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作物学报

作物学报 ›› 2024, Vol. 50 ›› Issue (7): 1669-1683.doi: 10.3724/SP.J.1006.2024.31073

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

两个RIL群体中小麦籽粒品质相关性状QTL定位及KASP标记开发

毕俊鸽1,2,**(), 曾占奎1,2,**(), 李琼1,2, 洪壮壮1,2, 颜群翔1,2, 赵越1,2, 王春平1,2,*()   

  1. 1河南科技大学农学院 / 河南省旱地作物种质资源利用工程研究中心, 河南洛阳 471000
    2神农种业实验室, 河南郑州 450000
  • 收稿日期:2023-11-30 接受日期:2024-01-31 出版日期:2024-07-12 网络出版日期:2024-03-02
  • 通讯作者: *王春平, E-mail: chunpingw@163.com
  • 作者简介:毕俊鸽, E-mail: bijungee@163.com;
    曾占奎, E-mail: zengzhankui0301@163.com

    **同等贡献

  • 基金资助:
    河南省重大科技专项“小麦营养基因组学解析及功能食品创制与产业化”项目(231100110300);神农种业实验室“一流课题”项目(SN01-2022-01)

QTL mapping and KASP marker development of grain quality-relating traits in two wheat RIL populations

BI Jun-Ge1,2,**(), ZENG Zhan-Kui1,2,**(), LI Qiong1,2, HONG Zhuang-Zhuang1,2, YAN Qun-Xiang1,2, ZHAO Yue1,2, WANG Chun-Ping1,2,*()   

  1. 1College of Agriculture, Henan University of Science and Technology / Henan Dryland Crop Germplasm Resources Utilization Engineering Research Center, Luoyang 471000, Henan, China
    2Shennong Laboratory, Zhengzhou 45000, Henan, China
  • Received:2023-11-30 Accepted:2024-01-31 Published:2024-07-12 Published online:2024-03-02
  • Contact: *E-mail: chunpingw@163.com
  • About author:

    **Contributed equally to this work

  • Supported by:
    Major Science and Technology of Henan Province Project “The Wheat Nutrigenomics Analysis and Functional Food Creation and Industrialization Fund Project”(231100110300);Shennong Laboratory “First-Class Subject” project(SN01-2022-01)

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摘要：

本研究利用小麦55K SNP (55K single-nucleotide polymorphism)芯片和DArT (diversity array technology)标记对Avocet/Chilero和Avocet/Huites构建的两个F6重组自交系群体(recombinant inbred line, RIL)进行了小麦籽粒蛋白质含量(grain protein content, GPC)、湿面筋含量(wet gluten content, WGC)和沉降值(sedimentation value, SV)的QTL (quantitative trait loci)定位。共鉴定到68个与小麦籽粒蛋白质含量、湿面筋含量和沉降值相关的QTL, 表型贡献率为3.60%~22.53%, 其中, 位于3A(2)、4D、5D(2)、6A(8)和7B染色体上的14个QTL可在多环境下被重复检测到。此外, 在3A、3D、4B、5D、6A(2)和7B染色体上检测到7个QTL簇, 位于3AS染色体9.32~60.01 Mb和6AS染色体38.47~82.95 Mb的稳定QTL簇C3A和C6A.2, 同时与小麦籽粒蛋白质含量、湿面筋含量和沉降值显著相关, 分别解释了6.55%~14.21%和3.83%~22.53%的表型变异。同时, 在2个QTL簇中筛选到16个可能与籽粒蛋白质含量相关的候选基因, 并根据候选基因开发了可供育种利用的KASP标记CGPC-6A-KASP-1和CGPC-6A-KASP-2。本研究为小麦籽粒品质相关性状的遗传改良提供了新的QTL位点和KASP标记, 为分子标记辅助育种提供依据与参考。

关键词: 小麦, 籽粒蛋白质含量, QTL定位, KASP标记

Abstract:

