Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research
Year:2018 | Volume:25 | Issue:2 (50) |Papers Count:18

Real-time PCR technique is available for determining changes of gene expression. However, normalizing gene expression data and obtaining reliable results depends on the selection of appropriate reference genes which are stably expressed under different experimental conditions. In this study, we investigated suitability of two commonly used reference genes [elongation factor 1 alpha (OvEF1alpha) and 18S ribosomal RNA (18S rRNA)] in seven Thyme populations and under exogenously applied gibberellic acid (GA3) hormone (control, 30, 60 and 90ppm), plant age (young plants and middle-aged plants and temperature/light stress. Results showed that expression levels of both genes in all populations were affected significantly. Plant age did not significantly effect on expression of the two genes. Also, ovEF1alpha unaffected by exogenously applied GA3. Under temperature/light stress, expression of OvEF1alpha increased in Babgorgy population but in Babzangy decreased. In contrast, expression of 18S rRNA in Babgorgy was significantly decreased but has stable expression in Thyme and Babzangy. Thus, these results showed that ovEF1alpha for comparison of gene expression between Thyme, Hanza, Sirch and Zarand and 18S rRNA for Babzangy, Sirch and Hanza or Zarand and Thyme populations are suitable as internal control gene. Also, for temperature/light stress both genes are suitable in Thymus vulgaris L. but only 18SrRNA is suitable in Thymus caramanicus Jalas.

In this study, an indirect organogenesis regeneration protocol for Salvia officinalis L. is described. Leaf (0. 5 × 0. 5 cm) and internode (1 cm in length) explants sections, from 45 day old in vitro germinated plant, were cultured in Murashige and Skoog (MS) basal medium supplemented with 12 different combinations of 2, 4-dichlorophenoxy acetic acid (2, 4-D) or 1-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP). The best calli induction response was observed when 1 mg/l NAA and 0. 5 mg/l BAP combination was applied to the internode explants. Also, results showed an interesting possibility for the stimulation of shoot regeneration (5. 66) through internode segments cultured on MS medium containing 50 mg/l putrescine and 0. 5 mg/l TDZ. Shoot elongation and rooting were achieved on MS medium with 1mg/l IBA. Excellent acclimatization to greenhouse conditions was observed for all transferred plantlets. By this procedure no morphological differences were observed between the regenerated and mother plants. This protocol may be also utilized to carry out plant regeneration after genetic transformation, in order to develop transformed plants without the presence of chimeric zones.

Fennel (Foeniculum vulgare Mill. ) is one of the well-known medicinal and aromatic plants. To determine variability between 16 Iranian fennel accessions based on a molecular marker and morphological traits an experiment was carried out based on a completely randomized design with three replications at Payame Noor University of Kermanshah, Iran, during 2014. Eleven morphological traits were recorded. Genotypes were evaluated based on 10 SCoT primer markers as well. Analysis of variance showed significant differences among the genotypes for the studied characters. Mean comparison according to DMRT was carried out and significant differences were revealed for all of the studied traits (p≤ 0. 05). Cluster analyses classified the accessions into four groups. Principle components analysis revealed that 81. 4 percent of variance explained by four PCAs. The strongest heritability (%71. 37) belonged to main stem. SCot primers showed 55 visible bands. SC5 and SC29 primers had the most number of bands with 10 and 9 bands respectively. Means of polymorphic information content (PIC=0. 36), marker index (MI= 1. 82), effective multiplex ratio (EMR= 4. 8) and resolving power (RP= 5. 86) indices were calculated for all primers. Total genetic similarity based on the primers was 0. 66 percent. Cluster analysis classified the accessions into 3 groups. The clustering results was confirmed by principal coordinate analysis.

Savory (Satureja khuzistanica) is one of the nine endemic species which have phenolic compounds such as carvacrole and thymol in its essential oil and rosmarinic and other phenolic acids in its extract, has strong antioxidant and antimicrobial properties and remarkable effects on several diseases. Drought stress was induced by stopping irrigation at flowering stage. Samples were taken five times with three interval days and several physiological traits were measured. For oil extraction, Clevenger apparatus was used. For determination of essential oil’ s components and methanolic extract compounds, GC, GC/MS and HPTLC were utilized. Profiling of volatiles using GC/MS, showed an increasing-decreasing trend at major phenolic and terpenes compounds such as thymol, γ-Terpinene, p-Cymene, Rosmarinic acid and Caffeic acid. Drought stress also led to a significant increase in oil yield, soluble sugars and proline as well as a significant reduction in leaf water potential, relative water content and pigments. Metabolite profiling of Satureja khuzistanica represents the strategies employed by savory in generating different biochemical phenotype. Applicable results of this study for increasing product quality is effective application of drought stress before harvesting. The results verified that drought stress affected physiological characteristics and secondary metabolism of savory which were useful for future work by using metabolic engineering focusing on important species to increase main compounds, such as carvacrol and rosmarinic acid and caffeic acid.

