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烟草磷转运蛋白基因NtPHT2;1克隆和表达分析

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[目的]植物对磷酸盐的吸收与利用主要依靠磷转运蛋白,其中PHT2家族编码的低亲和磷转运蛋白主要负责植物在正常供磷条件下磷酸盐的吸收、转运与再利用,为了探究低亲和磷转运蛋白基因NtPHT2;1在烟草转运磷酸盐中的作用和表达模式,通过NtPHT2;1培育烟草磷素高效利用新品种.[方法]以普通烟草K326的cDNA为模板,克隆得到NtPHT2;1,对该基因进行生物信息学分析和蛋白质的亚细胞定位,并通过荧光定量PCR技术对该基因在低磷等非生物胁迫下的基因表达模式进行分析.[结果](1)NtPHT2;1基因全长1 764 bp,编码587个氨基酸;(2)亚细胞定位结果显示,NtPHT2;1蛋白定位于叶绿体上;(3)同源性比对发现,NtPHT2;1蛋白与辣椒CaPHT2;1蛋白的同源性最高达到91.00%;(4)NtPHT2;1启动子含有参与调控植物衰老、逆境胁迫相关的顺式作用元件;(5)NtPHT2;1在叶片表达量最高,新叶中的表达量比老叶中高;在低磷诱导条件下,该基因表达量与正常条件相比差异不显著;(6)非生物胁迫下的表达模式分析显示,NtPHT2;1基因表达量在盐胁迫和干旱胁迫下显著降低.[结论]NtPHT2;1基因主要是负责烟株正常生长发育条件下磷酸盐的转运与利用.
Cloning and expression analysis of the phosphorus transporter gene NtPHT2;1 in Nicotiana tabacum
[Objective]Phosphorus absorption and utilization by plants mainly depend on phosphorus trans-porters,among which the low-affinity phosphorus transporters encoded by the PHT2 family are responsi-ble for the absorption,transport and reuse of phosphate by plants under normal phosphorus conditions.We aim to explore the role and expression pattern of NtPHT2;1 gene in tobacco phosphate transport,and to cultivate new tobacco varieties with efficient phosphorus utilization through NtPHT2;1.[Methods]We cloned NtPHT2;1 using the cDNA of a common tobacco K326 as a template,and conducted bioinformat-ics analysis and subcellular localization analysis of the protein.The expression pattern of this gene under abiotic stress including low phosphorus were analyzed by fluorescence quantitative PCR.[Results](1)The total length of NtPHT2;1 gene was 1 764 bp,encoding 587 amino acids.(2)NtPHT2;1 protein was lo-calized to chloroplast.(3)The homology between NtPHT2;1 and CaPHT2;1 protein was 91%.(4)Nt-PHT2;1 promoter contains cis-acting elements involved in the regulation of plant aging and stress.(5)The expression of NtPHT2;1 was the highest in leaves,and expression in new leaves was higher than that in old leaves.There was no significant difference in the expression of this gene under low phosphorus compared with normal conditions.(6)The expression of this gene was decreased significantly under salt stress and drought stress.[Conclusion]NtPHT2;1 gene is involved in phosphate transport and utilization in growth and development of tobacco plants.

Nicotiana tabacumgene cloningNtPHT21expression patternabiotic stress

湛佳伟、朱紫童、李晨依、温海洋、贺帆、贾宏昉

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河南农业大学烟草学院,郑州 450002

烟草 基因克隆 NtPHT2 1 表达模式 非生物胁迫

河南省科技攻关项目中国烟草总公司贵州省公司重点研发项目河南农业大学高层次人才支持项目

2021021100082022XM0630501289

2024

西北植物学报
西北农林科技大学,陕西省植物学会

西北植物学报

CSTPCD北大核心
影响因子:1.031
ISSN:1000-4025
年,卷(期):2024.44(1)
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