小麦周8425B×小偃81重组自交系群体千粒重相关性状的QTL定位及单倍型分析

doi:10.3864/j.issn.0578-1752.2022.18.001

摘要/Abstract

摘要：

【目的】周8425B是中国小麦重要骨干亲本之一,小偃81是李振声院士选育的高产、优质、多穗型品种。千粒重是影响小麦产量的重要因素,发掘周8425B和小偃81的千粒重及相关性状的QTL,并分析不同生态区小麦品种所含QTL的单倍型与千粒重的关系,发掘优异单倍型,为不同生态区提高小麦产量及分子辅助育种提供基因型参考。【方法】以周8425B×小偃81衍生的重组自交系群体（F₈）为研究对象,分别于2015和2016年在陕西杨凌（早晚播）进行田间种植,收获后对籽粒相关性状进行测量。利用90K芯片标记构建的高密度遗传图谱对3个环境下的千粒重、籽粒长、籽粒宽和籽粒厚进行QTL定位。同时,针对稳定的主效QTL开发相应的KASP分子标记,并以479份国内外小麦种质组成的自然群体为材料进行分子检测,结合自然群体的千粒重等性状进行关联分析;此外,从479份小麦中挑选出106份含有660K芯片基因型数据的当前黄淮麦区推广的小麦品种,以主效QTL置信区间的差异SNP为基础,进行目标QTL定位区间的单倍型分析,从而判断黄淮麦区中陕西、河南和山东种质材料中的优势类群。【结果】QTL定位结果显示,3个环境下共在8条染色体上检测到22个QTL,表型变异解释率（PVE）范围为4.77%—19.95%,12个位点为主效QTL（PVE>10%）,其中Qkgw.nwafu-6B可能为新QTL。4A、6A、6B、7D染色体上的QTL在多个环境中被检测到,其中,4A和7D染色体处QTL与已报道的相关位点位置相同或接近。6A染色体上的QTL区间包含已知千粒重基因TaGW2-6A,根据TaGW2-6A的分子功能标记检测结果,周8425B和小偃81同时含有TaGW2-6A,此外,基于单倍型分析结果,二者存在于不同的类群中,因此,该位点不同于TaGW2-6A,也可能为新的QTL。单倍型分析结果显示,Qkgw.nwafu-6A总共分为5种单倍型,在不同产区占比超过20%的为6A_h1,其在3个地点千粒重数据均高于其他单倍型;Qkgw.nwafu-6B总共分为8种单倍型,在不同产区占比超过20%的为6B_h6,在河南两点千粒重数据较高,推测含有这两类单倍型的材料为优势类群。此外,针对Qkgw.nwafu-6B开发出共分离的KASP标记,并在479份材料组成的自然群体显著性检测中证明该位点与千粒重表型显著相关。【结论】Qkgw.nwafu-6A和Qkgw.nwafu-6B可能为新的与千粒重相关的主效QTL位点,6A_h1和6B_h6为优势单倍型,开发了一个与Qkgw.nwafu-6B共分离的分子标记KASP_IWA349,可用于分子标记辅助育种。

关键词: 小麦, 千粒重, 90K芯片, QTL定位, 单倍型分析, 标记开发

Abstract:

