JOURNAL OF AGRICULTURAL BIOTECHNOLOGY
Year:2019 | Volume:10 | Issue:4 |Papers Count:9

Objective Astragalus verus is one of best species for tragacanth production. The aim of this study was optimization of cell suspension culture in this plant. Materials and methods First, the callus production was optimized then optimization of cell suspension was performed. The callus production experiment was conducted in a factorial arrangement with explants of root, hypocotyl and cotyledon in MS medium containing BAP in combination with 2, 4-D and NAA. In experiment of cell suspension optimization, callus that derived from medium containing NAA and 2, 4-D were investigated using 16 hormonal treatments and three replications in a completely randomized design. Also, it was investigated the application of dark treatments, ascorbic acid, poly-vinyl pyrrolidone and EDTA iron chelate to avoid phenolic compounds. Results The results showed that the highest callus production percentage (average of 67%) was obtained in the medium of 2, 4-D with root explant. The best hormone treatment of cell suspension was 3 mg /L 2, 4-D in combination with 0. 5 mg/L Kin with an average of 0. 277 viability, and 0. 040 g dry weight (at 5ml). In the study of the effect of browning inhibitors, the highest dry weight was obtained with 100 mg/l ascorbic acid in cell culture medium with an average of 0. 76 g (at 5ml). This treatment was effective in prevention phenolic compounds. Conclusions The results of this study showed that 2, 4-D could be used alone for inducing proper callus and in combination with Kin for the production of cell suspension. It was also found that ascorbic acid can well control the process of browning cells of cell suspension. The results of this study can be used for in vitro tragacant production.

Objective Arbuscular mycorrhizal fungi (AMF) establish symbiosis with a great number of plant species. This group of fungi plays an important role in ecosystem stability. Some AMF species can quickly adapt to stress conditions. The diversity of native AMF is less studied in Iran. In the present study, we identified AMF symbionts of pistachio trees under salinity and drought stress. Furthermore, the results were compared with some native plants in Kerman province. Materials and methods In this study, 56 and 40 samples from the rhizosphere of pistachio trees and native plants were collected respectively. After root colonization assessment, 37 samples with more colonization percentages were selected as initial inoculums. Inoculums prepared with sorghum plant in a pot experiment. In the next step, molecular identification of eight isolates (four isolates from pistachio and four isolates from the native plant) was performed using qPCR method in root samples of Andropogon gerardii. Results The result showed that root colonization was lower in pistachio than sorghum plants which were in contrast to native plants. Morphological identification of AMF spores showed all samples were contained Rhizophagus except for sample ID 42 of pistachio orchards. Claroideoglomus, Funneliformis, and Rhizophagus were found as dominant species in pistachio orchards. Conclusions The present study shows that AMF diversity was higher in native plants than pistachio. Diversispora had the highest rate of root colonization followed by Funneliformis, Rhizophagus, Claroideoglomus, and Racocetra.

Objective Apple (Malus domestica Borkh) from Rosaceae family is one of the most important fruit trees in the world. Evaluation of genetic diversity is one of the basic steps for plant breeding and preserving of germ plasm through gene banks. Materials and methods The aim of this study was evaluation of the genetic diversity of 22 varieties of apples collected from Khorasan Aagricultural Research Center and some region of Jiroft using ISSR markers based on the polymerase chain reaction (PCR). Materials and methods DNA extraction was done and then PCR conducted by five ISSR-primers. Results Apple cultivars had 81. 48% polymorphism. Cluster analysis classified them into six groups. The first group had five apple cultivars from Khorasan Razavi including Mohali Gonabad, Tabasi number3, Ghasemabai, Mohamadi and Golshahi Oghaz as well as Golab Esfehan, Sibe Ghermez and Gerani Esmith from Jiroft. Genotypes of Zard Lobnan, Sib Ghermez Khoni, Mohali Sardo, Golab Sardo from Jiroft and Mashali, Morabei Mashhad, Ali Mori Dovom Ras, Arbabi, SIB number3 from Khorasan Razavi categorized into the second group. Genotypes of Shaikh Ahmad Tabriz and Shafiabadi were placed in the third and forth group respectively. The fifth and sixth groups include Gol Mashhad and Abasi Azghade, Atlasi Tabas from Khorasan Razavi. Also, ISSR primer 842 showed the highest polymorphism among apple cultivars. There is no correlation between the geographical origin and genetic grouping of apple cultivars. Probably, setting of different regions genotypes in the same genetic group is due to transport of genotypes among regions in past. Conclusions Results indicated that ISSR is an effective technique for identification of polymorphism, genetic distance and germplasm management among apple cultivars.

