利用等位基因特异扩增快速检测水稻香味基因

水稻香味受隐性基因控制, 杂合材料不表现出香味, 因此在香稻选育中需要寻找一种快速、简便的香味基因检测方法。本试验通过等位基因特异扩增(ASA)方法对9个香稻和3个非香稻品种 (品系), 以及3个香稻/非香稻组合的F1的香味基因, 进行快速检测。结果显示, 纯合非香稻可获得长度为585和355 bp的两条带, 纯合香稻可获得长度为577和257 bp的两条带, 杂种F1可获得长度为355、257和580 bp左右的3条带。供试材料的分子检测结果与香味基因型完全相符, 所用方法快速有效, 可应用于香稻育种实践。

Detection of Rice Fragrant Gene by Allele-Specific Amplification

The technically simplified approach of marker-assisted selection has directly and practically used in rice breeding. Fragrance in rice, a recessive trait, has been known to be due to an 8 bp deletion and 3 SNPs difference in a gene on chromosome 8. Rice breeders have an interest in gaining access to a simple and inexpensive method for distinguishing between fragrant and non-fragrant rice. Allele specific amplification (ASA) is a low-cost, robust technique that can be utilized to discriminate between the alleles. We used a single tube ASA assay which allows discrimination between 9 fragrant and 3 non-fragrant rice varieties and 3 F₁ plants to identify homozygous fragrant, homozygous non-fragrant, and heterozygous non-fragrant individuals. The research showed that using tetras-primer amplification refractory mutation system PCR could result in three possible outcomes. External primers generated a fragment of approximately 580 bp as a positive control for each sample. Internal and corresponding external primers produced a 355 bp fragment from a non-fragrant allele, a 257 bp fragment from a fragrant allele and both 355 bp and 257 bp fragment from a heterozygous allele allowing simple analysis on agarose gels. A single tube allele specific PCR which allows determination of the genotypic status of an individual rice plant, either homozygous fragrant, homozygous non-fragrant or heterozygous has practical utility for rice breeders worldwide.