We utilized a 55K single-nucleotide polymorphism (55K SNP) array and diversity array technology (DArT) to identify QTLs for grain protein content (GPC), wet gluten content (WGC), and sedimentation value (SV) in two F6 recombinant inbred lines derived from Avocet/Chilero and Avocet/Huites. Sixty-eight QTLs were identified related to grain protein content, wet gluten content, and settlement value, explaining 3.60%-22.53% of the phenotypic variances. Fourteen QTLs were found to be present in multiple environments, located on chromosomes 3A(2), 4D, 5D(2), 6A(8), and 7B, respectively. Additionally, seven QTLs clusters were detected on chromosomes 3A, 3D, 4B, 5D, 6A(2), and 7B, respectively. Two stable QTL clusters, C3A and C6A.2, identified in the physical intervals of 9.32-60.01 Mb and 38.47-82.95 Mb, respectively, were significantly associated with grain protein content and wet gluten content. These clusters accounted for 6.55%-14.21% and 3.83%-22.53% of the phenotypic variances for grain protein content, wet gluten content, and sedimentation value. Meanwhile, a total of 16 candidate genes associated with grain protein content were predicted within the two stable QTL clusters. Moreover, two KASP marker, CGPC-6A-KASP-1 and CGPC-6A-KASP-2, were developed based on the candidate genes. The findings of this study provide support for the identification of new QTL and KASP markers that can contribute to the genetic improvement of grain quality-related traits in wheat. These results also offer valuable insights for marker-assisted breeding in wheat.

Key words: wheat, grain protein content, QTL mapping, KASP marker

表1

不同环境下两个F6 RIL群体及亲本中籽粒品质相关性状表型变异分析"

群体
Population		 性状
Trait		 环境
Env.		 亲本Parents RIL家系 广义遗传力
H2		 基因型
Genotype
(G)		 环境
Environment
(E)		 基因型×
环境
G × E
Avocet Chilero Huites 范围
Range		 平均值±标准误
Mean ± SD		 偏度
Skewness		 峰度
Kurtosis		 变异系数
CV (%)
AC
Population		 GPC
(%)		 E1 13.88** 14.95** - 12.19-17.16 14.52±0.90 0.14 ‒0.06 6.20 0.82 431.42** 6,867.82** 97.12**
E2 13.94** 14.93** - 13.78-18.51 15.66±0.88 0.62 0.90 5.62
E3 14.90* 15.42* - 12.42-18.94 15.27±1.29 0.20 ‒0.21 8.45
E4 13.62** 16.05** - 12.49-16.80 14.87±0.86 ‒0.11 ‒0.20 5.78
WGC
(%)		 E1 29.57** 33.15** - 26.50-36.85 31.99±2.17 ‒0.38 ‒0.31 6.78 0.85 338.43** 3,464.10** 59.57**
E2 30.48** 33.39** - 29.57-39.80 34.24±2.01 0.27 0.33 5.87
E3 31.48* 34.29* - 26.04-40.40 32.78±2.86 0.03 ‒0.28 8.72
E4 30.53** 35.03** - 26.33-36.63 32.48±2.08 ‒0.55 ‒0.18 6.40
SV
(%)		 E1 25.74** 35.61** - 21.76-39.02 30.36±3.53 ‒0.18 ‒0.19 11.63 0.84 233.56** 1,648.79** 42.91**
E2 27.90** 34.13** - 24.27-43.47 32.95±3.22 0.12 0.39 9.77
E3 29.47** 38.65** - 22.29-46.82 33.21±4.86 0.06 ‒0.24 14.63
E4 31.81** 39.49** - 22.74-40.29 32.62±3.67 ‒0.49 ‒0.11 11.25
AH
Population		 GPC
(%)		 E5 14.68* - 15.29* 12.56-16.73 14.83±0.88 0.08 ‒0.58 5.93 0.88 952.79** 1,077.77** 132.19**
E6 15.12** - 16.04** 12.10-18.66 14.99±1.34 0.25 ‒0.08 8.94
E7 14.69** - 16.03** 13.13-17.89 14.83±0.92 0.33 ‒0.13 6.20
WGC
(%)		 E5 30.26 - 30.27 26.89-36.27 31.98±2.03 ‒0.04 ‒0.61 6.35 0.90 446.82** 285.39** 49.13**
E6 31.71** - 34.76** 25.64-39.26 32.06±2.80 0.13 ‒0.35 8.73
E7 31.06** - 34.86** 28.30-38.52 32.31±2.06 0.13 ‒0.39 6.38
SV
(%)		 E5 39.43* - 43.24* 19.39-43.24 31.68±3.79 0.04 0.80 11.96 0.83 272.28** 1,075.43** 54.59**
E6 34.52* - 36.38* 22.55-46.57 33.83±4.96 0.22 ‒0.28 14.66
E7 29.78* - 37.07* 24.81-43.81 33.89±3.57 ‒0.08 ‒0.14 10.53

