小麦单位面积穗数和粒长主效QTL紧密连锁KASP标记的开发及其效应评价

doi:10.3864/j.issn.0578-1752.2021.14.002

Abstract

Abstract:

【Objective】Spike numbers per unit area (SN) and kernel length (KL) are both important yield-related traits of wheat. Genetic improvement on SN and KL will help increase the yield of wheat. The corresponding KASP molecular markers were developed for the QSn.sau-2D.2 and the QTL Kl.sau-3D.2, which were identified in previous study for increasing SN and KL, respectively. Both KASP markers were verified and evaluated in the RIL population crossed by Chuannong 18 and T1208, which laid a foundation for better utilization of these two QTLs.【Method】The major QTL QSn.sau-2D.2 for SN and the major QTL QKl.sau-3D.2 for KL were previously identified in the RIL population crossed by Chuannong 18 and T1208. The SNP molecular marker sequence of Wheat 55K SNP array within the two QTLs were used to develop and design the KASP molecular markers. The polymorphic KASP markers were screened with wheat parents and then verified in the RIL population. The selected KASP molecular markers were genotyped and used to identify the corresponding phenotypic traits in Chuannong 18×T1208 RILs population. The effects of the two major QTLs on other agronomic traits were analyzed.【Result】KASP-AX-111151907 and KASP-AX-109962767 have polymorphic between the wheat parents. Subsequently, these two KASP markers were verified in the RIL population linkage with QSn.sau-2D.2 and QKl.sau-3D.2, respectively. KASP-AX-111151907 and KASP-AX-10996276 could divide the genotypes into two groups. According to the phenotype, the average selection rate of KASP-AX-111151907 for multi spike lines was 72.58%, the average selection rate of KASP-AX-111151907 for few spike lines was 71.68%, the average selection rate of KASP-AX -10996276 for long kernel lines was 69.86%, and the average selection rate of KASP-AX-10996276 for short kernel lines was 61.52%, indicating the reliability of the two KASP markers. Moreover, the genotyping results based on KASP molecular markers showed that the two QTLs had significant effects on plant height, 1000 kernel weight, kernel length, kernel width, kernel diameter ratio, spike numbers per unit area and kernel weight per spike, respectively. The validation in the RIL population of Chuannong17×Chuannong11 also indicated that these two KASP markers had a certain effect on the selection of the corresponding traits.【Conclusion】This study developed two KASP markers for two major QTLQSn.sau-2D.2 and QKl.sau-3D.2, respectively. Both KASP markers can be used for the selection of corresponding traits. These two QTLs which are tightly linked to these two KASP markers could significantly improve the SN and KL, respectively. In addition, QSn.sau-2D.2had negative effects on plant height, 1000 kernel weight, kernel length, kernel width, kernel diameter ratio and kernel weight per spike. On the other hand,QKl.sau-3D.2 had positive effect on plant height, 1000 kernel weight, kernel width, kernel diameter ratio and kernel weight per spike, but had negative effects on SN. These two QTLs and the developed KASP markers can be used in the future high yield wheat breeding program.

Key words: wheat, spike numbers per unit area, kernel length, QTL, KASP, agronomic traits

FAN Tao,LI Zhi,JIANG Qing,CHEN ShuLin,OU Xia,CHEN YongYan,REN TianHeng. Development and Effect Evaluation of KASP Markers Closely Linked to Major QTLs of Spike Number Per Unit Area and Grain Length in Wheat[J].Scientia Agricultura Sinica, 2021, 54(14): 2941-2951.

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References 35

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SNP标记 SNP marker	染色体 Chromosome	变异碱基 Base	序列 Sequence (5′-3′)
AX-109283238	2D	C//T	CCCTTGTCTCTTCCATCTATTTTCCCACCATCGTA[C/T]GGTTCCTTCTTTAATATTGTGTCTCTTTCCTCGAT
AX-111151907	2D	G//T	GCCACCTACCTAGAAACCCTACCCAGCAGCCACAA[G/T]ATCCCCCAAAAATCGGATTCCTTTGCTGGACTAAC
AX-86163992	2D	A//G	TTGCTCAGCTTCTGGAAATACTGCTGCTTCTGAAC[A/G]AGCTTCTCGCGTGGGTAAAATGGTGCTTCTTCGAG
AX-108802182	2D	A//G	TGCAGGATAATCCTAGAGCCAGGGGGATGTACGGT[A/G]TTCCATTTCCGCATTTCGATGTGTTTGATGCGGTG
AX-110073027	2D	G//T	TCGATATGAAGTCTGGTGAAACCGATGGGTACGAC[G/T]CTGTTTTCTTGAGTCCTCACAAATTTGTCGGGGGA
AX-109962767	3D	A//G	AACTATCGGATTGGAAACAAATTGACTCACATACA[A/G]CAAATGATATGTGAGGCCGAGTAGCACAAGCTAGG
AX-108923200	3D	A//G	CTCGAACAAATGAGTTGAAGAAACTGCCATCATCA[A/G]AAGACTGAAGCAAAACAAGTTTGAGACAACCACTG

