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A method for predicting resistance to wheat sheath blight at seedling stage

A sheath blight and wheat technology is applied in the field of wheat plant protection and breeding to achieve the effects of reducing the influence of human visual error, accurate results and reducing influence

Active Publication Date: 2017-03-01
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the relationship between the DNA content of sheath blight in wheat stalks and the resistance of wheat itself to sheath blight has not been reported in the literature, nor has there been any report on the application of fluorescent quantitative PCR to predict the resistance of wheat sheath blight at seedling stage

Method used

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  • A method for predicting resistance to wheat sheath blight at seedling stage
  • A method for predicting resistance to wheat sheath blight at seedling stage
  • A method for predicting resistance to wheat sheath blight at seedling stage

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Example 1 Establishment and Application of Standard Curve for DNA Content of Rhizoctonia graminearum in Wheat Basal Stems

[0028] 1. Establishment of standard curve

[0029] (a) Wheat Ningmai 13 grains uninfected with Rhizoctonia graminearum were soaked at 20°C, germinated and cultured for 15 days. The base of the seedlings including the stem 1 cm of the leaf sheath was taken, and the total DNA was extracted by conventional CTAB method. Adjust to 50ng / μL;

[0030] (b) get the hyphae of Rhizoctonia graminearum R0301 grown on the PDA medium, adopt the conventional CTAB method to extract its total DNA, and adjust its concentration to 8ng / μL;

[0031] (c) The extracted Rhizoctonia graminearum DNA was mixed into the wheat Ningmai 13DNA solution according to the concentration gradient, so that the final concentration of the wheat Ningmai 13DNA in the mixed solution was 25ng / μL, and the final concentration of the sheath blight DNA was 40000 , 4000, 400, 40, 4pg / μL;

[0032...

Embodiment 2

[0047] Example 2 Examples of Prediction of Wheat Seedling Sheath Blight Resistance

[0048] In this example, the wheat varieties (lines) to be tested are: AHY11, BHY04, PH-82-2-2, 11E20, 11E29, Handan 6172, Huai 9720, Huai Mai 20, Huai Mai 21, Huai Mai 25, Jimai 20. Jimai 22, Jimai 25, Luomai 86036, Ning S1101, Ning S1129, Shaanxi 229, Shanyou 225, Wanmai 38, Wanmai 52, Wanmai 53, Wanmai 55, Yanyou 361, Xinzi 10 -911, Xu Mai 26, Xu Mai 27, Xu Mai 29, Yu Mai 18;

[0049] The control wheat variety was Huaimai 18.

[0050] 1. Soak the wheat grains of the tested wheat variety (line) and the control wheat at 20°C to make the wheat grains germinate 4-6mm young sprouts, and place the germinated young wheat grains on the hyphae of Rhizoctonia graminearum at 20°C Soak in the suspension for 5 minutes to inoculate, and continue to culture for 12 days after inoculation;

[0051] The preparation method of Rhizoctonia graminearum R0301 suspension and the cultivation method of wheat refer...

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Abstract

The invention provides a method for predicting sheath blight resistance of wheat at a seedling stage. The method comprises the following steps: (A) soaking and germinating wheat grains of to-be-detected wheat variety or strain, inoculating and cultivating for 12 days; (B) taking a to-be-detected wheat seedling base part including 1cm of stalk of a leaf sheath, weighing, marking as A, wherein the unit is mg, extracting total DNA of the stalk as a to-be-detected DNA sample, marking the volume as B, wherein the unit is muL; (C) with the to-be-detected DNA sample obtained from the step B as a template, SEQ.ID NO:1 as a positive primer, and SEQ.ID.NO:2 as a reverse primer, carrying out fluorescent quantitative PCR detection to obtain a CP value, repeatedly detecting twice again, so as to obtain a CP mean value C from detection by three times; (D) calculating the DNA content E of rhizoctonia cerealis of the stalk of the wheat seedling base part according to a formula E=[B*10(31.784-C) / 3.562] / A; (E) obtaining the DNA content E1 of the rhizoctonia cerealis of the stalk of the base part of a huai-wheat No.18 seedling employing the same steps A to D; and (F) predicting the sheath blight resistance of the wheat at the seedling stage according to the ratio of E to E1.

Description

technical field [0001] The invention relates to the field of wheat plant protection and breeding, in particular to a method for predicting resistance to wheat sheath blight at seedling stage. Background technique [0002] Wheat sharp eyespot, mainly caused by Rhizoctonia cerealis Vander hoeven, is a worldwide soil-borne fungal disease of wheat with great harm (Hamada MS, Yin YN, ChenHG, Ma Z. The escalating threat of Rhizoctonia cerealis, the causal agent of sharp eyespot in wheat. "Pest Managgment Science", 2011,67:P.1411-1419), has become a major disease in the Yangtze River Basin of my country, and gradually spread to the Huanghuai wheat region, every year Caused serious loss of wheat yield. [0003] Breeding and using resistant varieties of sheath blight is undoubtedly the most effective and economical way to control the disease, the most critical of which is the establishment of resistance identification methods for sheath blight. Currently, the reported identification m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/06C12R1/645
CPCC12Q1/6851C12Q2545/113C12Q2563/107
Inventor 周淼平姚金保马鸿翔余桂红
Owner JIANGSU ACAD OF AGRI SCI