N6-甲基腺苷(m6A)是mRNA最丰富的内部修饰之一,在基因表达调控中起着重要作用,和植物的生长发育。Vir样m6A甲基转移酶缔合物(VIRMA)用作桥接m6A甲基转移酶复合物的催化核心组分的支架。VIRMA在调节叶片发育中的作用及其相关机制尚未见报道。这里,我们鉴定并表征了两个陆地棉(陆地棉)VIRMA基因,命名为GhVIR-A和GhVIR-D,他们彼此共有98.5%的身份。GhVIR-A和GhVIR-D在不同组织中普遍表达,在叶片和主茎尖(MSA)中表达相对较高。通过病毒诱导的基因沉默(VIGS)系统敲低GhVIR基因的表达会影响叶片细胞大小,细胞形状,和细胞总数,从而确定棉花叶片形态发生。斑点印迹测定和比色实验表明,与对照植物相比,GhVIR-VIGS植物叶片中mRNA中m6A与A的比例较低。信使RNA(mRNA)高通量测序(RNA-seq)和qRT-PCR实验表明,GhVIR通过影响某些转录因子基因的表达来调节叶片发育,微管蛋白基因,和叶绿体基因,包括光系统,碳固定,和核糖体组装。叶绿体结构,叶绿素含量,与对照植物相比,GhVIR-VIGS植物的光合效率发生了变化,不适合叶片的生长和发育。一起来看,我们的结果表明GhVIR通过叶绿体依赖性和独立性途径在棉花叶片发育中发挥功能。
N6-methyladenosine (m6A) is one of the most abundant internal modifications of mRNA, which plays important roles in gene expression regulation, and plant growth and development. Vir-like m6A methyltransferase associated (VIRMA) serves as a scaffold for bridging the catalytic core components of the m6A methyltransferase complex. The role of VIRMA in regulating leaf development and its related mechanisms have not been reported. Here, we identified and characterized two upland cotton (Gossypium hirsutum) VIRMA genes, named as
GhVIR-A and
GhVIR-D, which share 98.5% identity with each other.
GhVIR-A and
GhVIR-D were ubiquitously expressed in different tissues and relatively higher expressed in leaves and main stem apexes (MSA). Knocking down the expression of
GhVIR genes by the virus-induced gene silencing (VIGS) system influences leaf cell size, cell shape, and total cell numbers, thereby determining cotton leaf morphogenesis. The dot-blot assay and colorimetric experiment showed the ratio of m6A to A in mRNA is lower in leaves of
GhVIR-VIGS plants compared with control plants. Messenger RNA (mRNA) high-throughput sequencing (RNA-seq) and a qRT-PCR experiment showed that GhVIRs regulate leaf development through influencing expression of some transcription factor genes, tubulin genes, and chloroplast genes including photosystem, carbon fixation, and ribosome assembly. Chloroplast structure, chlorophyll content, and photosynthetic efficiency were changed and unsuitable for leaf growth and development in GhVIR-VIGS plants compared with control plants. Taken together, our results demonstrate GhVIRs function in cotton leaf development by chloroplast dependent and independent pathways.