基于新遗传连锁图谱的豇豆抗豆象QTL定位

doi:10.3864/j.issn.0578-1752.2021.22.003

中国农业科学 ›› 2021, Vol. 54 ›› Issue (22): 4740-4749.doi: 10.3864/j.issn.0578-1752.2021.22.003

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

基于新遗传连锁图谱的豇豆抗豆象QTL定位

王彦¹(),范保杰¹,曹志敏¹,张志肖¹,苏秋竹¹,王珅¹,王学清²,彭秀国³,梅丽⁴,武玉华¹,刘少兴¹,田胜民¹,徐俊杰¹,蒋春志¹,王伟娟⁵,刘长友¹(),田静¹()

¹河北省农林科学院粮油作物研究所/河北省作物遗传育种实验室,石家庄 005035
²河北省农林科学院,石家庄 005031
³石家庄市植物园,石家庄 050000
⁴北京市农业技术推广站,北京 100029
⁵张家口市崇礼区植保站,河北张家口 076350

收稿日期:2021-05-06 接受日期:2021-07-02 出版日期:2021-11-16 发布日期:2021-11-19
通讯作者: 刘长友,田静
作者简介:王彦,Tel：0311-87670607;E-mail： 411678486@qq.com。
基金资助:
国家重点研发计划(2019YFD1000700);国家重点研发计划(2019YFD1000703);财政部和农业农村部：国家现代农业产业技术体系(CARS-08);河北省重点研发计划(19226353D);河北省现代农业产业技术体系(HBCT2018070203);河北省青年拔尖人才(2018);河北省农林科学院创新工程项目(2019-4-02-08)

Quantitative Trait Locus Mapping of Bruchids Resistance Based on A Novel Genetic Linkage Map in Cowpea (Vigna unguiculata)

WANG Yan¹(),FAN BaoJie¹,CAO ZhiMin¹,ZHANG ZhiXiao¹,SU QiuZhu¹,WANG Shen¹,WANG XueQing²,PENG XiuGuo³,MEI Li⁴,WU YuHua¹,LIU ShaoXing¹,TIAN ShengMin¹,XU JunJie¹,JIANG ChunZhi¹,WANG WeiJuan⁵,LIU ChangYou¹(),TIAN Jing¹()

¹Institute of Cereal and Oil Crops, Hebei Academy of Agricultural and Forestry Sciences/Hebei Laboratory of Crop Genetic and Breeding, Shijiazhuang 050035
²Hebei Academy of Agricultural and Forestry Sciences, Shijiazhuang 050031
³Shijiazhuang Botanical Garden, Shijiazhuang 050000
⁴Beijing Agricultural Technology Extension Centre, Beijing 100029
⁵Plant Protection Department of Chongli District, Zhangjiakou 076350, Hebei

Received:2021-05-06 Accepted:2021-07-02 Online:2021-11-16 Published:2021-11-19
Contact: ChangYou LIU,Jing TIAN

摘要/Abstract

摘要：

【目的】豆象是危害豇豆最主要的仓储害虫。发掘豇豆抗豆象基因为抗性品种的选育,以及减少豆象对豇豆生产的危害奠定基础。【方法】以中豇1号（感）和Pant-lobia-1（抗）为亲本构建的包含282个株系的RIL群体为研究材料,利用人工接种法分别对282个株系接种绿豆象和四纹豆象,进行抗豆象表型鉴定,并利用两亲本对3 992个来源于绿豆、小豆和豇豆的SSR标记进行多态性筛选,然后利用筛选到的多态性标记对282个株系进行基因型分析,最后结合RIL群体各株系抗豆象表型鉴定数据和基因型分型数据,采用完备区间作图法（ICIM-ADD）进行抗豆象QTL定位分析,在此基础上构建遗传连锁图谱,并定位豇豆抗豆象基因。【结果】中豇1号和F₁籽粒的被害率均为100%,Pant-lobia-1籽粒的被害率分别为22.5%和42.5%。推测Pant-lobia-1对绿豆象和四纹豆象的抗性均为隐性遗传;筛选到182个多态性标记,利用这些多态性标记构建了一个包含11个连锁群的豇豆遗传连锁图谱,图谱总长1 065.23 cM,相邻标记间平均遗传距离为5.85 cM;经2种豆象处理,分别在连锁群1和连锁群5上检测到2个稳定的QTL位点,暂定名为vubr1-1和vubr5-1,其中vubr1-1位于标记XD11-44和HAAS_VR_2274之间,标记间的遗传距离为7.6 cM,在2种豆象处理中分别可以解释表型变异的7.16%和6.92%;vubr5-1位于标记XD1-14和CP185之间,标记间的遗传距离为2.90 cM,在2种豆象处理中分别可以解释表型变异的6.96%和6.37%。【结论】构建了一个包含11个连锁群、182个多态性标记的豇豆遗传连锁图谱,检测到2个与抗豆象相关的QTL位点。

关键词: 豇豆, SSR, 抗豆象, 遗传图谱, QTL定位

Abstract:

