Method for authenticating rice indica subspecies and special primer composition thereof

Abstract

The invention discloses a method for authenticating rice indica subspecies and a special primer composition thereof. The primer composition provided by the invention comprises a primer pair RM130 which is formed by DNA shown as a sequence 1 of a sequence table and DNA shown as a sequence 2 of the sequence table, a primer pair RM7009 which is formed by DNA shown as a sequence 3 of the sequence table and DNA shown as a sequence 4 of the sequence table and a primer pair RM7337 which is formed by DNA shown as a sequence 5 of the sequence table and DNA shown as a sequence 6 of the sequence table. The primer composition can be used for assisting to authenticate the rice indica subspecies, has the advantages of high speed, high efficiency, high sensitivity, high resolution, accurate and reliableresult, and the like and is suitable for application in indica attribute authentication, rice breeding, plant evolution research, and the like of rice species.

Description

technical field [0001] The invention relates to a method for identifying rice indica and japonica subspecies and a special primer composition thereof. Background technique [0002] Indica-japonica differentiation is the mainstream of genetic differentiation in Asian cultivated rice, and indica-japonica subspecies show great differences in many aspects. Therefore, the utilization of heterosis between indica and japonica subspecies is a very important means for super-high-yielding rice breeding, and the direct use of heterosis between indica and japonica has become the main direction of hybrid breeding. In order to carry out super-high-yield breeding of heterosis between indica and japonica subspecies, the first step is to correctly identify indica and japonica. Indica-japonica intermediate type or atypical indica-japonica may originate from natural hybridization or artificial hybridization of indica-japonica, and may also originate from primitive transitional varieties befor...

AI Technical Summary

Problems solved by technology

[0005] However, most of the classification standards used in traditional classification methods are quantitative traits, which require not only experienced appraisers with accurate observation ability, but also the correct choice of appraisal period, and the use of standards is difficult to grasp

RFLP method: Southern hybridization is required, the operation is complicated and the workload is heavy; single-copy probes are generally used, the number of markers is relatively small, and the polymorphism is low; the amount of DNA is large, and relatively complete DNA is required; enzyme digestion reaction requires DNA higher purity

RAPD method: the template DNA is very sensitive to the template concentration during amplification, that is, different concentrations will produce different amplification patterns; the amplification reaction requires the template to contain as little impurities as possible; use low temperature renaturation, usually at 36°C (temperature at Below 36°C, a change of less than 1°C will result in a completely different amplification pattern), low specificity, and a high mismatch rate between primers and templates; low repeatability, even in the same laboratory, it is not easy to repeat, different There is even a lack of comparability between laboratories

Therefore, there is no classification system that can be directly used in actual work so far.

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