图1

AC群体(a)和AH群体(b)籽粒品质相关性状的频率分布 不同的填充形状和线条表示不同的环境; BLUE为最佳线性无偏估计。缩写同表1。"

图2

AC群体(a)和AH群体(b)中不同环境下籽粒品质相关性状的相关系数 白色到黑色的渐变表示从正相关到负相关; E1GPC、E1WGC、E1SV、E2GPC、E2WGC、E2SV、E3GPC、E3WGC、E3SV、E4GPC、 E4WGC、E4SV、E5GPC、E5WGC、E5SV、E6GPC、E6WGC、E6SV、E7GPC、E7WGC和E7SV分别为在E1、E2、E3、E4、E5、 E6和E7环境下的籽粒蛋白质含量、湿面筋含量和沉降值; *表示在P < 0.05水平差异显著。"

表S1

AC群体籽粒品质相关性状的QTL"

性状
Trait		 QTL 环境
Environment		 遗传位置
Genetic position (cM)		 侧翼标记
Flanking marker		 LOD值
LOD value		 表型贡献率
PVE (%)		 加性效应
Additive		 物理区间
Physical intervals (Mb)		 优势等位基因
Favorable allele
GPC QGPC.haust-AC-1B E4 6.00 AX-94871279-AX-111525685 6.20 15.70 ‒0.33 670.39-670.78 Chilero
QGPC.haust-AC-2B E4 46.00 AX-109885265-AX-108951217 4.76 11.77 ‒0.27 782.02-799.25 Chilero
QGPC.haust-AC-3A BLUE 271.00 AX-111543258-AX-111142459 3.54 8.77 ‒0.22 27.88-57.73 Chilero
QGPC.haust-AC-3D.1 E1 94.00 AX-108729781-AX-110834100 8.83 13.98 ‒0.44 537.29-552.24 Chilero
QGPC.haust-AC-3D.2 E3 189.00 AX-110808052-AX-95632197 2.69 5.57 ‒0.33 533.94-2.63 Chilero
QGPC.haust-AC-6A.1 E1 12.00 AX-109042664-AX-108939533 6.11 8.95 0.40 61.56-61.89 Avocet
E4 13.00 AX-109042664-AX-108939533 4.08 9.88 0.30 61.56-61.89 Avocet
QGPC.haust-AC-6A.2 E3 42.00 AX-94591230-AX-111122628 9.74 21.05 0.61 38.47-71.43 Avocet
BLUE 42.00 AX-94591230-AX-111122628 8.92 22.53 0.35 38.47-71.43 Avocet
QGPC.haust-AC-7B E3 54.00 AX-110412930-AX-108833738 3.82 7.52 ‒0.38 643.48-653.98 Chilero
BLUE 54.00 AX-110412930-AX-108833738 3.70 8.67 ‒0.22 643.48-653.98 Chilero
WGC QWGC.haust-AC-3A.1 E2 274.00 AX-111543258-AX-111142459 4.22 10.12 ‒0.77 27.88-57.73 Chilero
QWGC.haust-AC-3A.2 E3 237.00 AX-89583101-AX-109397508 5.98 10.64 ‒0.96 29.03-25.05 Chilero
BLUE 237.00 AX-89583101-AX-109397508 5.50 14.21 ‒0.62 29.03-25.05 Chilero
QWGC.haust-AC-3D.1 E1 96.00 AX-108729781-AX-110834100 5.91 16.34 ‒0.87 537.29-552.24 Chilero
QWGC.haust-AC-3D.2 E3 163.00 AX-110056468-AX-109831081 3.60 6.50 ‒0.74 349.68-342.70 Chilero
QWGC.haust-AC-4A E4 78.00 AX-110907767-AX-109440196 3.45 5.32 0.54 618.93-619.31 Avocet
QWGC.haust-AC-4B E3 24.00 AX-109986770-AX-110076607 5.11 9.22 ‒0.94 12.49-11.43 Chilero
QWGC.haust-AC-5D.1 E4 1.00 AX-108775585-AX-111011707 7.74 13.80 ‒0.83 556.11-547.81 Chilero
QWGC.haust-AC-5D.2 E4 76.00 AX-110476332-AX-109394742 7.62 13.32 ‒0.81 6.39-12.58 Chilero
QWGC.haust-AC-6A E3 42.00 AX-94591230-AX-111122628 9.39 17.62 1.22 38.47-71.43 Avocet
E4 42.00 AX-94591230-AX-111122628 4.05 6.24 0.54 38.47-71.43 Avocet
BLUE 42.00 AX-94591230-AX-111122628 7.65 20.45 0.74 38.47-71.43 Avocet
QWGC.haust-AC-6B E2 161.00 AX-109306114-AX-110400029 3.30 8.79 0.71 11.98-681.42 Avocet
SV QSV.haust-AC-1D E1 47.00 AX-111561523-AX-111535968 3.70 9.10 1.19 418.20-484.43 Avocet
QSV.haust-AC-2B.1 E4 134.00 AX-110064360-AX-111012832 5.19 7.22 ‒1.21 569.13-554.92 Chilero
QSV.haust-AC-2B.2 BLUE 94.00 AX-108837623-AX-89735721 4.69 11.69 ‒0.96 705.68-725.96 Chilero
QSV.haust-AC-3A.1 E2 278.00 AX-111543258-AX-111142459 3.17 10.27 ‒0.98 27.88-57.73 Chilero
QSV.haust-AC-3A.2 E3 236.00 AX-110520445-AX-89583101 3.10 9.30 ‒1.23 37.35-29.03 Chilero
BLUE 237.00 AX-89583101-AX-109397508 4.74 11.92 ‒0.98 29.03-25.05 Chilero
QSV.haust-AC-4B E3 24.00 AX-109986770-AX-110076607 3.52 10.64 ‒1.39 12.49-11.43 Chilero
QSV.haust-AC-6A.1 E1 42.00 AX-94591230-AX-111122628 5.50 13.30 1.21 38.47-71.43 Avocet
E2 42.00 AX-94591230-AX-111122628 3.46 9.92 0.94 38.47-71.43 Avocet
E3 42.00 AX-94591230-AX-111122628 4.70 14.38 1.52 38.47-71.43 Avocet
BLUE 42.00 AX-94591230-AX-111122628 5.91 15.09 1.09 38.47-71.43 Avocet
QSV.haust-AC-6A.2 E4 11.00 AX-111504079-AX-109974361 4.35 5.72 1.22 59.69-60.97 Avocet
QSV.haust-AC-7A.1 E4 173.00 AX-109473518-AX-109364614 8.14 11.43 ‒1.56 692.77-701.31 Chilero
QSV.haust-AC-7A.2 E4 204.00 AX-109947373-AX-111089494 11.07 16.16 1.78 731.14-708.47 Avocet
QSV.haust-AC-7B E1 17.00 AX-109306944-AX-95098493 5.07 12.30 1.17 54.79-708.12 Avocet
QSV.haust-AC-7D E2 55.00 AX-109301921-AX-111607406 2.68 7.56 ‒0.83 12.57-16.88 Chilero

