Development of Useful Recombinant Promoter and Its Expression Analysis in Different Plant Cells Using Confocal Laser Scanning Microscopy

BACKGROUND: Designing functionally efficient recombinant promoters having reduced sequence homology and enhanced promoter activity will be an important step toward successful stacking or pyramiding of genes in a plant cell for developing transgenic plants expressing desired traits(s). Also basic knowledge regarding plant cell specific expression of a transgene under control of a promoter is crucial to assess the promoter's efficacy. METHODOLOGY/PRINCIPAL FINDINGS: We have constructed a set of 10 recombinant promoters incorporating different up-stream activation sequences (UAS) of Mirabilis mosaic virus sub-genomic transcript (MS8, -306 to +27) and TATA containing core domains of Figwort mosaic virus sub-genomic transcript promoter (FS3, -271 to +31). Efficacies of recombinant promoters coupled to GUS and GFP reporter genes were tested in tobacco protoplasts. Among these, a 369-bp long hybrid sub-genomic transcript promoter (MSgt-FSgt) showed the highest activity in both transient and transgenic systems. In a transient system, MSgt-FSgt was 10.31, 2.86 and 2.18 times more active compared to the CaMV35S, MS8 and FS3 promoters, respectively. In transgenic tobacco (Nicotiana tabaccum, var. Samsun NN) and Arabidopsis plants, the MSgt-FSgt hybrid promoter showed 14.22 and 7.16 times stronger activity compared to CaMV35S promoter respectively. The correlation between GUS activity and uidA-mRNA levels in transgenic tobacco plants were identified by qRT-PCR. Both CaMV35S and MSgt-FSgt promoters caused gene silencing but the degree of silencing are less in the case of the MSgt-FSgt promoter compared to CaMV35S. Quantification of GUS activity in individual plant cells driven by the MSgt-FSgt and the CaMV35S promoter were estimated using confocal laser scanning microscopy and compared. CONCLUSION AND SIGNIFICANCE: We propose strong recombinant promoter MSgt-FSgt, developed in this study, could be very useful for high-level constitutive expression of transgenes in a wide variety of plant cells

Stage-specific, Nonlinear Surface Ozone Damage to Rice Production in China

China is one of the most heavily polluted nations and is also the largest agricultural producer. There are relatively few studies measuring the effects of pollution on crop yields in China, and most are based on experiments or simulation methods. We use observational data to study the impact of increased air pollution (surface ozone) on rice yields in Southeast China. We examine nonlinearities in the relationship between rice yields and ozone concentrations and find that an additional day with a maximum ozone concentration greater than 120 ppb is associated with a yield loss of 1.12% ± 0.83% relative to a day with maximum ozone concentration less than 60 ppb. We find that increases in mean ozone concentrations, SUM60, and AOT40 during panicle formation are associated with statistically significant yield losses, whereas such increases before and after panicle formation are not. We conclude that heightened surface ozone levels will potentially lead to reductions in rice yields that are large enough to have implications for the global rice market

A common genetic mechanism underlies morphological diversity in fruits and other plant organs

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