小麦HBD基因家族全基因组鉴定及功能分析

doi:10.7668/hbnxb.20195577

摘要/Abstract

摘要：

为探究HBD家族各成员在重要粮食作物小麦中的潜在生物学功能,首先基于生物信息学分析普通六倍体小麦HBD家族成员及其序列特性,其次通过转录组和荧光定量分析其表达模式及功能。结果显示,共鉴定到90个小麦HBD基因,分别包含2~18个外显子和编码111~1 863个氨基酸,据进化关系可分为6个亚组。组织表达模式的结果显示,多数HBD基因在植株的根、茎、穗和籽粒中相对高表达,体现其生物学功能的多样性。这些HBD成员的启动子区域共包含62种顺式作用元件,主要涉及参与胁迫响应的光和激素调控元件。HBD不同成员分别响应多种非生物胁迫,包括磷、盐、低温、高温、干旱和高温干旱协同胁迫,其中,TaHBD23、TaHBD28、TaHBD67、TaHBD78和TaHBD85等在多种胁迫下均显著差异表达,作为胁迫响应的重要候选基因。针对生物胁迫,HBD基因家族响应假禾谷镰刀菌和半透明黄单胞菌胁迫的数量明显少于响应禾谷镰刀菌,暗示其在赤霉菌抗性应答中的关键作用。通过转录组分析在禾谷镰刀菌和水处理的小麦Bobwhite材料中共鉴定到41个表达量发生显著变化的HBD基因,其中10个基因与表达数据库的结果重合。定量分析与转录组数据趋势一致,显示TaHBD28/17、TaHBD67和TaHBD90/84负向响应赤霉菌,而TaHBD39/37、TaHBD45和TaHBD68/79正向响应赤霉菌。

关键词: 小麦, HBD基因家族, 生物信息学分析, 表达分析

Abstract:

To explore the potential biological functions of HBD family members in the important cereal crop wheat,it first conducted a bioinformatic analysis of the HBD family members and their sequence characteristics in common hexaploid wheat.Subsequently,transcriptome and Real-time Quantitative PCR(qRT-PCR)analyses were performed to assess their expression patterns and functions.The results identified a total of 90 wheat HBD genes,which contained between 2 and 18 exons and comprised 111 to 1 863 amino acids;they could be divided into six subgroups based on their evolutionary relationships.The tissue expression pattern results showed that most HBD genes were relatively highly expressed in the roots,stems,spikes,and grains of the plants,while their expression in leaves was relatively low,reflecting the diversity of their biological functions.The promoter regions of these HBD members contain 62 types of cis-acting elements,mainly involved in light and hormone regulatory elements that participated in stress responses.Different members of the HBD gene family responded to various abiotic stresses,including phosphorus,salinity,low temperature,high temperature,drought,and heat-drought synergistic stress.Among these,TaHBD23,TaHBD28,TaHBD67,TaHBD78,and TaHBD85 showed significant differential expression under various stresses,serving as important candidate genes for stress response.In response to biotic stress,the number of HBD family genes responding to Fusarium pseudograminearum and Pseudomonas translucens was fewer than those responding to F.graminearum, suggesting their critical role in the response to F. graminearum resistance.Further research on the transcriptome data from wheat Bobwhite materials infected with F.graminearum and treated with water identified 41 HBD genes with significantly changed expression levels.Among them,10 genes overlapped with the database,and quantitative analysis was consistent with the trends in transcriptome data,indicating that TaHBD28/17,TaHBD67,and TaHBD90/84 negatively responded to Fusarium head blight,while TaHBD39/37,TaHBD45,and TaHBD68/79 positively responded to Fusarium head blight.

Key words: Wheat, HBD gene family, Bioinformatics analysis, Expression analysis

中图分类号:

S512.1小麦

姜昊, 张琳杰, 蔡嘉茹, 王雪晴, 卢杰, 周毅, 朱玉磊, 王升星. 小麦HBD基因家族全基因组鉴定及功能分析[J]. 华北农学报, 2025, 40(4): 29-40. doi: 10.7668/hbnxb.20195577.