Objective The recognition of pathways involved in salt tolerance mechanisms is one of the interesting topics in plant sciences and the new data-mining techniques provide a new insight for researchers. In this research, various attribute weighting and decision tree (DT) algorithms were executed to discover biochemical markers linked to salinity tolerance. Materials and methods In this regard, to assess some biochemical markers such as protein, superoxide dismutase, peroxidase, catalase, ascorbate peroxidase, proline, sodium and potassium in wheat and its wild relative Aegilops crassa, a factorial experiment was conducted in a completely randomized design with three replications. The factors in the study were genotypes (Arg (salt tolerance) and Alamout (salt sensitive), and a wild relative (Ae. crassa)) and salinity (sodium chloride 0 mM and 150 mM). Results According to these approaches, proline, superoxide dismutase and ascorbate peroxidase can be used as biochemical markers to screen wheat genotypes for salt tolerance. Feature selection and decision tree results showed that hierarchy combination of proline, superoxide dismutase and ascorbate peroxidase can be used as biochemical markers for selection of salinity tolerant wheat. The DT Parallel Gain Ratio model when run on the Info Gain Ratio dataset and the DT Gain Ratio model when run on the Rule dataset were the best model in distinguishing sensitive and tolerant genotypes with 97. 5% and 91. 67% performances, respectively. Conclusions Overall, the results showed that combining bioinformatics, and laboratory studies can lead to identifying pathways associated with salinity and provide a better understanding of the mechanisms underpinning stress tolerance.

Objective St. John’ s wort belongs to Hypericaceae family and has been used since ancient Greeks as a herb for the treatment of anxiety, depression, wounds and burns. In present study, the effects of different concentrations of CPPU and BA (each in three concentrations) in combination with two concentrations of IAA on callus production and shoot regeneration of Hypericum perforatum from leaf and stem explants were evaluated to identify the best growth regulator combination for obtaining the highest yield of in vitro regenerated shoots. Materials and methods This was implemented in a CRD-based factorial experiment with three replicates and five samples in each replicate and the traits including callus fresh weight, number of shoots, shoot length, length of the tallest shoots, and the number of nodes of the tallest shoots were measured and examined. Results Results showed that the most effective combination for regeneration was 1. 5 mg/l CPPU and 1 mg/l IAA, which may be a proof for more effectiveness of CPPU in comparison with BA as cytokinin source on shoot regeneration in Hypericum perforatum. Conclusions The use of 1 mg/l IAA in combination with 1. 5 mg/l CPPU or BA resulted in a significant increase in shoot regeneration. It was also showed that 1 mg/l IAA in combination with 0. 5 mg/l BA could cause a significant increase in callus fresh weight.

Objective The olive is considered as one of the most important horticultural crops, with more abundant number of cultivars. Thus, the aim of this research was to study the genetic diversity in a set of olive cultivars in Taroum station of Tran. Materials and methods In this research, 25 CBDP markers were used to study the genetic diversity and relationships of 20 promising and commercial Iranian and foreign varieties of olive. Results The 25 used primers amplified a total of 755 polymorph alleles, and the mean of PIC and MI indices were 0. 941 and 5. 64, respectively, indicating the high efficiency and differentiation power of CBDP markers. The genetic parameters of Nei and Shannon indices were higher in foreign cultivars than those of Iranian cultivars. The high gene flow (4. 46) was observed among olive cultivars, which confirmed the low values of the inter-population differentiation (Gst) and FsT indices. Analysis of molecular variance (AMOVA) differentiated the total genetic variation into inter-group (Iranian and foreign cultivars) (17%) and within-group (83%) diversity. The cluster analysis using Dice coefficient and UPGMA method divided the Iranian and foreign cultivars into five groups, which the promising cultivar T24 and Koroneiki with the maximum difference with others were placed in separate groups. The Principal Coordinate Analysis (PCoA) was able to distinguish promising Iranian cultivars from the other commercial cultivars. The population structure analysis confirmed the results of the cluster and PCoA analysis and clearly separated the two groups of Iranian and foreign cultivars. Conclusions In conclusion, it was observed that the promising cultivar T24 with having a genetic similarity to both groups of Iranian and foreign cultivars can be a suitable candidate for commercialization with complementary characteristics of both groups. Also, it is recommended to use this marker to study the genetic diversity of other olive varieties.

