Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (22): 4740-4749.doi: 10.3864/j.issn.0578-1752.2021.22.003

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

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()   

  1. 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 E-mail:411678486@qq.com;35931915@qq.com;nkytianjing@163.com

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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

Fig. 1

Seed of two parents and F1 plant"

Table 1

The seed damage rate of two parents and F1 plant"

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

Fig. 2

Frequency distributions of seed damage rate within the RIL population ♀: Zhongjiang No.1; ♂: Pant-lobia-1. The same as below"

Table 2

Analysis of polymorphic SSR markers used in this study"

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%)

Fig. 3

SSR profiles generated by marker JD6-28 in a part of RIL individuals of cowpea M: Marker"

Table 3

Markers of linkage groups and their distribution"

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

Fig. 4

A genetic linkage map of cowpea *, **, and ***: Indicated markers exhibiting significant deviation from the expected segregation ratio at the 0.05, 0.01 and 0.001 levels, respectively"

Table 4

QTL mapping of bruchids resistance in cowpea RIL population"

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 Thresholda
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

Fig. 5

QTL mapping of bruchids resistance in cowpea A: Location map of QTL vubr1-1; B: Location map of QTL vubr5-1"

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