Molecular marker primer combination for rapid high-throughput identification of sex traits of torreya grandis and application thereof

Abstract

The invention discloses a molecular marker primer combination for rapid high-throughput identification of sex traits of torreya grandis and application thereof, so as to be applied to assisting screening of early molecular markers for torreya grandis breeding. The molecular marker primer combination is JY1754, a PCR amplification product of the JY1754 has a length of only 97 bp, and the JY1754 isparticularly suitable for high-throughput detection platforms such as fluorescent quantitative PCR instruments and the like, and can be used for rapid identification of sex traits of torreya grandis germplasm resources. The invention provides a method for assisting screening of torreya grandis germplasm resources with different sexes based on the developed molecular marker primer combination, hasimportant guiding significance in predicting phenotype of sex traits of torreya grandis, and has advantages of simple operation, high throughput, universal multi-detection analysis platform, accurateresult and the like.

Description

technical field [0001] The invention relates to a set of molecular marker primer combinations for rapid and high-throughput identification of the sex traits of Torreya torreya, and provides a method for assisting in identifying or screening the traits of different sex germplasm resources of Torreya torreya, belonging to the field of molecular biology. Background technique [0002] Torreya grandis (Torreya grandis Fort.ex Lindl.cv.Merrillii Hu) is a relict plant of the Tertiary Period, mainly dioecious, and occasionally monoecious. It is a rare plant unique to my country. The precious dried fruit combined with oil extraction is now mainly distributed in Zhejiang, Jiangxi, Jiangsu, Anhui, Fujian and Hunan provinces, and its cultivation area is still expanding year by year. Torreya torreya grows slowly in the early stage, and the gender is difficult to distinguish. The seedlings generally enter the flowering and fruiting period in 15 to 20 years. It is often the phenomenon that ...

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Problems solved by technology

As of the present invention, although there have been research reports on the early identification of Torreya sex from aspects such as RAPD and AFLP markers, its operation steps involve processes such as enzyme digestion, connection, and gel electrophoresis, which are very cumbersome, and the technical threshold and detection cost are very high. High, poor practicability, and its detection accuracy is unknown

In addition, the invention patent with the authorized announcement number CN201810144040.1 discloses a specific molecular marker TGMI001 for the sex identification of Torreya torreya seedlings, wherein TGMI001, a male-specific molecular marker of Torreya torreya, is provided, which can be used to identify the sex of Torreya torreya, but the The invention does not mention whether the monoecious plant type of Torreya chinensis can be identified; in addition, the PCR product fragment of the molecular marker TGMI001 is relatively long (461bp), which is not suitable for high-throughput detection technology platforms such as fluorescent quantitative PCR instrument genotyping (It is usually required that the product fragment length is <400bp, and the amplification effect of the product fragment length is about 100bp is the best); moreover, the molecular marker TGMI001 requires relatively long PCR amplification time and subsequent traditional gel electrophoresis Detecting the size of the product, etc., the sensitivity is lower than that of a fluorescent quantitative PCR instrument, and the operation and technical links are relatively cumbersome, time-consuming and labor-intensive

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