Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research
Year:2019 | Volume:26 | Issue:2 (52) |Papers Count:18

MicroRNAs (miRNAs) are a group of 18_22 nucleotides long noncoding small endogenous that derived from its precursor sequence and evolutionary conserved post-transcriptional regulatory RNAs, which show an enormous role in various biological and metabolic processes in both animals and plants. MiRNA detection methods including microarray, qRT-PCR, northern blot and bioinformatics methods in which the easiest and cheapest way to identify miRNA is bioinformatics method. A bioinformatics approach was used to identify potential miRNA in red clover. EST-based homology search was applied to find potential miRNA of red clover. We blasted publicly available EST sequences obtained from NCBI GenBank against previously known plant miRNAs. A total of six miRNA target genes based on their complementary sequences were also detected. Target genes encoding mono dehydro ascorbate reductase 1 play an important role in maintaining balance in the cycle of reactive oxygen species ascorbate-glutathione, FES1 proteins that contain zinc finger domain binding DNA, RHF2A gene encoding E3 ubiquitin-protein ligase involved in the positive regulation of the gametogenesis progression, RNA helicase ATP-dependent is involved in the RNA repair and metabolism, Major facilitator superfamily (MFS) is a superfamily of membrane transport proteins that facilitate movement of small solutes across cell membranes in response to chemiosmotic gradients, AT-hook DNA binding motif protein.

Poppy (Papaver somniferum L. ) is one of the oldest medicinal plant in the world. It produces several important narcotic benzylisoquinoline alkaloids. In this study, 4’ OMT2 (3′-hydroxy-Nmethylcoclaurine 4′-O-methyltransferase) gene was isolated, sequenced and characterized. After amplification of genomic and coding sequence of 4’ OMT2 gene using PCR, the fragments were cloned into pTZ57R/T plasmid and sequenced. Sequencing results showed two fragments of 1074 and 1189bp for coding and genomic sequences, respectively. For the first time, results showed one intron of 115bp in the genomic sequence of 4’ OMT2 in the species. Isolated gene from Iranian genotypes was similar to Papaver genus with high identity, and had 67% identity to Ranunculales plants. Further bioinformatics analyses revealed, 4’ OMT2 gene produces a stable enzyme without signal peptide that localize in the cytoplasm. For gene expression analysis, five poppy RNA-seq (mRNA type) libraries (belonging to flower bud, leaf, developing fruit, stem and root tissues) were retrieved from SRA database and analysis of 4’ OMT2 gene expression were done using IDEG6 software and statistical tests. Results showed that 4’ OMT2 has differential expression among different tissues and has maximum expression in stem tissues. Identification and characterization of biosynthetic genes is the first step in genetic manipulation and metabolic engineering. In present study, for the first time, full length coding sequence of 4’ OMT2 gene was isolated from poppy plant and presence of one intron in its sequence was determined.

This study was conducted to investigate intra-and inter-population genetic diversity of 15 populations of Pistacia atlantica from Zagros forests, represented by 181 genotypes, using 15, 12 and 13 primers based on three molecular marker including inter simple sequence repeats (ISSR), inter retrotransposal amplified polymorphism (IRAP) and start codon targeted (SCoT) markers, respectively. ISSR, IRAP and SCoT primers produced a total of 186, 143 and 136 bands, of which 162 (87. 0%), 130 (90. 9%) and 120 (88. 0%) showed polymorphisom, respectively. Results showed that all marker systems revealed relatively high genetic variation at population level (hISSR = 0. 22, PPLISSR = 61. 92%; hIRAP = 0. 25, PPLIRAP = 74. 19%; hSCoT = 0. 20, PPLSCoT = 64. 09%). According to ISSR, IRAP and SCoT based genetic differentiation coefficient (Φ ST; 0. 28, 0. 23, and 0. 22, respectively), a higher level of genetic variation was recorded within populations than that of among populations, that could be due to wind-based extensive gene flow among the populations. UPGMA cluster analysis based on three used molecular systems, devided the studied populations into distinct clusters. According to Mantel test, no significant correlation was recorded between genetic and geographic distances of the populations.

