Development and Application of High Efficiency KASP Markers of Wheat Fusarium Head Blight Resistance Genes Based on High Throughput Chip

doi:10.7668/hbnxb.20195775

Abstract

Abstract:

Fusarium head blight(FHB)is a devastating fungal disease that seriously threatens the safety of wheat production.Marker-assisted selection(MAS)and pyramiding of resistance genes represent efficient strategies for FHB-resistant breeding.To establish a high-throughput screening system for FHB resistance genes and enhance wheat resistance in Sichuan Province,we performed genome-wide genotyping using a 100K SNP array on 14 Sichuan wheat varieties(lines)along with three FHB-resistant genetic materials.Based on the reported genetic linkage intervals of major FHB resistance genes(Fhb2,Fhb4,Fhb5),we identified SNPs co-segregating with Fhb5 or linked to Fhb2,Fhb4,and subsequently developed kompetitive allele-specific PCR(KASP)markers.Results showed that the genetic relationship of 17 wheat varieties(lines)could be clustered into two major groups:two northern wheat-derived resistant materials(NMAS070 and NMAS069)formed an independent cluster distinct from the Sichuan varieties(lines)while the remaining 15 varieties(lines)were clustered together and subdivided into two subgroups.Functional gene profiling revealed FHB-resistant parents carried superior resistance loci,whereas agronomic parents harbored favorable alleles for yield and quality traits.Through SNP screening,we identified 8 critical SNPs within the linkage intervals of Fhb2,Fhb4 and the co-segregation region of Fhb5.These SNPs enabled the successful development of 4,2,and 2 high-specificity KASP marker systems for Fhb2,Fhb4 and Fhb5,respectively.Validation experiments confirmed all KASP markers achieved precise genotyping and were effectively implemented in molecular breeding for FHB-resistance.This study established a high-efficiency KASP marker system for Fhb2,Fhb4 and Fhb5,providing a robust technical platform for improving FHB resistance breeding of wheat varieties in Southwest China.

Key words: Wheat, Fusarium head blight(FHB), KASP marker, Marker selection, FHB-resistance breeding

CLC Number:

DENG Qingyan, LUO Jiangtao, ZHENG Jianmin, PU Zongjun. Development and Application of High Efficiency KASP Markers of Wheat Fusarium Head Blight Resistance Genes Based on High Throughput Chip[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(4): 175-184. doi: 10.7668/hbnxb.20195775.

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Figures/Tables 8

References 38

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性状 Trait type	基因/QTL Gene/QTL	NMAS069	NMAS070	NMAS071	川麦625 Chuanmai 625	21pin3-9	川麦704 Chuanmai 704	21pin4-3	21pin6-5	川麦614 Chuanmai 614	川麦618 Chuanmai 618	川麦620 Chuanmai 620	川麦623 Chuanmai 623	川麦702 Chuanmai 702	川麦914 Chuanmai 914	绵麦60 Mianmai 60	内麦175 Neimai 175	蜀麦1925 Shumai 1925
赤霉病	Fhb1^†	+	+	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Fusarium	Fhb2^†	+	-	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-
head blight	Fhb4^†	+	+	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Fhb5^†	-	+	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Qfhb.caas-3BL	-	-	-	-	+	+	-	-	+	+	-	+	+	+	-	+	-
	QFhb.hbaas-5AS	-	-	-	+	+	-	+	-	-	-	+	-	-	-	-	+	-
	QFhb.hbaas-5AL	-	-	+	-	-	-	+	-	+	-	-	-	+	+	-	-	-
	QFhb.caas-5AL	+	-	-	+	-	+	-	+	+	+	+	+	+	+	+	-	+
千粒质量	TaSus2-2B	高	低	高	高	高	低	高	高	低	低	低	低	低	高	高	高	高
1000-grain	Tabas1	高	低	低	高	低	高	低	高	高	高	高	高	高	高	高	低	高
weight	TaGS5-A1	杂合	高	低	高	低	高	高	高	高	高	高	高	高	高	高	低	高
	TaGW2-6A	高	高	高	高	高	高	高	NA	高	高	高	高	高	高	NA	NA	高
	TaCwi-A1	低	低	高	高	低	高	高	高	高	高	高	高	高	高	高	高	高
	TaT6P	低	低	低	低	低	高	高	高	高	高	低	高	低	低	低	低	低
品质	Glu-B3h	-	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+
Quality	TaLCYE-B1	低	高	低	高	低	高	低	高	高	高	高	高	高	高	高	高	低
株高	Rht-B1	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	+	-	NA
Plant	Rht-D1	+	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA
height	RHT-8	+	+	-	+	-	-	+	-	-	-	+	-	-	-	-	-	+
	Rht24_AP2	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+
	αPht-2D	-	-	+	+	+	-	+	+	+	-	+	-	+	+	-	+	-

