Correlation between gene expression levels under drought stress and synonymous codon usage in rice plant by in-silico study.

We studied the correlation of synonymous codon usage (SCU) on gene expression levels under drought stress in rice. Sixty genes related to drought stress (with high, intermediate and low expression) were selected from rice meta-analysis data and various codon usage indices such as the effective number of codon usage (ENC), codon adaptation index (CAI) and relative synonymous codon usage (RSCU) were calculated. We found that in genes highly expressing under drought 1) GC content was higher, 2) ENC value was lower, 3) the preferred codons of some amino acids changed and 4) the RSCU ratio of GC-end codons relative to AT-end codons for 18 amino acids increased significantly compared with those in other genes. We introduce ARSCU as the Average ratio of RSCUs of GC-end codons to AT-end codons in each gene that could significantly separate high-expression genes under drought from low-expression genes. ARSCU is calculated using the program ARSCU-Calculator developed by our group to help predicting expression level of rice genes under drought. An index above ARSCU threshold is expected to indicate that the gene under study may belong to the "high expression group under drought". This information may be applied for codon optimization of genes for rice genetic engineering. To validate these findings, we further used 60 other genes (randomly selected subset of 43233 genes studied for their response to drought stress). ARSCU value was able to predict the level of expression at 88.33% of the cases. Using third set of 60 genes selected amongst high expressing genes not related to drought, only 31.65% of the genes showed ARSCU value of higher than the set threshold. This indicates that the phenomenon we described in this report may be unique for drought related genes. To justify the observed correlation between CUB and high expressing genes under drought, possible role of tRNA post transcriptional modification and tRFs was hypothesized as possible underlying biological mechanism.

Selection of ideotype to increase yield potential of GM and non-GM rice cultivars.

Using classical breeding, plant breeders envision a plant type whose yield they aim to enhance by selecting for individual traits to create model/ideal plants or ideotypes. To achieve this, those factors restricting yield need to be identified and controlled through the use of new technologies to achieve the desired ideotype. This study aimed to determine the ideotype of seven genetically modified (GM) and non-GM rice (Oryza sativa L.) cultivars. Field experiments were carried out in three isolated regions in the north of Iran under the Iranian bio-safety standard protocol. Four of the GM cultivars carried the cry1Ab gene in the vegetative stage while three non-GM cultivars served as the control. R2 values showed that five, six and seven variables in Sari, Amol and Rasht regions accounted for 63 %, 52 % and 74 % of paddy yield variation, respectively. In the same three regions, paddy yield variation due to white heads accounted for 28.38 %, 8.45 % and 3.95 % of the total variation in paddy yield, respectively. The total estimated variation in paddy yield in Sari, Amol and Rasht was 1810.50, 2377.6 and 2176.47 kg ha-1, respectively. Average data over the three regions indicated that highest loss in paddy yield was observed in non-GM 'Nemat', 'Khazar' and 'Tarom Hashemi'. GM cultivars derived from 'Khazar' showed significantly lower paddy yield loss than the non-GM parent. Dead heart, a condition that occurs in the vegetative stage in which the stem borer larva enters the stem and feeds on the growing shoot, causing the central shoot to dry, as well as white heads, which is a condition in which whole ear heads of adult plants become dry and yield chaffy grains, in all three regions were important variables contributing to paddy yield loss. In the future, producing GM rice resistant to striped stem borer with an active promoter in the reproductive growth stage might allow farmers to reduce a significant part of paddy yield loss resulting from white heads, which is directly negatively correlated with filled spikelets per panicle (R2 = -0.57**), in order to achieve an ideotype.

Field trial evidence of non-transgenic and transgenic Bt. rice genotypes in north of Iran.