表S2

AH群体籽粒品质相关性状的QTL"

性状
Trait		 QTL 环境
Environment		 遗传位置
Genetic position (cM)		 侧翼标记
Flanking marker		 LOD值
LOD value		 表型贡献率
PVE (%)		 加性效应
Additive		 物理区间
Physical interval (Mb)		 优势等位基因
Favorable allele
GPC QGPC.haust-AH-2A E6 800.00 DArT3958592-SNP1019498 3.16 5.98 ‒0.37 605.66-625.28 Huites
QGPC.haust-AH-2B.1 E7 647.00 DArT1128457-DArT3533008 4.24 9.29 ‒0.31 784.26-784.04 Huites
QGPC.haust-AH-2B.2 BLUE 242.00 DArT3021488-DArT1079407 2.64 5.44 ‒0.27 97.34-55.23 Huites
QGPC.haust-AH-3A.1 E5 530.00 DArT3064883-DArT1223327 5.61 15.48 ‒0.41 476.31-435.80 Huites
QGPC.haust-AH-3A.2 E6 581.00 DArT1254451-SNP980942 3.56 6.55 ‒0.39 46.17-46.00 Huites
QGPC.haust-AH-3A.3 E6 774.00 DArT3958575-DArT3946190 3.60 7.00 0.41 32.97-9.32 Avocet
QGPC.haust-AH-3A.4 E7 519.00 SNP1213892-SNP100357542 4.39 9.53 ‒0.31 521.30-544.19 Huites
QGPC.haust-AH-3A.5 BLUE 570.00 DArT100082954-DArT1013788 4.96 8.35 ‒0.34 60.01-54.94 Huites
QGPC.haust-AH-4D E7 25.00 DArT1107261-DArT1233229 2.71 5.34 ‒0.23 207.70-319.32 Huites
QGPC.haust-AH-5A.1 E5 394.00 SNP998276-SNP993328 3.41 9.48 0.32 569.66-570.19 Avocet
QGPC.haust-AH-5A.2 E6 117.00 DArT1205298-DArT1120172 3.76 7.03 ‒0.40 29.63-26.75 Huites
QGPC.haust-AH-6A.1 E6 363.00 SNP4910893-SNP1096119 4.98 10.15 0.54 76.63-77.89 Avocet
BLUE 364.00 SNP4910893-SNP1096119 6.99 12.26 0.45 76.63-77.89 Avocet
QGPC.haust-AH-6A.2 E7 313.00 DArT3946508-SNP1151487 4.81 11.45 0.41 586.30-202.91 Avocet
QGPC.haust-AH-6A.3 E7 431.00 SNP2358794-SNP1094001 2.83 6.23 ‒0.25 12.23-12.89 Huites
BLUE 426.00 DArT1233578-DArT4544113 3.51 5.84 ‒0.29 11.01-12.08 Huites
QGPC.haust-AH-7B BLUE 61.00 DArT1218440-SNP2253713 6.36 10.63 ‒0.38 1.38-1.44 Huites
WGC QWGC.haust-AH-1A BLUE 137.00 SNP1218247-DArT1665599 2.61 3.60 ‒0.48 500.84-499.81 Huites
QWGC.haust-AH-2A BLUE 394.00 SNP1240003-DArT2277504 4.36 8.01 ‒0.74 13.12-581.96 Huites
QWGC.haust-AH-2B E7 645.00 DArT1218607-DArT1128457 5.00 7.47 ‒0.65 776.57-234.76 Huites
QWGC.haust-AH-3A E7 519.00 SNP1213892-SNP100357542 6.70 9.47 ‒0.74 521.30-544.19 Huites
QWGC.haust-AH-3B BLUE 394.00 DArT1138136-DArT1695648 2.86 4.02 ‒0.50 123.60-113.98 Huites