性状 Trait type	基因/QTL Gene/QTL	NMAS069	NMAS070	NMAS071	川麦625 Chuanmai 625	21pin3-9	川麦704 Chuanmai 704	21pin4-3	21pin6-5	川麦614 Chuanmai 614	川麦618 Chuanmai 618	川麦620 Chuanmai 620	川麦623 Chuanmai 623	川麦702 Chuanmai 702	川麦914 Chuanmai 914	绵麦60 Mianmai 60	内麦175 Neimai 175	蜀麦1925 Shumai 1925
赤霉病	Fhb1^†	+	+	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Fusarium	Fhb2^†	+	-	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-
head blight	Fhb4^†	+	+	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Fhb5^†	-	+	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Qfhb.caas-3BL	-	-	-	-	+	+	-	-	+	+	-	+	+	+	-	+	-
	QFhb.hbaas-5AS	-	-	-	+	+	-	+	-	-	-	+	-	-	-	-	+	-
	QFhb.hbaas-5AL	-	-	+	-	-	-	+	-	+	-	-	-	+	+	-	-	-
	QFhb.caas-5AL	+	-	-	+	-	+	-	+	+	+	+	+	+	+	+	-	+
千粒质量	TaSus2-2B	高	低	高	高	高	低	高	高	低	低	低	低	低	高	高	高	高
1000-grain	Tabas1	高	低	低	高	低	高	低	高	高	高	高	高	高	高	高	低	高
weight	TaGS5-A1	杂合	高	低	高	低	高	高	高	高	高	高	高	高	高	高	低	高
	TaGW2-6A	高	高	高	高	高	高	高	NA	高	高	高	高	高	高	NA	NA	高
	TaCwi-A1	低	低	高	高	低	高	高	高	高	高	高	高	高	高	高	高	高
	TaT6P	低	低	低	低	低	高	高	高	高	高	低	高	低	低	低	低	低
品质	Glu-B3h	-	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+
Quality	TaLCYE-B1	低	高	低	高	低	高	低	高	高	高	高	高	高	高	高	高	低
株高	Rht-B1	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	+	-	NA
Plant	Rht-D1	+	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA	NA
height	RHT-8	+	+	-	+	-	-	+	-	-	-	+	-	-	-	-	-	+
	Rht24_AP2	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+
	αPht-2D	-	-	+	+	+	-	+	+	+	-	+	-	+	+	-	+	-

基因 Gene	SNP位置 SNP position	NMAS069	NMAS070	NMAS071	川麦614 Chuanmai 614	21pin 3-9	21pin 4-3	川麦702 Chuanmai 702	21pin 3-3	绵麦60 Mianmai 60	內麦175 Neimai 175	川麦620 Chuanmai 620	川麦623 Chuanmai 623	川麦618 Chuanmai 618	21pin 4-1	21pin 6-5	蜀麦1925 Shumai 1925	川麦914 Chuanmai 914
Fhb2	chr6B:118879886	GG	GG	CC	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG
Fhb2	chr6B:152096128	CC	TT	CT	TT	TT	TT	TT	TT	TT	TT	TT	TT	TT	TT	TT	TT	TT
Fhb2	chr6B:165725747	TT	CC	TT	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC
Fhb2	chr6B:169959743	AA	CC	AA	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC
Fhb4	chr4B:463122524	AA	GG	GG	GG	GG	AA	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG
Fhb4	chr4B:545511172	TT	TT	TT	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC
Fhb5	chr5A:191466469	GG	AA	AA	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG
Fhb5	chr5A:209001461	GG	AA	AA	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG

基因 Gene	SNP位置 SNP position	NMAS069	NMAS070	NMAS071	川麦614 Chuanmai 614	21pin 3-9	21pin 4-3	川麦702 Chuanmai 702	21pin 3-3	绵麦60 Mianmai 60	內麦175 Neimai 175	川麦620 Chuanmai 620	川麦623 Chuanmai 623	川麦618 Chuanmai 618	21pin 4-1	21pin 6-5	蜀麦1925 Shumai 1925	川麦914 Chuanmai 914
Fhb2	chr6B:118879886	GG	GG	CC	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG
Fhb2	chr6B:152096128	CC	TT	CT	TT	TT	TT	TT	TT	TT	TT	TT	TT	TT	TT	TT	TT	TT
Fhb2	chr6B:165725747	TT	CC	TT	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC
Fhb2	chr6B:169959743	AA	CC	AA	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC
Fhb4	chr4B:463122524	AA	GG	GG	GG	GG	AA	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG
Fhb4	chr4B:545511172	TT	TT	TT	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC	CC
Fhb5	chr5A:191466469	GG	AA	AA	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG
Fhb5	chr5A:209001461	GG	AA	AA	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG	GG

标记 Marker	扩增基因 Amplified gene	染色体位置 Chrmosome position	引物名 Primer name	引物序列(5'-3') Primer sequence(5'-3')	扩增片段/bp Amplified fragment
DQK004	Fhb2	chr6B:165725747	Forward-FAM	GAAGGTGACCAAGTTCATGCTACAGAACTTCCTAGATCATTTGT	100
			Forward-HEX	GAAGGTCGGAGTCAACGGATTACAGAACTTCCTAGATCATTTGC
			Reverse	GGAACCAGATGTTTTGGACTCA
DQK008	Fhb2	chr6B:169959743	Forward-FAM	GAAGGTGACCAAGTTCATGCTAAGGAATCAGCTGGGCAATT	70
			Forward-HEX	GAAGGTCGGAGTCAACGGATTAAGGAATCAGCTGGGCAATG
			Reverse	GCTTGCGAAAGAAGAGAGAAGG
DQK009	Fhb2	chr6B:118879886	Forward-FAM	GAAGGTGACCAAGTTCATGCTGTCCAATGTTGTCCGTTCTAC	121
			Forward-HEX	GAAGGTCGGAGTCAACGGATTGTCCAATGTTGTCCGTTCTAG
			Reverse	GTAACTGCGTGCCTCCTC
DQK011	Fhb2	chr6B:152096128	Forward-FAM	GAAGGTGACCAAGTTCATGCTGACCAGTAAAAAATCCGTCG	68
			Forward-HEX	GAAGGTCGGAGTCAACGGATTGACCAGTAAAAAATCCGTCA
			Reverse	CAGAAGGGTGGTTAGACTGC
DQK005	Fhb4	chr4B:463122524	Forward-FAM	GAAGGTGACCAAGTTCATGCTGACATGATCACGGCCCGA	63
			Forward-HEX	GAAGGTCGGAGTCAACGGATTGACATGATCACGGCCCGG
			Reverse	ACCACCTAGGGATAGTTGCAC
DQK007	Fhb4	chr4B:545511172	Forward-FAM	GAAGGTGACCAAGTTCATGCTGCTGACACCAACGTAGCTGAT	100
			Forward-HEX	GAAGGTCGGAGTCAACGGATTGCTGACACCAACGTAGCTGAC
			Reverse	AGAGATTCAACGACGGAAAGTC
DQK001	Fhb5	chr5A:191466469	Forward-FAM	GAAGGTGACCAAGTTCATGCTTCCAAGAAAAAGAGAAACAGAAACT	97
			Forward-HEX	GAAGGTCGGAGTCAACGGATTTCCAAGAAAAAGAGAAACAGAAACC
			Reverse	AGGATTACTTGCTGCTGCTTTTA
DQK002	Fhb5	chr5A:209001461	Forward-FAM	GAAGGTGACCAAGTTCATGCTAAGCGCGCGAGAACCCGT	81
			Forward-HEX	GAAGGTCGGAGTCAACGGATTAAGCGCGCGAGAACCCGC
			Reverse	GAGCTCCGCTTGGAACGT

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