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作物学报 ›› 2019, Vol. 45 ›› Issue (6): 818-828.doi: 10.3724/SP.J.1006.2019.84133

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

甘蓝型油菜茎高QTL定位及株高相关位点整合

魏丽娟1,2,*,刘瑞影1,2,*,张莉1,2,陈志友1,2,杨鸿1,2,霍强1,2,李加纳1,2,*()   

  1. 1 西南大学农学与生物科技学院 / 油菜工程研究中心, 重庆 400715
    2 西南大学现代农业科学研究院, 重庆400715
  • 收稿日期:2018-10-23 接受日期:2019-01-19 出版日期:2019-06-12 网络出版日期:2019-06-12
  • 通讯作者: 魏丽娟,刘瑞影,李加纳
  • 作者简介:魏丽娟, E-mail: lijuan525888@163.com|刘瑞影, E-mail: 1243480719@qq.com
  • 基金资助:
    本研究由中国博士后科学基金第62批面上项目(2017M622945);博士后创新人才计划项目(BX201700201)(BX201700201);博士后创新人才计划项目(BX201700201)(Chongqing Graduate Student Research Innovation Project CYS17078);重庆市研究生科研创新项目(CYS17078)(973 Program 40203917);国家重点基础研究发展计划(973计划)项目(40203917)和国家自然科学基金项目(31701460)资助

Detection of stem height QTL and integration of the loci for plant height- related traits in B. napus

Li-Juan WEI1,2,*,Rui-Ying LIU1,2,*,Li ZHANG1,2,Zhi-You CHEN1,2,Hong YANG1,2,Qiang HUO1,2,Jia-Na LI1,2,*()   

  1. 1 College of Agronomy and Biotechnology, Southwest University / Chongqing Engineering Research Center for Rapeseed, Chongqing 400715, China
    2 Academy of Agricultural Sciences, Southwest University, Chongqing 400715, China
  • Received:2018-10-23 Accepted:2019-01-19 Published:2019-06-12 Published online:2019-06-12
  • Contact: Li-Juan WEI,Rui-Ying LIU,Jia-Na LI
  • Supported by:
    This study was supported by the China Postdoctoral Science Foundation Funded Project(2017M622945);National Postdoctoral Program for Innovative Talents(BX201700201);National Postdoctoral Program for Innovative Talents(Chongqing Graduate Student Research Innovation Project CYS17078);the National Basic Research Program of China(973 Program 40203917);National Natural Science Foundation of China (31701460).

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摘要：

甘蓝型油菜主茎高度(茎高)是株型的构成因子之一, 研究其遗传机理对油菜株型改良具有重要的理论指导意义。目前对甘蓝型油菜茎高研究的报道较少。本研究以2个油菜茎高差异较大的亲本构建的重组自交系群体为材料, 利用SNP高密度遗传图谱, 2年共检测到11个茎高QTL, 分布在A04、A06、C04、A08和C01染色体上, 位点的表型贡献率为7.25%~19.61%。同时, 以455份来源不同的甘蓝型油菜为材料, 结合重测序产生的SNP标记, 对茎高进行全基因组关联分析, 2年共检测到5个SNP与茎高性状显著关联, 分布在A08、A10、C02和C06染色体上。根据茎高定位结果, 找到一些与激素途径(生长素、赤霉素和油菜素内酯)、光形态建成及植物生长发育相关的候选基因。在此基础上, 结合国内外株高相关性状定位研究结果, 将株高相关性状位点整合到甘蓝型油菜参考基因组上, 发现4个以上群体都在A01、A03、A07、C03和C06染色体上找到株高定位的区间, 2个群体在A10染色体上找到主花序长度共同定位的区间, 在A02和C03染色体上找到一次分枝高度共同定位的区间。本研究中的茎高定位结果与整合后的株高相关性状QTL定位区间有部分重叠, 位于A04、A06、A08、C04和C06染色体上。上述结果为甘蓝型油菜理想株型育种提供了理论依据。

关键词: 甘蓝型油菜, 茎高, QTL定位, GWAS, 位点整合

Abstract:

