四倍体小麦成株抗白粉病基因定位

doi:10.7668/hbnxb.20195107

摘要/Abstract

摘要：

摘要:四倍体小麦是普通小麦的祖先种,也是重要的粮食作物。发掘四倍体小麦抗病种质并鉴定其携带的抗病基因,旨在为小麦新品种选育提供新抗源。TDI-1是栽培二粒小麦,在多年的田间种植过程中表现出对白粉病免疫的表型。为了确定TDI-1携带的抗病基因,为小麦白粉病抗性遗传提供理论依据,首先将其与表现为感白粉病表型的硬粒小麦TDU-1进行杂交并构建了遗传群体,然后对亲本及其杂交F₁、F₂、F_2:3群体进行了白粉菌小种E09接种试验和抗病性分析,最后利用混合分离群体分析法(BSA)对抗白粉病基因进行了定位。结果表明,TDI-1在苗期对E09表现为感病,在成株期对E09表现为抗病;TDI-1和TDU-1的F₁植株在成株期对E09表现为抗病;F₂群体中,表现为抗病的单株数和感病的单株数的比例符合3:1($χ_{3:1}^{2}$= 0.11,P=0.74);F_2:3家系中,纯合抗病、抗病性分离、纯合感病的家系数目之比符合1:2:1($χ_{1:2:1}^{2}$= 0.47,P=0.79),表明TDI-1成株期白粉病抗性由1个显性基因控制,暂将其命名为PmTDI-1。对TDI-1和TDU-1及其杂交后代F₂群体进行分子标记筛选,发现4个位于2A染色体短臂上的分子标记Xwmc407、NRM-2AS29、NRM-2AS45和NRM-2AS84与PmTDI-1紧密连锁,其中,NRM-2AS45和NRM-2AS84位于两侧,遗传距离分别为1.8,4.6 cM。由此,将成株期抗白粉病基因PmTDI-1初步定位于2A染色体短臂上。研究结果显示,从四倍体小麦TDI-1中鉴定到1个新的显性成株抗白粉病基因PmTDI-1。

关键词: 四倍体小麦, 白粉病, 抗病基因, 混合分离群体分析法, 基因定位

Abstract:

Abstract: Tetraploid wheat is the ancestor specie of common wheat and an important food crop.Aiming to provide new resistance sources for wheat variety breeding,the resistance tetraploid wheat germplasm was explored and their resistance genes were identified.TDI-1 is a cultivated emmer wheat that has been immune to powdery mildew in the field for many years.To determine the resistance genes carried by TDI-1,and provide a theoretical basis for genetic improvement of wheat resistance,a durum wheat TDU-1 that was susceptible to powdery mildew was used to hybridize with TDI-1,and their F₁ plants,F₂ population,and F_2:3 lines were obtained.Genetic analysis of resistance was conducted on parents TDI-1,TDU-1,and their hybrid offspring that were inoculated with powdery mildew isolate E09.Then,bulked segregant analysis method combined with molecular markers was used to map the resistance gene.The results showed that TDI-1 was susceptible to E09 during the seedling stage but immune during the adult stage.F₁ plants derived from the cross of TDI-1 and TDU-1 were immune to E09 during the adult stage.The resistance of adult F₂individuals was separated,and the ratio of resistant and susceptible plants was 3:1($χ_{3:1}^{2}$=0.11,P=0.74);the ratio of the number of homozygous resistant,separated resistant,and homozygous susceptible F_2:3 lines was 1:2:1($χ_{1:2:1}^{2}$=0.47,P=0.79),indicating that the resistance to powdery mildew in the adult stage of TDI-1 was controlled by one dominant gene,temporarily named PmTDI-1.Subsequently,a set of molecular markers was used to amplify the parents and their F₂ population,and then four markers on chromosome 2A,including Xwmc407,NRM-2AS29,NRM-2AS45 and NRM-2AS84, confirmed to be linked to PmTDI-1. PmTDI-1 was between the flanking markers NRM-2AS45 and NRM-2AS84,with genetics distances of 1.8 cM and 4.6 cM,respectively.Therefore,the adult stage powdery mildew resistance gene PmTDI-1 was preliminarily localized on chromosome 2A.This study identified a novel dominant adult-plant-resistance powdery mildew gene PmTDI-1 from tetraploid wheat TDI-1.