Sumac (Rhus coriaria L. ) as a forest shrub has been dispersed in Mediterranean and Eastern Asia specially Iran and has several medicinal and industrial applications. In this research, intermicrosatellites markers were used to study genetic diversity of sumac genotypes collected from five natural habitats located in North West of Iran (West and East Azerbaijan provinces). Fifteen samples were select randomly from each population and genomic DNA was fingerprinted by 18 ISSR markers. Results revealed existence of suitable genetic variation among the studied sumac genotypes and portion of inter population (79. 8%) variation were higher than intra population (20. 2%) variation. Minimum value of genetic distance (14. 18) was observed between Aghberaz-Horand (East Azerbaijan province) and Nir-Arasbaran (East Azerbaijan province) populations and the maximum one (31. 08) was observed between Kachelleh-Urmia (West Azerbaijan) and Nir-Arasbaran populations. According to classification of genotypes based on molecular data, Aghberaz-Horand and Nir-Arasbaran populations located in the same group and populations from other habits located in distinguished groups. Classification of genotypes via ISSR markers was in agreement with their geographical distributions. ISSR markers could be effectively applicable in genetic evaluation of sumac germplasm and also in identification of genetic relatedness among known and unknown sumac samples.

Lolium is an important cool season grass species growing in different climates. To identify variation among nine Lolium multiflorum and nine Lolium perenne genotypes, this experiment was carried out based on 15 SCoT molecular markers. All of the SCoT primers showed 86 visible bands. Seventy four out of 86 bands were polymorphs. Primers SC35, SC36 and SC26 (with 9 and 8 bands respectively) showed the highest number of bands and SC10 and SC44 (with 2 and 3 bands) showed the lowest number of bands. The greatest number of band for polymorphic information content (PIC) belonged to SC5، SC40، SC35، SC63، SC13, C44، SC26، SC21and SC10. The greatest Resolving Power (RP), Marker Index (MI), Effective Multiplex Ratio (EMR) belonged to SC35 andSC26 primers. Genetic similarity based on Jaccard coefficient ranged from 0. 35 to 0. 91. Mean similarity between the genotypes was 0. 64. Cluster analysis grouped genotypes into two groups. The results of clustering was confirmed by principal coordinate (PCo) analysis. Analysis of molecular variance of annual and perennial species revealed that variation within species was 51% and variation between species was 49%. Based on heterogeneity (He) and Shannon indices (I), the highest intraspecific variability and gene variation belonged to L. Multiflorum.

Ferula pseudalliacea (bitter assafoetida) is a rangeland plant which its gum has a lot of medicinal properties. Since there is no report about evaluating genetic diversity of the valuable species in Iran, this is the first study to investigate genetic diversity of the species in its habitats in Yazd province using several morphological traits and ISSR markers. Among 9 traits, there was significant differences between habitats in terms of canopy cover, stem diameter and weight of 1000 seed. Cluster analysis by morphological traits on the basis of Ward's method classified the samples into three main groups. Among 22 primers, 8 primers reproduced the pattern DNA well and revealed 224 sharp bands. ISSR-16 and ISSR-55 primers had better performance according to the number of bands, PIC and Marker Index. Cluster analysis using Jacquard's coefficient and UPGMA method, grouped two populations into 10 clusters. Principal coordinates analysis showed three components explained 57. 94 percent of total variance and separated the populations from each other. These grouping matched relatively with the geographical regions. AMOVA showed that within group genetic diversity (92%) was more than between group diversity (8%). Comparing morphological and molecular dendrograms revealed that there was no similarity between them. The results showed that ISSR marker was suitable to investigate genetic diversity among the studied populations of the species.

Iranian mountain ash (Sorbus persica Hedl. ) is one of the endangered species with great values in terms of soil conservation, climate stratification, medicinal properties, high resistance to frost, and importance of habitat for wildlife. Present research was conducted with the aim of achieving to an appropriate method of micropropagation of the species. Nodal segments were taken from stem axis of in vitro elongated plants. The segments were cultured on WPM medium, supplemented with 27 hormonal composition of BAP, IBA and TDZ. Then the resulting shoots were transferred to a media supplemented with various concentrations of IBA or NAA for rooting induction, and finally they were moved to a greenhouse. Percentages of regeneration of the explants were 98-100%. The highest number of shoots was obtained by TDZ (0. 23 μ M) + IBA (0. 1 μ M) and BA (4. 4 μ M), the highest shoot length (5. 2 cm) was obtained by TDZ (0. 0 μ M) + IBA (0. 0 μ M) and BA (4. 4 μ M) and the highest number of nodes (8) was achieved by TDZ (0. 23 μ M) + IBA (0. 0 and 0. 1 μ M) and BA (2. 2 and 4. 4 μ M). There was a significant difference between IBA concentrations in different treatments for rooting percentage. Maximum rooting percentage was 14. 7% in IBA concentration of 5 μ m. The rooted segments were successfully acclimated to greenhouse conditions. Results of the experiment showed that in the absence of BAP hormone, umber of shoots produced by the segments significantly were decreased for all of the tested treatments. Fifteen percent of the transferred explants to the rooting media produced one to three roots per explant.

