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作物学报 ›› 2021, Vol. 47 ›› Issue (11): 2091-2098.doi: 10.3724/SP.J.1006.2021.01083

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

黄淮麦区小麦粒重基因等位变异的分子鉴定及育种应用

张福彦1(), 程仲杰1, 陈晓杰1, 王嘉欢1, 陈锋2, 范家霖1, 张建伟1,*(), 杨保安1,*()   

  1. 1河南省科学院同位素研究所有限责任公司 / 河南省核农学重点实验室, 河南郑州 450015
    2河南农业大学农学院 / 省部共建小麦玉米作物学国家重点实验室, 河南郑州 450002
  • 收稿日期:2020-11-03 接受日期:2021-03-19 出版日期:2021-11-12 网络出版日期:2021-04-14
  • 通讯作者: 张建伟,杨保安
  • 作者简介:E-mail: zhangfuyan704@163.com
  • 基金资助:
    河南省小麦产业技术体系建设专项(Z2010-01-04);河南省重点研发与推广专项(科技攻关)(202102110028);河南省科学院基本科研业务专项(190604015);省部共建小麦玉米作物学国家重点实验室开放课题(30500884)

Molecular identification and breeding application of allelic variation of grain weight gene in wheat from the Yellow-Huai-River Valley

ZHANG Fu-Yan1(), CHENG Zhong-Jie1, CHEN Xiao-Jie1, WANG Jia-Huan1, CHEN Feng2, FAN Jia-Lin1, ZHANG Jian-Wei1,*(), YANG Bao-An1,*()   

  1. 1Isotope Institute Co., Ltd, Henan Academy of Sciences / Henan Key Laboratory of Nuclear Agricultural Sciences, Zhengzhou, 450015, Henan, China
    2Agronomy College, Henan Agricultural University / National Key Laboratory of Wheat and Maize Crop Science in Henan Province, Zhengzhou, 450002, Henan, China
  • Received:2020-11-03 Accepted:2021-03-19 Published:2021-11-12 Published online:2021-04-14
  • Contact: ZHANG Jian-Wei,YANG Bao-An
  • Supported by:
    Henan Wheat Research System(Z2010-01-04);Key Research and Development and Promotion Program of Henan Province(202102110028);Basic Scientific Research Project of Henan Academy of Sciences(190604015);National Key Laboratory of Wheat and Maize Crop Science(30500884)

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摘要：

采用特异性引物PCR扩增方法对183份黄淮海麦区的小麦品种(系)的粒重基因TaCwi-A1、TaGw8-B1和TaGS-D1的等位变异进行分子鉴定, 并结合2016—2017和2017—2018年度的千粒重表型数据, 分析不同等位变异类型对小麦粒重的影响, 从而找出优势基因型组合。结果表明, 不同年份间参试品种(系)的千粒重差异达到极显著水平(P<0.01); TaCwi-A1位点上发现TaCwi-A1a和TaCwi-A1b两种等位变异, 其分布频率分别为66.7%和33.3%; TaGw8-B1位点上TaGw8-B1a等位变异分布频率较高, 为94.5%, 而TaGw8-B1b等位变异分布频率极低, 仅为5.5%; TaGS-D1位点上发现TaGS-D1a和TaGS-D1b两种等位变异, 其分布频率分别为79.8%和20.2%。不同等位变异组合的品种千粒重存在显著差异(P<0.05), 其中具有3个高千粒重等位变异组合TaCwi-A1a/TaGS-D1a/TaGw8-B1a品种的平均千粒重最高, 与具有TaCwi-A1b/TaGS-D1a/TaGw8-B1a品种的平均千粒重差异不显著, 但是显著高于其他组合(P<0.05)。TaCwi-A1a/TaGS-D1a/TaGw8-B1b基因型组合小麦品种(系)的平均千粒重最低。TaCwi-A1、TaGw8-B1和TaGS-D1位点上的不同等位变异均会导致小麦千粒重的显著变化, 其中TaGw8-B1和TaGS-D1位点上的等位变异对小麦粒重的影响更为重要。在参试材料中没有发现具有3个低千粒重等位变异组合TaCwi-A1b/TaGS-D1b/TaGw8-B1b的品种, 在7种不同等位变异组合中, 具有3个高千粒重等位变异组合TaCwi-A1a/TaGS-D1a/TaGw8-B1a品种的平均千粒重最高, 是优势基因型组合。