Stem height (SH) is important for the plant architecture and yield in Brassica napus. To date, the research about stem height in B. napus is very less. In the study, QTL mapping of stem height in B. napus was performed using the high density SNP genetic map constructed from the recombinant inbred lines (RIL) population with Illumina 60K SNP array. Totally 11 QTLs were detected in two years on A04, A06, C04, A08, and C01, which explained 7.25%-19.61% of the phenotypic variation. In addition, genome-wide association analysis (GWAS) of stem height in the 455 B. napus genotyped by resequencing was performed, and five significantly associated SNP loci on A08, A10, C02, and C06 were identified. And the candidate genes involved in photomorphogenesis, plant growth and development, and hormone pathway such as auxin, gibberellin and brassinosteroid were identified. We also integrated the loci for plant height-related traits including plant height (PH), main inflorescence length (MIL), branch segment (BS) and branch initiation height (BIH) in B. napus, in which four loci for PH on A01, A03, A07, C03, and C06 were identified in at least four populations, one locus for MIL on A10 and two loci for BIH on A02 and C03 were identified in two populations. The loci for stem height in our study were consistent with pervious QTL mapping results for plant height, branch initiation height and main inflorescence length. This study lays a foundation for the breeding of plant architecture in B. napus.

Key words: Brassica napus, stem height, QTL mapping, GWAS, loci integration

表1

重组自交系和自然群体甘蓝型油菜茎高的表型变异"

年份
Year		 平均值±标准偏差
Mean±SD (cm)		 范围
Range (cm)		 变异系数
CV (%)
父本ZY821 Male parent ZY821 2016 138.00±5.51
2017 131.90±12.30
母本GH06 Female parent GH06 2016 159.60±6.46
2017 145.60±8.77
重组自交系RIL 2016 150.64±11.49 119.00-174.25 7.63
2017 148.53±10.91 114.40-175.00 7.34
自然群体 Natural population 2016 156.58±17.11 93.86-200.10 10.93
2017 143.01±21.20 62.00-202.60 14.82

图1

重组自交系(a)和自然群体(b)茎高频率分布图"

表2

甘蓝型油菜茎高与株型和经济产量性状相关系数"

性状
Trait		 重组自交系 RIL 自然群体 Natural population
2016SH 2017SH 2016SH 2017SH
主花序长度 Main infloresences length (cm) -0.403** -0.355** -0.089 -0.306**
株高 Plant height (cm) 0.738** 0.617** 0.865** 0.865**
结荚高度 Height of podding (cm) 0.638** 0.581** 0.745** 0.850**
结荚厚度 Thickness of pod canopy (cm) 0.089 0.024 0.185 0.101
一次有效分枝起点 Branch initiation height (cm) 0.610** 0.589** 0.621** 0.772**
经济产量 Yield (g) 0.341** 0.037 0.255** 0.286**

表3

甘蓝型油菜茎高QTL结果"

位点
QTL		 染色体
Chr.		 LOD阈值
LOD score		 标记区间
SNP interval		 加性效应a
Additive effect		 贡献率
R2 (%)		 物理区间
Physical position (bp)
q2016SH-A04-1 A04 3.72 SNP42542-SNP9367 6.21 13.70
q2016SH-A04-2 5.61 SNP9354-SNP9343 7.17 19.52 18264803-18455956
q2016SH-A04-3 2.90 SNP42544-SNP9263 5.61 10.78 17846583-18115323
q2016SH-A06-1 A06 4.33 SNP13035-SNP13048 -4.72 14.46 1469838-1480581
q2016SH-A06-2 5.50 SNP13263-SNP34194 -5.45 19.61 1860393-2106757
q2016SH-A06-3 4.56 SNP12979-SNP13009 -4.88 15.07 1360428-1442742
q2016SH-A06-4 2.83 SNP14849-SNP14835 3.65 8.88 6967630-7274382
q2016SH-C04 C04 3.29 SNP49157-SNP49146 3.85 9.97 7352388-7867004
q2017SH-A08 A08 3.94 SNP18566-SNP18577 2.95 7.25 18081289-18861275
q2017SH-C01-1 C01 6.11 SNP2398-SNP37156 3.80 11.86
q2017SH-C01-2 6.02 SNP37161-SNP38210 3.79 11.70 12068324-12309768

图2

甘蓝型油菜茎高QTL在连锁图上的位置 QTL方框为99%置信区间的左右边界, 其上“T”为95%置信区间的边界。"

图3

甘蓝型油菜群体结构图"

图4

甘蓝型油菜茎高全基因组关联分析Manhattan图和QQ图SH: 茎高。SH: stem height."

