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作物学报 ›› 2022, Vol. 48 ›› Issue (4): 791-800.doi: 10.3724/SP.J.1006.2022.14062

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

大豆突变体ygl2黄绿叶基因的精细定位

王好让1(), 张勇2, 于春淼2, 董全中, 李微微1,3, 胡凯凤, 张明明2, 薛红, 杨梦平2, 宋继玲, 王磊2, 杨兴勇, 邱丽娟2,*()   

  1. 1中国农业科学院作物科学研究所, 北京 100081
    2黑龙江省农业科学院克山分院, 黑龙江齐齐哈尔 161606
    3东北农业大学农学院, 黑龙江哈尔滨 150030
  • 收稿日期:2021-04-16 接受日期:2021-07-12 出版日期:2022-04-12 网络出版日期:2021-08-06
  • 通讯作者: 邱丽娟
  • 作者简介:王好让, E-mail: wanghaorang1018@163.com第一联系人：**同等贡献
  • 基金资助:
    国家自然科学基金项目(31630056);中央级公益性科研院所基本科研业务费专项资助(S2021ZD02)

Fine mapping of yellow-green leaf gene (ygl2) in soybean (Glycine max L.)

WANG Hao-Rang1(), ZHANG Yong2, YU Chun-Miao2, DONG Quan-Zhong, LI Wei-Wei1,3, HU Kai-Feng, ZHANG Ming-Ming2, XUE Hong, YANG Meng-Ping2, SONG Ji-Ling, WANG Lei2, YANG Xing-Yong, QIU Li-Juan2,*()   

  1. 1Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
    2Keshan Branch of Heilongjiang Academy of Agricultural Sciences, Qiqihar 161606, Heilongjiang, China
    3College of Agriculture, Northeast Agricultural University, Harbin 150030, Heilongjiang, China
  • Received:2021-04-16 Accepted:2021-07-12 Published:2022-04-12 Published online:2021-08-06
  • Contact: QIU Li-Juan
  • About author:First author contact:**Contributed equally to this work
  • Supported by:
    National Natural Science Foundation of China(31630056);Central Public-intreest Scientific Institution Basal Research Fund(S2021ZD02)

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摘要：

叶片是大豆进行光合碳同化的主要器官, 其颜色与光能的捕获力和转化效率有关, 也与大豆的产量密切相关。因此, 大豆叶色相关基因的挖掘对从光合碳同化途径解析大豆产量问题具有重要意义。黄绿叶是区别于大豆普通绿色叶片的突变类型, 是研究大豆叶色相关基因的重要遗传材料。本研究发现了一个黄绿叶突变体ygl2 (yellow-green leaf 2), 该突变体是由大豆品系GL11自然突变而来, 其黄绿叶表型可以稳定遗传。与绿叶野生型GL11相比较, 突变体ygl2叶片中叶绿素含量极显著降低, 株高、百粒重、蛋白含量均存在显著差异。利用GL11和ygl2构建分离群体, 遗传分析表明, ygl2的黄绿叶表型受1对隐性核基因控制, 利用分离群体将黄绿叶基因ygl2定位于2号染色体末端SSR标记02_104到02_107之间, 区间物理距离为56.1 kb, 包含9个基因。本研究结果为大豆黄绿叶基因图位克隆及分子标记辅助育种奠定了基础。

关键词: 大豆, 黄绿叶突变体, 遗传分析, 精细定位

Abstract:

Leaf is the main organ of photosynthetic carbon assimilation in soybean, and its color is not only related to the trapping power and conversion efficiency of light energy, but also closely related to the yield of soybean. Therefore, the mining of soybean leaf color-related genes is of great significance to analyze the yield of soybean from the pathway of photosynthetic carbon assimilation. Yellow-green leaf is a mutation type different from common green leaves of soybean, and it is an important genetic material to explore the genes related to leaf color of soybean. In this study, we found a yellow-green leaf mutant ygl2 (yellow-green leaf 2), which was naturally mutated from soybean strain GL11, and its yellow-green leaf phenotype could be stably inherited. Compared with the green leaf wild type GL11, the leaf chlorophyll content of mutant ygl2 decreased significantly, and there were significant differences in plant height, 100-grain weight, and protein content. The segregated population was constructed by GL11 and ygl2. Genetic analysis showed that the yellow-green leaf phenotype of ygl2 was controlled by a pair of recessive nuclear genes. The yellow-green leaf gene ygl2 was located between SSR markers 02_104 and 02_107 at the end of chromosome 2 using the isolated population, with an interval physical distance of 56.1 kb, and contained nine genes. These results laid a solid foundation for map-based cloning and molecular marker-assisted breeding of yellow and green leaf genes in soybean.

