Gene network mediated by WRKY13 to regulate resistance against sheath infecting fungi in rice (Oryza sativa L.).

OsWRKY13 TF gene is known to play a regulatory role of signaling in physiological pathways related to either development or disease resistance in rice plants. Rice cultivars IR 50 and TRY 3, resistant and susceptible respectively to sheath blight, TRY 3 and CO 43 resistant and susceptible respectively to sheath rot were challenged with fungal pathogens and disease scoring was carried out. Percent Disease Index (PDI) was significantly higher in susceptible varieties than resistant varieties. RT-PCR and qPCR analyses of WRKY13 using RNA extracted from the plant tissues revealed higher WRKY13 expression in resistant varieties (both diseases) upon pathogen challenge compared to uninfected control and also the susceptible varieties. To compute and evaluate the possible molecular mechanism for observed resistance correlated to WRKY13 gene expression, rice gene expression profiles against bacterial leaf blight and leaf blast disease from ROAD database were used to prioritize the locus IDs that were used as input in RiceNet v2 tool. The expression of WRKY13-regulated TIFY9 gene was predicted and validated using RT-PCR and qRT-PCR along with WRKY12 and PR2. All three genes showed induced expression in R. solani challenged sheath blight resistant variety. WRKY12 and PR2 expression in S. oryzae challenged sheath rot resistant variety was higher. Agrobacterium mediated transformation was carried out in rice plants using overexpression construct of WRKY13 (agroinfection in seeds of varieties susceptible to sheath blight and sheath rot, followed by selection in antibiotic media, germinating and hardening of putative transgenic lines). Based on qPCR analysis, the expression level of WRKY13 and the co-expression levels of WRKY12, TIFY9 and PR2 were found higher in PCR-positive T1 plants compared to wild-type. Infection bioassays in the transgenic plants of both varieties revealed enhanced resistance to the pathogens. A mechanism by which WRKY13 would influence the MAPK cascade with TIFY9 acting as a mediator, is proposed.

Prospects and progress in the production of valuable carotenoids: Insights from metabolic engineering, synthetic biology, and computational approaches.

Carotenoids are isoprenoid pigments synthesized exclusively by plants and microorganisms and play critical roles in light harvesting, photoprotection, attracting pollinators and phytohormone production. In recent years, carotenoids have been used for their health benefits due to their high antioxidant activity and are extensively utilized in food, pharmaceutical, and nutraceutical industries. Regulation of carotenoid biosynthesis occurs throughout the life cycle of plants, with vibrant changes in composition based on developmental needs and responses to external environmental stimuli. With advancements in metabolic engineering techniques, there has been tremendous progress in the production of industrially valuable secondary metabolites such as carotenoids. Application of metabolic engineering and synthetic biology has become essential for the successful and improved production of carotenoids. Synthetic biology is an emerging discipline; metabolic engineering approaches may provide insights into novel ideas for biosynthetic pathways. In this review, we discuss the current knowledge on carotenoid biosynthetic pathways and genetic engineering of carotenoids to improve their nutritional value. In addition, we investigated synthetic biological approaches for the production of carotenoids. Theoretical biology approaches that may aid in understanding the biological sciences are discussed in this review. A combination of theoretical knowledge and experimental strategies may improve the production of industrially relevant secondary metabolites.

Proteome-level investigation of Brassica carinata-derived resistance to Leptosphaeria maculans.

Plants resistant to the fungal pathogen Leptosphaeria maculans were generated by an interspecific cross between the highly susceptible Brassica napus (canola) and the highly resistant Brassica carinata. Changes in the leaf protein profiles of these lines were investigated in order to understand the biochemical basis for the observed resistance. Two-dimensional electrophoresis followed by tandem mass spectrometry led to the identification of proteins unique to the susceptible (5 proteins) and resistant genotypes (7 proteins) as well those that were differentially expressed in the resistant genotype 48 h after challenge with the pathogen (28 proteins). Proteins identified as being unique in the resistant plant material included superoxide dismutase, nitrate reductase, and carbonic anhydrase. Photosynthetic enzymes (fructose bisphosphate aldolase, triose phosphate isomerase, sedoheptulose bisphosphatase), dehydroascorbate reductase, peroxiredoxin, malate dehydrogenase, glutamine synthetase, N-glyceraldehyde-2-phosphotransferase, and peptidyl-prolyl cis-trans isomerase were observed to be elevated in the resistant genotype upon pathogen challenge. Increased levels of the antioxidant enzyme superoxide dismutase were further validated and supported by spectrophotometric and in-gel activity assays. Other proteins identified in this study such as nitrate reductase and peptidylprolyl isomerase have not been previously described in this plant-pathogen system, and their potential involvement in an incompatible interaction is discussed.