Because of adaptation to different ecological conditions, cross pollination and heterozygosity, Populus euphratica contains a large amount of genetic diversity, which could have an important role in superior phenotypes selection, in inter and intra specific hybridization processes. The main aim of this study was evaluation of leaf morphologic and Isoenzyme characteristics in several P. euphratica genotypes, produced by seed regeneration. A total of 50 superior seedlings from superior phenotypes of 12 parent trees of the species were selected. Seedlings evaluation was performed in a nursery located at Karaj research station, a city close to Tehran, Iran, during 2011 to 2015. In order to determine existing genetic variation, leaf morphology of seedlings and analysis of peroxidase isoenzyme were conducted. Peroxidase isoenzyme analysis was conducted on leaf extracts using polyacrylamid gel electrophoresis (PAGE ). Analysis of variance showed significant differences among the seedlings of the parent trees for leaf morphologic traits. The most values of leaf length, leaf width and leaf area were observed on Gotvand and Lorestan originated seedlings, and the lowest value of the characters was observed on seedlings originated from Zabol habitat. Analysis of variance of superior genotypes from different habitats revealed within superior genotypes genetic variations (69%) and among habitats genetic variations (31%). Variation index (Phi) showed significant differences within progenies and superior parental trees. The most part of heterozygosity observed in Dezfol, Sarakhs and Zabol trees, with 1, 0. 83, 0. 83 values respectively. Maximum number of effective alleles and Shannon index belonged to Zabol habitat with 2. 93 and 1. 12 respectively.

Phalaenopsis amabilis is one of the best and well-known orchid species that is easy to grow and has good compatibility. This study was done for induction of polyploidy in Phalaenopsis amabilis by in vitro conditions. The study was conducted as factorial based on a completely randomized design with three replicates and each replicate including three explants of auxiliary buds (approximately 3 mm ) containing two leaves using seven different concentrations of colchicine (0, 0. 01, 0. 05, 0. 1, 0. 15, 0. 2 and 0. 25 percent) in two time periods (24 and 48 hours). Explants were cultured on MS medium without hormones after treatment with colchicine. After 10 weeks sevral morphological traits that were under influence of polyploidy, including number and size of leaves, number and size of roots, length and width of stomata guard cells, number of chloroplast, and survival rate of seedlings were evaluated. Cytological observations of new developed leaves were evaluated by flow cytometry to assess the number of chromosomes in plants. The results showed that the highest percentage of polyploidy was obtained using 0. 15 and 0. 2 percent of colchicine concentrations. Correlation of some morphological traits with polyploidy levels showed that in addition to flow cytometry method, some important morphological traits such as stomata characteristics and its density can be used to recognize polyploidy in orchid plant. Polyploidy was resulted to enhancement in size and reduction in density of guard cells. Colchicine is usable to produce ornamental polyploid orchid.

With many applications in treating and relieving of illnesses, lemon balm (Melissa officinalis L. ) is considered as a valuable medicinal plant; as a result, sufficient knowledge about its genetic variation and its classification is necessary for selecting suitable genotypes for breeding purposes. Therefore, present study was to investigate genetic diversity and relationships between several traits of 11 populations of lemon balm consisted of nine Iranian plant populations and two foreign plant populations using randomized completely block design with three replications during 2011 and 2012 in research farm of University of Tabriz. Results of analysis of variance showed that genetic diversity for most of the traits was significant. Study of genetic parameters revealed that percentage and yield of essential oil, number of stems per plant, fresh weight of stem and citral concentration had the highest amounts of genetic advance, genetic and phenotypic coefficient of variations. The highest heritability was observed on fresh weight of stems per plant (h2=0. 71), essential oil percentage (h2=0. 59) and number of stem per plant (h2=0. 58). Ardabil population was superior in terms of citral concentration and percentage and yield of essential oil among the populations. Pearson correlation coefficients indicated a positive and significant relationship between essential oil yield with chlorophyll index, leaf length and width and essential oil percentage and also a significant negative correlation with number of nodes. In factor analysis based on principal component analysis method with eigenvalues greater than one, five factors were determined that a total of 87. 40% of the total variance was explained by them. Cluster analysis divided the studied populations into three groups. Due to the observed variability in the present study, selection of desirable populations for breeding programs regarding to improvement of agronomic traits and essential oil of the species is possible.

