Journal of Plant Molecular Breeding
Volume:7 Issue: 1, Winter and Spring 2019

Regarding high potential of green plants for development of recombinant vaccines, this research was conducted to evaluate expression of a novel recombinant vaccines against Foot and Mouth Disease (FMDV) in tobacco plant. For this purpose, a synthetic gene encoding 129-169 amino acids of foot and mouth disease virus capsid protein VP1 was transferred to tobacco plant via Agrobacterium-mediated genetic transformation. Considering codon usage optimized for tobacco, ribosome binding site and endoplasmic reticulum signal peptide were included in the synthetic gene to enhance expression level. Expression of the synthetic gene in tobacco seedlings was analyzed at transcription and translation levels and production of recombinant protein was quantified. Moreover, an in vivo immunization assay was carried out to verify immunogenicity of the expressed peptide in model animals. Results showed that expression of the recombinant protein in two lines of transgenic plants was a high as 0.65% and 0.72% of total soluble protein. The recombinant protein was able to induce immunogenic response when parenterally administered in rabbit.

One of the most effective biotechnological techniques for producing plant metabolites is the hairy roots (HRs) culture system. HRs are genetically and biologically stable and able to produce secondary metabolites in a short time. Ficus carica L. is one of the most important plant sources of valuable medicinal compounds, especially polyphenolic compounds. The aim of this study was to investigate the growth and morphological features of HRs, methyl jasmonate (MeJA) elicitation and plant regeneration potential of HRs induced by different strains of Agrobacterium rhizogenes on Ficus carica cv. Siah. Four bacterial strains (A4, A7, A13 and ATCC 15834) were used for HR induction in leaf and shoot samples. The MS medium containing 2 mg/l of 2,4-D in combination with 1 mg/l of TDZ or BAP was used to induce callus from HRs, and then the callus regeneration ability was evaluated in MS medium containing TDZ and NAA. Depending on explant type and bacterial strain, the roots were induced directly or indirectly (via callus formation) from the wound sites, and exhibited different morphology. The HRs showed high levels of phenolic compounds. A 4-day elicitation with MeJA, in dependence on the concentration, enhanced the phenolic capacity and antioxidant capacity of HRs. The calli obtained from HRs showed root (70-80%) and bud (23.33%) regeneration potential. The current study described that the HR culture systems, in addition to providing the possibility of plant regeneration from transgenic roots, could be a promising in vitro technique for high production of secondary metabolites through elicitation.

Grain and cooking qualities in rice are measured by several physical and chemical traits, each of them controlled by several genes. Molecular markers have become fast and reliable tools for screening genotypes for grain quality. However, As different populations may carrydifferent genes, the efficacy of previously developed markers in new populations should be tested. In order to assess the effectiveness of molecular markers in predicting grain quality in Iranian rice genotypes, a total of 38 genotypes from three different backgrounds were fingerprinted by 9 grain quality specific molecular markers and 10 laboratory traits.A total of 31 alleles were detected with an average of 3.1 alleles per locus and the polymorphic information content values ranged from 0.245 to 0.74. Cluster analysis based on molecular markers divided the rice genotypes into three major clusters and effectively differentiated between various genotypes. However, the dendrogram based on the common set of qualitative traits didn't succeed in discriminating between original groups. There were significant associations between molecular markers and quality traits except for milling factor. However, these associations weren't necessarility specific to their supposed traits. Therefore, association of markers developed in other rice populations, specially those developed in Japonioca populations should be tested prior to application in Iranian rice marker assisted breeding programs.

Sclerotinia stem rot disease caused by Sclerotinia sclerotiorum is one of the most important diseases of sunflower. Telomeres are nucleoprotein structures at the ends of chromosomes that are essential for maintaining the integrity of the genome. The aim of this study was to determine the alteration of telomerase enzyme gene (tert) expression under fungal infection stress. The expression of tert gene in both susceptible (SDR19) and resistant (LC1064-C) genotypes of sunflower was evaluated by qRT-PCR after infection with A37 isolate of S. sclerotiorum. The results showed significant and drastic decreased levels of tert expression in both susceptible and resistant genotypes of sunflower, immediately after fungal infection. This depletion followed by mild variation when infection continued for more hours, which was more constant in resistant line, compared with the susceptible one. In conclusion, the expression of tertgene in sunflower is downregulated in response to Sclerotinia rot disease.