【Objective】Zhou 8425B is one of the most important founder parents in China and Xiaoyan 81 is an elite cultivar with high yield and good quality. Thousand-grain weight (TGW) is an important factor that affects wheat yield. Identification of QTL associated with grain related traits from Zhou 8425B and Xiaoyan 81, and haplotype analysis of these QTL in wheat cultivars from different ecological regions would be beneficial for yield improvement by molecular marker-assisted selection.【Method】In this study, a RIL population (F₈) derived from Zhou 8425B × Xiaoyan 81 was planted in Yangling during 2015 and 2016 cropping seasons to evaluate grain related traits. Using a high-density genetic map constructed by 90K SNP markers, QTL mapping of thousand-grain weight, grain length, grain width and thickness was performed under three environments. Simultaneously, the KASP markers linked to the identified QTL were developed and molecular detection was carried out among 479 wheat accessions worldwide. Moreover, haplotype analysis of target QTL was performed in 106 current wheat commercial cultivars from Yellow and Huai River Valley Winter Wheat Region selected from 479 wheat accessions.【Result】A total of 22 QTL on 8 chromosomes were detected, and the phenotypic variation explanation (PVE) ranged from 4.77% to 19.95%. Among them, 12 QTL are major QTL (PVE>10%) and Qkgw.nwafu-6B is a new QTL. QTL on chromosomes 4A, 6A, 6B, and 7D were detected in multiple environments, of which, the QTL on chromosomes 4A and 7D are same as previously reported ones. Compared to TaGW2-6A using molecular detection, both Zhou 8425B and Xiaoyan 81 carried the same allele of TaGW2. Based on haplotype result of Qkgw.nwafu-6A, Zhou 8425B and Xiaoyan 81 were placed in different groups. Therefore, Qkgw.nwafu-6A tends to be a new one. Haplotype analysis showed that there were five haplotypes for Qkgw.nwafu-6A and there were eight haplotypes for Qkgw.nwafu-6B. 6A_h1 and 6B_h6 accounted for over 20% in different ecological regions. In addition, a co-segregated KASP marker was developed for Qkgw.nwafu-6B and was significantly associated with the grain weight in the 479 wheat accessions.【Conclusion】Qkgw.nwafu-6A and Qkgw.nwafu-6B are possible new QTL associated with thousand-grain-weight, and 6A_h1 and 6B_h6 are likely favorable haplotypes. A molecular marker KASP_IWA349 co-segregated with Qkgw.nwafu-6B was developed and will be useful for marker assisted selection.

Key words: wheat, thousand-grain-weight, 90K gene chip, QTL mapping, haplotype analysis, development of molecular markers

邹林翰,周新颖,张泽源,蔚睿,袁梦,宋晓朋,简俊涛,张传量,韩德俊,宋全昊. 小麦周8425B×小偃81重组自交系群体千粒重相关性状的QTL定位及单倍型分析[J]. 中国农业科学, 2022, 55(18): 3473-3483.

LinHan ZOU,XinYing ZHOU,ZeYuan ZHANG,Rui YU,Meng YUAN,XiaoPeng SONG,JunTao JIAN,ChuanLiang ZHANG,DeJun HAN,QuanHao SONG. QTL Mapping of Thousand-Grain-Weight and Its Related Traits in Zhou 8425B × Xiaoyan 81 Population and Haplotype Analysis[J]. Scientia Agricultura Sinica, 2022, 55(18): 3473-3483.

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链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2022.18.001

https://www.chinaagrisci.com/CN/Y2022/V55/I18/3473

图/表 8

表1

表2

图1

表3

籽粒相关性状定位结果"