QWGC.haust-AH-4D E5 63.00 DArT100270810-DArT1101297 3.80 12.80 ‒0.76 456.68-464.36 Huites
E7 75.00 DArT988923-DArT1111555 4.33 6.16 ‒0.59 469.43-479.62 Huites
BLUE 72.00 DArT1239874-DArT988923 5.08 7.57 ‒0.68 456.05-469.43 Huites
QWGC.haust-AH-5A BLUE 175.00 SNP4910949-DArT3941315 3.40 4.78 ‒0.55 415.19-430.25 Huites
QWGC.haust-AH-5B BLUE 242.00 DArT1100263-DArT1112093 6.61 9.71 0.78 658.56-658.45 Avocet
QWGC.haust-AH-5D E5 3.00 DArT3944272-DArT1695607 3.12 10.60 ‒0.71 5.79-25.49 Huites
E7 2.00 SNP1234827-DArT3944272 7.80 11.28 ‒0.81 4.36-5.79 Huites
BLUE 2.00 SNP1234827-DArT3944272 8.77 13.60 ‒0.94 4.36-5.79 Huites
QWGC.haust-AH-6A.1 E7 426.00 DArT1233578-DArT4544113 5.62 8.18 ‒0.70 11.01-12.08 Huites
QWGC.haust-AH-6A.2 E6 369.00 DArT3026247-DArT100000998 3.00 11.06 0.98 82.95-72.49 Avocet
E7 369.00 DArT3026247-DArT100000998 2.94 3.83 0.48 82.95-72.49 Avocet
QWGC.haust-AH-7B E7 61.00 DArT1218440-SNP2253713 6.32 8.79 ‒0.70 1.38-1.44 Huites
SV QSV.haust-AH-1A E7 123.00 DArT1207393-DArT4993415 2.60 8.90 ‒1.03 503.46-514.10 Huites
QSV.haust-AH-1B BLUE 825.00 SNP1090153-DArT3021227 3.04 8.08 ‒1.43 521.00-2.92 Huites
QSV.haust-AH-2B BLUE 29.00 SNP4988941-SNP1100485 2.59 9.29 ‒1.54 289.16-13.91 Huites
QSV.haust-AH-3B BLUE 150.00 SNP100526939-SNP4990292 3.03 5.61 ‒1.18 75.71-61.62 Huites
QSV.haust-AH-4A E6 664.00 DArT4537961-DArT1254189 3.04 7.98 ‒1.59 140.70-90.59 Huites
QSV.haust-AH-5D E7 0 SNP1234827-DArT3944272 3.54 11.89 ‒1.22 4.36-5.79 Huites
E5 4.00 DArT3944272-DArT1695607 4.22 9.91 ‒1.46 5.79-25.49 Huites
QSV.haust-AH-6A.1 E6 427.00 DArT1233578-DArT4544113 2.78 5.99 ‒1.41 11.01-12.08 Huites
QSV.haust-AH-6A.2 E6 365.00 SNP1096119-SNP100562777 6.33 14.90 2.93 77.89-138.23 Avocet
BLUE 364.00 SNP4910893-SNP1096119 3.81 6.90 1.43 76.63-77.89 Avocet
QSV.haust-AH-6B.1 E6 855.00 DArT3955224-SNP100542305 4.18 9.43 ‒1.75 298.01-715.70 Huites
QSV.haust-AH-6B.2 E5 53.00 DArT1159426-DArT4543423 3.59 10.77 ‒1.48 221.72-708.71 Huites
QSV.haust-AH-7B E5 75.00 SNP2253713-SNP2276097 2.81 15.01 ‒1.74 1.44-16.53 Huites