QTL	分子标记 Molecular marker	引物序列 Primer sequence (5′-3′)
QSn.sau-2D.2	KASP-AX-109283238-1	GAAGGTGACCAAGTTCATGCTCCATCTATTTTCCCACCATCGTAC
	KASP-AX-109283238-2	GAAGGTCGGAGTCAACGGATTCCATCTATTTTCCCACCATCGTAT
	KASP-AX-109283238-3	ACCTTTATCGAGGAAAGAGACACA
	KASP-AX-111151907-1	GAAGGTGACCAAGTTCATGCTCCTACCCAGCAGCCACAAG
	KASP-AX-111151907-2	GAAGGTCGGAGTCAACGGATTCCTACCCAGCAGCCACAAT
	KASP-AX-111151907-3	AAGGCCAGTGTTAGTCCAGC
	KASP-AX-86163992-1	GAAGGTGACCAAGTTCATGCTGGAAATACTGCTGCTTCTGAACA
	KASP-AX-86163992-2	GAAGGTCGGAGTCAACGGATTGGAAATACTGCTGCTTCTGAACG
	KASP-AX-86163992-3	TTGTATGCAGGATGACGCTC
	KASP-AX-108802182-1	GAAGGTGACCAAGTTCATGCTCATCGAAATGCGGAAATGGAAT
	KASP-AX-108802182-2	GAAGGTCGGAGTCAACGGATTCATCGAAATGCGGAAATGGAAC
	KASP-AX-108802182-3	GTGTCTTTGTGTTGATTTGTTTCA
	KASP-AX-110073027-1	GAAGGTGACCAAGTTCATGCTTGTGAGGACTCAAGAAAACAGC
	KASP-AX-110073027-2	GAAGGTCGGAGTCAACGGATTTGTGAGGACTCAAGAAAACAGA
	KASP-AX-110073027-3	ATCCTTTCTGCAGTGGACCC
QKl.sau-3D.2	KASP-AX-109962767-1	GAAGGTGACCAAGTTCATGCTTGGAAACAAATTGACTCACATACAA
	KASP-AX-109962767-2	GAAGGTCGGAGTCAACGGATTTGGAAACAAATTGACTCACATACAG
	KASP-AX-109962767-3	GTGCTACTCGGCCTCACATA
	KASP-AX-108923200-1	GAAGGTGACCAAGTTCATGCTGTTGAAGAAACTGCCATCATCAA
	KASP-AX-108923200-2	GAAGGTCGGAGTCAACGGATTGTTGAAGAAACTGCCATCATCAG
	KASP-AX-108923200-3	CGTCCAGTGGTTGTCTCAAAC

QTL	环境 Environment	基因型A Genotype A	基因型B Genotype B	差值 Difference	P值 P value
QSn.sau-2D.2	2015	311.60（n=158）	275.56（n=211）	36.04	<0.001
	2016	303.41（n=158）	263.75（n=211）	39.66	<0.001
	2017	290.50（n=158）	234.36（n=211）	56.14	<0.001
QKl.sau-3D.2	2016	7.11（n=188）	7.43（n=175）	0.32	<0.001
	2018	6.98（n=188）	7.45（n=175）	0.47	<0.001
	2019	7.01（n=188）	7.64（n=175）	0.63	<0.001

QTL	基因型 Genotype	株高 Plant height (cm)	千粒重 1000 kernel weight (g)	粒长 Kernel length (mm)	粒宽 Kernel width (mm)	粒径比 Kernel diameter ratio	单位面积穗数 Spike numbers per unit area/m²	每穗粒重 Kernel weight per spike (g)
QSn.sau-2D.2	A	78.85***	45.77***	6.89***	3.45***	1.99***	290.67***	2.26***
	B	88.47	49.58	7.30	3.50	2.09	234.49	2.62
QKl.sau-3D.2	A	80.25	45.89	6.90	3.46	1.99	281.48	2.29
	B	88.26***	49.95***	7.34***	3.50***	2.10***	235.13***	2.62***

Development and Effect Evaluation of KASP Markers Closely Linked to Major QTLs of Spike Number Per Unit Area and Grain Length in Wheat

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