Salinity is one of the main environmental factors limiting production and distribution of plants worldwide. Aegilops is a wild species from grasses family with useful genes such as environmental stress tolerance. Two genotypes, 575 from Aegilop cylindrica as resistant and 675 from Aegilop crassa as sensitive to salinity were used base on previous experiments in salinity stress. After seed germination, the seedlings were transferred to a sandy environment under controlled light and temperature conditions and irrigation was performed under two normal conditions and 200 mM salinity with Hoagland solution. Leaf tissue samples were taken at 0, 6, 24, 48 and 72 hours after salinity treatments. After extracting the RNA and the cDNA synthesis, its accuracy was verified by 18S rRNA. The expression of the three transcription factors of CBF14, NAC2 and MYB was investigated by using Real Time-PCR. Results showed that NAC2 and CBF14 transcription factors had the highest expression at 48 hours after salinity stress, but the MYB gene had the highest expression at 24 hours after stress. Overall, the expression of CBF14, NAC2, and MYB genes in the 575 genotype compared with genotype 675 was in higher level in all hours. The results showed that the three mentioned genes play important role in salt tolerance in the tolerant genotype, therefore, considering the high genetic similarity between Aegilops and wheat, the genes can be as candidate genes for using in wheat breeding programs.

This experiment was conducted to investigate the effect of drought stress on photosynthetic pigments contents, biomass of roots and shoots, total protein content and antioxidant enzymes activity and non-enzymatic antioxidant of six species of Aegilops genus along with two drought resistant and susceptible wheat cultivars as checks in a factorial based on randomized complete block design with three replications. Drought stress treatments were applied at three levels of non-stress (FC=100%), moderate stress (FC=50%) and severe stress (FC=25%) conditions. Ae. caudata, Ae. triuncialis and Ae. cylindrica were ranked higher than resistance cultivar, Sirvan, as check in term of root fresh and dry weights under stresses. The highest total chlorophyll content was related to Ae. cylindrica, Ae. umbellulata and Ae. caudata in stresses. Carotenoid showed decreasing trend in all studied species except for Ae. umbellulata, and Ae cylindrica had the highest amount at both stress and non-stress conditions. The highest SOD under severe stress was observed in Ae. crassa and Sirvan as resistant cultivar. Maximum APX under severe stress was observed in Ae. triuncialis and Ae. caudata and the lowest activity was in Ae. neglecta. The highest level of GPX was observed in Ae. triuncialis, Ae. caudata and Sirvan cultivar under drought stress. Also the maximum CAT enzyme under severe stress was observed in Ae. triuncialis, Ae. caudata and Ae. crassa, respectively, and Ae. umbellulata and Darya cultivar had the lowest amount of CAT enzyme. In conclusion, because the three species Ae. caudata, Ae. triuncialis and Ae. cylindrica had the highest amounts for the traits associated with increased drought tolerance, including root volume, carotenoid, CAT, GPX and APX enzymes, and also these species were same in having CC genome. Therefore, it seems that the CC genome could have a high potential for future wheat breeding programs in increasing of drought tolerance.

Tanacetum polycephalum from Anthemideae tribe of Asteraceae family is a rangeland species with high morphological diversity and vast dispersal in Northwest, East and centre parts of Iran. The purpose of this research was studying genetic diversity of 20 populations of T. polycephalum in West Azarbayjan using ISSR molecular marker. Ten ISSR primers out of 14 primers could be scored based on presence or absence of each band. Over all 208 bands were produced. The range number of bands was between 15(IS24) to 28 (IS20) and the average was 20. 8. Regarding the polymorphism bands percentage, IS8, IS1, IS10, IS13, IS20 markers had high percentage (100) and IS24 marker had the lower percentage (80) and mean percentage of polymorphism of the primers was %96. 03. PIC, RP, EMR and MI means were 0. 30, 6. 06, 19. 5 and 6. 13 respectively. Over all the most effective markers were IS8, IS1 and IS10. Results of cluster analysis revealed the grouping of studied populations in 2 main and 4 subgroups, in which the different populations collected from the same locality were located in the same group. Therefore, a close relationship was characterized between geographic location and genetic diversity and it may be concluded that geographic and climatic factors could be one of the responsible main factor for genetic diversity.