表2

两个F6 RIL群体QTL簇分析"

QTL簇
QTL cluster		 性状
Trait		 群体
Population		 QTL 连锁标记
Linkage marker		 平均LOD值
Average LOD value		 平均表型贡献率
Average PVE (%)		 环境
Environment		 优势等位基因
Favorable allele		 物理区间
Physical intervals (Mb)
C3A GPC/WGC/SV AC Pupulation QGPC.haust-AC-3A AX-111543258-AX-111142459 3.54 8.77 BLUE Chilero 9.32-60.01
AC Pupulation QWGC.haust-AC-3A.1 AX-111543258-AX-111142459 4.22 10.12 E2 Chilero
AC Pupulation QWGC.haust-AC-3A.2 AX-89583101-AX-109397508 5.74 12.43 E3/BLUE Chilero
AC Pupulation QSV.haust-AC-3A.1 AX-111543258-AX-111142459 3.17 10.27 E2 Chilero
AC Pupulation QSV.haust-AC-3A.2 AX-109397508-AX-110520445 3.92 10.61 E3/BLUE Chilero
AH Pupulation QGPC.haust-AH-3A.2 DArT1254451-SNP980942 3.56 6.55 E6 Huites
AH Pupulation QGPC.haust-AH-3A.3 DArT3958575-DArT3946190 3.60 7.00 E6 Avocet
AH Pupulation QGPC.haust-AH-3A.5 DArT100082954-DArT1013788 4.96 8.35 BLUE Huites
C3D GPG/WGC AC Pupulation QGPC.haust-AC-3D.1 AX-108729781-AX-110834100 8.83 13.98 E1 Chilero 537.29-552.24
AC Pupulation QWGC.haust-AC-3D.1 AX-108729781-AX-110834100 5.91 16.34 E1 Chilero
C4B WGC/SV AC Pupulation QWGC.haust-AC-4B AX-109986770-AX-110076607 5.11 9.22 E3 Chilero 11.43-12.49
AC Pupulation QSV.haust-AC-4B AX-109986770-AX-110076607 3.52 10.64 E3 Chilero
C5D WGC/SV AC Pupulation QWGC.haust-AC-5D.2 AX-110476332-AX-109394742 7.62 13.32 E4 Chilero 4.36-25.49
AH Pupulation QWGC.haust-AH-5D SNP1234827-DArT1695607 6.56 11.83 E5/E7/BLUE Huites
AH Pupulation QSV.haust-AH-5D SNP1234827-DArT1695607 3.88 10.90 E5/E7 Huites
C6A.1 GPC/WGC/SV AH Pupulation QGPC.haust-AH-6A.3 DArT1233578-SNP1094001 3.17 6.04 E7/BLUE Huites 11.01-12.89
AH Pupulation QWGC.haust-AH-6A.1 DArT1233578-DArT4544113 5.62 8.18 E7 Huites
AH Pupulation QSV.haust-AH-6A.1 DArT1233578-DArT4544113 2.78 5.99 E6 Huites
C6A.2 GPC/WGC/SV AC Pupulation QGPC.haust-AC-6A.1 AX-109042664-AX-108939533 5.10 9.42 E1/E4 Avocet 38.47-82.95
AC Pupulation QGPC.haust-AC-6A.2 AX-94591230-AX-111122628 9.33 21.79 E3/BLUE Avocet
AC Pupulation QWGC.haust-AC-6A AX-94591230-AX-111122628 7.03 14.77 E3/E4/BLUE Avocet
AC Pupulation QSV.haust-AC-6A.1 AX-94591230-AX-111122628 4.89 13.17 E1/E2/E3/BLUE Avocet
AC Pupulation QSV.haust-AC-6A.2 AX-111504079-AX-109974361 4.35 5.72 E4 Avocet
AH Pupulation QGPC.haust-AH-6A.1 SNP4910893-SNP1096119 5.99 11.21 E6/BLUE Avocet
AH Pupulation QWGC.haust-AH-6A.2 DArT3026247-DArT100000998 2.97 7.45 E6/E7 Avocet
AH Pupulation QSV.haust-AH-6A.2 SNP4910893-SNP1096119 3.81 6.90 BLUE Avocet
C7B GPC/WGC/SV AH Pupulation QGPC.haust-AH-7B DArT1218440-SNP2253713 6.36 10.63 BLUE Huites 1.38-16.53
AH Pupulation QWGC.haust-AH-7B DArT1218440-SNP2253713 6.32 8.79 E7 Huites
AH Pupulation QSV.haust-AH-7B SNP2253713-SNP2276097 2.81 15.01 E5 Huites

