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水稻白叶枯病广谱抗性基因Xa7共显性功能标记的开发与应用

彭建 肖友伦 于江辉 张阳军 贾琳 唐小美 刘俊 周小平 余成 刘佳

彭建,肖友伦,于江辉,等. 水稻白叶枯病广谱抗性基因Xa7共显性功能标记的开发与应用 [J]. 福建农业学报,2023,38(4):444−453 doi: 10.19303/j.issn.1008-0384.2023.04.008
引用本文: 彭建,肖友伦,于江辉,等. 水稻白叶枯病广谱抗性基因Xa7共显性功能标记的开发与应用 [J]. 福建农业学报,2023,38(4):444−453 doi: 10.19303/j.issn.1008-0384.2023.04.008
PENG J, XIAO Y L, YU J H, et al. Codominance Functional Marker of Bacterial Blight Resistant Xa7 in Rice [J]. Fujian Journal of Agricultural Sciences,2023,38(4):444−453 doi: 10.19303/j.issn.1008-0384.2023.04.008
Citation: PENG J, XIAO Y L, YU J H, et al. Codominance Functional Marker of Bacterial Blight Resistant Xa7 in Rice [J]. Fujian Journal of Agricultural Sciences,2023,38(4):444−453 doi: 10.19303/j.issn.1008-0384.2023.04.008

水稻白叶枯病广谱抗性基因Xa7共显性功能标记的开发与应用

doi: 10.19303/j.issn.1008-0384.2023.04.008
基金项目: 常德市科技转化与推广服务项目(2019〔常财农〕0051-000)
详细信息
    作者简介:

    彭建(1980−),男,农艺师,主要从事水稻遗传育种研究(E-mai:haishipengjian@126.com);共同第一作者

    肖友伦(1980−),男,博士,副研究员,主要从事水稻分子育种研究(E-mai:732427358@qq.com

    通讯作者:

    于江辉(1986−),男,硕士,助理研究员,主要从事水稻分子育种研究(E-mail:yujianghui2022@163.com

    张阳军(1984−),男,博士,高级农艺师,主要从事水稻遗传育种研究(E-mail:zyj020116@126.com

  • 中图分类号: S511.032

Codominance Functional Marker of Bacterial Blight Resistant Xa7 in Rice

  • 摘要:   目的  对水稻白叶枯病广谱抗性基因Xa7进行精准检测和世代跟踪,通过分子标记检测Xa7基因材料的纯合或杂合型。  方法  根据Xa7xa7和无等位基因型序列的差异,通过Premier 5软件设计了含4条引物Xa7-F、Xa7-R、Xa7null-F、Xa7null-R的功能标记Xa7fun,且采用PCR方法分别对不同遗传资源材料进行分子标记特异性检测和验证,自然高温下于孕穗期对含Xa7基因的3份杂交改良系及亲本采用剪叶接种法接种7个白叶枯病菌菌株进行抗性鉴定,并于成熟期考察记录其农艺性状。  结果  分子标记特异性检测结果表明,Xa7基因纯合型材料R084可扩增出大小为91 bp的条带,无等位基因型材料Nip可扩增出大小为153 bp的条带,杂合体Nip/R084可扩增出大小为91 bp、153 bp的条带,共显性标记Xa7fun得到的电泳条带与引物设计时预测的目标片段完全吻合;18份不同类型的种质资源均未扩增出91 bp大小的功能条带,说明这些材料均不含Xa7基因;杂交改良系Ry-1、Ry-2、Ry-3均仅含有91 bp大小的功能条带,表明Xa7基因纯合;在高温下,华占对7个菌株表现为高感、中感或感病,R084除对菌株PX099感病外,对其余6菌株均为高抗、中抗或抗病,Ry-1对GDA2、HNA1-4、FuJ、GD1358、YN24等5个菌株为高抗或中抗,Ry-2对GDA2、GD1358、HNA1-4、PXO86、YN24等5个菌株为高抗或抗病,Ry-3对HNA1-4、FuJ、GDA2、GD1358、PXO86、YN24 等6个菌株均为高抗或中抗。因此,Xa7基因的渗入对华占改良系Ry-1、Ry-2、Ry-3的白叶枯病抗性有大幅度的提高;通过对3个改良系和亲本等农艺性状分析表明,3个改良系的生育期、株高、穗长、单株总粒数、结实率、千粒重介于R084和华占之间,Ry-1和Ry-3单株有效穗数显著高于华占和R084,Ry-2与华占、R084差异不显著,单株产量与华占或R084相比差异不显著。  结论  本研究开发的功能标记Xa7fun能够准确、高效地识别水稻Xa7基因纯合型、杂合型等,而且Xa7基因的渗入不会造成杂交改良系重要农艺性状变差,可在水稻白叶枯病抗性分子育种中推广应用。
  • 图  1  共显性功能标记Xa7fun引物设计策略

    Figure  1.  Design of primers for Xa7fun

    图  2  梯度PCR检测Xa7fun分子标记

    M为DNA marker(DL2000), 1~3分别为R084、 Nip、Nip/R084。

    Figure  2.  Detection of Xa7fun by gradient PCR

    M: DNA marker (DL2000); 1-3: R084, Nip, and Nip/R084, respectively.