表4

甘蓝型油菜茎高显著关联位点"

位点
Locus		 环境
Environment		 模型
Model		 位点
SNP		 染色体
Chr.		 位置
Position		 阈值
-lg (P)		 贡献率
R2(%)
2016SH-1 2016 Naive S8_12459044 A08 12459044 5.90 15.90
2016SH-2 S10_16428321 A10 16428321 5.62 15.64
2017SH-1 2017 K+PCA S12_13075515 C02 13075515 7.62 10.08
2017SH-2 S12_13075493 C02 13075493 5.64 6.91
2017SH-3 S16_28288339 C06 28288339 5.70 6.86

图5

不同群体株高相关性状位点整合。 PH: 株高; MIL: 主花序长度; BS: 分枝区段长度; BIH: 一次分枝高度; SH: 主茎高度。"

表5

茎高候选基因鉴定"

位点名称
Name		 基因
Genes		 物理位置
Physical position		 拟南芥同源基因
Homologs in A. thaliana		 功能注释
Functional annotation
q2016SH-A04-2 BnaA04g25550D A04: 18331184-18331589 AT2G44080/ARL 调控细胞大小
Regulated cell expansion
q2016SH-A04-3 BnaA04g24760D A04: 18025141-18026983 AT2G43010/PIF4 编码转录因子bHLH, 参与生长素生物合成, 调控下胚轴伸长
Encoded the bHLH transcription factor, involving in the auxin biosynthesis and regulating hypocotyl elongation
BnaA04g24770D A04: 18030030-18033008 AT2G43020/PAO2 参与ABA介导的植物生长
Involved in plant development mediated by ABA
BnaA04g24970D A04: 18140559-18140805 AT2G43280/FAR1 调控光敏色素信号传导
Involved in phytochrome A signaling
BnaA04g24980D A04: 18141000-18144940 AT2G43280/FAR1 调控光敏色素信号传导
Involved in phytochrome A signaling
q2016SH-A06-2 BnaA06g03090D A06: 1869158-1871192 AT1G49770/RGE1 编码转录因子bHLH,影响植物生长发育
Encoded the bHLH transcription factor, regulating plant growth and development
q2016SH-A06-4 BnaA06g13320D A06: 6972730-6976892 AT1G19220/ARF19 参与生长素和乙烯信号传导过程
Involved in auxin and ethylene signaling
q2016SH-C04 BnaC04g10190D C04: 7847368-7848334 AT2G35150/EXL7 调控植物生长发育
Regulated plant growth and development
q2017SH-A08 BnaA08g28200D A08: 18580209-18583117 AT1G05180/AXR1 参与生长素抵抗反应
Involved in auxin resistance
q2017SH-C01-2 BnaC01g17440D C01: 12009590-12010572 AT4G25560/MYB18 调控光形态建成
Regulated photomorphogenesis in the light
2016SH-1 BnaA08g15400D A08: 12809202-12810946 AT4G36730/GBF1 调控光形态建成
Regulated photomorphogenesis in the light
2016SH-2 BnaA10g24860D A10: 16153991-16157850 AT5G05690/CPD 编码CP90A蛋白, 参与油菜素内酯生物合成
Encoded a member of the CP90A family, and involved in the brassinosteroid biosynthesis
2017SH-1 BnaC02g17570D C02: 13199941-13200513 AT5G13790/AGL15 MADS家族成员, 参与赤霉素代谢
Encoded a member of the MADS family, and involved in the gibberellin metabolic process
2017SH-2 BnaC06g27170D C06: 28615555-28619464 AT5G37020/ARF8 参与生长素稳态
Involved in auxin homeostasis
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