JIANG Hao, ZHANG Linjie, CAI Jiaru, WANG Xueqing, LU Jie, ZHOU Yi, ZHU Yulei, WANG Shengxing. Genome-wide Identification and Functional Analysis of HBD Gene Family in Wheat[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(4): 29-40. doi: 10.7668/hbnxb.20195577.

http://www.hbnxb.net/CN/Y2025/V40/I4/29

图/表 11

表1 qRT-PCR的引物碱基信息

Tab.1 Primers used in this study for qRT-PCR

基因ID或基因名 Gene ID or gene name	正向引物(5'-3') Forward primer (5'-3')	反向引物(5'-3') Reverse primer (5'-3')
Actin	CTATGTTCCCGGGTATTGCT	AAGGGAGGCAAGAATCGAC
TraesCS2D02G387300	TTCCAGAACAAGCGCGCG	GAGGGCGAGCTTCTCTTG
TraesCS2D02G473700	GAGAGCGGCGACAACAAC	CTCCTGCTCCGCGTTGTC
TraesCS2D02G515600	CATGCGCCGTCCTACCTG	GGGAGATCTTGGGCGCCA
TraesCS6A02G120300	CGCTAGTAGGGAGTCGCTG	TCCTTCCAGTATTTCCCGGC
TraesCS6A02G120600	CGCAGGAAGTAGAGGAGCTC	TCCCTTGACTCCCTTCCTCT
TraesCS6A02G240400	GAGGTGGACGACAAGAGCA	TGTCATACTCGTACCAGCCC
TraesCS6B02G148700	TACCGTGACGACCATAAGCA	CCGATATTTCCCGCCACATG
TraesCS6D02G222600	TGAAGGCGAAGGATGAGAGC	ATGACGTCTCCCACAGCTC
TraesCS2A02G389400	ACTACGACGCCCTCCACT	TTGCAGCTGCTGCTGCT
TraesCS4B02G084700	GGCATGCAAACCGCCAA	CTTCTTTGCATTGCGTCG
TraesCS5A02G043400	CCTCTGCAACATCGAGC	GGAGGACTCCTTGCGCT
TraesCS6A02G240400	GAGTCGCTCAAGCAGGAA	CTTGCGCTCTGGTCCTGC
TraesCS6A02G255800	CATGCAGTTCCCTTACG	CGACCGGCTGTCGTTCT
TraesCS7B02G208600	CTGGTTCCAGAACAAGC	AGTGCCGTGTTCTCCTG

图1 拟南芥和小麦HBD基因家族系统进化树

Fig.1 Phylogenetic tree of HBD gene families in Arabidopsis thaliana and Triticum aestivum L.

图2 小麦HBD基因结构及其启动子顺式作用元件分布数字1-24分别表示不同类型的启动子顺式作用元件:1.厌氧诱导,2.光响应,3.低温响应,4.脱落酸响应,5.茉莉酸响应,6.赤霉素响应,7.生长素响应,8.玉米醇溶蛋白代谢相关,9.昼夜节律相关,10.分生组织相关,11.干旱诱导,12.水杨酸响应,13.DNA结合蛋白,14.缺氧特异性诱导,15.种子特异性,16.防御压力相关,17.胚乳表达,18.最大刺激物介导,19.胚乳特异负调控,20.α-淀粉酶启动子保守序列,21.分生组织特异活性,22.根特异,23.细胞周期调节相关,24.光敏色素下调表达。罗马数字Ⅰ和Ⅱ分别表示非翻译区和编码区。字母A-Q分别表示不同类型的结构域:A.START_ArGLABRA2_like,B.Homeobox,C.COG5576,D.bZIP,E.SRPBCC,F.SbcC,G.MEKHLA,H.BRLZ,I.HALZ,J.EEP,K.WAD,L.DDT,M.HARE-HTH,N.WHIM1,O.PTZ00121,P.TPH,Q.PHA03247.