BACKGROUND: Field-testing genetically modified crops provides scientists with an opportunity to collect information on environmental interactions and agronomic performance, which is critical to a full environmental safety assessment as required by regulatory authorities. As a result, the goal of this research was field trial of transgenic and conventional rice genotypes. The experiment was carried out in a randomized complete blocks design (RCBD) with four replications and seven genotypes in three isolated regions under the biosafety standard protocol in north of Iran in 2016. In this study, four transgenic lines with an active cry1Ab gene in the vegetative stage and three conventional genotypes (control) as treatment were assessed. RESULTS: The findings demonstrated that in all three regions, transgenic lines derived from Khazar cultivar, were similar to their parent(s) in terms of growth phenology, agronomical traits, grain amylose content, gel consistency, and gelatinization temperature. In all the three regions, the highest number of panicle per m2, number of filled spikelet per panicle, and filled spikelet percentage per panicle were obtained for transgenic lines. Khazar cultivar compared to the transgenic lines showed lower paddy yield. In all the three regions, transgenic lines had lower yield loss than their non-transgenic parent. The lowest number of white heads belonged to transgenic Tarom Molaii and transgenic lines. The most positive direct effect on paddy yield was related to the number of filled spikelet per panicle. Thus, this can be a good trait to achieve higher yield derived from reducing the negative indirect effect of dead heart and white heads. CONCLUSION: It can be concluded that by producing transgenic rice, which is resistant to stem borer with an active promoter in the reproductive stage, farmers can reduce significant part of yield loss resulting from dead heart and white heads directly correlating with the number of filled spikelet per panicle and paddy yield.

Effects of Environmental Conditions on High-Yield Magnetosome Production by Magnetospirillum gryphiswaldense MSR-1

Background: Magnetotactic bacteria are a heterogeneous group of Gram-negative prokaryote cells that produce linear chains of magnetic particles called magnetosomes, intracellular organelles composed of magnetic iron particles. Many important applications have been defined for magnetic nanoparticles in biotechnology, such as cell separation applications, as well as acting as carriers of enzymes, antibodies, or anti-cancer drugs. Since the bacterial growth is difficult and the yield of magnetosome production is low, the application of magnetosome has not been developed on a commercial scale. Methods: Magnetospirillum gryphiswaldense strain MSR-1 was used in a modified current culture medium supplemented by different concentrations of oxygen, iron, carbon, and nitrogen, to increase the yield of magnetosomes. Results: Our improved MSR-1 culture medium increased magnetosome yield, magnetosome number per bacterial cell, magnetic response, and bacterial cell growth yield significantly. The yield of magnetosome increased approximately four times. The optimized culture medium containing 25 mM of Na-pyruvate, 40 mM of NaNO3, 200 µM of ferrous sulfate, and 5-10 ppm of dissolved oxygen (DO) resulted in 186.67 mg of magnetosome per liter of culture medium. The iron uptake increased significantly, and the magnetic response of the bacteria to the magnetic field was higher than threefold as compared to the previously reported procedures. Conclusion: This technique not only decreases the cultivation time but also reduces the production cost. In this modified method, the iron and DO are the major factors affecting the production of magnetosome by M. gryphiswaldense strain MSR-1. However, refining this technique will enable a further yield of magnetosome and cell density.

Identification of differentially accumulated proteins associated with embryogenic and non-embryogenic calli in saffron (Crocus sativus L.).

BACKGROUND: Somatic embryogenesis (SE) is a complex biological process that occurs under inductive conditions and causes fully differentiated cells to be reprogrammed to an embryo like state. In order to get a better insight about molecular basis of the SE in Crocus sativus L. and to characterize differentially accumulated proteins during the process, a proteomic study based on two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time of flight mass spectrometry has been carried out. RESULTS: We have compared proteome profiles of non-embryogenic and embryogenic calli with native corm explants. Total soluble proteins were phenol-extracted and loaded on 18 cm IPG strips for the first dimension and 11.5% sodium dodecyl sulfate-polyacrylamide gels for the second dimension. Fifty spots with more than 1.5-fold change in abundance were subjected to mass spectrometry analysis for further characterization. Among them 36 proteins could be identified, which are classified into defense and stress response, protein synthesis and processing, carbohydrate and energy metabolism, secondary metabolism, and nitrogen metabolism. CONCLUSION: Our results showed that diverse cellular and molecular processes were affected during somatic to embryogenic transition. Differential proteomic analysis suggests a key role for ascorbate metabolism during early stage of SE, and points to the possible role of ascorbate-glutathione cycle in establishing somatic embryos.

