research Institute of Northwest and West region of Iran-Tabriz. Correlation coefficients analysis showed that dry forage yield in A. elongatum, had significant positive correlation with canopy diameter and negative correlation with number of days to pollination. For A. desertorum and A. cristatum dry forage yield had also significant and positive correlation with number of stems and negative correlation with flag leaf width. Using stepwise regression analysis, in A. elongatum, canopy diameter and number of days to pollination and in A. desertorum and A. cristatum, number of stem and panicle length were entered into regression model. Path analysis for dry forage yield as a dependent variable, revealed that in A. elongatum canopy diameter had positive direct effect (0. 61) and number of days to pollination had negative direct effect (-0. 46) and in A. desertorum and A. cristatum, number of stems had positive direct effect (0. 73) and panicle length had negative direct effect (-0. 44) on dependent variable. Results showed that canopy diameter and the number of days to pollination in A. elongatum, and the number of stem in the A. desertorum and A. cristatum, had great effect in determining forage dry matter yield and therefore, could be recommended as indices in screening suitable genotypes.