关键词: 小麦, 粒重基因, 功能标记, 等位变异, 千粒重

Abstract:

Grain weight is one of the most important yield traits in wheat. To accelerate the application process of dominant allelic variation of grain weight gene in wheat breeding, the allelic variation of grain weight gene was identified by using functional markers and the combinations of dominant grain weight genotypes were investigated the allelic variations of grain weight genes TaCwi-A1, TaGw8-B1, and TaGS-D1 in 183 wheat varieties (lines) from the Yellow-Huai-River Valley were identified by PCR amplification with specific primers. To identify the dominant genotype combinations, the effects of different allelic variation genotypes on wheat grain weight were studied by combining the phenotypic data of the 1000-grain weight (TGW) from 2016-2017 and 2017-2018. The results showed that the difference of TGW between different years was highly significant at P < 0.01. Two alleles, TaCwi-A1a and TaCwi-A1b, were detected at TaCwi-A1 locus, with the frequencies of 66.7% and 33.3%, respectively. The frequency of TaGw8-B1a allele on TaGw8-B1 locus was up to 94.5%, while the frequency of TaGw8-B1b allele was only 5.5%. In addition, two alleles, TaGS-D1a and TaGS-D1b, were found in TaGS-D1 locus, and their frequencies were 79.8% and 20.2%, respectively. Further results indicated that there were significant differences in TGW among different allelic variation combinations at P < 0.05. Among them, the average TGW of varieties with TaCwi-A1a/TaGS-D1a/TaGw8-B1a genotype was the highest. There was not significant difference in TGW between TaCwi-A1a/TaGS-D1a/TaGw8-B1a and TaCwi-A1b/ TaGS-D1a/TaGw8-B1a genotype, but it was significantly higher than other genotypes at P < 0.05. The average TGW of TaCwi-A1a/TaGS-D1a/TaGw8-B1b genotype was the lowest. The allelic variations at the loci of TaCwi-A1, TaGw8-B1, and TaGS-D1 all led to significant changes in TGW, and the allelic variations of TaGw8-B1 and TaGS-D1 loci were more important to TGW in wheat. There were no varieties with three low TGW allelic variation combinations TaCwi-A1b/TaGS-D1b/TaGw8-B1b in the tested materials. Among the seven different allele combinations, the average TGW of three high TGW allelic variation combinations TaCwi-A1a/TaGS-D1a/TaGw8-B1a was the highest, which was the dominant genotype combination.

Key words: wheat, grain weight gene, functional marker, allelic variation, TGW

表1

检测目的基因等位变异的功能标记及其扩增信息"

基因
Genes		 等位变异
Alleles		 标记引物
Mark primer		 引物序列
Primer sequences (5′-3′)		 退火温度
Annealing
temperature (℃)		 扩增片段
Targeted
fragment (bp)		 参考文献
Reference
TaGS-D1 TaGS-D1a GS7D-F AACTTAGGGAGCGAAAACAA 58 562 [7]
TaGS-D1b GS7D-R CACCAAGACTGGAGATGAAA 522 [7]
TaCwi-A1 TaCwi-A1a CWI22-F GGTGATGAGTTCATGGTTAAT 56 402 [5]
CWI22-R AGAAGCCCAACATTAAATCAAC
TaCwi-A1b CWI21-F GTGGTGATGAGTTCATGGTTAAG 56 404 [5]
CWI21-R AGAAGCCCAACATTAAATCAAC
TaGw8-B1 TaGW8-B1a TaGW8-7B-F CGCTCATCCATTCCTTCATCG 60 1097 [15]
TaGW8-B1b TaGW8-7B-R GCTATATGGGTTGGTGTCGC 1373 [15]

