荒漠植物花花柴HTR基因家族表达模式分析及在细菌中抗逆性分析
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1.塔里木大学;2.塔里木大学生命科学与技术学院

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新疆生产建设兵团财政科技计划项目—“强南”科技创新骨干人才项目“沙漠植物花花柴耐高温相关基因的克隆及转化棉花的研究”(2021CB013)第一作者徐靖辰(1997-),女,博士研究生,研究方向为植物逆境生物学。E-mail760997181qq.com*信作者王彦芹(1974-),女,博士,教授,研究方向为植物逆境生物学。E-mailwyqwxf@126.com


Expression pattern analysis of HTR Gene family of desert plant Karelinia caspica and stress resistance analysis in bacteria
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    摘要:

    摘要: 植物常遭受许多非生物胁迫的危害,温度、高盐是影响植物生长发育的重要非生物因子之一,因此研究其响应逆境胁迫的分子机理显得尤为重要。本研究从荒漠植物花花柴高温转录组数据中筛选并克隆了HTR基因家族的7个成员,通过构建原核表达载体并诱导表达,明确其cDNA长度在708~789 bp之间,所表达的蛋白质分子量在27~29 kD之间,编码基因和蛋白质大小相符;通过实验探索,发现在大肠杆菌BL21最适生长温度37℃下该家族蛋白的最佳诱导表达体系为OD600 为0.8、IPTG诱导浓度为0.5 mmol/L、诱导时间为8~10 h。通过对重组大肠杆菌的模拟高温、低温、盐胁迫处理,结果显示KcHTRs能够在一定程度上增强宿主对高温、低温和高盐的耐受性,该结果与本研究中花花柴KcHTRs的非生物胁迫表达模式分析结果一致。 为后期进一步验证基因功能以及荒漠植物抗逆相关基因资源的挖掘利用提供了研究基础和依据。

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    Abstract: Plants are often subjected to many abiotic stresses. Temperature and high salt are one of the important abiotic factors affecting plant growth and development. Therefore, it is particularly important to study the molecular mechanism of plant response to stress. In this study, 7 members of KcHTR gene family were screened and cloned from the high-temperature transcriptome data of desert plant Karelinia caspica. By constructing prokaryotic expression vectors and inducing expression, it was clear that the length of their cDNA was 708 ~ 789 bp, the molecular weight of the expressed protein was 27 ~ 29 kD, and the encoding gene was consistent with the size of the protein. Through experimental exploration, it was found that under the optimal growth temperature of 37℃, the optimal expression system of this family protein was OD600 0.8, the induction concentration of IPTG was 0.5 mmol/L, and the induction time was 8 ~ 10 h. Through simulated high temperature, low temperature and salt stress treatment of recombinant E.coli, the results showed that KcHTRs could enhance the host tolerance to high temperature, low temperature and high salt to a certain extent, which was consistent with the results of abiotic stress expression pattern analysis of KcHTRs in this study. It provides the research basis and basis for further verification of gene function and mining and utilization of desert plant stress-resistance related gene resources.

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徐靖辰,王彦芹.荒漠植物花花柴HTR基因家族表达模式分析及在细菌中抗逆性分析[J].西北植物学报,2024,44(1):13-21

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  • 收稿日期:2023-07-10
  • 最后修改日期:2023-08-28
  • 录用日期:2023-10-11
  • 在线发布日期: 2024-01-02
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