大豆引进种质抗胞囊线虫病、抗花叶病毒病和耐盐基因型鉴定及优异等位基因聚合种质筛选

doi:10.3724/SP.J.1006.2018.01263

作物学报 ›› 2018, Vol. 44 ›› Issue (9): 1263-1273.doi: 10.3724/SP.J.1006.2018.01263

• 研究论文 • 下一篇

大豆引进种质抗胞囊线虫病、抗花叶病毒病和耐盐基因型鉴定及优异等位基因聚合种质筛选

叶俊华^1,²(),杨启台^2,³,刘章雄²,郭勇²,李英慧²,关荣霞²,邱丽娟^2,^*

1 东北农业大学农学院, 黑龙江哈尔滨 150030
2 国家农作物基因资源与遗传改良重大科学工程 / 农业部种质资源利用重点实验室 / 中国农业科学院作物科学研究所, 北京 100081
3 蚌埠医学院生物科学系, 安徽蚌埠 233000

收稿日期:2018-03-04 接受日期:2018-06-12 出版日期:2018-09-10 网络出版日期:2018-07-02
通讯作者: 邱丽娟
基金资助:
本研究由国家重点研发计划项目(2016 YFD0100304);本研究由国家重点研发计划项目(2016YFD0100304);中国农业科学院科技创新工程项目资助

Genotyping of SCN, SMV Resistance, Salinity Tolerance and Screening of Pyramiding Favorable Alleles in Introduced Soybean Accessions

Jun-Hua YE^1,²(),Qi-Tai YANG^2,³,Zhang-Xiong LIU²,Yong GUO²,Ying-Hui LI²,Rong-Xia GUAN²,Li-Juan QIU^2,^*

1 College of Agriculture, Northeast Agricultural University, Harbin 150030, Heilongjiang, China
2 National Key Facility for Gene Resources and Genetic Improvement / Key Laboratory of Crop Germplasm Utilization, Ministry of Agriculture/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
3 Department of Biotechnology, Bengbu Medical College, Bengbu 233000, Anhui, China

Received:2018-03-04 Accepted:2018-06-12 Published:2018-09-10 Published online:2018-07-02
Contact: Li-Juan QIU
Supported by:
This study was supported by the National Key R&D Program(2016 YFD0100304);This study was supported by the National Key R&D Program(2016YFD0100304);the Agricultural Science and Technology Innovation Program of CAAS

摘要/Abstract

摘要：

我国从美国、俄罗斯、日本等26个国家或地区共引进大豆种质3218份, 仅对部分种质进行了大豆胞囊线虫病(Soybean cyst nematode, SCN)、大豆花叶病毒病(Soybean mosaic virus, SMV)和盐敏感性的抗性鉴定, 但基因型的系统分析尚未见报道。本研究针对大豆抗胞囊线虫病3个基因(rhg1、Rhg4、SCN3-11)和耐盐基因(GmSALT3)开发KASP标记5个, 结合与大豆花叶病毒抗性相关的1个SCAR标记(SCN11), 对1489份大豆引进种质进行基因型鉴定。结果表明, 具有优异等位基因的种质共1084份; 携带3个位点优异等位基因的种质19份, 包括抗胞囊线虫病3个位点(rhg1、Rhg4、SCN3-11)叠加(Peking型)种质3份, 聚合抗胞囊线虫病基因和抗花叶病毒病标记7份, 聚合抗胞囊线虫病和耐盐基因2份, 聚合抗胞囊线虫病、抗花叶病毒病和耐盐基因7份; 携带4个位点优异等位基因的种质9份, 包括聚合抗胞囊线虫病基因和抗花叶病毒病标记6份, 聚合抗胞囊线虫病和耐盐基因2份, 聚合抗胞囊线虫病、抗花叶病毒病和耐盐7份; 携带5个位点优异等位基因8份, 聚合了抗胞囊线虫病、抗花叶病毒病和耐盐优异等位变异。在这些携带优异等位变异的种质中, 44份已由前人证明具有相应的抗性。携带3个或3个以上优异等位基因的36份种质中, 有52.78%种质的一种或两种特性已被报道。在不携带抗性优异等位变异的种质中, 93份已证明有耐盐性或对SMV3号株系抗性, 这些种质可能存在新的抗性(等位)基因。本研究利用高通量分子标记筛选出的携带抗病、抗逆优异等位基因的种质为我国大豆资源表型鉴定、抗源的快速筛选及利用提供理论依据和新思路。

关键词: 大豆种质, 分子标记, KASP技术, 基因型鉴定

Abstract:

China has introduced 3218 soybean accessions from 26 countries such as the United States, Russia and Japan, and some of them have been carried out soybean cyst nematode (SCN), soybean mosaic virus (SMV) and salinity tolerance resistance evaluation. However, the systematic genotyping of these accessions has not been reported yet. In this study, five robust functional markers have been developed for KASP assays, three SCN loci (rhg1, Rhg4, SCN3-11) and salinity tolerance gene (GmSALT3) included. A total of 1489 introduced soybean accessions were genotyped by these markers with high-throughput assay as well as a SCAR marker (SCN11) which is related to soybean mosaic virus resistance. The results showed that there were 1084 accessions detected with favorable alleles; where accessions detected with resistant alleles at three loci were as much as 19, including three pyramiding SCN genes (rhg1, Rhg4, SCN3-11) which were Peking type and seven pyramiding SCN and SMV , two pyramiding SCN and salinity favorable alleles, as well as seven pyramiding SCN, SMV and salinity favorable alleles; and accessions detected with four favorable alleles were as much as nine accessions, including six pyramiding SCN and SMV resistance alleles, one accession detected with SCN and salinity tolerance and two detected with SCN, SMV and salinity favorable alleles, eight detected with all the favorable alleles in this study. Among the elite accessions mentioned above, it has been proved that 44 accessions resistant to SCN, SMV-3 or tolerant to salinity. Among the 36 accessions with three or more favorable alleles, 52.78% had been reported of one or two characteristics. Among the accessions without resistance or tolerance alleles, it has been reported that 93 accessions were tolerant to salinity or resistant to SMV-3, where new resistance or tolerance genes could be found. Screening out the accessions with high-throughput SNP detection assays for resistance and tolerance alleles in soybean provides information for their further phenotyping, screening and breeding.

Key words: soybean germplasm, molecular markers, KASP, genotyping

叶俊华,杨启台,刘章雄,郭勇,李英慧,关荣霞,邱丽娟. 大豆引进种质抗胞囊线虫病、抗花叶病毒病和耐盐基因型鉴定及优异等位基因聚合种质筛选[J]. 作物学报, 2018, 44(9): 1263-1273.

Jun-Hua YE,Qi-Tai YANG,Zhang-Xiong LIU,Yong GUO,Ying-Hui LI,Rong-Xia GUAN,Li-Juan QIU. Genotyping of SCN, SMV Resistance, Salinity Tolerance and Screening of Pyramiding Favorable Alleles in Introduced Soybean Accessions[J]. Acta Agronomica Sinica, 2018, 44(9): 1263-1273.

图/表 6

表1

图1

表2

图2

表3

Table 4

Thirty-six accessions with three or more resistance alleles"