Key words: Tetraploid wheat, Powdery mildew, Resistance gene, Bulked segregant analysis(BSA), Gene mapping

闫贵云, 古春霞, 王敏, 谭丹, 刘晓宇, 卢成达, 左静静. 四倍体小麦成株抗白粉病基因定位[J]. 华北农学报, 2024, 39(4): 187-193. doi: 10.7668/hbnxb.20195107.

YAN Guiyun, GU Chunxia, WANG Min, TAN Dan, LIU Xiaoyu, LU Chengda, ZUO Jingjing. Molecular Mapping of Adult Plant Resistance Gene to Powdery Mildew in Tetraploid Wheat[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(4): 187-193. doi: 10.7668/hbnxb.20195107.

http://www.hbnxb.net/CN/Y2024/V39/I4/187

图/表 8

表1 与PmTDI-1连锁的分子标记

Tab.1 Molecular markers linked to PmTDI-1

引物名称 Primer name	染色体位置/Mb Chromosome location	正向引物(5'—3') Forward sequence (5'—3')	反向引物(5'—3') Reverse sequence(5'—3')	退火温度/℃ Tm
Xwmc407	2A/7.81	GGTAATTCTAGGCTGACATATGCTC	CATATTTCCAAATCCCCAACTC	61
NRM-2AS29	2A/16.90	GGTCTGCGAGCGTTTACT	GCTTTCTTCCTTTCTGCC	58
NRM-2AS45	2A/10.60	GCTAATAACCCGCACTAAA	GCAAACTACCTTACAGAGCA	58
NRM-2AS84	2A/15.36	TGTTGGAGAGAAGGAAAGG	ACGGGAGATGAGGTGTAGT	58

图1 TDI-1和TDU-1的白粉病害反应型 A.TDI-1和TDU-1幼苗白粉病害反应型;B,C.TDI-1和TDU-1成株白粉病害反应型。

Fig.1 The powdery mildew infection types of TDI-1 and TDU-1 A.Infection types of Bgt isolate E09 on seedlings of TDI-1 and TDU-1;B,C.Infection type of Bgt isolate E09 on TDI-1 and TDU-1 adult plant,respectively.

表2 TDI-1和TDU-1植株苗期对白粉菌株E09的分级分析

Tab.2 Grading analysis of TDI-1 and TDU-1 plants to Bgt isolate E09 at seedling stage

亲本 Parents	株数 Number of plants				感病株数Susceptible plants
亲本 Parents	株数 Number of plants	0;	1	2	3	4
TDI-1	10	0	0	0	0	10
TDU-1	10	0	0	0	0	10

图2 TDI-1×TDU-1 F₂群体成株期对白粉病的抗性反应

Fig.2 Resistance reaction to powdery mildew of adult plants of TDI-1 × TDU-1 F₂ population

图3 TDI-1×TDU-1 F_2:3家系成株期对白粉病的抗性反应

Fig.3 Resistance reaction to powdery mildew of adult plants of TDI-1×TDU-1 F_2:3 lines

表3 TDI-1×TDU-1 遗传群体抗病性卡方分析

Tab.3 Chi square analysis of disease resistance in TDI-1×TDU-1 genetic populations

亲本/群体 Parents/ population	株/系数 Number of plants/lines	抗病株/系 Resistant plants/lines	杂合抗病株/系 Heterozygous plants/lines	感病株/系 Susceptible plants/lines	理论比值 Theoretical ratio	χ²	P
TDI-1	10	10	0	0
TDU-1	10	0	0	10
F₂	187	138	0	49	3:1	0.11	0.74
F_2:3	187	51	91	45	1:2:1	0.47	0.79

图4 PmTDI-1的遗传图谱和标记引物扩增电泳带型 M.Ladder Marker;PR.抗病亲本TDI-1;PS.感病亲本TDU-1;BR.抗病池;BS.感病池;R1—R5.纯合抗病单株;H1—H5.杂合抗病单株;S1—S5.纯合感病单株。

Fig.4 Genetic map and marker electrophoretic bands of PmTDI-1 M.Ladder Marker;PR.Resistant parent TDI-1;PS.Susceptible parent TDU-1;BR.Resistant bulk;BS.Susceptible bulk;R1—R5.Homozygous resistant individuals;H1—H5.Heterozygous resistant individuals;S1—S5.Homozygous susceptible individuals.

图5 PmTDI-1候选区间高置信注释基因

Fig.5 High confidence annotated genes in the candidate region of PmTDI-1

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