Method for identifying clubroot resistance of radishes

A radish root and resistance technology is applied in the field of vegetable crop genetics and breeding, and achieves the effects of simple and convenient detection method, clear and simple band pattern of amplified products, and low requirements for equipment and technology.

Active Publication Date: 2020-07-17
NANJING AGRICULTURAL UNIVERSITY +1
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

However, there are few reports on the identification and evaluation of radish clubroot resistance, and no resistance-related genes have been isolated and identified

Method used

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  • Method for identifying clubroot resistance of radishes
  • Method for identifying clubroot resistance of radishes
  • Method for identifying clubroot resistance of radishes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Identification of Radish Germplasm Clubroot Resistance

[0033] Radish germplasm seeds are sown in plug trays equipped with vermiculite: peat = 1:1 sterilized matrix after being sterilized, washed and accelerated germination. Sow 6 seeds for each germplasm, 1 seed for each hole, repeat 3 times, and plant a protective line around the tested materials to prevent marginal effects.

[0034] After sowing, inject 2 mL of 'XY-2' Plasmodium brassica bacteria solution into each hole for inoculation (spore concentration 1×10 7 mL -1 ).

[0035] After 40 days of inoculation, investigate, identify and analyze the diseased conditions of the roots of the tested plants, identify and grade them one by one.

[0036] Refer to as figure 1 The grading standards for the identification of radish clubroot at the seedling stage are as follows: Grade 0, each root system grows normally, without galls; Grade 1, the main root does not expand, and there are small galls on the fibrous...

Embodiment 2

[0042] Example 2: Development and application of radish clubroot resistance marker based on RsNBS135 gene

[0043] The whole genome nucleotide sequence, annotated CDS sequence and amino acid sequence of radish were downloaded from Radish Genome Database (http: / / radish.kazusa.or.or.jp / ). According to the NBS protein domain (NB-ACR, PF00931) (http: / / pfam.xfam.org / family / PF00931) using the Markov model (HMM, Hidden MarkovModel) to search the radish genome protein sequence, E-value is set to 0.01. All sequences were submitted to Pfam, NCBI and SMART to verify the reliability of the sequences. After removing the non-NBS domain and incomplete sequences, the identified 188 NBS-containing disease resistance candidate genes were named RsNBS001-RsNBS188, respectively. The TIR-NBS-LRR gene RsNBS135 was closely related to the resistance to clubroot disease (race 4) through the sequence difference analysis of the clubroot disease-resistant germplasm. Using RT-qPCR to analyze the expression...

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Abstract

The invention discloses a method for identifying clubroot resistance of radishes. The turnip clubroot resistance functional marker RsNBS135C is obtained by amplifying turnip genome DNA (deoxyribonucleic acid) by virtue of an upstream primer RsNBS135C-F and a downstream primer RsNBS135C-R. A nucleotide sequence obtained by amplification of the marker in radish genome DNA presents a specific bandingpattern after agarose gel electrophoresis, can quickly and directly identify a target resistance gene locus in radish germplasm, is beneficial to efficient and reliable utilization of a clubroot-resistant target gene, and accelerates a clubroot resistance genetic improvement process.

Description

technical field [0001] The invention belongs to the field of genetic breeding of vegetable crops, and relates to a method for identifying resistance to clubroot of radish. Background technique [0002] Radish (Raphanus sativus L.) is an important cruciferous root vegetable crop native to my country. It is rich in nutrition, resistant to storage and transportation, and has high therapeutic value. It is deeply loved by people. Radish has a long history of cultivation in my country (Jia Sixie (Northern Wei Dynasty), Qimin Yaoshu Jinshi [M]. Zhonghua Book Company 2007), and the planting area is wide. [0003] Clubroot mainly occurs in cruciferous plants and is a soil-borne disease caused by Plasmodiophora brassicae. The pathogen infects the infected plants and causes the roots to swell abnormally, resulting in a decline in the quality and commercial value of vegetable products, which seriously restricts the industrialization of cruciferous crops. Due to the strong vitality and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 柳李旺孟玥徐良王燕张婉婷张晓莉
Owner NANJING AGRICULTURAL UNIVERSITY
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