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作物学报 ›› 2021, Vol. 47 ›› Issue (11): 2099-2110.doi: 10.3724/SP.J.1006.2021.04245

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

甘蓝型油菜容重及其相关性状的全基因组关联分析

雷维1,2(), 王瑞莉1, 王刘艳1, 袁芳1,2, 孟丽姣1,2, 邢明礼1,2, 徐璐1,2, 唐章林1,2, 李加纳1,2, 崔翠1,*(), 周清元1,2,*()   

  1. 1西南大学农学与生物科技学院, 重庆 400715
    2重庆市油菜工程技术研究中心, 重庆 400715
  • 收稿日期:2020-10-17 接受日期:2021-03-19 出版日期:2021-11-12 网络出版日期:2021-04-01
  • 通讯作者: 崔翠,周清元
  • 作者简介:E-mail: 1305600171@qq.com
  • 基金资助:
    国家重点研发计划项目(2018YFD0100500);国家现代农业产业技术体系建设专项(CARS-12);重庆市技术创新与应用发展项目(cstc2019jscx-msxmX0383)

Genome-wide association study of seed density and its related traits in Brassica napus L.

LEI Wei1,2(), WANG Rui-Li1, WANG Liu-Yan1, YUAN Fang1,2, MENG Li-Jiao1,2, XING Ming-Li1,2, XU Lu1,2, TANG Zhang-Lin1,2, LI Jia-Na1,2, CUI Cui1,*(), ZHOU Qing-Yuan1,2,*()   

  1. 1College of Agronomy and Biotechnology, Southwest University, Chongqing 400715, China
    2Chongqing rape Engineering Technology Research Center, Chongqing 400715, China
  • Received:2020-10-17 Accepted:2021-03-19 Published:2021-11-12 Published online:2021-04-01
  • Contact: CUI Cui,ZHOU Qing-Yuan
  • Supported by:
    National Key Research and Development Program of China(2018YFD0100500);China Agriculture Research System(CARS-12);Chongqing Technology Innovation and Application Development Project(cstc2019jscx-msxmX0383)

摘要:

种子容重大小反映了作物光合产物在籽粒中的积累特性, 是油菜千粒重重要的组成部分, 筛选高容重种质资源, 研究容重的遗传特性在油菜遗传育种中具有非常重要的作用。本文以不同遗传背景的187份甘蓝型油菜品种(系)构成的自然群体为研究对象, 进行2年种子的容重及其相关性状(千粒重、体积)测定和资源评价, 基于最优模型对各性状进行全基因组关联分析(genome-wide association analysis, GWAS)和候选基因预测。结果显示, 187份材料在2年中容重及其关联性状在品种(系)间差异均达到显著水平(P<0.05), 筛选出3个种子千粒重较大的高容重种质资源。全基因组关联分析共检测到24个与种子容重及其相关性状显著关联的SNP位点, 可解释表型变异的8.21%~10.40%。通过单倍型分析确定关联SNP位点的Block区间, 其所在的Block覆盖了12个与容重、粒重和体积有关的候选基因, 主要编码转录因子(如WOX8、HAIKU1、AP2/ERF转录因子、Dof家族-Zinc finger超家族和BZR1转录因子)、酶类(如BKI1、KAT2、CEL1和UBP15)、DNA结合蛋白和激素响应蛋白(如ARF2和J3)。本研究结果将为进一步解析油菜千粒重的遗传机制、培育高容重油菜品种及后续基因的功能研究提供理论依据。

关键词: 甘蓝型油菜, 容重, 千粒重, 体积, 全基因组关联分析

Abstract:

Seed density reflects the accumulation characteristics of crop photosynthetic products in the grains, which plays an important role in the thousand-seed weight of rape. Selecting high seed density germplasm resources and studying the genetic characteristics of seed density are very important in the breeding of rapeseed. A natural population containing 187 Brassica napus L. varieties (lines) with different genetic backgrounds was used as plant materials to determine the seed density and its related traits (thousand-seed weight and seed volume) in the two environments. Genome-wide association study was carried out based on the optimal model and the candidate genes associated with seed density, thousand-seed weight, and seed volume was predicted. In the two years, there were significant differences in the seed density and its related traits among 187 materials at P < 0.05, and three materials with high seed density or thousand-seed weight were selected. A total of 24 SNP loci, that were significantly associated with seed density, seed weight, and seed volume, were identified by GWAS, which explained the phenotypic variation of 8.21%-10.40%. Haplotype analysis was used to determine the block interval of the SNP sites. The blocks containing 11 SNPs covered 12 candidate genes, which mainly encoded transcription factors such as WOX8, HAIKU1, AP2/ERF transcription factors, Dof family-zinc finger superfamily, BZR1 transcription factors, enzymes such as BKI1, KAT2, CEL1, UBP15, DNA binding proteins, and hormone response proteins such as ARF2 and J3. These results provide the theoretical basis for the development of high seed density rape varieties and the functional research of subsequent genes.

Key words: Brassica napus L., seed density, thousand-seed weight, seed volume, genome-wide association study (GWAS)

图1

群体容重、千粒重和体积在2种环境下的频率分布"

表1

187份甘蓝型油菜品种(系)籽粒容重、千粒重和体积的表型统计"

性状
Trait
年份
Year
最大值
Max.
最小值
Min.
均值
Average
标准差
SD
变异系数
CV (%)
籽粒容重Seed density (kg m-3) 2018-2019 969.05 527.31 859.96** 61.16 7.11
2019-2020 946.00 433.93 804.35** 99.47 12.37
体积Seed volume (×10-9 m3) 2018-2019 7.30 2.58 4.60** 0.89 19.38
2019-2020 11.97 1.28 3.52** 1.27 35.93
千粒重Thousand-seed weight (×10-3 kg) 2018-2019 5.75 1.73 3.95** 0.76 19.24
2019-2020 7.39 0.70 2.80** 0.89 31.79

表2

81份高容重甘蓝型油菜品种(系)体积和千粒重的表型统计"

性状
Trait
年度
Year
最大值
Max.
最小值
Min.
均值
Average
标准差
SD
变异系数
CV (%)
籽粒容重Seed density (kg m-3) 2018-2019 969.05 860.09 901.76** 26.41 2.93
2019-2020 946.00 806.90 875.16** 31.35 3.58
体积Seed volume (×10-9 m3) 2018-2019 6.36 2.69 4.51** 0.78 17.30
2019-2020 6.01 1.28 3.19** 0.87 27.26
千粒重Thousand-seed weight (×10-3 kg) 2018-2019 5.75 2.36 4.06** 0.69 16.87
2019-2020 5.01 1.16 2.78** 0.74 26.54

表3

油菜3种种子各性状间的相关性分析"

性状
Trait
容重
Seed density
体积
Seed volume
千粒重
Thousand-seed weight
容重Seed density 1 -0.29** 0.1
体积Seed volume -0.03 1 0.90**
千粒重Thousand-seed weight 0.31** 0.93** 1

图2

两年度中容重、千粒重和体积在各模型下的QQ图 A: 2018-2019年度容重; B: 2019-2020年度容重; C: 2018-2019年度千粒重; D: 2019-2020年度千粒重; E: 2018-2019年度体积; F: 2019-2020年度体积。"

图3

两年度中容重及其相关性状的Manhattan图 A: 2018-2019年度容重; B: 2019-2020年度容重; C: 2018-2019年度千粒重; D: 2019-2020年度千粒重; E: 2018-2019年度体积; F: 2019-2020年度体积。"

表4

容重、千粒重、体积相关性状的显著关联标记"