Validation of EST-derived STS markers localized on Qfhs.ndsu-3BS for Fusarium head blight resistance in wheat using a 'Wangshuibai' derived population.

A few EST-derived STS markers localized on Qfhs.ndsu-3BS, a major QTL for resistance to Fusarium head blight (FHB) in wheat, have been previously identified in the 'Sumai 3'/'Stoa' population. In this study, we used a 'Wangshuibai' (resistant) /'Seri82' (susceptible) derived population, linkage group, QTL, and quantitative gene expression analysis to assess the genetic background dependence and stability of the EST-derived STS markers for use in marker aided selection to improve FHB resistance in wheat. Based on our results, a QTL in the map interval of Xsts3B-138_1-Xgwm493 on chromosome 3BS was detected for FHB resistance, which accounted for up to 16% of the phenotypic variation. BLASTN analysis indicated that Xsts3B-138_1 sequence had significant similarity with the resistance gene analogue. Real-time quantitative PCR showed that the relative expression of Xsts3B-138_1 in 'Wangshuibai' at 96 h after inoculation was 2.6 times higher than 'Seri82'. Our results underlined that EST-derived STS3B-138 markers could be predominantly used in marker aided selection to improve FHB resistance in wheat.

Evaluation of Bt (Bacillus thuringiensis) rice varieties against stem borer (Chilo suppressalis).

Three transgenic rice varieties namely Khazar, Neda and Nemat, all containing a cry1Ab gene, were evaluated through PCR analysis and field examinations for their resistance at natural infestation of insect pests during 2007. The results showed that all transgenic varieties produced 1.2 kb PCR product derived from application of cry1Ab gene. In field conditions, transgenic varieties exhibited high levels of resistance against natural infestation of stem borer and the damaged plants based on dead heart or white heat for them were less than 1%. Moreover, in stem-cut bioassay 100% of released larvae died within four days after infestation. These results demonstrate that expression of cry1Ab gene in the genome of transgenic varieties provided season-long protection from the natural infestation of lepidopteran insects.

Proteomic analysis of rice leaves during drought stress and recovery.

Three-week old plants of rice (Oryza sativa L. cv CT9993 and cv IR62266) developed gradual water stress over 23 days of transpiration without watering, during which period the mid-day leaf water potential declined to approximately -2.4 MPa, compared with approximately -1.0 MPa in well-watered controls. More than 1000 protein spots that were detected in leaf extracts by proteomic analysis showed reproducible abundance within replications. Of these proteins, 42 spots showed a significant change in abundance under stress, with 27 of them exhibiting a different response pattern in the two cultivars. However, only one protein (chloroplast Cu-Zn superoxide dismutase) changed significantly in opposite directions in the two cultivars in response to drought. The most common difference was for proteins to be up-regulated by drought in CT9993 and unaffected in IR62266; or down-regulated by drought in IR62266 and unaffected in CT9993. By 10 days after rewatering, all proteins had returned completely or largely to the abundance of the well-watered control. Mass spectrometry helped to identify 16 of the drought-responsive proteins, including an actin depolymerizing factor, which was one of three proteins detectable under stress in both cultivars but undetectable in well-watered plants or in plants 10 days after rewatering. The most abundant protein up-regulated by drought in CT9993 and IR62266 was identified only after cloning of the corresponding cDNA. It was found to be an S-like RNase homologue but it lacked the two active site histidines required for RNase activity. Four novel drought-responsive mechanisms were revealed by this work: up-regulation of S-like RNase homologue, actin depolymerizing factor and rubisco activase, and down-regulation of isoflavone reductase-like protein.