Plantago ovata is from "Plantaginaceae" family which has several species. It is an annual herbaceous plant. Its height reaches to 10-40 cm. Its primary origin is India and Pakistan and it has wide distribution in Iranian flora. In order to evaluate responses of Plantago ovata to salinity an experiment was conducted using in vitro selection and tissue culture technique as well as somaclonal variation for identification and production of tolerant genotypes. The experiment was performed based on a completely randomized design with 12 treatments and 15 replications in each treatment. Hypocotyl and root explants showed the highest and the lowest callus induction respectively in non-saline (control) condition. Regeneration percentage of hypocotyl calli at all NaCl concentrations was more than cotyledon calli. Increasing concentrations of NaCl led to reduce percentage of regeneration. Cotyledon and root explants showed the highest and the lowest dry and fresh weight at all concentrations of NaCl. Also increasing of NaCl concentrations led to increment of sodium and decrement of potassium ingredients of callus. Amount of calcium varied by increasing NaCl concentration. Rooting happened a week after transferring regenerated plants to rooting medium. Then suitable seedlings were transferred to soil.

Sainfoin (Onobrychis viciifolia Scop. ), one of the main perennial legumes forages, is cultivated in both dry land farming and irrigated area for grazing and hay production for feeding of livestock. It is a perennial legume widely adapted to environmental conditions and has been successfully used as a pasture and hay forage. In order to evaluate and select the best ecotypes of sainfoin for a synthetic variety development program, a field experiment was conducted during 2011 to 2014 cropping seasons, in Tikmadash research station, East Azarbaijan Agricultural and Natural Resources Research and Education Center, Tabriz, Iran. Quantitative and qualitative traits of 25 sainfoin ecotypes were evaluated. Analysis of variance showed no significant differences between the ecotypes for day's to 10% flowering, but for other quantitative traits such as plant height, shoot fresh and dry weight, leave fresh and dry weight, fresh and dry forage yield were significant. Crude protein, crude fiber and dry matter digestible were significantly different among the studied sainfoin ecotypes. Fresh and dry protein yield indices were instructed by multiplying rude protein by fresh and dry yield, respectively, and used as criteria for ecotype selection. Considering quantitative and qualitative traits, specially fresh and dry protein yield indices, four superior ecotypes were selected to be used in synthetic variety (Syn1) development.

Arbuscular mycorrhizal fungi are one of the most important and the most abundant microorganisms in soil. The fungi have symbiotic relation with 80 to 90 percent of plant species. Arbuscular mycorrhizal symbiosis is able to alter pattern of dehydrin expression under drought stress. During period of drought acclimation that causes dehydration of cells, dehydrin proteins accumulate inside cells and protect membranes and macromolecules structures. All seedlings were irrigated three times per week during two years then were exposed to severe drought stress for nine days until wilt symptoms appeared on their leaves, then irrigated normally. It was evaluated whether Funneliformis mosseae is able to alter expression functions of dehydrin in the mycorrhiza seedlings of Pistacia vera under drought stress. Results showed that mycorrhizal colonization percentage in roots of P. vera declined significantly due to water stress treatments. The result revealed that the expression of dehydrin was increased significantly in nonmycorrhizal seedlings at 9 days exposure to the drought stress compared to the mycorrhizal seedlings, but the levels of dehydrin accumulation in mycorrhizal treatment subjected to drought were considerably lower than that of corresponding non-mycorrhiza seedlings, indicating that the accumulation of dehydrin proteins is not a mechanism by which the mycorrhizal symbiosis protects their host seedlings and suggesting that mycorrhizal seedlings were less strained by drought due to primary drought-avoidance mechanisms.

