玉米生物钟基因ZmPRR1-2的克隆及表达分析
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国家自然科学基金河南省联合基金(U2004153);国家自然科学基金青年基金(31901548,31901561);河南省重点研发与推广专项(科技攻关)(202102110010)


Cloning and Expression Analysis of Circadian Gene ZmPRR1-2 in Maize
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    摘要:

    为探究玉米生物钟基因ZmPRR1-2的功能及表达特性,解析玉米光周期途径调控开花的机理,该研究以玉米骨干自交系‘黄早4’为材料,克隆ZmPRR1-2基因的cDNA序列并进行生物信息学分析;利用qRT-PCR技术对该基因进行组织特异性表达分析和48 h的昼夜节律表达分析。结果表明:(1) 成功克隆获得ZmPRR1-2基因的编码区全长1 554 bp,编码517个氨基酸,编码的蛋白属于PRR基因家族,含有1个REC结构域和1个CCT结构域,多序列比对和系统进化分析显示ZmPRR1-2基因在禾本科植物中高度保守;ZmPRR1-2蛋白属亲水性蛋白,不包含跨膜结构域和信号肽。(2)ZmPRR1-2基因在玉米叶片中的表达量最高,显著高于其他7个组织,表明该基因主要在叶片中发挥功能,而在果穗和花丝中表达量相对较低,且显著低于雄穗中的表达量。(3)昼夜节律表达分析显示,在短日照条件下,ZmPRR1-2基因的表达量于光照3 h后开始逐渐上升,在光照结束后3 h时达到表达高峰;在长日照条件下,于光照6 h后ZmPRR1-2基因的表达量才开始逐渐上升,且在光照结束时达到表达高峰。研究认为,ZmPRR1-2基因在玉米开花转化过程中对雄穗发育和雌穗发育的调控功能存在差异,推测ZmPRR1-2基因可能在玉米生物钟调控中发挥重要作用。

    Abstract:

    In order to explore the function and expression characteristics of ZmPRR1-2 gene and analyze the mechanism of the flowering regulation by photoperiod pathway in maize, we cloned cDNA sequence of the ZmPRR1-2 gene in the maize backbone inbred line ‘Huangzao 4’ and analyzed by bioinformatics. Tissue-specific expression analysis and 48 h circadian rhythm expression analysis of the gene were performed using qRT-PCR. The results showed that: (1) the full-length coding region of ZmPRR1-2 gene was 1 554 bp, encoding 517 amino acids. The encoded protein belonged to the PRR gene family, containing one REC domain and one CCT domain. Multiple sequence alignment and phylogenetic analysis showed that ZmPRR1-2 gene was highly conserved in Poaceae plants. ZmPRR1-2 is a hydrophilic protein without transmembrane domain and signal peptide. (2) Tissue-specific expression analysis showed that the expression of ZmPRR1-2 gene in leaves was significantly higher than that of the other seven tissues, which indicated that ZmPRR1-2 gene mainly functions in the leaves, while the expression level in ear and silk was relatively low, and significantly lower than that in tassel. (3) Circadian rhythm expression analysis showed that under short-day conditions, the expression level of ZmPRR1-2 gene began to increase gradually after 3 h of light, and reached the peak at 3 h after the end of light. Under long-day conditions, the expression level of ZmPRR1-2 gene began to increase gradually after 6 h of light, and reached the peak at the end of light. It is suggested that ZmPRR1-2 gene has different regulatory functions on tassel development and ear development during maize flowering transformation. ZmPRR1-2 gene may play an important role in the regulation of maize circadian clock.

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董柯清,王雷立,刘青青,等.玉米生物钟基因ZmPRR1-2的克隆及表达分析[J].西北植物学报,2023,43(1):21-28

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  • 在线发布日期: 2023-02-17
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