Downregulation of the DST Transcription Factor Using Artificial microRNA to Increase Yield, Salt and Drought Tolerance in Rice

Abstract

Abiotic stresses like salinity and drought directly affect plant growth and water availability, resulting in lower yield in rice. So, a combination of stress tolerance along with enhanced grain yield is a major focus of rice breeding. It was reported earlier that loss in function of the drought and salt tolerance ( DST) gene results in increase in grain production through downregulating Gn1a/OsCKX 2 expression. Moreover, dst mutants also showed enhanced drought and salt tolerance in rice by regulating genes involved in ROS homeostasis. In the present study, we proceeded to test these reports by downregulating DST using artificial microRNA technology in the commercial but salt sensitive, high-yielding, BRRIdhan 28 (BR28). This cultivar was transformed with DST _artificial microRNA ( DST _amiRNA) driven by the constitutive CaMV 35 S promoter using tissue culture independent Agrobacterium mediated in planta transformation. DST _amiRNA transgenic plants were confirmed by artificial microRNA specific PCR. Transformed plants at T0 generation showed vigorous growth with significantly longer panicle length and higher primary branching resulting in higher yield, compared to the wild type (WT) BR28. Semi-quantitative RT PCR confirmed the decrease in DST expression in the BR28 transgenic plants compared to WT. T1 transgenic plants also showed improvement in a number of physiological parameters and greater growth compared to WT after 14 days of 120 mM salt (NaCl) stress at seedling stage. Therefore, DST downregulated transgenic plants showed both higher stress tolerance as well as better yields. Furthermore, stable inheritance of the improved phenotype of the DST _amiRNA transgenics will be tested in advanced generations.