携带抗性位点数 Number of resistant loci		统一编号 Accession No.	名称 Name		优异等位基因 Favorable alleles		抗性表型 Resistance
3个位点 Three loci		WDD00828	Centennial		rhg1, Rhg4, SCN3-11		SCN^*
		WDD01994	M87-1569		rhg1, Rhg4, SCN3-11		SCN^*
		WDD00691	AGS272⁺		rhg1, Rhg4, SCN3-11		SCN^*
		WDD03200	LINE272H Xuan^#		Rhg4, SCN3-11, SALT3(H1)		SCN, Salt
		WDD00289	T221		rhg1, Rhg4, SALT3(H1)		SCN^*, Salt
		WDD00858	N80-2317^#		rhg1, SCN3-11, SCN11		SCN, SMV
		WDD00859	N80-50232^#		rhg1, SCN3-11, SCN11		SCN, SMV
		WDD00666	AGS65⁺		rhg1, SCN3-11, SCN11		SCN, SMV
携带抗性位点数 Number of resistant loci	统一编号 Accession No.			名称 Name		优异等位基因 Favorable alleles		抗性表型 Resistance
	WDD00683			AGS175⁺		rhg1, SCN3-11, SCN11		SCN, SMV
	WDD02019			9234		Rhg4, SCN3-11, SCN11		SCN^*, SMV
	WDD01607			Delsoy4900		rhg1, Rhg4, SCN11		SCN^*, SMV
	WDD03086			G04-Ben229lR-M		rhg1, Rhg4, SCN11		SCN, SMV
	WDD02211			H6255		rhg1, SALT3(H1), SCN11		SCN, Salt, SMV
	WDD03205			M017-1^#		rhg1, SALT3(H1), SCN11		SCN, Salt, SMV
	WDD00916			SRE-D-14A^#		Rhg4, SALT3(H1), SCN11		SCN, Salt, SMV
	WDD01640			Lamar		Rhg4, SALT3(H1), SCN11		SCN^*, Salt, SMV
	WDD00334			Clark-G		SCN3-11, SALT3(H1), SCN11		SCN, Salt, SMV
	WDD02252			P951341		SCN3-11, SALT3(H1), SCN11		SCN, Salt, SMV
	WDD00335			Clark-S		SCN3-11, SALT3(H1), SCN11		SCN, Salt, SMV
4个位点 Four loci	WDD02102			PI90763		rhg1, Rhg4, SCN3-11, Salt3(H1)		SCN^*, Salt
	WDD00926			TGX814-26D^#		rhg1, Rhg4, SCN3-11, SCN11		SCN, SMV
	WDD01583			Newton		rhg1, Rhg4, SCN3-11, SCN11		SCN^*, SMV
	WDD00595			Custer		rhg1, Rhg4, SCN3-11, SCN11		SCN^*, SMV
	WDD00602			Franklin		rhg1, Rhg4, SCN3-11, SCN11		SCN^*, SMV
	WDD02015			TBD		rhg1, Rhg4, SCN3-11, SCN11		SCN, SMV
	WDD01538					rhg1, Rhg4, SCN3-11, SCN11		SCN, SMV
	WDD03083			S01-9364		rhg1, SCN3-11, SALT3(H1), SCN11		SCN^, Salt^, SMV
	WDD00774			S-10-1		Rhg4, SCN3-11, SALT3(H1), SCN11		SCN, Salt, SMV
5个位点 Five loci	WDD03084			S01-9391		rhg1, Rhg4, SCN3-11, SALT3(H1), SCN11		SCN^*, Salt, SMV
	WDD01632			Bryan		rhg1, Rhg4, SCN3-11, SALT3(H1), SCN11		SCN^*, Salt, SMV
	WDD01614			Rhodes		rhg1, Rhg4, SCN3-11, SALT3(H1), SCN11		SCN^*, Salt, SMV
	WDD00721			Forrest		rhg1, Rhg4, SCN3-11, SALT3(H1), SCN11		SCN^*, Salt, SMV
	WDD01971			D83-3349		rhg1, Rhg4, SCN3-11, SALT3(H1), SCN11		SCN^*, Salt, SMV
	WDD00661			A-66 Jia⁺		rhg1, Rhg4, SCN3-11, SALT3(H1), SCN11		SCN, Salt, SMV
	WDD02989			Pin-din-guan		rhg1, Rhg4, SCN3-11, SALT3(H1), SCN11		SCN^, Salt^, SMV
	WDD03094			S99-2281		rhg1, Rhg4, SCN3-11, SALT3(H1), SCN11		SCN^*, Salt, SMV

Table 4

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性状 Trait	标记 Marker	标记类型 Type	基因 Gene	引物序列信息 Primer sequence (5°-3°)	等位基因 Allele	参考文献 Reference
大豆胞囊线虫抗性 SCN resistance
	Rhg4-389	KASP	Glyma08g11490	Rhg4-COM: CTACACCGCCGTCCTCAAC		[17]
	Rhg4-389	KASP	Glyma08g11490	Rhg4-FAM: GAGGTGGCCGCCGGAGC	G/G
				Rhg4-HEX: GAGGTGGCCGCCGGAGG	C/C
	rhg1	KASP	Glyma18g02590	rhg1-COM: CTGGCATCTGCCAACTCTGTAAAGA		[18]
			Glyma18g02590	rhg1-FAM: TTCTAATGCATTGGTTATAGCAACAACG	C/C	[18]
				rhg1-HEX: TTCTAATGCATTGGTTATAGCAACAACC	G/G
	SCN3-11	KASP	Glyma11g35820	SCN3-11-COM: CAGAAGTCGATTGAGATTTACGAAGAGATA		[9]
				SCN3-11-FAM: ATTATTGTTGAGGGATTGGCGAGC	C/C
				SCN3-11-HEX: AAATTATTGTTGAGGGATTGGCGAGT	T/T
大豆耐盐性 Soybean salinity tolerance
	SALT3	KASP	Glyma03g32900	GmSALT3-COM: GATCATTGGATGTAATTGGGTGGAGAA		[14]
			Glyma03g32900	GmSALT3-FAM: GATACCAGCAAATATTAAATGTGTGTTTTT	T/T
				GmSALT3-HEX: GATACCAGCAAATATTAAATGTGTGTTTTA	-/-
	Ncl-5	KASP	Glyma03g32900	Ncl-5-COM: AGGTACTTACCCTTATGAAGAAAACA		[14]
			Glyma03g32900	Ncl-5-FAM: AGAACTCGTATTTTATTTTGGTTGAC	G/G
				Ncl-5-HEX: AGAACTCGTATTTTATTTTGGTTGAT	A/A
大豆花叶病毒抗性 SMV resistance
	SCN11	SCAR	—	SCN11-F: TTCACGTGGCCCTCCTATC	—	[12]
				SCN11-R: CGCCGCAAACTCACAGGAC	—

