人工合成小麦SHW-L1高硒含量KASP分子标记开发及其应用

doi:10.3864/j.issn.0578-1752.2020.20.001

中国农业科学 ›› 2020, Vol. 53 ›› Issue (20): 4103-4112.doi: 10.3864/j.issn.0578-1752.2020.20.001

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

人工合成小麦SHW-L1高硒含量KASP分子标记开发及其应用

魏广辉^1,²(),李执²,陈强²,李阳²,陈诗豪²,裴英²,周勇³,程梦萍³,唐豪³,王际睿^1,³,魏育明^1,³,刘登才^1,³,陈黎⁴,郑有良^1,³,蒲至恩^1,²()

¹西南作物基因资源发掘与利用国家重点实验室,成都611130
²四川农业大学农学院,成都611130
³四川农业大学小麦研究所,成都611130
⁴宜宾市产品质量监督检验所,四川宜宾644000

收稿日期:2019-12-02 接受日期:2020-03-07 出版日期:2020-10-16 发布日期:2020-10-26
通讯作者: 蒲至恩
作者简介:魏广辉,E-mail: 18437957800@163.com。
基金资助:
国家重点研发计划(2017YFD0100900);四川省教育厅重点项目(17ZA0315)

Development and Utilization of KASP Marker for Se Concentration in Synthetic Wheat SHW-L1

WEI GuangHui^1,²(),LI Zhi²,CHEN Qiang²,LI Yang²,CHEN ShiHao²,PEI Ying²,ZHOU Yong³,CHENG MengPing³,TANG Hao³,WANG JiRui^1,³,WEI YuMing^1,³,LIU DengCai^1,³,CHEN Li⁴,ZHENG YouLiang^1,³,PU ZhiEn^1,²()

¹State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Chengdu 611130
²College of Agronomy, Sichuan Agricultural University, Chengdu 611130
³Triticeae Research Institute, Sichuan Agricultural University, Chengdu 611130
⁴Yibin Product Quality Supervision and Inspection Institute, Yibin 644000, Sichuan

Received:2019-12-02 Accepted:2020-03-07 Online:2020-10-16 Published:2020-10-26
Contact: ZhiEn PU

摘要/Abstract

摘要：

【目的】硒作为人体必不可少的元素之一,在疾病预防与抗氧化方面具有重要作用。小麦作为主要作物之一,是人类获取植物性硒源的主要来源,因此,提高小麦籽粒硒含量是人体补充硒的一条高效、低廉、简单易行的途径。拟通过对已定位到的SNP位点开发KASP标记用于开展高硒小麦选育。【方法】利用已经完成的660K小麦SNP标记的人工合成小麦SHW-L1重组自交系（recombinant inbred lines,RILs）群体,将前期定位的籽粒硒含量相关的QTL区间进一步缩小,开发稳定基因内的SNP位点为KASP标记。【结果】成功开发出2个KASP标记AX-1和AX-2,标记能够在群体中进行基因型分型和鉴定材料籽粒硒含量的相对高低。2个标记都可将供试材料按照基因型分为2类,即高硒和低硒材料,符合1:1的等位基因分离比例。但按照表现型来划分,标记对低硒材料的筛选率（大于90%）要远高于高硒材料（小于30%）。具有高硒含量的表现型材料中,有81%的材料具有高硒的基因型,表明了标记的可靠性。同时,在以人工合成小麦SHW-L1为亲本的其他衍生后代株系中也能用该KASP标记进行基因型分型,分型结果与表型结果一致,说明了标记的有效性,也表明可通过开发到的KASP标记来提升选择效率。【结论】AX-1和AX-2可作为一个实用的分子标记用于SHW-L1为亲本的高硒小麦品种选育和种质资源创制。

关键词: 人工合成小麦, 硒含量, KASP标记

Abstract:

【Objective】As one of the essential elements of the human body, selenium plays an important role in disease prevention and antioxidants. Wheat, as one of the main crops, is the main source for humans to obtain plant selenium sources, so increasing the selenium content of wheat grains is an efficient, inexpensive and easy way to supplement selenium. This study intends to develop KASP markers for the identified SNP loci for the breeding of high-selenium wheat.【Method】Using the completed 660K wheat SNP markers of synthetic wheat SHW-L1 recombinant inbred lines population ( recombinant inbred lines, RILs), scanning the RILs population, further narrowing the QTL interval related to the selenium content of the previously located grains, and developing the SNP loci in the stable gene as KASP markers.【Result】Two successfully developed KASP markers AX-1 and AX-2, markers can be used for genotyping and identification of selenium content in the grain of the material. Both markers can be used to classify test materials according to genotype into two categories, namely high-selenium and low-selenium materials, meets the 1:1 allele separation ratio. However, according to the phenotype, the screening rate (greater than 90%) for low-selenium materials is much higher than that of high-selenium materials (less than 30%), and it should be clear that 81% of the phenotype materials with high selenium content have high selenium genotypes, indicating the reliability of the label. Therefore, the KASP markers developed can be used to improve selection efficiency. At the same time, the KASP marker can also be used for genotyping in other derived offspring lines with artificial wheat SHW-L1 as the parent, and the typing results are consistent with the phenotypic results, indicating the validity of the marker.【Conclusion】AX-1 and AX-2 can be used as a practical molecular marker for breeding and germplasm resource creation of high selenium wheat varieties with SHW-L1 as the parent.

Key words: synthetic wheat, selenium content, KASP

魏广辉,李执,陈强,李阳,陈诗豪,裴英,周勇,程梦萍,唐豪,王际睿,魏育明,刘登才,陈黎,郑有良,蒲至恩. 人工合成小麦SHW-L1高硒含量KASP分子标记开发及其应用[J]. 中国农业科学, 2020, 53(20): 4103-4112.

WEI GuangHui,LI Zhi,CHEN Qiang,LI Yang,CHEN ShiHao,PEI Ying,ZHOU Yong,CHENG MengPing,TANG Hao,WANG JiRui,WEI YuMing,LIU DengCai,CHEN Li,ZHENG YouLiang,PU ZhiEn. Development and Utilization of KASP Marker for Se Concentration in Synthetic Wheat SHW-L1[J]. Scientia Agricultura Sinica, 2020, 53(20): 4103-4112.

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变异来源 Source of variation	平方和 Type Ⅲ SS	自由度 df	均方 MS	F值 F value	Sig.
V	0.02995031	114	0.00026272	13.410	**
T	0.00009015	1	0.00009015	0.4890
V×T	0.02100010	110	0.00019091
误差Error	0.00415800	110	0.00003780

籽粒硒含量 Grain selenium content	编号 Number	染色体 Chromosome	标记 Marker	位置 Position (cM)	R²	加性效应 Additive	LOD
Grain se-1	1	5A	22	43.62	0.0765	0.0003	2.5551
Grain se-2	1	5A	13	29.54	0.0654	0.0003	2.7476
Grain se-2	2	5A	16	32.66	0.0774	0.0003	3.4722
Grain se-2	3	5A	21	35.58	0.0877	0.0004	3.9723

编号 Number	染色体 Chromosome	变异碱基 Base	序列 Sequence (5′-3′)
AX-1	5A	A/G	CGTACCATGCGATCACACTTGGAGATGCGTGTGAG[A/G]GAAAAAGAGTACTCCATCCATCCAAATTCCATCCC
AX-2	5A	A/G	TGTAAATTAATTTGTTGTGGGTAGGGCATCTTGAG[A/G]AATGTTTACCTTCTTGCTGCTGCCTGGTTCGGTTT
AX-3	5A	A/C	GGGTGATAGCAAGGAAGCTGAAGAGCATGATGGAC[A/C]AGGGACAGACGGAAAAGAGTTGCAGGATGGACAAG
AX-4	5A	A/G	GATGACTTTGCGGCAATATTAAGAGATGTGCATCC[A/G]TTAGCAAGCAAGCTAGCGGACAAAACCAGCTTAAC
AX-5	5A	T/C	ACCCCTTGTTTAGTACCACCTCACCAAGTGAGAGT[C/T]ATTTCAACCGGTTTATAAGATTCAGCGTTCCAACC

品种或自交系 Cultivar or inbred lines	籽粒硒含量 Selenium content in grains (mg·kg^-1)	SNP基因分型 SNP genotyping
蜀麦969 Shumai 969	0.020	高硒High Se
蜀麦580 Shumai 580	0.011	低硒Low Se
蜀麦980 Shumai 980	0.009	低硒Low Se
蜀麦830 Shumai 830	0.017	高硒High Se
蜀麦114 Shumai 114	0.013	高硒 High Se

人工合成小麦SHW-L1高硒含量KASP分子标记开发及其应用

Development and Utilization of KASP Marker for Se Concentration in Synthetic Wheat SHW-L1

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