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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (12): 1875-1881.doi: 10.3724/SP.J.1006.2018.01875

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Genetic Mapping of Bruchid Resistance Gene in Mungbean V1128

Chang-You LIU1,Qiu-Zhu SU1,Bao-Jie FAN1,Zhi-Min CAO1,Zhi-Xiao ZHANG1,Jing WU2,Xu-Zhen CHENG2,Jing TIAN1,*()   

  1. 1 Institute of Food and Oil Crops, Hebei Academy of Agricultural and Forestry Sciences / Hebei Laboratory of Crop Genetic and Breeding, Shijiazhuang 050031, Hebei, China
    2 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2018-05-08 Accepted:2018-08-20 Online:2018-12-12 Published:2018-09-18
  • Contact: Jing TIAN E-mail:nkytianjing@163.com
  • Supported by:
    This work was supported by the National Natural Science Foundation of China(31601367);the China Agriculture Research System(CARS-08);the Science and Technology Program of HebeiProvince (F18R494004-01).(16227508D);the Modern Agricultural Science and Technology Innovation Program of Hebei Province(F18R494004-01)

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Abstract:

It is an urgent research topic to map the bruchid resistance gene and to carry out bruchid resistance breeding using molecular marker-assisted method in mungbean. This study was carried out to identify a F2 isolated group formed by the hybrid of a bruchid-resistant cultivar “V1128” and a bruchid-susceptible cultivar “Jilyu 7”, and to analyze the genetic regularity of V1128 in resistance to bruchids. The bulked segregant analysis (BSA) method was used for screening the polymorphic markers. Genetic linkage map construction and quantitative trait locus (QTL) mapping were conducted using software QTL IciMapping 4.0. The results showed that the bruchid resistance of V1128 was controlled by a dominant gene with main effect. According to previous naming rules, the bruchid resistance gene of V1128 was temporarily named as “Br3”. When we treated bruchid-resistance as a quality trait, Br3 was used as a marker for linkage map construction and was positioned between the markers DMB158 and VRBR-SSR033 (VRID5, VRBR-SSR032, and VRBR-SSR033 are located at the same map position). The genetic distances of Br3 away from the two markers were 4.4 cM and 5.8 cM, respectively. Br3 was positioned on chromosome 5 in the physical range of about 288 kb. By using the inclusive composite interval mapping (ICIM) to locate the seed damage rate, a main QTL locus with LOD score of 38.04 was identified in the marker intervals from DMB158 to VRBR-SSR033, contributing 71.64% of the observed phenotypic variation. The allele of male parent V1128 had a significant effect on reducing the rate of seed damage. The results can provide useful information for the molecular marker-assisted breeding of mungbean, and the fine localization and cloning of Br3.

Key words: mungbean, bruchid resistance, V1128, Br3, QTL

Table 1

Information of polymorphic DNA markers used in this study"