QTL	环境 Environment	染色体 Chromosome	位置 Position (cM)	物理位置 Physical position (Mb)	标记区间Marker interval		LOD	表型贡献率 PVE (%)	加性效应 Add	置信区间<BOLD>C</BOLD>onfidence interval
QTL	环境 Environment	染色体 Chromosome	位置 Position (cM)	物理位置 Physical position (Mb)	左侧Left	右侧Right	LOD	表型贡献率 PVE (%)	加性效应 Add	LeftCI	RightCI
Qgt.nwafu-1B	E2	1B	93.00	564.91—566.60	RAC875_c55891_659	RAC875_c55891_712	2.94	5.52	-0.04	92.50	94.50
Qgw.nwafu-1D	E1	1D	5.50	7.83—7.94	Kukri_c2464_560	CAP7_c3533_280	2.66	11.78	0.13	0.00	11.32
Qgt.nwafu-2B	E1	2B	0.00	1.60—10.17	RAC875_c40246_71	BobWhite_c39433_450	2.84	7.20	0.09	0.00	0.50
Qgt.nwafu-2D	E2	2D	175.00	592.62—641.96	wsnp_JD_c19101_17343310	RAC875_c23815_716	4.02	7.36	0.05	174.50	175.00
Qkgw.nwafu-3A	E3	3A	269.00	700.69—700.74	Kukri_rep_c89509_83	IAAV1134	2.52	4.77	-2.59	268.53	270.78
Qgt.nwafu-3B.1	E2	3B	13.00	6.39—8.97	tplb0043c20_1046	Tdurum_contig34149_219	6.13	11.85	-0.06	12.50	14.50
Qgt.nwafu-3B.2	E2	3B	371.00	759.17—766.64	BS00057988_51	wsnp_CAP7_c5097_2266314	4.04	8.46	0.05	361.54	380.55
Qkgw.nwafu-4A	E3	4A	156.00	625.86—626.02	Kukri_c77040_87	RAC875_c56535_256	2.67	5.08	2.40	153.50	156.50
Qgt.nwafu-4A	E2	4A	156.00	625.86—626.02	Kukri_c77040_87	RAC875_c56535_256	4.56	8.73	0.05	153.50	156.50
Qgw.nwafu-5B	E1	5B	274.00	663.52—667.68	Kukri_c32825_95	RAC875_c29488_56	3.20	12.34	0.12	273.14	275.31
Qkgw.nwafu-6A.1	E3	6A	45.00	100.26—103.15	Ku_c1021_1642	RAC875_c64560_111	3.89	7.81	2.98	44.50	46.50
Qkgw.nwafu-6A.2	E1	6A	47.00	105.17—107.11	BS00044311_51	Kukri_c10611_632	4.73	17.24	3.45	46.50	47.50
Qgt.nwafu-6A	E2	6A	46.00	100.26—127.19	RAC875_c64560_111	Excalibur_c49419_202	8.14	16.98	0.07	45.50	46.50
Qkgw.nwafu-6B	E2	6B	240.00	688.20—690.73	wsnp_BE518379B_Ta_2_2	TA005327-0480	3.30	13.92	3.29	239.87	240.50
Qgl.nwafu-6B	E1	6B	239.00	686.80—690.73	RFL_Contig2206_1694	D_F5XZDLF01B7ECX_54	2.75	12.70	0.11	238.61	240.40
Qgt.nwafu-7A	E1	7A	333.00	679.90—680.34	Excalibur_c17899_352	wsnp_Ku_rep_c103889_90513365	3.13	8.08	0.09	332.87	333.91
Qkgw.nwafu-7B	E3	7B	36.00		GENE-3775_392	RAC875_c43810_265	8.60	14.55	-20.54	35.50	38.50
Qgw.nwafu-7B	E2	7B	175.00	684.42—710.11	Excalibur_c6330_1158	Kukri_rep_c68594_530	33.04	10.57	-2.47	173.60	178.90
Qgl.nwafu-7B.1	E2	7B	175.00	684.42—710.11	Excalibur_c6330_1158	Kukri_rep_c68594_530	40.38	12.34	-4.40	173.60	178.90
Qgl.nwafu-7B.2	E1	7B	0.00	1.25—1.26	Excalibur_c53111_144	Kukri_rep_c71778_644	3.96	17.26	-0.12	0.00	2.50
Qgt.nwafu-7D.1	E2	7D	174.00	548.80—553.22	Ku_c32426_324	RAC875_rep_c106588_205	4.88	9.19	-0.05	172.99	174.56
Qgt.nwafu-7D.2	E1	7D	173.00	548.80—553.22	Ku_c32426_324	RAC875_rep_c106588_205	7.08	19.95	-0.14	172.99	174.56

表3

图2

图3

图4

图5

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引物 Primer	引物序列 Primer sequence (5′-3′)
TaGW2-6A-KASP	F：GAAGGTGACCAAGTTCATGCTTCCCGCTCCAGCTATCTGGTGAAC H：GAAGGTCGGAGTCAACGGATTTCCCGCTCCAGCTATCTGGTGAAA C：TTCCCAGTCTTTGACATGTTCCGCC
1B/1R-KASP	F：GAAGGTGACCAAGTTCATGCTGGAGCAGGTCCAGATCGCG H：GAAGGTCGGAGTCAACGGATTCGGAGCAGGTCCAGATCGCA C：GAAGCTCCGGTAGATGGAGGCTA
Qkgw.nwafu-6B (KASP_IWA349)	F：GAAGGTGACCAAGTTCATGCTGGGCACAAAAATTAACTGGCCTA H：GAAGGTCGGAGTCAACGGATTGGGCACAAAAATTAACTGGCCTG C：GTCCCCCACCACACAGCTATCCT

环境 Environment	亲本Parent		最小值 Min	最大值 Max	均值 Mean	标准差 SE	偏度 Skewness	峰度 Kurtosis	遗传力 Heritability
环境 Environment	Z8425B	XY81	最小值 Min	最大值 Max	均值 Mean	标准差 SE	偏度 Skewness	峰度 Kurtosis	遗传力 Heritability
E1	54.55	40.75	19.95	61.25	40.63	9.84	-0.12	-0.69	0.74
E2	61.36	42.66	30.81	61.36	49.33	6.01	-0.32	0.30
E3	60.67	43.43	25.66	63.64	49.00	6.64	-0.42	0.31