Quercus infectoria G. Oliver is one of the valuable and main species of Zagros forests of Iran. The species is important in environmental, economic, genetic and medicinal aspects. Cytogenetic studies on plant species and their populations provide quantitative information on evolution history and determine relationships between the species and karyological characteristics. Our study aimed to determine karyotypic and chromosomal constitution of Quercus infectoria and its karyological diversity among four populations of the species in western Iran (Sardasht, Marivan, Shine-Ghalaei and Kakasharaf) using root meristem and visual analyses. After pretreatment, stabilization, hydrolysis and staining, microscopic stages were performed and karyotypic parameters were investigated. Results showed that chromosome number of most the observed cells were 2n =2x= 24 but 2n=2x=25 and 2n=2x=26 and triploidy were also observed in several cells. Analysis of variance and mean comparisons were performed on 3 to 10 cells of 3 to 7 genotype of each population. Results showed significant differences among the studied populations in properties such as chromosome length, long and short arm length. Symmetric karyotype indices are indicative of the relative asymmetry of chromosomes in the studied populations. Principal components analysis based on karyotypic parameters showed that chromosome length, long arm length and short arm length, played the most important role in the first component. The first component contained more than 75% of total karyotypic variation. Cluster analysis classified the populations into two classes. Shine-Ghalaei and Kakasharaf was assigned to one cluster and Marivan and Sardasht, allocated to the second one.

Hawthorn (Crataegus spp) is one of the important forest genera in Iran. It is high compatibel to cold and drought and usefull as a rootstock for quince, pear and apple. This research was carried out in order to assess shoots and rooting of three species of hawthorn (C. aronia, C. pseudohetrophylla, C. meyeri). For shoot production, different concentrations of plant growth regulators and single-node were used. The explants were cultured on different concenterations of BAP and NAA on MS medium. On average after 6 weeks maximum number of shoots in concenteration of 1mg/l NAA plus 4 mg/l BAP (4. 33 branch) was observed. The highest percentage of explants (89. 67 %) was observed on C. aronia and maximum shoot length (22. 67 mm) was observed on C. meyeri using 1 mg/l NAA and 1 mg/l BAP. For rooting stage, we used MS, 1/2MS and rock wool with proliferated explants derived from suker and in vitro proliferated shoots. The explants were cultured on 16 different media treatments of MS, 1/2MS and rock wool medium. Root was not formed in none of the species, treatments and media. Maximum shoot length (22. 67 mm) was observed on C. meyeri, and maximum number of branches was observed on C. aronia.

Hybridization is one of the most important methods in tree improvement. This method can helps plant breeders to increase yield of tree species. Adaptability investigation of interspecific hybrids for planting in different regions is necessary because of different climate conditions in the regions. Populus species were planted in Iran for wood production from many years ago. In this research clones of P. alba× P. euphratica and P. euphratica × P. alba were selected and saplings were supplied from them. Saplings from P. alba× P. euphratica and P. euphratica×P. alba with P. alba and P. nigra as a native species were planted in Boldaji station in Chaharmahal and Bakhtiari province based on a Randomized Complete Block Design. Quantitative and qualitative characteristics such as total height, trunk height, diameter (DBH), disease and cold tolerance were recorded. Results showed that plant height, trunk height and DBH growth of P. alba×P. euphratica were significantly more than other treatments (p<0. 01). Quantitative and qualitative characteristics of P. alba were similar to P. alba×P. euphratica. P. euphratica × P. alba clone was intolerant against cold.

Dry forage yield of 24 accessions of Agropyron trichophorum were investigated by a randomized complete block design with three replications at two environments of normal irrigation and rainfed conditions for two years at Islam Abade Gharb research station, Kermanshah, Iran. Analysis variance showed significant variation (P<0. 01) between the studied accessions and the environments. Interaction effect of accession×environment was significant at 1% level. Two, interaction principal components from AMMI, analysis expressed 96. 77% of the interaction variance and were significant at 1% level. The accessions of 12-13, 313, 314, 8-4, 10-7 and 7-5 were classified as superior accessions for dry forage yield based on Duncan's test (p<0. 05) and cluster analysis and located in the same group. Accessions 8-4 and 10-7 had the highest general stability for forage yield based on interaction principal components and stability parameters, being suitable to be introduced for breeding programs in Kermanshah climate. Accessions 7-5, 313, 12-13 showed specific adaptation with normal irrigation conditions and accession 314 had specific adaptation to stress conditions, being sutable to be introduced in breeding programs, especially with emphasis on Kermanshah rainfed conditions.