图3

在两个RIL群体中关于籽粒蛋白质含量、湿面筋含量和沉降值的QTL簇 两条染色体上的蓝色区段表示QTL的物理区间, 实心矩形表示优势等位基因来自父本Chilero或Huites, 空心矩形表示优势等位基因来自母本Avocet。用不同的颜色表示来自不同群体不同性状的QTL。"

表S3

筛选获得的候选基因信息"

QTL簇
QTL cluster		 基因
Gene		 物理位置
Physical position (bp)		 基因注释
Gene annotation		 表达部位
expression location
C3A TraesCS3A02G027500 14764482-14766910 E3泛素蛋白连接酶 E3 ubiquitin-protein ligase 穗部 Spike
TraesCS3A02G028600 15294546-15298077 E3泛素蛋白连接酶 E3 ubiquitin-protein ligase 籽粒 Grain
TraesCS3A02G028800 15302528-15304140 E3泛素蛋白连接酶 E3 ubiquitin-protein ligase 籽粒 Grain
TraesCS3A02G029300 15477245-15478646 E3泛素蛋白连接酶 E3 ubiquitin-protein ligase 穗部 Spike
TraesCS3A02G029500 15603607-15604989 E3泛素蛋白连接酶 E3 ubiquitin-protein ligase 穗部 Spike
TraesCS3A02G030000 15779481-15780912 E3泛素蛋白连接酶 E3 ubiquitin-protein ligase 穗部 Spike
TraesCS3A02G031400 17257262-17258628 E3泛素蛋白连接酶 E3 ubiquitin-protein ligase 穗部 Spike
TraesCS3A02G032200 17621292-17623525 E3泛素蛋白连接酶 E3 ubiquitin-protein ligase 穗部 Spike
TraesCS3A02G073400 45592602-45603876 E3泛素蛋白连接酶 E3 ubiquitin-protein ligase 籽粒 Grain
TraesCS3A02G077900 50382036-50384496 含NAC结构域的蛋白质 NAC domain-containing protein 籽粒 Grain
TraesCS3A02G079200 51280020-51291788 泛素家族蛋白 Ubiquitin family protein 籽粒 Grain
C6A.2 TraesCS6A02G086800 555455195-55550804 E3泛素蛋白连接酶 E3 ubiquitin-protein ligase 籽粒 Grain
TraesCS6A02G087100 55709408-55710595 多泛素 Polyubiquitin 籽粒 Grain
TraesCS6A02G089000 57680849-57684135 核糖体生物发生调节蛋白样蛋白 Ribosome biogenesis regulatory protein-like protein 籽粒 Grain
TraesCS6A02G108300 77097709-77100361 含NAC结构域的蛋白质，推定 NAC domain-containing protein, putative 叶部 Leaf
TraesCS6A02G112400 81257411-81259132 多泛素 Polyubiquitin 籽粒 Grain

