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作物学报 ›› 2018, Vol. 44 ›› Issue (12): 1875-1881.doi: 10.3724/SP.J.1006.2018.01875

• 研究简报 • 上一篇    下一篇

栽培绿豆V1128抗豆象基因定位

刘长友1,苏秋竹1,范保杰1,曹志敏1,张志肖1,武晶2,程须珍2,田静1,*()   

  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, 河北石家庄 050031
    2 中国农业科学院作物科学研究所, 北京 100081
  • 收稿日期:2018-05-08 接受日期:2018-08-20 出版日期:2018-12-12 网络出版日期:2018-09-18
  • 通讯作者: 田静
  • 基金资助:
    本研究由国家自然科学基金项目(31601367);国家现代农业产业技术体系建设专项(CARS-08);河北省科技计划项目(16227508D);河北省现代农业科技创新工程项目资助(F18R494004-01)

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 Published:2018-12-12 Published online:2018-09-18
  • Contact: Jing TIAN
  • 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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摘要：

对抗豆象资源中蕴藏的抗豆象基因进行定位, 是对其充分利用的前提和基础。本研究通过对抗豆象栽培绿豆V1128和感豆象栽培绿豆冀绿7号杂交形成的F2分离群体进行抗豆象鉴定, 分析V1128抗豆象遗传规律; 并利用混合群体分离分析法(BSA法)筛选抗感池间的多态性标记, 进而利用QTL IciMapping 4.0对V1128抗豆象基因进行染色体定位分析。结果表明, V1128对绿豆象的抗性由具有主效作用的显性单基因控制, 暂命名其为“Br3”。在将抗豆象性状作为质量性状的条件下, 按照显性单基因的定位方法, 将抗豆象基因Br3定位在绿豆染色体5上, 位于标记DMB158和VRBR-SSR033 (标记VRID5、VRBR-SSR032与VRBR-SSR033的连锁群位置相同)之间, 两侧遗传距离分别为4.4 cM和5.8 cM, 所在物理区间约288 kb。将抗豆象性状作为数量性状, 采用完备区间作图法(ICIM)对种子被害率进行QTL定位, 同样在标记DMB158和VRBR-SSR033之间检测到1个主效QTL, 其LOD值为38.04, 可以解释表型变异(PVE)的71.64%, 来自父本V1128的等位基因具有明显减少种子被害率的效应。该研究结果可以为绿豆抗豆象分子标记辅助育种及抗豆象基因Br3的精细定位和克隆提供有用信息。

关键词: 绿豆, 抗豆象, V1128, Br3, QTL

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

表1

多态性标记信息"

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]

图1

F2群体种子被害率分布图"

图2

抗豆象基因Br3连锁定位分析 利用QTL IciMapping 4.0软件进行标记连锁分析, LOD=5; 连锁群左右两侧分别标示图距(单位为cM)和标记名称。"

图3

抗豆象QTL qBr3的定位图谱 利用QTL IciMapping 4.0软件的完备区间作图法(ICIM)进行QTL 定位分析; 连锁群左右两侧分别标示图距(单位为cM)和标记名称; 竖线标示LOD阈值(3.18)。"

图4

QTL定位结果共线性比较分析 基于7个共有标记, 利用MapChart 2.2软件[30]作本研究与Kaewwongwal等[21]构建的连锁图谱的共线性图谱; 左侧标尺标示连锁群长度, 单位为cM; “LG1”为本研究构建的连锁图谱, “LG2”为Kaewwongwal等构建的连锁图谱; QTL名字用斜体字标示。"

表2

V1128抗豆象基因位点范围内基因注释"

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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