In the present experiment, the in vitro callus production from tea leaf explants was induced on three different basal media. Then, the efficiency of tea callus proliferation was enhanced through application of thiourea to the proliferation medium. Furthermore, the caffeine and Epigallocatechin Gallate content of the callus and leaf tissues were estimated through HPLC method. The calli mass volume in WPM (woody plant medium) supplemented with thiourea (either 0.1 or 1.0 mM) was significantly higher than other basal media. The callus tissue collected from media enriched with thiourea had significantly more caffeine content but the level of Epigallocatechin Gallate was not statistically affected by thiourea treatments. The amount of these two compounds was also estimated in young and old leaves of the tea mother plants and it was found that the callus tissue had little amount of caffeine and Epigallocatechin Gallate as compared to maternal tissues. It is concluded that, in species with low rate callus proliferation, addition of thiourea to the medium may be followed as an effective and low cost option for callus proliferation improvement.

Wheat is one of the most important edible crops used in several food products. The success of plant breeding programs depends on the availability of parents with desired genetic diversities. In this context, we evaluated the genetic parameters in eight wheat genotypes namely Kohdashet, Morvaread, N-80-19, Darya, UR-82-17, Ehsan, Bacounoura and Atrack as well as theirs F1 progenies in term of grain yield and the relevant traits. Randomized complete block design with three replications was used. Data analysis indicated that genotypes and their F1 hybrids significantly affected grain yield, biological yield, thousand-grain weight, grains per spike, grain weight in spike, plant height, peduncle length and second internode length. The Model I of Griffing's method II was used in SAS software to determine GCA, SCA and other genetic parameters. Among parents, Ehsan cultivar exhibited the highest GCA for grain yield, thousand-grain weight, grain weight in spike, plant height and biological yield. P6×P7 cross was found to have the highest SCA effects on grain yield, thousand-grain weight and plant height. In addition, among examined characteristics, the thousand-grain weight and the peduncle length had the highest broad sense (h2= 0.98) and narrow sense heritability (H2= 0.62), respectively. These findings indicated that the distinctive crosses engaged in progenitors belongs to high×low GCA; therefore, it is inessential to set crosses with high×high GCA. As a result, potential homozygous lines can be selected from transgressive segregations to increase yield. Such crosses can be applied for exploiting heterosis.

Low temperature is a major abiotic stress which can significantly affect the grape production. microRNAs play an important role in the control of plant development and response to adverse environmental conditions. Although miRNAs and their targets have been identified in several Vitis species, their participation during cold accumulation remains largely unknown. One such microRNA is miR395, which is conserved and regulates sulfate assimilation and distribution in plants. In this study, the possible role of miR395 in cold stress response was investigated. Identification of target genes, gene ontology and biological system analysis were performed to identify the major networks in which this miRNA is involved. Finally, the effects of gradual chilling and also a shock chilling on the expression of miR395 were investigated. In total, five target genes were identified, which all of them are targeted by miR395s a to m, whereas of the five target genes, only one is identified by miR395n as a target. Three of these genes, including ATP sulfurylase, sat-1 and, LAST3-like are involved in the control of sulfur metabolism and transport. Pathway analysis showed that miR395 was involved in response to cold stress in grape through cellular response to sulfate and phosphate deficiency. Based on RT-PCR results, contrary expression patterns of miR395 under gradual (up-regulated) and shock chilling stress (down-regulated) were observed. The changes of sulfate assimilation process would influence the formation of sulfur-containing antioxidant compounds. These results provide an insight into the regulatory roles of miR395 in response to low-temperature stress in V. viniferae.

Hypericum perforatum is a medicinal plant which Hypericin, Hyperforin and phenolic compounds are its active secondary metabolites. Hairy root induction by Agrobacterium rhizogenes in this plant is difficult and has low efficiency. In the present study two inoculation methods, immersion in bacterial suspension and direct injection of A. rhizogenes has been compared. For this purpose, the best conditions for H. perforatum hairy root induction including A. rhizogenes strains (A4, LBA9402, NCPPB2656), plant explants (Stem, Apical bud, leaves), co-cultivation media (MS, ½MS, B5, and ½B5) and Acetosyringone (AS) concentration (0 and 100 µM) were specified and used for comparative analysis. It was found that strain A4, Stem explants, ½MS co-cultivation medium without AS constitute the best conditions for hairy root induction of H. perforatum. Transgenic nature of the potential hairy roots was confirmed using PCR and specific rolB and rolC genes primers. The results showed that the efficiency of applying direct injection method is four times higher than immersion in bacterial suspension in H. perforatum hairy root induction. In general, the results indicate that direct injection can be the method of choice to successful hairy root induction in H. perforatum.