标记名称
Marker		 标记类型
Type		 引物序列
Primer sequence (5′-3′)		 退火温度
Tm		 文献来源
Reference
Mchr5-17 SSR F: GCTTGCTTATGCTCAAAACT; R: TACAGATAAACCCAAGCCAT 55 [23]
Mchr5-20 SSR F: TCAAAACTTTCACTGGACCT; R: GCTGTTTGTCACATGCATAA 55 [23]
Mchr5-23 SSR F: ATCTTAATCCCATCCTTGGT; R: AACTGGCTTGTAAGGTGAGA 55 [23]
Mchr5-25 SSR F: CCGTTGTGAATCAACTTTTC; R: GAGTCGTCGTGTAATCCTTC 55 [23]
Mchr5-32 SSR F: ACTTGTAGGTGGAAGAGATGA; R: GATTAAGGGCGTGTTTTGT 55 [23]
Mchr5-33 SSR F: CTAATGAAACAGGACAAGGG; R: CTCACTCTTCTCATTCCACC 55 [23]
Mchr5-36 SSR F: ACCTACTGATTGGTGTTTGG; R: CAGTGAATGCTGACAGTGAC 55 [23]
Mchr5-37 SSR F: AACGGTTGGAGTTAGGAAAT; R: TGAACATCACCAACTATCAC 55 [23]
Mchr3-85 SSR F: AACAGCGTTGATTTATGGAC; R: AATCATGTTGGTGTGTTGTG 55 [23]
Mchr3-86 SSR F: TGCCTAAGGGTCAATTTCTA; R: ATGCACCAGAAGACAAAAAC 55 [23]
Mchr3-87 SSR F: AATGAGGTAATGCAGAGGTG; R: GACAAGGGTTGTTGTTCACT 55 [23]
Mchr3-89 SSR F: GAATTAAAGCCCTTGTTCTG; R: GGTGAATTTTCTGTTTCCAC 55 [23]
Mchr3-96 SSR F: CTGATGCTATTTCCATCCAT; R: TAACCTTTTGCATTTGGTGC 55 [23]
MUS150 SSR F: GCTGTTTGTCACATGCATAA; R: TCAAAACTTTCACTGGACCT 55 [23]
MUS365 SSR F: TGAGCCCGATTTTTATCTC;R: GCTCACAGATAACTGACACAAC 55 [23]
MUS569 SSR F: GGAGGGGATTTTTAAGATTG; R: GGATACGATTTTGTCGTGTT 55 [23]
HAAS_VR_379 SSR F: CCTATCCGAATCGACACCAC; R: GTAGCAATAGCAGCCCAAGG 55 [22]
HAAS_VR_106 SSR F: ACGGCTATTCATCGTTTTGC; R: CAACCCGAAGCCAAAAACTA 55 [22]
HAAS_VR_89 SSR F: GCTGGAAGGATCCAATTTCA; R: TCGCCATTCCCAAGATAAAG 55 [22]
HAAS_VR_1701 SSR F: CCGGGGTGAAATTGATACAC; R: CAAAGGGGCTATGAACAGGA 55 [22]
VRID1 Indel F: TCGGTTTCAGCTCGATAGATTC; R: GATGTTGTCTGAAGTAGTGGTA 55 [21]
VRID5 Indel F: AGAATAAAATGAATCTAGAAGACCA; R: TGAATTAATTTTCTTACCCTTGT 50 [21]
VRBR-SSR016 SSR F: GACGGCTAGGTACAACACTGC; R: TTTAGAGCAATTGGGTGGATTT 55 [20]
VRBR-SSR024 SSR F: TTTTGTGGACACTCCTTCCA; R: AAGCGTCACACCCTCAATTC 55 [20]
VRBR-SSR030 SSR F: CAGCTAGGAAACTCACCAAACC; R: CAGCTGGCAGGTGAAATATG 55 [20]
VRBR-SSR032 SSR F: GGTTATTTTGATCTAAAGGGCCA; R: GTGTGAGAAGATTTGGGAATGTAA 55 [20]
VRBR-SSR033 SSR F: CTCAAGTCTTATGTTTCCCCCTAT; R: GCACTAAAGGACTTTCCTTGAAC 55 [20]
VRBR-SSR039 SSR F: AAGTTGGTGTAGCACTTGCAGA; R: AATGAATTAAAAGAAAACACTACTG 55 [20]
GBssr-MB87 SSR F: TCCCTTGTGGGAGATCCT; R: CTTTGCCACACTCCTTGC 55 [24]
DMB158 SSR F: TGGAAAATTTGCAGCAGTTG; R: ATTGATGGAGGGCGGAAGTA 55 [25]
PVBR201 SSR F: GTGATGGTGCTGCTTTTCAA; R: ATGCGTGGGGAGAAGTAAGA 50 [26]

Fig. 1

Distribution of seed damage rate in F2 population"

Fig. 2

Positioning analysis of the bruchids resistance gene Br3 Linkage map constructed by using QTL IciMapping 4.0 software, LOD=5; map distances (cM) and marker names are shown on the left and right sides of the linkage group, respectively."

Fig. 3

Location of QTL qBr3 controlling resistance to bruchids QTL mapping based on inclusive composite interval mapping method (ICIM) using QTL IciMapping 4.0 software; map distances (cM) and marker names are shown on the left and right sides of the linkage group, respectively; the vertical line shows the LOD threshold (3.18)."

Fig. 4

Collinearity comparison analysis of QTL mapping results A synteny map between the genetic linkage map from this study and the linkage map constructed by Kaewwongwal et al. [21], this map was drown using MapChart 2.2 software [30] based on seven common markers; the scale bars on the left of the figure indicate the length of each linkage group in cM; “LG1” is the linkage group constructed in this study, “LG2” was constructed by Kaewwongwal et al.; the names of QTLs are shown in italics."

Table 2

Genes annotation in the range of bruchids resistance site of V1128"

基因序号
Gene ID		 基因位置
Gene location		 基因注释
Gene annotation
LOC106760111 5 338 753…5 340 435 RD22类蛋白 RD22-like protein
LOC106761406 5 346 646…5 349 613 未知蛋白 Uncharacterized protein
LOC106760112 5 361 741…5 362 985 未知蛋白 Uncharacterized protein
LOC106761824 5 367 108…5 367 592 RD22类蛋白 RD22-like protein
LOC106761559 5 389 064…5 390 972 RD22类蛋白 RD22-like protein
LOC111241568 5 403 161…5 404 109 未知蛋白 Uncharacterized protein
LOC106760880 5 409 223…5 413 311 RD22类蛋白 RD22-like protein
LOC106761219 5 511 814…5 514 459 RD22类蛋白 RD22-like protein
LOC106760236 5 561 973…5 563 546 多聚半乳糖醛酸酶抑制剂类蛋白(VrPGIP1) Polygalacturonase inhibitor-like (VrPGIP1)
LOC106760237 5 590 847…5 591 984 多聚半乳糖醛酸酶抑制剂类蛋白(VrPGIP2) Polygalacturonase inhibitor-like (VrPGIP2)
LOC106760238 5 590 527…5 598 757 假基因 Pseudogene
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