Abstract
Glycerol-3-phosphate acyltransferases (GPATs), critical for multiple biological processes like male fertility, have been extensively characterized. However, their precise functions and underlying regulatory mechanism in cotton anther development are unclear. This research demonstrated the importance of GhGPAT12/25 (a paralogs pair on A12/D12 sub-chromosome of cotton) to regulate the degradation of tapetum, anther cuticle formation, and pollen exine development. GhGPAT12 and GhGPAT25 exhibited specifically detected transcripts in tapetum and pollen exine during the early anther developmental stages. GhGPAT12/25 are sn-2 glycerol-3-phosphate acyltransferases and can transfer the acyl group of palmitoyl-CoA to glycerol-3-phosphate (G3P). CRISPR/Cas9-mediated knockout identified the functional redundancy of GhGPAT12 and GhGPAT25. Knockout of both genes caused completely male sterility associated with abnormal anther cuticle, swollen tapetum, and inviable microspores with defective exine and irregular unrestricted shape. RNA-seq analysis showed that the loss of function of GhGPAT12/25 affects the processes of wax metabolic, glycerol monomer biosynthesis, and transport. Consistently, cuticular waxes were dramatically reduced in mutant anthers. Yeast one-hybrid system (Y1H), virus-induced gene silencing (VIGS), and dual-luciferase (LUC) assays illustrated that GhMYB80s are likely to directly activate the expression of GhGPAT12/25. This study provides important insights for revealing the regulatory mechanism underlying anther development in cotton.
Highlights
In flowering plants, male reproductive development is a complex biological process involving the differentiation of stamen primordium to produce anther, the transition from sporophytic to gametophytic generation, and the formation of pollen grains (Scott et al, 2004)
Based on the combination analysis of these data, GhGPAT12/25
The results suggested that GhGPAT12/25 were anther-specific expressed and showed negligible expression in all the other tissues tested
Summary
Male reproductive development is a complex biological process involving the differentiation of stamen primordium to produce anther, the transition from sporophytic to gametophytic generation, and the formation of pollen grains (Scott et al, 2004). Mutations in genes responsible for the formation or transport of these lipid components usually cause abnormal programmed cell death (PCD) in the tapetum Examples of such genes are AMS/TDR, MYB80, MS1/PTC1, MS2/DPW, CYP703A2/CYP703A3, and CYP704B1/CYP704B2 (Wilson et al, 2001; Sorensen et al, 2003; Li et al, 2006, 2010, 2011; Morant et al, 2007; Dobritsa et al, 2009; Chen et al, 2011; Phan et al, 2011; Shi et al, 2011; Yang et al, 2014; Pan et al, 2020)
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