Fig.2 HBD gene structure,and distribution of promoter cis-acting elements in Triticum aestivum L. Figures 1-24 represent different types of promoter cis-acting elements:1.Anaerobic induction,2.Light response,3.Low temperature response,4.Abscisic acid response,5.Jasmonic acid response,6.Gibberellic acid response,7.Auxin response,8.Zein metabolism,9.Circadian rhythm,10.Meristem,11.Drought induction,12.Salicylic acid response,13.DNA binding protein,14.Hypoxia-specific induction,15.Seed-specific,16.Defense stress-related,17.Endosperm expression,18.Maximal stimulus-mediated,19.Endosperm-specific negative regulation,20.The conserved sequence of α-amylase promoter,21.Meristem-specific activity,22.Root-specific activity,23.Cell cycle regulation-related activity,24.Photochrome down-regulated expression.The Roman numerals Ⅰ and Ⅱ represent the untranslated region(UTR)and the coding region(CDS),respectively.The letters A-Q represent different types of domains:A.START_ArGLABRA2_like,B.Homeobox,C.COG5576,D.bZIP,E.SRPBCC,F.SbcC,G.MEKHLA,H.BRLZ,I.HALZ,J.EEP,K.WAD,L.DDT,M.HARE-HTH,N.WHIM1,O.PTZ00121,P.TPH,Q.PHA03247.

图3 小麦HBD基因家族染色体定位分析

Fig.3 Chromosome localization of HBD gene family in Triticum aestivum L.

图4 小麦HBD基因家族共线性分析

Fig.4 Collinearity analysis of HBD gene family in Triticum aestivum L.

图5 小麦HBD基因家族的GO分析

Fig.5 GO analysis of HBD gene family in Triticum aestivum L.

图6 HBD基因家族在小麦不同组织中的表达

Fig.6 Expression of HBD gene family in different tissues in Triticum aestivum L.

图7 非生物胁迫下HBD基因家族在小麦中的转录组表达磷胁迫(P).磷胁迫10 d的根与0 d的根的比值;盐胁迫(S).处理24 h与水处理24 h的比值;低温胁迫(LT).4 ℃胁迫与23 ℃胁迫的比值;高温胁迫(HT).高温胁迫6 h与对照组的比值;干旱胁迫(D).干旱胁迫6 h与对照组的比值;干旱高温(D&HT).干旱与高温协同胁迫6 h与对照组的比值。

Fig.7 Transcriptome expression of the HBD gene family under abiotic stresses in Triticum aestivum L. Phosphorus stress(P).The ratio of roots under phosphorus stress for 10 days to roots under phosphorus stress for 0 days;Salt stress(S).The ratio of salt stress to water treatment after 24 h;Low temperature stress(LT).The ratio of 4 ℃ stress to 23 ℃ stress;High temperature stress(HT).The ratio of high temperature stress to the control group after 6 h;Drought stress(D).The ratio of drought stress 6 h to the control group;Drought and high temperature stress(D&HT).The ratio of drought and heat stress to the control group after 6 h.

图8 盐胁迫下HBD部分基因在小麦中的表达 S_CS.Chinese spring转录组数据的盐胁迫表达;S_AN188.Annong 188荧光定量数据的盐胁迫表达;S_HJ18.Hengjinmai 18荧光定量数据的盐胁迫表达。不同小写字母表示差异显著(P<0.05)。图10同。

Fig.8 The expression of some HBD genes under salt stresses in Triticum aestivum L. S_CS.Salt stress expression in CS transcriptome data;S_AN188.Salt stress expression in Annong 188 quantitative data;S_HJ18.Salt stress expression in Hengjinmai 18 quantitative data.Different lowercase letters indicate significant differences(P<0.05).The same as Fig.10.

图9 生物胁迫下HBD基因家族在小麦中的转录组表达 Fhb.禾谷镰刀菌胁迫;Fp.假禾谷镰刀菌胁迫;Xt.半透明黄单胞菌侵染。

Fig.9 Transcriptome expression of the HBD gene family under biological stresses in Triticum aestivum L. Fhb.Fusarium graminearum stress;Fp.F.pseudograminearum stress;Xt.Xanthomonas stress.

图10 禾谷镰刀杆菌胁迫下HBD部分基因在小麦中的表达 Fhb_F.Fielder转录组数据的禾谷镰刀菌胁迫表达;Fhb_BW.Bobwhite转录组数据的禾谷镰刀菌胁迫表达;Fhb_BWq.Bobwhite荧光定量数据的禾谷镰刀菌胁迫表达。

Fig.10 The expression of some HBD genes under Fusarium graminosarum stresses in Triticum aestivum L. Fhb_F.The stress of Fusarium graminearum in the Fielder transcriptome data;Fhb_BW.The stress of Fusarium graminearum in Bobwhite transcriptome data;Fhb_BWq.The stress of Fusarium graminearum in Bobwhite quantitative data.

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