性状 Trait	标记名称 Marker	种质有效鉴定数目 Accessions effectively identified No.	等位基因 Allele	对应表型¹⁾ Corresponding phenotype ¹⁾	种质Accessions
性状 Trait	标记名称 Marker	种质有效鉴定数目 Accessions effectively identified No.	等位基因 Allele	对应表型¹⁾ Corresponding phenotype ¹⁾	数量 Size (No.)	比例 Ratio (%)
大豆胞囊线虫抗性 SCN resistance	rhg1	1383	G/G	R	31	2.24
			C/C	S	1352	97.76
	Rhg4-389	1480	G/G	R	37	2.50
			C/C	S	1441	97.37
	SCN3-11	1356	T/T	R	66	4.87
			C/C	S	1290	95.13
大豆耐盐性 Soybean salinity tolerance	SALT3	1380	-/-	—	236	17.10
			T/T	S	1127	81.67
	Ncl-5	1479	G/G	—	1384	93.58
			A/A	S	85	5.75
	SALT3, Ncl-5	1380	SALT3(H1)	T	235	17.03
大豆花叶病毒抗性 SMV resistance	SCN11	1489	—	R	960	64.47
大豆花叶病毒抗性 SMV resistance			—	S	529	35.53

	抗性特性 Resistance	优异基因型 Favorable allele	携带优异等位基因的种质 Germplasm with favorable alleles
	抗性特性 Resistance	优异基因型 Favorable allele	数目 Size (No.)	比例 Ratio (%)	代表性种质 Representative germplasm
单位点 Single locus			901	60.51
	SCN	SCN3-11	15	1.01	中特1号 Zhongte 1
	SMV	SCN11	796	53.46	Provar
	Salt	SALT3(H1)	90	6.04	Harosoy 71
双位点 Two loci			147	9.87
	SCN	rhg1/Rhg4	1	0.07	M044
	SCN	Rhg4/SCN3-11	2	0.13	PR 149-3
	SCN, SMV	rhg1/SCN11	2	0.13	PI576145
	SCN, SMV	Rhg4/SCN11	5	0.34	Mack
	SCN, SMV	SCN3-11/SCN11	12	0.81	CA50
	SCN, Salt	SCN3-11/SALT3(H1)	8	0.54	PI590579
	SCN, Salt	Rhg4/SALT3(H1)	2	0.13	D85-10412
	SMV, Salt	SALT3(H1)/SCN11	115	7.72	Lee 68
3个位点 Three loci			19	1.28
	SCN	rhg1/Rhg4/SCN3-11	3	0.20	Centennial
	SCN, SMV	rhg1/Rhg4/SCN11	2	0.13	Delsoy4900
	SCN, SMV	rhg1/SCN3-11/SCN11	4	0.27	AGS175
	SCN, SMV	Rhg4/SCN3-11/SCN11	1	0.07	9234
	SCN, Salt	rhg1/Rhg4/SALT3(H1)	1	0.07	T221
	SCN, Salt	Rhg4/SCN3-11/SALT3(H1)	1	0.07	LINE272H选 LINE272H Xuan
	SCN, SMV, Salt	rhg1/SALT3(H1)/SCN11	2	0.13	H6255RR
	SCN, SMV, Salt	Rhg4/SALT3(H1)/SCN11	2	0.13	Lamar
	SCN, SMV, Salt	SCN3-11/SALT3(H1)/SCN11	3	0.20	P951341RR
4个位点 Four loci			9	0.60
	SCN, SMV	rhg1/Rhg4/SCN3-11/SCN11	6	0.40	Newton
	SCN, Salt	rhg1/Rhg4/SCN3-11/SALT3(H1)	1	0.07	PI90763
	SCN, SMV, Salt	rhg1/SCN3-11/SALT3(H1)/SCN11	1	0.07	S01-9364
	SCN, SMV, Salt	Rhg4/SCN3-11/SALT3(H1)/SCN11	1	0.07	S-10-1
5个位点 Five loci			8	0.54
	SCN, SMV, Salt	rhg1/Rhg4/SCN3-11/SALT3(H1)/SCN11	8	0.54	Pin-din-guan

大豆引进种质抗胞囊线虫病、抗花叶病毒病和耐盐基因型鉴定及优异等位基因聚合种质筛选

Genotyping of SCN, SMV Resistance, Salinity Tolerance and Screening of Pyramiding Favorable Alleles in Introduced Soybean Accessions

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