The members of the fourth subgroup of R2R3-MYB(Sg4 members)are well-known inhibitors of phenylpropanoid and lignin synthesis pathways.The C2 domain is closely related to the transcriptional inhibitory activity of Sg4 members.Phosphorylation modification enhances the transcriptional inhibitory activity of Sg4 members.Here,we identified a phosphorylation site on the C2 domain of Cs MYB4a from tea plants(Camellia sinensis).A mitogen-activated protein kinase(MAPK),named Cs MPK3-2,phosphorylated this site on the C2 domain of Cs MYB4a.Further experiments revealed that phosphorylation of Cs MYB4a weakened its ability to inhibit the gene expression of PAL,C4H,and 4CL in the phenylpropanoid pathway and activated the expression of transcription factor YABBY5,maintaining the adaxial-abaxial polarity of the leaf.Knocking out Nt YAB5 in Cs MYB4a transgenic tobacco partially repaired the leaf wrinkling phenotype caused by Cs MYB4a.The C1 domain exhibited an activation function when the C2 domain of Cs MYB4a was phosphorylated by Cs MPK3-2,causing this reversal phenomenon.These results enrich our understanding of the regulatory diversity of Sg4 members.
为了阐明卵黄蛋白原基因对溞属枝角类生殖和胚胎发育的影响机制,研究了RNA干扰下中华拟同形溞(Daphnia sinensis)的卵黄蛋白原基因表达及产幼溞数变化。结果表明,RNA干扰后,与EGFP(Enhanced green fluorescent protein)组相比,2%大肠杆菌浓度下中华拟同形溞在Vtg组的首次产幼溞数和Vtg基因相对表达量没有显著差异;5%大肠杆菌浓度下中华拟同形溞前四个成龄在Vtg组的产幼溞数均显著小于EGFP组,且中华拟同形溞的第一成龄、第四成龄时在Vtg组的Vtg基因相对表达量均显著小于EGFP组,分别下降了5.25%和78.03%。此外,RNA干扰后Vtg组的中华拟同形溞还出现了胚胎消解和后代发育不全的现象。结果表明,高强度、长时间的RNA干扰能够显著降低中华拟同形溞Vtg基因的相对表达水平,也能够显著抑制其生殖量和胚胎发育。因此,Vtg基因对中华拟同形溞的生殖和胚胎发育起重要作用。