表3

KASP分子标记引物序列"

KASP标记
KASP tag		 引物
Primer		 引物序列
Primer sequence (5′-3′)
CGPC-6A-KASP-1 通用反向引物
Universal reverse primers		 CCT TAT GCG CCA ATC TTT TAT ACA
特异性正向引物
Specific forward primers		 GAA GGT GAC CAA GTT CAT GCT TAG GGA GGC GTC CGT GGA A
GAA GGT CGG AGT CAA CGG ATT TAG GGA GGC GTC CGT GGA C
CGPC-6A-KASP-2 通用反向引物
Universal reverse primers		 GCA TGG TAA TTG CAT ACA CGG G
特异性正向引物
Specific forward primer		 GAA GGT GAC CAA GTT CAT GCT GCA TAA GGG CCA CTC ACA TAA C
GAA GGT CGG AGT CAA CGG ATT GCA TAA GGG CCA CTC ACA TAA T

图4

CGPC-6A-KASP-1 (a)和CGPC-6A-KASP-2 (b)在自然群体的等位基因分离及效应分析 黑色表示空白对照; 图4-a中橙色表示A等位基因, 蓝色表示C等位基因; 图4-b中橙色表示C等位基因, 蓝色表示T等位基因; E8和E9分别表示自然群体在2020-2021年和2021-2022年种植于河南科技大学农学院试验田; *和**分别表示在P < 0.05和P < 0.01水平差异显著。"

表4

前人研究在3A、4D、5D和6A染色体上检测到的籽粒品质相关性状QTL"

性状
Trait		 染色体
Chr.		 QTL 连锁标记
Linkage marker		 物理位置
Physical position (Mb)		 平均表型贡献率
Average PVE (%)		 参考文献
Reference
GPC 6A QGpc.uhw-6A wsnp_Ex_c15268_23489498 535.52-583.20 — [15]
6A QGpc-6A-2 D-1112857-S-2362461 51.40-64.18 10.15 [16]
6A qGPC.XX-6A.1 AX-110368218-AX-110368140 595.06-595.18 4.19 [18]
6A QNGPC.cau-6A — 63.00-83.00 4.07 [44]
6A — BS00046964_51 4.42 [45]
WGC 3A QWGC.hau-3A.1 AX-111674404-AX-109456846 739.32-740.02 1.98 [17]
3A QLWGC.cau-3A.1 wsnp_BE426222A_Ta_2_1 191.53 2.77 [44]
3A QLWGC.cau-3A.2 wsnp_Ku_c44089_51445136 401.71 3.92 [44]
3A QLWGC.cau-3A.3 BobWhite_c28464_814 484.64 3.06 [44]
3A QNWGC.cau-3A BS00078430_51 374.87 3.40 [44]
4D qWGC.XX-4D.1 AX-109282550-AX-110572006 16.43-19.18 12.50 [18]
5D QWGC.hau-5D.1 AX-109684926-AX-109455090 98.26-100.94 3.06 [17]
6A QWGC.hau-6A.1 AX-109418600-AX-110992859 547.81-549.05 4.42 [17]
6A QWGC.hau-6A.2 AX-111504079-AX-109418640 59.69-81.31 6.92 [17]
6A QNWGC.cau-6A — 73.00-83.00 3.98 [44]
6A — BS00046964_51 4.42 — [45]
SV 3A Qsdss-3A AX-110439700-AX-109956297 580.2-649.4 2.47 [46]
5D Qsdss-5D INDEL1489469-INDEL8181683 1.50-8.20 8.11 [46]
6A QSv-6A-2.1 wPt-731556-D-3952327 50.35-50.34 13.34 [16]
6A QSv-6A-2.2 D-1112857-D-1092061 51.40-63.19 11.08 [16]
6A QSV.hau-6A.1 AX-109843320-AX-111089021 8.61-14.39 11.57 [17]
6A QSV.hau-6A.2 AX-109934181-AX-110547197 580.32-598.05 12.18 [17]
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