黄瓜AHP基因家族的鉴定及其非生物胁迫表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2021-1338

摘要/Abstract

摘要：

组氨酸磷酸转运蛋白（AHP）是细胞分裂素信号转导途径中重要蛋白之一,在植物生长发育和抵御系列非生物胁迫过程中发挥着重要作用。探讨黄瓜AHP的分子特征与表达模式,可为进一步探究黄瓜AHP基因功能和提高植物的非生物胁迫耐受性提供理论依据。通过对黄瓜AHP基因家族成员的数量、进化关系、顺式作用元件、基因结构、发育及非生物胁迫下基因表达模式进行分析。结果表明,黄瓜AHP基因家族有7个成员,不均匀分布在6条染色体上;系统进化分析发现黄瓜AHP基因家族成员与冬瓜和甜瓜亲缘关系较近;其启动子序列主要含有光响应、低温响应、干旱响应、激素响应和防御应激等顺式作用元件;基因表达谱分析发现CsAHP1、CsAHP3和CsAHP4在非生物胁迫下显著上调或下调,且CsAHP1在开花当天表达量最高;实时荧光定量PCR（qRT-PCR）结果显示,CsAHP1和CsAHP3均在干旱、高温胁迫下显著上调表达,CsAHP1、CsAHP2和CsAHP5在低温弱光胁迫下显著上调表达。黄瓜AHP基因家族可能与植物发育和非生物胁迫响应有着密切的关系。

关键词: 黄瓜, 组氨酸磷酸转运蛋白, 非生物胁迫, 基因表达

Abstract:

Histidine-phosphate transporter（AHP）is one of the important proteins in cytokinin signal transduction pathway,which plays an important role in plant growth and development and resistance to a series of abiotic stresses. Exploring the molecular characteristics and expression pattern of AHP in C. sativus can provide theoretical basis for further exploring the function of AHP gene in C. sativus and improving abiotic stress tolerance of plants. The number,evolutionary relationship,cis-acting elements,gene structure,development and expression patterns of AHP gene family members under abiotic stress in C. sativus were analyzed. The results showed that there were a total of 7 members of AHP gene family in C. sativus,which were unevenly distributed on 6 chromosomes. Phylogenetic analysis indicated that the AHP gene family members of C. sativus were closely related to Benincasa hispida and Citrullus vulgaris Schrad L. Its promoter sequence mainly contained cis-acting elements including light response,low temperature response,drought response,hormone response and defense stress. Transcriptome analysis showed that CsAHP1,CsAHP3 and CsAHP4 were significantly up-regulated or down-regulated under abiotic stress and the expressions of CsAHP1 was the highest on flowering day. Furthermore,real-time fluorescence quantitative PCR（qRT-PCR）results indicated that CsAHP1 and CsAHP3 were significantly up-regulated under drought and high temperature stress,and CsAHP1,CsAHP2 and CsAHP5 were significantly up-regulated under low temperature and low light stress. The AHP gene family of C. sativus might be closely related to plant development and abiotic stress response.

Key words: C. sativus L., histidine phosphate transporter, abiotic stress, gene expression

周国彦, 银珊珊, 高佳鑫, 武春成, 闫立英, 谢洋. 黄瓜AHP基因家族的鉴定及其非生物胁迫表达分析[J]. 生物技术通报, 2022, 38(6): 112-119.

ZHOU Guo-yan, YIN Shan-shan, GAO Jia-xin, WU Chun-cheng, YAN Li-ying, XIE Yang. Identification of AHP Gene Family in Cucumis sativus and Its Expression Analysis Under Abiotic Stress[J]. Biotechnology Bulletin, 2022, 38(6): 112-119.

图/表 7

表1 qRT-PCR引物设计

Table 1 qRT-PCR primer design

引物名称Primer name	引物序列Primer sequence（5'-3'）
CsAHP1-F	TTGTTGAAGTGGTGTCTC
CsAHP1-R	CCTATGCTTGAACTGCTT
CsAHP2-F	TACTACAGAGACTCATCCA
CsAHP2-R	GGCATCCAACTTATTGAAG
CsAHP3-F	CCAATTCCTTCGCTACAA
CsAHP3-R	ATCAATACACTTCTGCTCAA
CsAHP4-F	AAGACAGTGGATTCAGTAT
CsAHP4-R	TAGCGATTCTCAGTTTATTG
CsAHP5-F	TTTGAGCAGGGTTATCTTG
CsAHP5-R	GTATGAGTCTGGCTGAATC
CsAHP6-F	GAACCGATTACTTGCCTTA
CsAHP6-R	TCATCTTGGAGTTGTTGTAG
CsAHP7-F	ATTGTTGAGGCAGATGAA
CsAHP7-R	CTATATGAATCACGGTAGCA
Cs-actin-F	ATTGTTCTCAGTGGTGGTTCTAC
Cs-actin-R	CCTTTGAGATCCACATCTGCT

图1 黄瓜AHP基因家族成员染色体定位分布图

Fig. 1 Chromosome location distribution map of AHP gene family members in C. sativus

图2 黄瓜、拟南芥和葫芦科AHP家族成员进化树分析

Fig. 2 Phylogenetic tree analysis of AHP family members in C. sativus,Arabidopsis thaliana and Cucurbitaceae Cs：黄瓜Cucumis sativus L.;At：拟南芥Arabidopsis thaliana L.;Cmo：南瓜Cucurbita moschata Duch L.;Cla：西瓜Citrullus vulgaris Schrad L.;MEL：甜瓜Cucumis melo L.;Bhi：冬瓜Benincasa hispida L.;Cp：西葫芦Cucurbita pepo L.;Lsi：葫芦 Lagenaria siceraria L.

图3 黄瓜AHP家族基因启动子序列顺式作用元件分析

Fig. 3 Analysis of cis-acting element in promoter sequence of AHP family gene in C. sativus

图4 黄瓜AHP家族基因结构与保守结构域分析

Fig. 4 Gene structure and conservative domain analysis of AHP family in C. sativus

图5 黄瓜非生物胁迫与不同发育时期AHP家族基因表达模式分析 a：盐胁迫;b：硅胁迫;c：温光胁迫;d：不同组织中的表达模式;e：开花期基因的表达模式

Fig. 5 Expression pattern analysis of AHP family in abiotic stress and different developmental stages of C. sativus a：Salt stress. b：Silicon stress. c：Temperature and light stress. d：Expression patterns in different organizations. e：Expression patterns of genes during anthesis

图6 在干旱、高温和低温弱光胁迫下黄瓜AHP家族基因qRT-PCR分析

Fig. 6 qRT-PCR analysis of AHP family gene under drou-ght,high temperature,and low temperature and low light stress in C. sativus

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