表2

不同年份中供试材料的千粒重方差分析"

年份
Year		 千粒重
Thousand-grain weight (g)		 变幅
Range		 方差
Variance		 标准差
Standard deviation		 变异系数
Coefficient of variation (%)
2016-2017 44.11 34.50-53.35 14.00** 3.74 8.48
2017-2018 46.29 34.20-58.45 16.42** 4.05 8.75

图1

标记CWI21 (A)和CWI22 (B)在部分小麦品种中扩增结果 M: DL2000; 1: 石4185; 2: 郑品麦22; 3: 商麦167; 4: 郑育麦16; 5: 豫丰11; 6: 中育1220; 7: 涡麦66; 8: 泛育麦17; 9: 豫农186; 10: 赛德麦1号; 11: 郑麦369; 12: 淮麦19。"

图2

TaGW8-7B标记在部分小麦品种中扩增结果 1: 郑品麦22; 2: 商麦167; 3: 温麦6号; 4: 郑麦518; 5: 豫农202; 6: 中新16; 7: 豫丰11; 8: 皖科06725; 9: 豫麦2号; 10: 冀麦38; 11: 石84-7111; 12: 丰德存麦5号; 13: 良星99; 14: 洛麦28; 15: 淮麦19; 16: 百农3217; M: DL2000。"

图3

GS7D标记检测部分小麦品种 1: 豫农186; 2: 赛德麦1号; 3: 鲁原502; 4: 郑麦366; 5: 中育9302; 6: 龙科1221; 7: 濮兴0369; 8: 豫丰11; 9: 农大2011; 10: 郑品麦8号; 11: 高麦6号; 12: 丰德存麦5号; M: DL2000。"

表3

粒重基因的不同等位变异对小麦千粒重影响"

基因位点
Gene locus		 等位变异
Allele		 品种数量
Number of varieties		 频率
Frequency (%)		 2016年千粒重
TGW in 2016 (g)		 2017年千粒重
TGW in 2017 (g)		 均值
Mean (g)
TaCwi-A1 TaCwi-A1a 122 66.7 44.46* 46.61* 45.54*
TaCwi-A1b 61 33.3 43.40 45.65 44.52
TaGw8-B1 TaGw8-B1a 173 94.5 44.27** 46.51** 45.39**
TaGw8-B1b 10 5.5 41.34 42.46 41.90
TaGS-D1 TaGS-D1a 146 79.8 44.44** 46.71** 45.58**
TaGS-D1b 37 20.2 42.79 44.63 43.71

表4

TaCwi-A1、TaGS-D1和TaGw8-B1位点上不同等位变异组合对小麦千粒重的影响"

等位变异组合
Allelic variation combination		 品种数量
Number of varieties		 频率
Frequency (%)		 表型†
Phenotype†		 2016年千粒重
2016 TGW (g)		 2017年千粒重
2017 TGW (g)		 均值‡
Mean (g)‡
TaCwi-A1a/TaGS-D1a/TaGw8-B1a 96 52.5 H 43.67 45.71 44.69 a
TaCwi-A1b/TaGS-D1a/TaGw8-B1a 43 23.5 H 43.28 45.73 44.50 a
TaCwi-A1b/TaGS-D1b/TaGw8-B1a 13 7.1 M 41.56 43.53 42.54 b
TaCwi-A1a/TaGS-D1b/TaGw8-B1a 21 11.5 M 41.61 43.39 42.50 b
TaCwi-A1b/TaGS-D1a/TaGw8-B1b 5 2.7 ML 41.69 42.90 42.30 b
TaCwi-A1a/TaGS-D1b/TaGw8-B1b 3 1.6 ML 41.13 42.92 42.03 bc
TaCwi-A1a/TaGS-D1a/TaGw8-B1b 2 1.1 L 41.18 41.60 41.39 c
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