【Objective】Bruchids are the main storage pests of cowpea. The discovery of bruchids resistance genes is helpful for breeding resistant varieties and reducing the harm of bruchids to cowpea production.【Method】In this study, a RIL population of 282 lines derived from the cross between Zhongjiang No.1 (a bruchid-susceptible cultivar) and Pant-lobia-1 (a bruchid-resistant cultivar) was used for phenotype identification of bruchids resistance by artificial inoculation of Callosobruchus chinensis and Callosobruchus maculates. The two parents were used to screen polymorphic markers from 3 992 SSR markers of mung bean, adzuki bean and cowpea. Genotypes of 282 lines were analyzed using the polymorphic SSR markers. Based on the phenotype identification and genotype analysis, a genetic linkage map was constructed and the bruchids resistance gene(s) of cowpea was located using Inclusive Composite Interval Mapping (ICIM-ADD). 【Result】The results showed that Zhongjiang No. 1 and F₁ seeds were 100% susceptible to Callosobruchus chinensis and Callosobruchus maculates, and the damage rates of Pant-lobia-1 were 22.5% and 42.5%, respectively. It was speculated that the resistance of Pant-lobia-1 to bruchids was recessive inheritance. 182 polymorphic markers were obtained from 3 992 SSR markers of mung bean, adzuki bean and cowpea. Using those polymorphic SSR markers, a genetic linkage map with 11 linkage groups was constructed. The map covered a total length of 1 065.23 cM with an average interval of 5.85 cM between adjacent markers. Two bruchid-resistant QTLs from linkage groups 1 and 5 were discovered, which were temporarily named as vubr1-1 and vubr5-1. QTL vuvr1-1 was located between markers XD11-44 and HAAS_VR_2274, which genetic distance was 7.6 cM, explaining 7.16% and 6.92% of the phenotypic variation in the two bruchid-resistant tests respectively. QTL vubr5-1 was located between markers XD1-14 and CP185, which genetic distance was 2.9 cM, explaining 6.96% and 6.37% of the phenotypic variation in the two bruchid-resistant tests respectively. 【Conclusion】A genetic linkage map containing 11 linkage groups and 182 polymorphic markers was constructed. Two QTLs linked with bruchids resistance were identified on linkage groups 1 and 5.

Key words: Vigna unguiculata, SSR, bruchid resistance, genetic mapping, quantitative trait locus

王彦,范保杰,曹志敏,张志肖,苏秋竹,王珅,王学清,彭秀国,梅丽,武玉华,刘少兴,田胜民,徐俊杰,蒋春志,王伟娟,刘长友,田静. 基于新遗传连锁图谱的豇豆抗豆象QTL定位[J]. 中国农业科学, 2021, 54(22): 4740-4749.

WANG Yan,FAN BaoJie,CAO ZhiMin,ZHANG ZhiXiao,SU QiuZhu,WANG Shen,WANG XueQing,PENG XiuGuo,MEI Li,WU YuHua,LIU ShaoXing,TIAN ShengMin,XU JunJie,JIANG ChunZhi,WANG WeiJuan,LIU ChangYou,TIAN Jing. Quantitative Trait Locus Mapping of Bruchids Resistance Based on A Novel Genetic Linkage Map in Cowpea (Vigna unguiculata)[J]. Scientia Agricultura Sinica, 2021, 54(22): 4740-4749.

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参考文献 35

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品种名称 Name	处理Ⅰ被害率 Seed damage rate (Ⅰ) (%)	处理Ⅱ被害率 Seed damage rate (Ⅱ) (%)
中豇1号 Zhongjiang No.1	100.0	100.0
Pant-lobia-1	22.5**	42.5**
F₁	100.0	100.0

物种 Species	标记数 No. of marker	有效扩增数（个/比率） No. of effective amplified	多态性标记数（个/比率） No. of polymorphic marker	标记来源 Source of marker
绿豆Mung bean	1972	1606(81.44%)	62(3.86%)	转录组Transcriptome
小豆Adzuki bean	1100	717(65.18%)	18(2.51%)	基因组Genome
豇豆Cowpea	920	862(93.70%)	102(11.83%)	基因组、CGKB Genome,CGKB
合计Total	3992	3185(80.11%)	182(6.07%)

连锁群 Linkage group	标记数 No. of marker	连锁群长 Length (cM)	平均距离 Average interval (cM)	偏分离标记数 No. of distorted marker
LG1	26	185.91	7.15	0
LG2	28	133.21	4.76	16
LG3	20	110.87	5.54	8
LG4	18	127.32	7.07	11
LG5	18	109.25	6.07	0
LG6	13	79.74	6.13	2
LG7	16	86.65	5.42	3
LG8	4	29.71	7.43	0
LG9	9	51.43	5.71	0
LG10	14	84.79	6.06	4
LG11	16	66.35	4.15	1
总计Total	182	1065.23	5.58	45

性状 Trait	连锁群 Linkage group	位置 Position (cM)	标记区间 Marker interval	LOD	贡献率 PVE(%)	加性效应 Add	LOD阈值 Threshold^a
CP-Br-C	LG5	57	XD1-14—CP185	4.58	6.96	-5.30	2.69
CP-Br-C	LG1	146	XD11-44—HAAS_VR_2274	3.86	7.16	-4.20	2.69
CP-Br-M	LG1	146	XD11-44—HAAS_VR_2274	3.91	6.92	-4.29	2.68
CP-Br-M	LG5	57	XD1-14—CP185	4.44	6.37	-3.70	2.68

基于新遗传连锁图谱的豇豆抗豆象QTL定位

Quantitative Trait Locus Mapping of Bruchids Resistance Based on A Novel Genetic Linkage Map in Cowpea (Vigna unguiculata)

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