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Method for determining genetic differentiation between rice mixed interplanting varieties by SSR markers

A genetic differentiation and rice technology, applied in biochemical equipment and methods, microbe determination/testing, DNA/RNA fragments, etc., can solve problems such as low efficiency, time-consuming and labor-intensive problems

Active Publication Date: 2020-09-29
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, traditional methods are often inefficient, and a large number of field experiments in the early stage are time-consuming and labor-intensive.

Method used

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  • Method for determining genetic differentiation between rice mixed interplanting varieties by SSR markers
  • Method for determining genetic differentiation between rice mixed interplanting varieties by SSR markers
  • Method for determining genetic differentiation between rice mixed interplanting varieties by SSR markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] One, a kind of SSR marker determines the method for genetic differentiation between rice mixed interplanting varieties, comprising the following steps:

[0041] (1) Extraction of rice genomic DNA

[0042] ①Reduce the lower opening of the washed 96-well plate. The size of the caliber is suitable for not dropping the rice seeds. Put the seeds into the 96-well plate one by one, and then place the 96-well plate on the suction plate to absorb water. Move it to a light incubator (28° C., humidity 90%, light for 12 hours), germinate after three days, and when it grows to 10-20 cm, it can be used as a material for extracting DNA.

[0043] ②Take a rice seedling, cut it into pieces and put it in a 1.5ML centrifuge tube, usually add to 1 / 3 of the volume of the centrifuge tube, add liquid nitrogen, and grind it into powder repeatedly with a grinding rod.

[0044] ③ Add 700 μL of 2% CTAB buffer [100mM Tris-HCl (pH8.0), 20mM EDTA (pH8.0), 1.4mol / L NaCl, 2% hexadecyl trimethyl bromide ...

Embodiment 2

[0088] 1.1 Materials and methods

[0089] 1.1.1 Experimental materials

[0090] Based on the rice varieties that have been used for large-scale planting under the mixed interplanting mode in Yunnan Province in the past 10 years, 22 rice varieties representing different ecological planting areas were selected, including traditional varieties and modern improved varieties. See Table 5 for species names, types and sources. The method of variety DNA extraction is consistent with embodiment 1. According to the genetic differentiation index (GDI) value obtained among each variety, combined with examples of main planting and interplanting rice varieties widely used in mixed interplanting in 10 years, the variety combination pairs used in field experiments were screened (see Table 6).

[0091] Table 5 Information table of rice samples in this study

[0092]

[0093] 1.1.2 Genetic differentiation index (GDI)

[0094] The calculation of the genetic differentiation index is based...

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Abstract

The invention relates to a method for determining genetic differentiation between rice mixed interplanting varieties by SSR markers, and belongs to the technical field of molecular markers. The methodcomprises the following steps: extracting total DNA from to-be-detected rice, performing amplifying by using 48 pairs of SSR specific primers, separating a PCR product by using a capillary electrophoresis method, reading the molecular weight of the PCR product, and carrying out datamation treatment on SSR molecular fingerprints, and calculating a genetic differentiation index (GDI) according to agenetic differentiation formula; and along with the increase of the genetic differentiation index GDI, in the diversified mixed interplanting planting, the overall rice blast control effect of the variety matching combination being better. According to the method, the genetic differentiation level among varieties can be rapidly and accurately identified by using a trace amount of DNA samples, thecontrol effect of diversity mixed interplanting assembled varieties on rice blast is predicted, and the method has important significance in research of variety matching selection in diversity mixedinterplanting.

Description

technical field [0001] The invention relates to the technical field of molecular markers, and more specifically relates to a method for determining genetic differentiation among mixed rice varieties by SSR markers. Background technique [0002] Rice blast is one of the major diseases threatening rice production and an important factor restricting high and stable rice yields. With the mechanization of rice and the large-scale promotion of high-yield varieties, large-scale long-term single planting of rice varieties has resulted in continuous mutation of the rice blast fungus and the rapid emergence and proliferation of dominant races, resulting in increasingly serious occurrence of rice blast, and the epidemic cycle is becoming more and more serious. Shorter and shorter, eventually leading to the rapid decline in the rice blast resistance of popularized rice varieties, the extensive use of pesticides and chemical fertilizers, resulting in the fragility and degradation of farm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11C12Q1/6858
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2531/113C12Q2537/165Y02A40/10
Inventor 韩光煜王哲张文龙王云月李勇成罗琼杨楠苏正亮
Owner YUNNAN AGRICULTURAL UNIVERSITY
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