基于KASP技术开发检测水稻耐高温TT1基因型的分子标记

Development of Molecular Markers for Genotyping Heat-tolerant Gene TT1 in Rice Based on KASP Technology

高温是对水稻产量和品质有重要影响的环境因素之一,随着全球气候逐渐变暖,高温热害对水稻生产构成了严重的威胁。研究发现非洲栽培稻CG14中的OgTT1等位基因可以显著提高水稻耐高温特性。为了更高效利用OgTT1等位基因选育耐高温水稻品种,根据OgTT1等位基因的功能性SNP位点设计了一个高通量分子标记TT1-KASP。利用该分子标记对来源于NIL(CG14)分别与申恢26、申6B杂交的F_4代重组自交系进行检测,检测结果表明在F_4代重组自交系的65个株系中,4个株系不含OgTT1等位基因,53个株系OgTT1等位基因纯合,而其余的8个株系为杂合基因型。对65个F_4代重组自交系株系进行基因测序后发现,得到的基因序列与TT1-KASP标记所得序列完全一致。该结果表明TT1-KASP分子标记可以用于耐高温水稻新品种的标记辅助选择育种工作。

High temperature is one of the environmental factors that have an important impact on the yield and quality of rice,with the gradual warming of the global climate,heat stress poses a serious threat to rice production.It has reported that the OgTT1 allele in African cultivated rice CG14 can significantly improve the heat tolerance of rice.In order to use OgTT1 alleles to breed heat-tolerant rice varieties more efficiently,a high-throughput molecular marker TT1-KASP was designed based on the functional SNP sites of OgTT1 alleles.This molecular marker was used to detect two F4generation recombinant inbred line(RIL)populations derived from the cross of NIL(CG14)×Shenhui26 and NIL(CG14)×Shen6B.The results showed that among the 65 lines of RIL,4 lines did not contain OgTT1 alleles,53 strains were homozygous for OgTT1 alleles,while the remaining 8 lines were heterozygous genotypes.After sequencing the genes of 65 F4recombinant inbred lines,it was found that the obtained gene sequences were completely consistent with the sequences obtained from the TT1-KASP marker.These results indicate that the TT1-KASP molecular marker can be used for marker-assisted selection breeding of heat-tolerant rice varieties.