    图  3  Xa7fun功能标记对部分水稻品种的分子检测

    A中,M:DNA marker(DL2000),+:R084,1~18对应水稻材料为长粒香、吉粳88、中花11、越光、绥粳4号、昆明小白谷、金23B、野香B、中浙2B、荃9311B、华占、农香39、五山丝苗、广恢390、R900、望恢006、黄华占、湘早籼45号。B中19~38为Y629的20个单株,C中39~58为Y192对应20个单株。

    Figure  3.  Molecular detection of Xa7fun in different rice varieties

    In A, M: DNA marker(DL 2000), 1–18: Changlixiang, Jijing 88, Zhonghua 11, Yueguang, Suijing 4, Kunmingxiaobaigu, Jin 23B, Yexiang B, Zhongzhe 2B, Quan 9311B, Huazhan, Nongxiang 39, Wushansimiao, Guanghui 390, R900, Wanghui 006, Huanghuazhan, and Xiangzaoxian 45, respectively (A); 19-38: 20 individual plants of Y629 (B); 39-58: 20 individual plants of Y192 (C).

    图  4  Xa7基因高世代杂交后代的选育和分子检测

    M:DNA marker(DL2000),+:R084。A为Ry-1、Ry-2、Ry-3选育图,B为连锁标记M5检测结果,C为共显性功能标记Xa7fun检测结果。

    Figure  4.  Breeding of filial generations and molecule detection of Xa7

    M:DNA marker(DL2000), +:R084. A: Breeding diagrams on Ry-1, Ry-2, and Ry-3; B: detection of linkage marker M5; C: detection of codominance functional marker Xa7fun.

    图  5  白叶枯病菌株诱发期间的温度

    Figure  5.  Temperature at time of bacterial blight pathogen induction

    表  1  Xa7fun标记引物序列

    Table  1.   Sequences of Xa7fun primers

    引物名称
    Primer name
    引物序列
    Primer sequence(5′-3′)
    碱基数
    Base number/bp
    Xa7-RGCTCTTCAAGTGTGCGATGC20
    Xa7-FAGCCCTGACTGCTAAAACCA20
    Xa7null-RGTACATCATTGTCCCCACGG20
    Xa7null-FAACCGAGGGAGGTGATTTGC20
    下载: 导出CSV

    表  2  Xa7基因高世代杂交后代对7种白叶枯病菌株的抗性

    Table  2.   The filial generations of Xa7 gene to 7 bacterial leaf blight strains

    品系 Lines FuJ GDA2 GD1358 HNA1-4 PXO86 PXO99 YN24
    病斑长度
    Lesion
    length/
    cm
    抗性
    Resistance
    病斑长度
    Lesion
    length/
    cm
    抗性
    Resistance
    病斑长度
    Lesion
    length/
    cm
    抗性
    Resistance
    病斑长度
    Lesion
    length/
    cm
    抗性
    Resistance
    病斑长度
    Lesion
    length/
    cm
    抗性
    Resistance
    病斑长度
    Lesion
    length/
    cm
    抗性
    Resistance
    病斑长度
    Lesion
    length/
    cm
    抗性
    Resistance
    Ry-1 3.7 MR 0.5 HR 5 MR 0.9 HR 10.2 MS 13.7 S 3.6 MR
    Ry-2 5.9 MS 0.4 HR 0.5 HR 0.5 HR 4.7 MR 16.3 S 4.1 MR
    Ry-3 3.5 MR 4.8 MR 4.5 MR 0.7 HR 4.9 MR 15.2 S 4.9 MR
    R084 2.1 R 0.2 HR 0.2 HR 0.3 HR 4.1 MR 15.3 S 1.7 R
    华占
    Huazhan
    24.6 HS 26.6 HS 12 MS 10.8 MS 8.9 MS 9.7 MS 19 S
    下载: 导出CSV

    表  3  Xa7基因对高世代杂交后代农艺性状的影响

    Table  3.   Effects of Xa7 on agronomic traits of filial generation hybrids

    品系
    Lines
    播始历期
    Days to flower/d
    株高
    Plant height/cm
    单株有效穗数
    Panicles per plant
    穗长
    Panicle length/cm
    单株总粒数
    Grains per plan
    结实率
    Seed set/%
    千粒重
    1000-grain weight/g
    单株产量
    Yield per plant/g
    Ry-173117.3±2.08 Bc15.5 ±1.00 Aa22.2±0.05 BCb1820.3±108.55 ABCbc90.7±0.28 Aa22.6±0.34 Bb37.3±2.70 Aa
    Ry-273119.3±2.52 ABbc11.3 ±0.58 Bb22.8±0.72 Bb2336.0±321.38 Aa89.1±0.79Aa21.5±0.28 Bb39.6±0.68 Aa
    Ry-377123.0±2.65 Aab15.3 ±0.50 Aa23.0±0.15 Bb1675.3±264.06 BCcd90.2±0.40Aa27.5±1.32 Aa41.8±6.95 Aa
    R08479124.7±1.53 Aa11.0 ±1.41 Bb21.2±0.54 Cc1312.3±128.39 Cd90.0±1.63Aa27.8±1.76 Aa38.4±1.58 Aa
    华占
    Huazhan
    7398.2±1.30 Cd13.0 ±2.45 ABb26.0±0.50 Aa2208.0±146.08 ABab84.2±1.05 Bb20.1±0.28 Bb39.7±1.37 Aa
    不同大、小写字母分别表示同一品种在不同氮素水平下差异极显著(P<0.01)或显著(P<0.05)。
    Data with different uppercase letters represent extremely significant differences (P<0.01); those with different lowercase letters, significant differences (P<0.05).
    下载: 导出CSV
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出版历程
  • 收稿日期:  2022-08-16
  • 修回日期:  2022-11-30
  • 网络出版日期:  2023-05-09
  • 刊出日期:  2023-04-28

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