Abstract
Whitefly infestation of cotton crop imparts enormous damage to cotton yield by severely affecting plant health, vigour and transmitting Cotton Leaf Curl Virus (CLCuV). Genetic modification of cotton helps to overcome both the direct whitefly infestation as well as CLCuV based cotton yield losses. We have constitutively overexpressed asparaginase (ZmASN) gene in Gossypium hirsutum to overcome the cotton yield losses imparted by whitefly infestation. We achieved 2.54% transformation efficiency in CIM-482 by Agrobacterium-mediated shoot apex transformation method. The relative qRT-PCR revealed 40-fold higher transcripts of asparaginase in transgenic cotton line vs. non-transgenic cotton lines. Metabolic analysis showed higher contents of aspartic acid and glutamic acid in seeds and phloem sap of the transgenic cotton lines. Phenotypically, the transgenic cotton lines showed vigorous growth and height, greater number of bolls, and yield. Among six representative transgenic cotton lines, line 14 had higher photosynthetic rate, stomatal conductance, smooth fiber surface, increased fiber convolutions (SEM analysis) and 95% whitefly mortality as compared to non-transgenic cotton line. The gene integration analysis by fluorescence in situ hybridization showed single copy gene integration at chromosome number 1. Collectively, asparaginase gene demonstrated potential to control whitefly infestation, post-infestation damages and improve cotton plant health and yield: a pre-requisite for farmer’s community.
Highlights
Infestation of cotton crop imparts enormous damage to cotton yield by severely affecting plant health, vigour and transmitting Cotton Leaf Curl Virus (CLCuV)
Plant asparaginases are further subdivided into two subclasses based on their dependency on potassium (K+): K+-dependent and K+-independent. These are isolated from various plants including Arabidopsis thaliana, Lupinus albus, Lupinus polyphyllus, Phaseolus vulgaris[4], Lupinus arboreus, Lupinus angustifolius[5], Lotus japonicus[6], Glycine max[7], Withania somnifera[8,9], and Pisum sativum[10]
On the basis of preliminary in vitro feed based bioassays, in the current study, we investigated the toxic effects of aspartic acid on whitefly
Summary
Infestation of cotton crop imparts enormous damage to cotton yield by severely affecting plant health, vigour and transmitting Cotton Leaf Curl Virus (CLCuV). We have constitutively overexpressed asparaginase (ZmASN) gene in Gossypium hirsutum to overcome the cotton yield losses imparted by whitefly infestation. Metabolic analysis showed higher contents of aspartic acid and glutamic acid in seeds and phloem sap of the transgenic cotton lines. Asparaginase gene demonstrated potential to control whitefly infestation, post-infestation damages and improve cotton plant health and yield: a pre-requisite for farmer’s community. Pest infestation activates the salicylic acid signaling pathway followed by jasmonic acid signaling pathway[20,21] Both these defense systems prepare the plant for upcoming challenges by modifying its carbon and nitrogen metabolism. To the best of our knowledge, no one has considered the role of aspartic acid/glutamic acid against the feeding-insect related plant defense systems. The pathway of aspartate, glutamate, glutamine, and asparagine were analyzed and their simultaneous effect on the plant’s health and insect resistance were evaluated
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