陆地棉GhPS2基因的克隆和表达分析
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河南科技大学农学院/河南省旱地作物种质资源利用工程研究中心

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Q785;Q786;Q789???

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国家自然科学(31101184)


Cloning and Expression Analysis of GhPS2 Gene in Gossypium hirsutum L.
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    摘要:

    卤代酸脱卤酶(HAD)在调节植物生长发育和响应磷缺乏胁迫方面具有重要作用。该研究基于前期陆地棉根部低磷胁迫基因差异表达序列数据分析,以陆地棉新陆早19为材料,对GhPS2基因进行克隆,并对其基因组DNA与cDNA测序分析,借助生物信息学方法分析GhPS2的基因结构和进化关系;采用荧光定量PCR(qRT-PCR)的方法检测该基因于根、茎、叶、花4个器官的基因表达量的变化和低磷胁迫下0,4,12,24,72 h的相对表达。结果表明:(1)成功获得陆地棉GhPS2基因,该基因的开放阅读框序列长度813 bp,编码270个氨基酸,存在3个内含子,属于HAD家族,其中存在1个保守结构域名为Put-Phosphatase。(2)序列比对和进化分析显示,陆地棉GhPS2与其他棉种PS2、榴莲PS2的相似性分别为93%和83.15%。(3)qRT-PCR结果表明,GhPS2基因在根中表达量最高,其次是茎和花,在叶中表达量最低,该基因在低磷胁迫4h时相对表达量达到最高值,低磷胁迫72h时是适磷处理的16.66倍。研究表明,GhPS2基因属于低磷胁迫响应基因,在棉花响应低磷胁迫过程中具有重要作用。

    Abstract:

    HAD-like hydrolase plays an important role in regulating plant growth and development and responding to phosphorus deficiency stress. Based on the genome-wide expression profile of cotton seedlings under low phosphorus stress in the early stage of our research group, related genes were excavated and their preliminary expression analysis was conducted. Genomic DNA and cDNA sequence of the gene were cloned and analyzed by bioinformatics method. Fluorescence quantitative PCR (qRT-PCR) were used to detect the changes of gene expression in root, stem, leaf and flower tissues.The relative expression patterns of GhPS2 under low phosphorus stress(0,4,12,24 and 72h) were analyzed by qRT-PCR technology.The results showed that: (1)GhPS2 gene of G. hirsutum L. was successfully cloned, and the full length of the open reading frame of the gene was 813bp, encoding 270 amino acids and containing 3 introns. Analysis revealed that this gene sequence belonged to the HAD family, with a conserved structure named PUT-Phosphatase. (2) Sequence alignment and evolution analysis showed that the similarity of GhPS2 to other cotton varieties PS2 and durian PS2 was 93% and 83.15%, respectively.(3)The results of qRT-PCR showed that GhPS2 gene was predominantly expressed in roots, followed by stems and flowers, with the lowest expression in leaves. The relative expression of GhPS2 gene was the highest in the roots treated with low phosphorus for 4 h, and it was 16.66 times that of suitable phosphorus treatment at 72 h under low phosphorus stress.This study demonstrated that GhPS2 gene belongs to low phosphorus stress response gene and may be involved in Gossypium hirsutum L. response low phosphorus stress.

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孟超敏,卿桂霞,耿翡翡,等.陆地棉GhPS2基因的克隆和表达分析[J].西北植物学报,2023,43(11):1820-1826

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  • 收稿日期:2023-06-07
  • 最后修改日期:2023-10-28
  • 录用日期:2023-10-30
  • 在线发布日期: 2023-11-14
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