Development and Utilization of KASP Marker LCd-38 for Cadmium Accumulation in Rice Grain

doi:10.13304/j.nykjdb.2021.1037

Abstract

Abstract:

Rice （Oryza sativa L.） is one of staple food in China， and the cadmium pollution of rice threatens food security. Screening and breeding of rice varieties with low-cadmium accumulation can effectively reduce the risk， and accurate molecular markers play vital roles in it. In this study， taking the rice mini-core collection as the material， a KASP molecular marker LCd-38 was developed based on the functional SNP site in the gene related to grain cadmium accumulation. The cadmium-polluted soil safe to use was chosen as the experimental area， and LCd-38 was used to genotype and screen rice varieties with low cadmium-accumulation in grain. The developed marker LCd-38 could effectively divide different rice varieties into high-cadmium accumulation genotypes （CC） and low-cadmium accumulation genotypes （TT）. 5 low-cadmium accumulation rice varieties and 5 high-cadmium accumulation rice varieties were quickly identified in the experimental area using leaves at seedling stage， which were consistent with the results of measured grain cadmium concentration at maturity stage. These results indicated that the molecular marker LCd-38 could efficiently and accurately predict the cadmium accumulation characteristics of different rice varieties and be used for early screening and molecular marker-assisted selection breeding of low-cadmium accumulation rice varieties.

Key words: Oryza sativa L., grain, cadmium, KASP molecular marker, breeding

摘要：

水稻（Oryza sativa L.）是我国主粮作物之一，稻米镉污染对我国粮食安全造成一定威胁。镉低积累水稻品种的选育可有效降低水稻籽粒镉污染风险，高效准确的镉积累相关分子标记在镉低积累水稻品种选育中具有至关重要的作用。以水稻微核心种质为材料，基于籽粒镉积累关联基因的功能性SNP位点开发了KASP分子标记LCd-38。选取安全利用类镉污染土壤为试验地，利用该标记对当地主栽水稻品种进行基因分型和镉低积累水稻品种筛选。开发的分子标记LCd-38能够有效地将不同水稻品种分为籽粒高镉积累基因型（CC）和低镉积累基因型（TT）。以水稻苗期叶片为材料，快速鉴定到试验区5个低镉积累水稻品种和5个高镉积累水稻品种，与成熟期实测籽粒镉含量结果一致。综上所述，分子标记LCd-38可高效准确地预测不同水稻品种的籽粒镉积累特性，可应用于镉低积累水稻品种的早期筛选和分子标记辅助选择育种。

关键词: 水稻, 籽粒, 镉, KASP分子标记, 育种

CLC Number:

S511

Jun XU, Ting LI, Minjun HU, Yugen JIANG, Huili YAN, Wenxiu XU, Yijun YU, Zhenyan HE. Development and Utilization of KASP Marker LCd-38 for Cadmium Accumulation in Rice Grain[J]. Journal of Agricultural Science and Technology, 2022, 24(3): 40-47.

徐君, 李婷, 胡敏骏, 蒋玉根, 闫慧莉, 许文秀, 虞轶俊, 何振艳. 水稻籽粒镉积累KASP分子标记LCd-38的开发与利用[J]. 中国农业科技导报, 2022, 24(3): 40-47.

Figures/Tables 8

Table 1 LCd?38 molecular marker primer sequence

引物 Primer	序列 Sequence
F1-FAM： Allele-T	5’-GAAGTGGTAAAAGCACCCACAT-3’
F2-HEX： Allele-C	5’-GAAGTGGTAAAAGCACCCACAC-3’
Common- primer-R	5’-ATTAACTTATGGAATGCTTAC CTACACA-3’

Fig. 1 Screening and genotyping of TagSNP?20089955Note： *** indicates significant differencescompared with TT genotype at P<0.001 level.

Table 2 Grain Cd content and genotypes of different genotype rice

基因型

Genotype

镉含量Cd content/（mg·kg^-1）

品种数

Number of varieties

均值Average

标准差Standard seviation

Fig. 2 KASP genotyping of different genotypes by LCd?38

Fig. 3 Results of LCd?38 markers on the grain Cd concentration of rice germplasm

Table 3 Grain Cd content and genotypes of different genotype rice verified

基因型 Genotype	镉含量Cd content/（mg·kg^-1）			品种数 Number of varieties
基因型 Genotype	均值Average	标准差Standard seviation	中位数Median	品种数 Number of varieties
TT	0.134	0.062	0.108	7
CT	0.436	0.327	0.436	2
CC	0.421	0.050	0.435	11

Fig. 4 KASP genotyping and grain Cd content of 19 rice varieties in experimental areaNote： * indicates significant differences with CC genotype at P<0.05 level； ** indicates significant differences with CC genotype at P<0.01 level.

Table 4 Grain Cd content and genotypes of different genotype rice in experimental area

基因型 Genotype	镉含量Cd content/（mg·kg^-1）			品种数 Number of varieties
基因型 Genotype	均值Average	标准差Standard seviation	中位数Median	品种数 Number of varieties
TT	0.048	0.019	0.050	5
CT	0.258	0.254	0.176	9
CC	0.672	0.336	0.817	5

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