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作物学报 ›› 2025, Vol. 51 ›› Issue (5): 1166-1177.doi: 10.3724/SP.J.1006.2025.44175

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

芥菜型油菜不同组织硫苷含量的QTL定位与候选基因分析

张金泽1(), 周庆国2, 肖莉晶1, 金海润1, 欧阳青静1, 龙旭1, 晏中彬1, 田恩堂1,*()   

  1. 1贵州大学农学院, 贵州贵阳 550025
    2安徽鼎科种业有限责任公司, 安徽合肥 230001
  • 收稿日期:2024-10-20 接受日期:2025-01-23 出版日期:2025-05-12 网络出版日期:2025-02-17
  • 通讯作者: *田恩堂, E-mail: erictian121@163.com
  • 作者简介:E-mail: a3184349049@163.com
  • 基金资助:
    贵州省科技支撑计划项目(黔科合支撑[2022]重点031);国家自然科学基金项目(32160483);国家自然科学基金项目(32360497);贵州大学国家自然科学基金后补助项目([2023] 093号)

QTL mapping and candidate gene analysis of glucosinolate content in various tissues of Brassica juncea

ZHANG Jin-Ze1(), ZHOU Qing-Guo2, XIAO Li-Jing1, JIN Hai-Run1, OU-YANG Qing-Jing1, LONG Xu1, YAN Zhong-Bin1, TIAN En-Tang1,*()   

  1. 1College of Agriculture, Guizhou University, Guiyang 550025, Guizhou, China
    2Anhui Dingke Seed Industry Co, Ltd., Hefei 230001, Anhui, China
  • Received:2024-10-20 Accepted:2025-01-23 Published:2025-05-12 Published online:2025-02-17
  • Contact: *E-mail: erictian121@163.com
  • Supported by:
    Guizhou Provincial Science and Technology Support Program(Guizhou Kehe Support [2022] Key 031);National Natural Science Foundation of China(32160483);National Natural Science Foundation of China(32360497);Post-Funded Project for National Natural Science Foundation of Guizhou University([2023] No. 093)

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摘要：

硫苷是重要的次生代谢物, 在油菜生长发育、病虫害防御等方面均具有重要作用。本研究以芥菜型油菜品系YufengZC和Qichi881为亲本创建的包含197个RIL-F8株系的作图群体(QY-RIL群体)为研究材料, 测定了全部株系的叶片、茎秆、花蕾和种子的硫苷含量。结果表明, 同一株系不同组织间硫苷含量存在显著差异, 而不同株系间相同组织的硫苷含量也存在较大变异, 且总体呈正态分布。相关性分析发现, 花蕾、茎秆与种子间硫苷含量呈显著正相关关系, 而叶片的硫苷含量仅与花蕾呈显著正相关关系。此外, 本研究还对叶片、茎秆、花蕾和种子硫苷含量的调控基因进行了QTL定位, 分别鉴定到9、10、9和18个QTL, 包含6个在多组织中共同检测到的cQTL (consensus QTL)。结合cQTL区间序列信息及所含基因的表达分析结果, 初步筛选出6个候选基因, 其中BjuB018426 (GTR1)和BjuB020498 (GTR2)参与硫苷从营养组织向生殖组织的转运, 可能是造成本研究中硫苷含量差异分布的重要调控基因。本研究结果可为解析芥菜型油菜不同组织的硫苷合成与分配机制及选育不同硫苷含量的多功能品种奠定基础。

关键词: 芥菜型油菜, 硫苷含量, QTL定位, 候选基因, 硫苷转运

Abstract:

Glucosinolates are critical secondary metabolites that play a pivotal role in the growth and development of rapeseed, as well as in its defense against diseases and pests. In this study, a mapping population comprising 197 recombinant inbred lines (RILs) of Brassica juncea was utilized to investigate glucosinolate distribution and genetic regulation. The glucosinolate contents in leaves, stems, buds, and seeds were quantified across all lines. The results revealed significant differences in glucosinolate content among different tissues within the same line and considerable variation across lines within the species, exhibiting an overall normal distribution. Correlation analysis showed a strong positive correlation in glucosinolate content among buds, stems, and seeds, as well as a notable positive correlation between leaves and flower buds. To further elucidate the genetic regulation of glucosinolate content, quantitative trait locus (QTL) mapping was conducted, resulting in the identification of 9, 10, 9, and 18 QTLs associated with glucosinolate content in leaves, stems, buds, and seeds, respectively. By integrating QTL interval sequence information with gene expression data, six candidate genes were preliminarily identified. Among them, BjuB018426 (GTR1) and BjuB020498 (GTR2) were implicated in the transport of glucosinolates from vegetative tissues to reproductive tissues, suggesting their potential roles as key regulatory genes underlying the differential distribution of glucosinolates observed in this study. These findings provide valuable insights into the mechanisms governing glucosinolate synthesis and distribution across various tissues in Brassica juncea. Moreover, they offer a foundational basis for breeding multifunctional varieties with tailored glucosinolate profiles to meet diverse agricultural and industrial needs.