Key words: soybean, yellow-green leaf mutant, genetic analysis, fine mapping

图1

突变体ygl2与野生型GL11表型 A: 突变体ygl2和野生型GL11苗期植株表型; B: 突变体ygl2和野生型GL11鼓粒期叶片表型。标尺为4 cm。"

图2

突变体ygl2与野生型GL11的主要农艺性状 误差线表示标准差; *和**分别表示在0.05、0.01水平显著差异。"

图3

苗期突变体ygl2与野生型GL11叶片中光合色素含量 野生型GL11和突变体ygl2在第4个三小叶展开时(V4期)叶片中光合色素含量及比值。误差线表示标准差; *和**分别表示在0.05、0.01水平显著差异。"

表1

突变体ygl2与正常绿色品种杂交F2的叶色分离"

组合名称
Combination		 总株数
Total number of plants		 绿叶植株数
No. of green leaf plants		 黄绿叶植株数
No. of yellow-green leaf plants		 期望比
Expected ratio		 卡方值
χ2 3:1		 P
ygl2×GL11 567 412 155 3:1 1.529 0.199

图4

利用ED关联方法鉴定黄绿叶基因候选区间 横坐标为染色体名称, 彩色的点代表每个SNP位点的ED值, 黑色的线为拟合后的ED值, 红色的虚线代表显著性关联阈值。"

表2

精细定位所用分子标记"

引物名称
Primer name		 正向引物
Forward primer (5°-3°)		 反向引物
Reverse primer (5°-3°)
02_55 GTGTTCCACTCCACGTTTCC CATTTCCCCTTTCACAATCG
02_81 AACCGAGTTTGGTTCGATTC TGCTGCTTGATGATGAGGAC
02_90 CCATCTTATGGACTTGTTTGGA GCCAAGAATGACCATTATGC
02_101 TCACTAATCACAACAACCCAAA CGACCGGTGTGTTTAAGGTC
02_103 TCAGTCGCAGATTGATCAGG CCCAATTGTATCCATCAACG
02_104 AACCTAGCATTGCAACCTGC TCATCACCCCTTATCCGTTC
02_107 AAAACGAGGCCTTAATCGAAA AAACCAAAGAATACCGTGAAAAA
02_110 CGAAATGCCACCTTTTCAAT AGCAAACTAAGGTCGTTTTCG
02_117 GCAGTTGTGCGTGGGAGAGAG GCGACATAGCTAATTAAGTAAGTT
02_122 GCGTGGTGCACGATCATATAGA GCGTCTCCTTCGCTATCTCAAAC
02_125 CCAGGAATGCAGGTTTCTCT CGTGACTCTTCTTCCTTTCCA
02_130 AATGGAGAGGGGAACACAACT CCTAACGCACGAAATTTTCTC
ID2192 GCCTAATTTTGGCACCTTCA CACTCCTCTGCTTTGTTTGCT

图5

ygl2基因在2号染色体上的精细定位 A: ygl2所在区域的6个交换单株(2001、2006、2027、2144、CJ50和1054)的基因型, 根据其后代的表型确认了这些交换单株基因型; B: ygl2定位区间内共9个基因。"

表3

定位区间内的编码基因及其推测功能"

基因名称
Gene name		 推测功能
Putative function
Glyma.02G304600 未知 Unknown
Glyma.02G304700 铁氧还蛋白氧化还原酶 Ferredoxin oxidoreductas
Glyma.02G304800 未知 Unknown
Glyma.02G304900 F-box相关 F-box-like
Glyma.02G305000 DNAj同源亚家族c成员 DNAj homolog subfamily c member
Glyma.02G305100 非特异性蛋白酪氨酸激酶/胞浆蛋白酪氨酸激酶/双特异性激酶
Non-specific protein-tyrosine kinase/cytoplasmic protein tyrosine kinase/dual-specificity kinase
Glyma.02G305200 核孔复合体蛋白Nup133 (NUP133) Nuclear pore complex protein Nup133 (NUP133)
Glyma.02G305300 钙转运ATP酶/钙转运p型 Calcium-transporting ATPase/calcium-translocating p-type
Glyma.02G305400 叶绿素a/b结合蛋白 Chlorophyll a/b binding protein

图6

2个候选基因表达分析 SAM: 茎顶端分生组织。误差线表示标准差; *和**分别表示在0.05、0.01水平显著差异。"

图7

9个候选基因的表达谱 数据来自Phytozome v12.1。"

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