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园艺学报 ›› 2024, Vol. 51 ›› Issue (3): 495-508.doi: 10.16420/j.issn.0513-353x.2023-0089

• 遗传育种·种质资源·分子生物学 • 上一篇    下一篇

菠萝AcZFP1的克隆及其在低温胁迫下的功能分析

罗健东1,邱梦青1,周慧敏1,解为玮1,黄海鑫1,刘洁琪1,张梓敏1,徐 健2,陈程杰1,何业华1,刘朝阳1,*   

  1. 1华南农业大学园艺学院,农业农村部华南地区园艺作物生物学与种质创制重点实验室,广州 510642;2广西南亚热带农业科学研究所,广西崇左 532415
  • 出版日期:2024-03-25 发布日期:2024-03-22
  • 基金资助:
    广东省自然科学基金面上项目(2022A1515010463);广州市科技计划项目(202102020226);广西科技重大专项(桂科AA22068096-2)

Cloning and Functional Analysis of Pineapple AcZFP1 Gene Under Low Temperature Stress

LUO Jiandong1,QIU Mengqing1,ZHOU Huimin1,XIE Weiwei1,Huang Haixin1,LIU Jieqi1,ZHANG Zimin1,XU Jian2,CHEN Chengjie1,HE Yehua1,and LIU Chaoyang1,*   

  1. 1Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in South China,Ministry of Agriculture and Rural Affairs,Colleage of Horticulture,South China Agricultural University Guangzhou 510642,China;2Guangxi South Subtropical Agricultural Science Research Insititute,Chongzuo,Guangxi 532415,China
  • Published:2024-03-25 Online:2024-03-22

摘要: 以‘神湾’菠萝为材料,克隆了1个C2H2型锌指蛋白基因,命名为AcZFP1,其开放阅读框为816 bp,编码271个氨基酸,含有2个典型的锌指蛋白结构域。进化树分析表明AcZFP1与水稻OsZFP252和OsMSR15的亲缘关系最近。基因表达分析发现,AcZFP1受低温、NaCl和聚乙二醇(PEG)干旱等多种逆境胁迫及外源ABA、SA和MeJA的诱导。试验表明,AcZFP1蛋白定位于细胞核中。AcZFP1过表达的拟南芥株系在低温处理后相比野生型对照有更高的成活率和Fv/Fm值,可溶性蛋白含量增加,SOD、POD酶活增强,MDA含量降低,植株的耐寒性显著增强。进一步的qRT-PCR分析显示,AcZFP1可以促进AtRD29A、AtCOR47等4个典型的拟南芥内源冷应答基因的表达,在低温胁迫中发挥正调控作用。

关键词: pineapple, AcZFP1, low temperature stress, function analysis

Abstract: In this study,a C2H2 zinc finger proteins gene,named AcZFP1,was obtained from ‘Shenwan’pineapple. The open reading frame of AcZFP1 was 816 bp,which encoded 271 amino acids,and contained two typical zinc finger protein domains. The phylogenetic tree analysis showed that AcZFP1 shared the closest relationship with OsZFP252 and OsMSR15. Gene expression pattern analysis showed that AcZFP1 could be induced by several kinds of stress and hormonal treatment like cold,NaCl,PEG,ABA,SA and MeJA. Subcellular localization assays showed that AcZFP1 located in the nucleus. The AcZFP1-overexpressing(OE)Arabidopsis lines have higher survival rate and Fv/Fm under low temperature stress compared to wild type Arabidopsis. Moreover,the OE lines also showed higher level of soluble protein content,higher activity of superoxide dismutase(SOD)and peroxidase(POD),lower level of malondialdehyde(MDA)content. Further qRT-PCR analysis indicated that AcZFP1 could promote the expression of four typical Arabidopsis endogenous cold-responsive genes such as AtRD29A and AtCOR47,and play a positive regulatory role in low temperature stress.

Key words: 菠萝, AcZFP1, 低温胁迫, 功能分析

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