Knowledge and understanding the genetic diversity of species is very important for adaptation to environmental changes and their long-term survival and also for the management of forest genetic resources conservation. Therefore, study of industrial or ecologically important species should be done in order of priority. The aim of this research was exploring the ecotypic diversity changes of Quercus castaneifolia populations in elevation classes of Golestan province (low, middle, and upper elevation) by the qualitative study of peroxidase biochemical marker in the biennial branches of six different populations. Qualitative study of peroxidase marker was done by vertical electrophoresis and PAGE method (Poly Acrylamide Gel Electrophoresis). Based on the results, populations of altitudinal classes were different regarding amount of diversity and ecotypes were separable from each other. Therefore, index bases in the seed production areas should be determined and their natural regeneration should be increased and gene flow should be improved through the seeds of index bases of the adjacent populations with similar ecological conditions with controlling the number of selected index bases considering the increased diversity in each habitat.

Cuminum cyminum is one of the oldest and most economically important species in the Apiaceae family that has the highest cultivated areas of Khorasan provinces in Iran. Genetic diversity of 20 ecotypes of Cuminum cyminum was investigated by protein markers. The ecotypes were randomly collected from three provinces of North, Razavi and South Khorasan provinces. Protein profile analysis was carried out by SDS-PAGE method. The method included: extracting protein from the sample seeds, determining samples concentration, and finally running on electrophoresis gel. Analysis of molecular variance showed that there was variation between and within the populations. Based on the protein markers %24 of the variations belonged to between populations and %76 belonged to within populations. Polymorphism percentage, polymorphic content mean, Shannon's information index and the average of Nei's information index were 27%, 0. 01, 0. 49, and 0. 68, respectively. Results of PCoA showed that the first three components accounted for 83% of the variation. Cluster analysis based on the similarity matrix classified the studied ecotypes in 5 clusters.

The objective of this study was to develop population hybrids by crossing five alfalfa cultivars of different geographic origin (Ghareh Yonjeh, Hamedani and Mahalie-Esfahan from Iran, Elchi from Turkey and Ordobad from Azerbaijan) and assessment of combining ability among them using diallel analysis. A half diallel was performed during 2012 between the selected cultivars. For each pairwise cross, ten plants were chosen at random to obtain F1 generation. Heterotic responses were determined by evaluation forage yield of the cultivars and their 10 half-diallel hybrids in seeded plots that were harvested three times in each of 2 years (2014 and 2015). Variation among crosses was attributed primarily to general combining ability (GCA) effects. However, specific combining ability (SCA) effects were also significant. Mid-parent heterosis ranged from 7. 1% in the Elchi x Hamedani to-9. 9 % in the Ghareh Yonjeh x Mahalie-Esfahan and high-parent heterosis ranged from 3. 4 % in the Elchi x Hamedani to-16. 7% Ghareh Yonjeh x Mahalie-Esfahan. Results indicated that Elchi cultivar could be considered as a member of a potential heterotic group for adapted cultivars (Ghareh Yonjeh and Hamedani) to the studied environment.

In order to identify tolerated populations of sainfoin to powdery mildew, seeds of ten populations of Onobrychis viciaefolia, originated from different regions of Iran were sown in Alborz research station during 2013. Tolerant and susceptible populations were cultivated in natural condition and evaluated in terms of contamination and other parameters as yield and quality traits. Result showed that two populations of 15353, 3001, Oshnaviaeh and Polycross had disease severity index less than 25% and nominated as tolerant populations. The rest of populations due to disease severity index between 25 to 50% and over than 50% were considered as semi tolerant and susceptible, respectively. The tolerant (15353, 3001, Oshnaviaeh and Polycross) populations had higher forage and seed yield than susceptible populations. They showed also higher values of quality traits (Protein content, water soluble carbohydrate, digestive material and ash content) than susceptible ones. In contrast the susceptible populations had higher crude fiber, ADF and NDF compared with tolerant populations. In principle component analysis (PCA), the first four components accounted for 83% of total variation. The first three components were known as yield, quality traits and disease severity index, respectively. Result of cluster analysis showed that population of 3001, 15353, 9263, 4083 and Oshnaviaeh and Polycross were located in cluster 2 that these popolations have lower mean values of disease severity index and higher forage and seed production.