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Peanut bacterial wilt resistance grading identification method and application thereof

A technology of peanut bacterial wilt disease and identification method, which is applied in the identification of peanut bacterial wilt resistance classification and the field of plant disease resistance identification, can solve the problems of no fixed standard, cumbersome classification and identification, etc., and achieves easy operation and efficient technical support , the result is accurate

Pending Publication Date: 2020-12-22
SHANDONG PEANUT RES INST
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The invention provides an identification method for the grading of resistance to peanut bacterial wilt and its application. The identification method is scientific, effective, and easy to operate, and can well solve the problems of cumbersome grading identification and no fixed standards in the prior art.

Method used

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  • Peanut bacterial wilt resistance grading identification method and application thereof
  • Peanut bacterial wilt resistance grading identification method and application thereof
  • Peanut bacterial wilt resistance grading identification method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Embodiment 1: Isolation of pathogenic bacteria of bacterial wilt

[0036] Bacteria wilt diseased peanut plants ( figure 1 ) roots, cut them crosswise with a knife, immersed them in sterile water for 6 h, and collected the extract; shake the extract evenly, and draw 200 μL and apply it on a TZC plate (1L plate medium formula: 10 g of bacto-peptone, 5 g of glucose , casamino acid 1g, yeast extract 1g, agar 15g, TZC 50mg, the rest was made up with sterile water, pH 7.2), streak culture at 28°C for 48 h, the colony morphology is as follows figure 2 shown. Pick a single red colony from the plate and put it in 200 μL LB liquid medium, culture it with shaking at 220 rpm for 48 h at 28 °C; centrifuge at 1500 rpm for 10 min to collect the bacteria and extract the DNA of the bacteria. Specific primers 759 / 760 for identification of R. solanacearum strains were used to amplify bacterial DNA, wherein the primer sequence was 759: 5'-gtcgccgtcaactcactttc-3'; 760: 5'-gtcgccgtcagcaa...

Embodiment 2

[0037] Example 2: Grading of resistance to bacterial wilt of high-yielding peanut varieties

[0038] Select 9 high-yielding peanut varieties: Huayu 665, Huayu 669, Huayu 960, Huayu 967, Huayu 968, Huayu 969, Huayu 6620, Huayu 9620, and Huayu 9625 for bacterial wilt resistance Graded identification test. The bacterial wilt resistant peanut variety "Rihua 1" and the bacterial wilt susceptible peanut variety "Huayu 40" were selected as controls. Plants of all peanut varieties were planted in the experimental base located in Laixi, Qingdao.

[0039] 1. The acquisition of peanut branches:

[0040] The seeds of all peanut varieties were sown in the soil at the same time in a conventional manner, and daily management was adopted. The individuals of each variety were randomly sampled on the 55th day after sowing, and long cuts were cut from the top of the second pair of side branches of the selected peanut plants. For the branches with leaves, put them into the sampling box and sto...

Embodiment 3

[0054] The disease-resistant variety "Rihua 1" was selected as the male parent (♂) and the highly susceptible variety "Huayu 967" was used as the female parent (♀) to cross the F3 generation, and the individuals in the recombinant inbred line (RIL) population were subjected to RIL Identification of disease resistance levels. All hybrid F3 generation seeds were planted in the experimental base located in Laixi, Qingdao.

[0055] Sampling in 55 days after sowing, two shoots are cut from the second pair of side branch tops of each plant, then according to the method described in embodiment 2, carry out the activation and inoculation of bacterial wilt disease bacterium, count hybridization F3 generation on the 5th day after inoculation The disease-resistant phenotype of the plants. At this time, the control variety "Rihua 1" showed no symptoms, and the susceptible varieties "Huayu 40" and "Huayu 967" had obvious symptoms. Identification results such as Figure 7 As shown, accord...

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Abstract

The invention discloses a peanut bacterial wilt resistance grading identification method and an application thereof. The identification method comprises the following steps of taking peanut bacterialwilt pathogenic bacteria extracted from peanuts suffering from bacterial wilt as inoculation bacteria, taking branches with leaves growing at the top ends of a second pair of side branches of a plant55 days after the peanuts are sown, and inserting bacterial wilt thalli with the concentration of 10<9>cfu / mL for culture; and counting the disease-resistant phenotype of each branch on the fifth dayafter inoculation, grading the bacterial wilt resistance of the peanut according to the phenotype of the branches, and evaluating the anti-infection characteristic of peanut varieties. The identification method is scientific, effective and simple to operate, and can provide a convenient and efficient technical support for screening peanut bacterial wilt resistant germplasm resources and breeding of high-yield and bacterial wilt resistant varieties.

Description

technical field [0001] The invention belongs to the field of agricultural plant protection, and relates to a method for identifying plant disease resistance, in particular to an identification method for grading peanut bacterial wilt resistance and an application thereof. Background technique [0002] Cultivated peanuts ( Arachis hypogaea L.) is an important economic crop and industrial raw material in the world. It has received more attention in the research of oil crops. However, the disease resistance and stress resistance of peanut have always been the key issues restricting the sustainable development of peanut industry. [0003] Peanut bacterial wilt (Bacterial Wilt) is recognized as the most harmful peanut bacterial disease so far in the world, and it is also an important factor restricting peanut production in my country and many Southeast Asian countries. The pathogen often causes large-scale reduction or even extinction of peanut production, which causes serious e...

Claims

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Application Information

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IPC IPC(8): C12Q1/18C12Q1/04C12Q1/689C12N15/11C12R1/01
CPCC12Q1/18C12Q1/04C12Q1/689
Inventor 吴琪王传堂王志伟王秀贞唐月异
Owner SHANDONG PEANUT RES INST
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