The genus Crataegus spp. (Hawthorn) is belongs to Rosaceae family. The hawthorn can be utilized as a rootstock as well as an ornamental plant. The fruits are sources of many essential nutrients and have beneficial effects on human health. The Crataegus genus has been founded in different areas such as Iran, especially in the Ardabil region. The collection and conservation of genetic material is important for future breeding programs. In this purpose, thirty hawthorn genotypes collected from several regions (Fandoghlo, Namin, Khalkhal, Aladizgeh, Germi) of Ardebil province and their genetic variations investigated with 15 ISSR markers. According to the results, all 15 primers amplified and produced total 128 bands (average = 8.53), as well as showed high polymorphism information content (average = %38.06). The results showed that there is a high level of genetic diversity and polymorphism ratio among the wild genotypes of hawthorn in the Ardabil region. Based on WARD technique, genotypes classified into three main clusters with several sub-clusters that exhibited high genetic diversity. The Germi genotypes completely isolated from the rest of the genotypes, which according to the structure analysis results, were similar to the Khalkhal genotypes.

There is no doubt that use of hybridization programs in the quinoa plant genotypes to induce genetic variation is difficult, however introducing the variations through mutation, to obtain promising genotypes, is much easier. In this research, quinoa seeds (Chipaya cv.) exposed to different doses of gamma rays and were cultivated in pots and open field under salinity stress. The results showed distinct differences at all studied traits in the native and mutant plants. Gamma ray’s irradiation caused genetic variations that was categorized based on studied traits, tolerance indices, cluster analysis of protein and ISSR data, which led to obtaining two promising mutations during M2. It should be noted that 90 and 120 Gy revealed the highest effects in producing desirable genetic variations. Also, the data resulting from the evaluation of phenotypic traits and tolerance indices of plants were confirmed by the biochemical and molecular analysis results. This research is providing new insights of using molecular breeding program for quinoa improvement to produce new promising genotypes powerfully face environmental stress and potential aid in future food shortage disasters.

Garlic rust is one of the most important diseases of garlic worldwide, which hardly can be controlled by applying fungicides while the weather condition goes on the favor of the disease progress. The NBS-profiling approach is one of the effective methods for separating the replicated parts of resistance gene analogues (RGA). In this study, 12 primers (NBS-LRR) were used on 16 Iranian garlic clones. Out of 499 scored marker sites in the range of 100 to 800 bp for NBS, from which 477 sites were multi-faceted (95.59 percent). The highest number of marker sites was for the primer combination NBS1-AluI and the lowest was for the primer combination NBS7-RsaI. The highest polymorphism occurred with combination NBS2-AluI and NBS1-AluI with 70 alleles and the lowest polymorphic composition occurred in NBS7- RsaI combination. The results of cluster analysis using UPGMA divided the clones into eight separate groups. This study showed that there is a significant diversity in the homologues of resistance genes in the Iranian garlic clones, which can be exploited in plant breeding programs. In addition, the results indicated that the NBS profiling technique is an efficient method for investigation on diversity of resistance genes in various plant species, including garlic. Using of NBS-profiling technique to study the diversity of resistance genes in garlic clones was addressed for the first time in the world in this study.

Silver nanoparticles are being extensively used in a broad range of applications in our daily routine life. In the present study, it was investigated if citric acid (CA) and hydrogen sulfide (H2S) can mitigate adverse effects of silver nanoparticles (AgNPs) in green bean plants. Green bean seedlings were applied with AgNPs either through soil drenching or foliar spray and were then treated with different concentrations of citric acid and NaHS, as H2S donor. Results indicated that AgNPs induced several stresses in green bean plants. Concomitant foliar and soil appliaction of nanoparticles caused adverse effects on photosynthetic pigments and reduced carotenoid and protein contents, while increasing H2O2 content and superoxide dismutase (SOD), and ascorbate peroxidase (APX) activities. It was revealed that citric acid and H2S application significantly alleviated adverse effects of AgNPs. In the plants challenged with AgNPs, the highest rates of catalase (CAT), glutathione S-transferase (GSTs), and malondialdehyde (MDA) activities were recorded, while these parameters were reduced when plants were also treated with H2S. Application of 1.5 g/L of citric acid caused sharp decreases in CAT, GST and MDA activities. Among the treatments, the highest levels of APX, SOD, and anthocyanin were observed in the plants treated with AgNPs trough both foliar and soil drench method without citric acid and H2S treatment. The findings of the present study would increase our knowledge of the interaction of plants with heavy metals and would be useful for designing sophisticated methods for reducing the damages in the stressed plants.