Key words: Brassica juncea, glucosinolate contents, QTL mapping, candidate genes, glucosinolate transport

表1

不同组织硫苷含量描述性统计分析"

性状
Trait		 组织
Tissue		 最小值
Min.		 最大值
Max.		 平均值
Mean		 标准差
SD		 变异系数
CV (%)		 偏度
Skewness		 峰度
Kurtosis
硫苷含量
Glucosinolate contents
(μmol g-1)		 叶片Leaves 17.62 82.36 37.70 9.74 25.84 1.08 1.93
茎秆Stems 0.09 12.97 5.50 3.36 60.65 -0.42 -0.18
花蕾Buds 27.21 85.39 60.30 10.36 17.18 0.36 -1.02
种子Seeds (S1) 81.09 190.25 134.75 20.88 15.50 0.07 -0.39
种子Seeds (S2) 50.70 197.10 130.99 20.13 15.37 -0.21 1.18

图1

不同组织硫苷含量的频数分布 S1为2023年收获的种子, S2为2024年收获的种子。"

图2

不同组织硫苷含量的相关性热图 * 和**分别表示在0.05和0.01水平上相关性显著。S1为2023年收获的种子, S2为2024年收获的种子。"

图3

不同组织的QTL在芥菜性油菜染色体上的分布 A和B分别为不同组织的QTL在A基因组和B基因组的分布。QTL名称表示同表2。"

表2

不同组织中与硫苷含量相关的QTL的相关信息"

组织
Tissue		 性状位点
Trait locus		 染色体
Chromosome		 QTL峰值
QTL peak (cM)		 QTL区间
QTL interval (cM)		 最近的SNP标记
Recent SNP marker		 LOD值
LOD score		 表型贡献率
PVE (%)
叶片
Leaves		 qL-A02 A02 35.86 31.97-38.91 Marker 4,424,668 3.44 7.85
qL-A03 A03 27.14 23.50-31.22 Marker 1,757,168 11.46 26.40
qL-A04 A04 33.40 28.29-37.03 Marker 4,955,778 4.14 9.60
qL-A05 A05 43.18 39.57-48.06 Marker 470,368 3.51 8.10
qL-B02 B02 54.16 49.39-58.75 Marker 51,065 3.51 8.10
qL-B03 B03 95.18 93.13-98.55 Marker 4,697,670 3.29 7.59
qL-B05 B05 15.33 11.89-19.87 Marker 3,595,171 3.15 7.29
qL-B06 B06 32.54 26.25-35.41 Marker 3,645,990 3.67 8.44
qL-B08 B08 92.18 87.52-93.50 Marker 982,815 3.45 7.80
茎秆
Stems		 qS-A02 A02 37.27 33.69-44.78 Marker 4,545,960 4.39 10.13
qS-A03 A03 26.36 21.44-32.09 Marker 1,838,334 7.02 16.20
qS-A04 A04 35.05 30.16-36.40 Marker 5,062,389 4.20 9.60
qS-A06 A06 76.01 73.67-80.43 Marker 1,382,718 3.45 7.80
qS-A08 A08 13.60 11.00-17.51 Marker 731,978 3.87 9.00
qS-A09 A09 0.53 0-7.19 Marker 4,053,639 3.00 6.90
qS-B04 B04 3.47 0-8.52 Marker 2,409,165 5.64 12.90
qS-B05 B05 13.22 9.48-17.74 Marker 3,360,685 3.81 8.70
qS-B06 B06 32.54 29.94-34.90 Marker 3,645,990 3.69 8.40
qS-B08 B08 90.87 86.75-96.93 Marker 920,085 4.20 9.60
花蕾
Buds		 qF-A02 A02 35.86 31.97-39.78 Marker 4,424,668 2.61 6.08
qF-A03 A03 26.11 21.69-31.48 Marker 1,944,874 9.39 22.35
qF-A04 A04 31.35 28.00-36.08 Marker 4,981,184 3.17 7.20
qF-A07 A07 38.85 33.93-44.88 Marker 2,098,338 3.42 7.80
qF-A09 A09 48.23 43.38-52.24 Marker 3,898,423 3.18 7.50
qF-B01 B01 33.91 29.17-37.58 Marker 1,517,061 5.31 12.00
qF-B05 B05 11.89 6.32-15.33 Marker 3,601,337 4.32 9.90
qF-B06 B06 31.51 29.09-34.90 Marker 3,631,838 3.90 8.85
qF-B07 B07 15.49 11.13-20.50 Marker 2,661,101 3.48 8.10
种子
Seeds (S1)		 qS1-A01 A01 96.91 91.17-100.11 Marker 3,090,840 3.10 7.44
qS1-A02 A02 33.69 28.47-39.78 Marker 4,441,071 4.25 10.13
qS1-A03 A03 23.76 20.92-27.14 Marker 1,809,778 8.11 19.20
qS1-A04 A04 34.19 32.26-37.03 Marker 4,999,692 2.95 9.00
qS1-A08 A08 16.73 13.09-23.10 Marker 602,965 3.11 7.38
qS1-B03 B03 94.66 91.57-98.55 Marker 4,754,822 3.00 7.20
qS1-B05 B05 14.01 9.48-17.74 Marker 3,441,132 3.72 8.70
qS1-B06 B06 28.09 25.21-35.16 Marker 3,847,963 3.33 7.89
qS1-B08 B08 91.13 79.77-92.72 Marker 1,000,761 3.15 7.35
种子
Seeds (S2)		 qS2-A02 A02 35.04 31.97-39.78 Marker 4,487,079 4.30 9.90
qS2-A03 A03 25.58 23.76-31.73 Marker 1,900,789 6.39 15.06
qS2-A04 A04 28.90 27.14-33.40 Marker 4,925,233 3.75 9.00
qS2-B01 B01 5.01 0.87-9.59 Marker 1,451,547 3.06 7.20
qS2-B03 B03 96.73 93.13-100.63 Marker 4,648,733 3.42 8.10
qS2-B04 B04 49.07 46.08-54.87 Marker 2,242,137 4.12 9.68
qS2-B05 B05 11.89 8.17-15.33 Marker 3,601,337 3.74 8.78
qS2-B06 B06 34.12 31.51-37.73 Marker 3,783,270 3.69 8.70
qS2-B08 B08 93.50 88.29-97.48 Marker 856,065 3.00 6.36