性状
Trait
环境
Environment
模型
Model
位点
SNP
染色体
Chr.
位置
Position
阈值
-log10(P)
贡献率
R2 (%)
基因型
Loci
容重
Seed density
2018-2019 K+PCA Bn-A01-p6774837 A01 6194252 5.12 9.52 T/C
Bn-scaff_17067_1-p175366 C02 27613356 5.06 8.64 T/C
Bn-A03-p8112016 C03 9899295 4.64 8.43 A/G
Bn-scaff_16069_1-p1916253 C07 38327703 4.59 9.39 A/G
Bn-scaff_21269_1-p313587 C08 37159594 4.53 8.54 A/G
2019-2020 K+PCA Bn-scaff_16394_2-p777469 C03 50479817 5.58 10.36 T/C
Bn-scaff_16394_2-p510062 C03 50830988 4.95 9.20 T/C
体积
Seed volume
2018-2019 K+PCA Bn-A10-p9973634 C09 40023501 4.41 8.22 A/C
2019-2020 K+PCA Bn-Scaffold000232-p58128 A07 826353 5.32 9.88 A/G
性状
Trait
环境
Environment
模型
Model
位点
SNP
染色体
Chr.
位置
Position
阈值
-log10(P)
贡献率
R2 (%)
基因型
Loci
Bn-A07-p3618045 A07 5593584 5.32 9.88 T/C
Bn-A10-p5210204 A10 4798717 4.43 8.25 T/G
Bn-A10-p12933288 A10 12981101 4.88 9.07 A/C
Bn-scaff_15838_5-p603655 C01 3432942 5.03 9.34 T/C
Bn-scaff_17731_1-p749457 C01 7033263 4.41 8.21 A/C
Bn-scaff_17731_1-p979512 C01 7260158 4.76 8.84 A/G
Bn-scaff_21186_1-p36313 C04 43747053 4.65 8.65 A/G
Bn-scaff_21566_1-p5088 C04 43859433 4.60 8.56 A/C
Bn-scaff_16770_1-p306276 C05 35633542 5.26 9.78 T/C
Bn-scaff_20270_1-p1211632 C05 41648492 5.04 9.37 T/C
Bn-scaff_18439_1-p633449 C06 12752920 5.22 9.70 T/C
Bn-scaff_15763_1-p595094 C06 20165181 4.65 8.64 T/G
Bn-A07-p16045526 C06 26606197 4.63 8.62 A/C
千粒重Thousand-seed weight 2018-2019 K
2019-2020 K Bn-scaff_21186_1-p36313 C04 43747053 4.55 8.43 A/G
Bn-scaff_21566_1-p5088 C04 43859433 5.62 10.40 A/C

图4

显著性标记所在的Block及候选基因"

表5

甘蓝型油菜容重相关性状的候选基因"

性状
Trait
物理区间
Physical interval
甘蓝型油菜基因编号
Gene ID in B. napus
拟南芥基因
Arabidopsis gene
基因
Gene
参考文献
Reference
SD 26393356-28833356 BnaC02g28720D AT5G42750 BKI1 [35]
9439295-10359295 BnaC03g18470D AT2G33150 KAT2 [36]
BnaC03g18850D AT5G45980 WOX8 [37]
BnaC03g19320D AT2G35230 IKU1 [38]
BnaC03g19550D AT2G35700 ERF38 [39]
37927703-38727703 BnaC07g36540D AT4G21030 ATDOF4.2 [40-41]
TSW 43147053-44459433 BnaC04g43460D AT1G70710 CEL1 [42]
SV 2632942-4232942 BnaC01g06800D AT1G17110 UBP15 [43]
6233263-7833263 BnaC01g11380D AT1G75080 BZR1 [35]
43147053-44459433 BnaC04g43460D AT1G70710 CEL1 [42]
12082920-13422920 BnaC06g10480D AT1G54060 ASIL1 [44]
39083501-40963501 BnaC09g35740D AT3G61830 ARF2 [45]
BnaC09g36560D AT3G44110 J3 [46]
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