图1 不同植物物种中miR396基因家族成熟区序列比对分析 A和B：植物体中的大多mat-miR396序列主要与拟南芥中的ath-miR396a-5p和ath-miR396b-3p两种成熟序列形式相同，ath-miR396a-5p和ath-miR396b-3p二者序列长度一致，但在末位具有一个差异碱基；C：植物中上述两种序列形式之外的其它mat-miR396序列比对结果发现各mat-miR396序列之间存在1个或多个碱基的差异

Fig.1 Alignment analysis of mature region sequences of miR396 gene family in different plant species A and B：The most mat-miR396 sequences in plants are mainly the same as the 2 mature sequence forms of ath-miR396a-5p and ath-miR396b-5p in Arabidopsis，ath-miR396a-5p and ath-miR396b-3p have the same sequence length，but they have a different base at the end. C：The alignment of other mat-miR396 sequences except above 2 sequences in plants revealed that there is one base difference or multiple base differences between mat-miR396 sequences

图2 不同植物物种中GRF蛋白家族氨基酸序列的保守结构域比对分析 不同植物中的GRF蛋白家族氨基酸序列比对分析发现，GRF蛋白的N端普遍包含QLQ和WRC两个保守功能结构域。A：GRF蛋白的QLQ结构域，在QLQ结构域中，除了保守的Gln-Leu-GLn（Q-L-Q）氨基酸外，还含有疏水性的酸性氨基酸残基 F/Phe、Y/Tyr、L/Leu、E/Glu和P/Pro等，或是含有其他理化性质类似的氨基酸；B：GRF蛋白的WRC结构域，在WRC结构域中，含有保守的Trp-Arg-Cys（W-R-C）氨基酸残基，并且含有由3个C/Cys和1个H/His组成的C3H motif。紧随C3H motif区域之后，有1个保守的“RSRK-VE区”，该区域对应的核苷酸序列即miR396在GRF基因上的作用靶点

Fig.2 Alignment analysis of the conserved domains of GRF protein family amino acid sequences in different plant species According to the amino acid sequence comparison analysis of GRF protein family in different plant species，it can be found that the N-terminal of GRF protein contained two conservative protein domain，QLQ and WRC. A：QLQ domain of GRF protein. In the QLQ domain，there are conserved Gln-Leu-GLn（Q-L-Q）amino acids and hydrophobic residues such as F/Phe，Y/Tyr，L/Leu，E/Glu and P/Pro，or other amino acids with similar physical and chemical properties. B：WRC domain of GRF protein. In the WRC domain，there are conserved Trp-Arg-Cys（W-R-C）amino acid residues and C3H motifs consisting of three C/Cys and an H/His. It is important to note that there is a conserved “RSRK-VE” region following the C3H motif region，its coded nucleotide sequence is the targets site of miR396 on the GRF gene

图3 不同植物物种中miR396在GRF基因序列上的作用位点及剪切位点 A：miR396在GRF基因上的作用位点，在GRF蛋白的WRC结构域中，有一个保守的“RSRK-VE”区，该区域在基因组中对应的核苷酸序列大多为高度保守的“CGTTCAAGAAAGCNTGTGGAA”序列；B：miR396在GRF基因的mRNA序列上的剪切位点，miR396在保守位点的“CGUUCAAGAA”和“AGCNUGUGGAA”之间剪切GRF基因的mRNA序列，进而负调控GRF基因的表达

Fig.3 Action sites and splicing sites of miR396 on GRF gene sequences in different plant species A：The action site of miR396 on GRF gene. In the WRC domain of GRF protein，there is a conserved “RSRK-VE ”region，their corresponding nucleotide sequences in the genome are mostly highly conserved “CGTTCAAGAACNTGTGAA” sequences. B：Splice site of miR396 on the mRNA sequence of GRF gene. miR396 cut the mRNA sequence of GRF gene between “CGUUCAAGAA” and “AGCNUGUGGAA” site，thus negatively regulating the GRF gene expression

图4 miR396-GRF模块调控植物生长发育的过程 miR396-GRF调控途径通过影响细胞扩增以及GA和IAA激素合成，从而影响植物营养生长、雄性生殖以及果实发育等生长发育过程。并且GIF蛋白与GRF蛋白会形成功能复合物从而增强GRF功能

Fig. 4 Process of miR396-GRF module regulating plant growth and development The miR396-GRF regulatory pathway affects vegetative growth，male reproduction and fruit development by affecting cell amplification and the synthesis of GA and IAA hormones. And GIF protein and GRF protein will form a functional complex to enhance GRF function

图5 miR396-GRF调控模块在胁迫响应过程中的可能调控途径 植物体中，miR396-GRF模块通过调控下游胁迫反应相关功能基因的表达和细胞增殖，参与着多种逆境胁迫相应过程

Fig. 5 Feasible regulation approach of miR396-GRF mod-ule in the process of stress response In plants, miR396-GRF module participates in a variety of stress response processes by regulating the expression of downstream stress response related functional genes and cell proliferation