表3

主效区间与硫苷含量相关的候选基因"

cQTL名称
cQTL name		 区间范围
Interval range (cM)		 候选基因ID
Candidate gene ID		 拟南芥同源基因信息
Information of
Arabidopsis
homologous genes		 基因功能
Gene function
cQTLA02 33.69-38.78 BjuA043503/BjuB010312 AT1G74080, MYB122 参与吲哚族硫苷生物合成与调控
Involved in indole glucosinolate biosynthesis and regulation
BjuA005541/BjuB010311 AT1G74100, SOT16 硫苷合成中硫基团转移
Sulfate group transfer in glucosinolate biosynthesis
cQTLA04 32.26-33.40 BjuA024283/BjuO013418 AT3G27785, MYB118 调控参与硫苷合成的下游基因
Regulates downstream genes involved in glucosinolate biosynthesis
BjuA015922/BjuB024226 AT2G22300, CAMTA3 响应钙信号调控硫苷合成
Responds to calcium signals to regulate glucosinolate biosynthesis
cQTLB05 11.89-15.33 BjuA002613 AT2G22330, CYP79B3 参与色氨酸途径, 生成吲哚硫苷的前体物质
Involved in the tryptophan pathway, producing precursor substances for indole glucosinolates
BjuO011663/BjuA003553 AT4G30530, GGP1 负责转化前体物质的酶
Enzyme responsible for converting precursor substances
cQTLB06 31.51-34.90 BjuA040529/BjuB003228 AT4G21990, APR3 提供硫苷合成所需的还原性硫
Provides reduced sulfur needed for glucosinolate biosynthesis
BjuA003915/BjuA040691 AT4G17880, MYC4 调节硫苷合成相关基因的表达
Regulates the expression of genes related to glucosinolate biosynthesis
cQTLA03 23.76-27.14 BjuB020498 AT5G62680, GTR2 参与硫苷从源到库组织的运输
Involved in transporting glucosinolates from source to sink tissues
BjuA009868 介导硫苷长距离运输
Mediates long-distance transport of glucosinolates
cQTLB08 88.29-92.72 BjuB018426/BjuA022496 AT3G47960, GTR1 将蛋氨酸和色氨酸衍生的硫苷转运到种子
Transports glucosinolates derived from methionine and tryptophan to seeds

图4

高低硫苷含量材料不同组织候选基因表达热图 纵坐标是候选基因及拟南芥中功能缩写, 横坐标表示高低硫苷含量材料不同组织样品缩写。其中H和L分别代表高和低硫苷含量的材料; F、L、S和Z07分别代表花蕾、叶片、茎秆和拨蕾自交7 d后的角果; R1、R2和R3代表3个重复, FPKM代表该基因在给定样本中的相对表达水平。"

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