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Rice purple leaf gene PL1 molecular marker, identification method thereof and application

A technology of molecular markers and identification methods, applied in the field of molecular biology, can solve the problems of long cycle, difficult to distinguish, and affect the effect of impurity removal, etc., to achieve stable molecular markers, speed up the breeding process, and improve the efficiency of selection or breeding efficiency Effect

Active Publication Date: 2017-05-10
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conventional artificial impurity removal technology is expensive and takes a long time, and many traits are affected by cultivation techniques and environmental factors and are not easy to distinguish, which affects the effect of impurity removal

Method used

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  • Rice purple leaf gene PL1 molecular marker, identification method thereof and application
  • Rice purple leaf gene PL1 molecular marker, identification method thereof and application
  • Rice purple leaf gene PL1 molecular marker, identification method thereof and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1: Genetic Analysis and Gene Mapping of Purple Leaf Characters

[0057] (1) F of Guangzi 06 / Sasanishiki 2 Construction of segregating populations and genetic analysis

[0058] F 1 Hybrids, planted F 1 Hybrids, selfed to get F 2 Populations were isolated for genetic analysis. According to the statistical analysis of field phenotype investigation, it was found that in F 2 Among the 542 individual plants in the isolated population, there were 411 individual plants with green leaves and 131 individual plants with purple leaves. Chi-square analysis showed that the ratio of green-leaf individual plants to purple-leaf individual plants conformed to the segregation ratio of 3:1 ( c 2 =0.199, R ˃0.05), which indicated that the purple leaf trait of "Guangzi 06" was controlled by a pair of recessive major nuclear genes.

[0059] (2) PL1 Molecular marker linkage analysis and preliminary localization

[0060] from F 2 Among the 542 individuals in the segregation ...

Embodiment 2

[0078] Example 2 Molecular labeling method

[0079] Using the whole genome DNA of the rice material to be identified as a template, one or more pairs of primers corresponding to the molecular markers P1, P2, P3, P4 and P5 described above are used for PCR amplification, and the amplified products are detected.

[0080] (1) Extraction of donor parent, recipient parent and F 2 Separate the DNA of each individual plant in the population;

[0081] (2) The reaction system of PCR is as follows:

[0082] Primer (4pmol / μL) 2.5μL

[0083] dNTP (2.5mmol / L) 2.0μL

[0084] DNA template (25ng / μL) 1.0μL

[0085] Taq DNA polymerase (5U / μL) 0.2μL

[0086] 10×PCR buffer 1.0μL

[0087] ddH2O 3.3 μL

[0088] The PCR reaction conditions were as follows: denaturation at 94°C for 5 minutes, 30s at 94°C, 30s at 56°C, 60s at 72°C, 35 cycles of amplification, and final extension at 72°C for 10 minutes.

[0089] The DNA amplification products were detected by polyacrylamide gel electrophoresis, ...

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Abstract

The invention discloses a rice purple leaf gene PL1 molecular marker, an identification method thereof. The rice purple leaf gene PL1 molecular marker is respectively provided with P1, P2, P3, P4 and P5. The method includes the steps: performing PCR (polymerase chain reaction) amplification for one pair or multiple pairs in primers corresponding to the molecular markers; detecting amplification products. The invention further provides an application of the molecular marker or the molecular marker identification method to positioning and cloning of rice purple leaf genes PL1. According to the molecular marker, purity of hybrid rice seeds or male sterile lines can be rapidly identified at seedling stage, selecting efficiency or breeding efficiency is improved, and breeding progress is accelerated.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a molecular marker of rice purple leaf gene PL1 and its identification method and application. Background technique [0002] Rice is one of the most important food crops for human beings, and it is also the largest food crop in my country. Therefore, how to increase the yield of rice is a key issue in my country's agricultural production. In the 1970s, Chinese rice breeders represented by Yuan Longping successfully bred three-line hybrid rice, which greatly increased the yield of rice and eased the contradiction between my country's rapid population growth and food shortage. However, in the production process of hybrid rice seeds, how to ensure the purity of rice hybrids and sterile lines is a difficult problem for breeders. Conventional artificial impurity removal technology is expensive and takes a long time, and many characters are not easy to distinguish due to the influence...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 覃宝祥李容柏刘芳李允振邱永福罗继景
Owner GUANGXI UNIV