Objective Cold is one of the important stresses in cold and temperate regions and improving cold tolerance is in the top priority of the lentil breeding programs. NAC transcription factors play an important role in response to various biological stresses, including cold stress. Materials and Methods: In this study, to evaluate the expression of MfNAC, MtNAC57 and GmNAC2 genes, the plants of Gachsaran lentil cultivar were subjected to five temprature treatments in a completely randomized design with three replications. Twenty-nine old plants were subjected to duration of cold stress treatments including 6h, 12h, 24h and 48h at 2°-4° c and plants grown under greenhouse conditions (23° C) were considered as controls. Total RNA extraction from tissues was performed using lithium chloride method and cDNA was synthesized using TaKara kit. For each treatment, real-time PCR was performed in both shoot and root in three biological and two technical replications. Actin gene was used as internal control. Results The results of analysis of variance of all studied genes showed that there was a significant difference between the different duration of cold treatments in both shoot and root. GmNAC2 gene expression decreased significantly in the shoot, 6 hours after stress, while in the root, the expression decreased 24 hours after stress. The expression of MtNAC57 gene in 6h, 12h, and 24h treatments decreased compared to control, but in the root in 12h treatment, a significant expression was observed. MfNAC gene in shoot showed a significant reduction in expression in all treatments, while in root a significant increase was only observed during long-term treatment of 48h. Conclusions In the present study, for first time, the expression pattern of some members of NAC gene family was examined in lentils under cold stress and the results could be useful for studies on this plant and other legumes.

Objective Homospermidine synthase (HSS) is the first enzyme in the specific pathway of pyrrolizidiene alkaloid (PA). HSS catalyzes the NAD1-dependent transfer of an aminobutyl group of spermidine to putrescine. Putrescine, spermidine and spermine are three polyamines which are naturally positively charged compounds found in virtually all living cells. These compounds bind to DNA and have been implicated in a number of crucial processes such as cell division, differentiation and membrane function. Very few studies on expression of homospermidine synthase gene in Senecio vulgaris, particularly in natural conditions have been reported. The objective of current research was to study homospermidine synthase gene expression in different tissues of S. vulgaris by real-time PCR. Material and methods The total RNA was extracted from roots, shoots (stems and leaves) and flowers, and was converted to cDNA. Specific primers were designed for HSS1 gene and also for the Pol II which was applied as a reference housekeeping gene in the experiment and expression level of HSS1 was assessed using real-time PCR. Results Melting curves of two genes showed the specificity of PCR amplification. Evaluation of normalized expression level of HSS1 gene showed that the gene was differentially expressed in different tissues of S. vulgaris. Maximum relative expression (>59-fold change relative to Pol II reference gene) was observed in roots where the homospermidine synthase is activated and homospermidine is produced. Interestingly, the gene is also expressed in flowers but with a lower extent (4-fold change relative to housekeeping Pol II gene). As expected the gene showed non-significant relative expression in shoots where the homospermidine is transported. Conclusions Expression of HSS1 gene in S. vulgaris is tissue-specific and is higher in the roots relative to other tissues.

Objective Optogenetics, as a new scientific branch, uses the photoreceptors to conduct basic and applied research. Identification of the novel photoreceptors and their characteristics is a way to develop the optogenetics. Cryptochrome/Photolyase protein family DASH crypthochromes have DNA repair ability and play role in signaling pathways. Current research goal was the adequate production of the Volvox predicted DASH1 cryptochrome protein (VcCryDASH1) to use in spectroscopy studies as well as DNA repair assay. Materials and methods First, the coding sequence of VcCryDASH1 was isolated from Volvox carteri cDNA library using the specific primers and cloned into pGEX-2TK expression vector. The recombinant protein production was analyzed using two different strains of E. coli under different incubation times, using IPTG to induce the protein expression and then its solubility was investigated by SDS-PAGE. DNA binding ability of the protein was studied by alignment method. Results The appropriate production of the protein obtained in Rosetta strain at 32° C under more than 3 hours incubation. Approximately, 50 percent of the protein was insoluble under 1 and 3 hours incubations. VcCryDASH1 alignment analysis showed that there is high percentage of amino acids in this protein with DNA binding ability. Conclusion These results revealed that under optimum conditions, VcCryDASH1 has dual solubility but our approach can produce the adequate quantity of the protein in the Rosetta. Also, because of having conserved amino acids, the protein can bind to DNA, nominating it as a possible resource to design the DNA-light mediator molecules in optogenetics.