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A method for identification of radish clubroot resistance

A radish root and resistance technology, which is applied in the field of vegetable crop genetics and breeding, and achieves the effects of low equipment and technical requirements, good stability and product separation effect, and simple and convenient detection method.

Active Publication Date: 2022-07-19
NANJING AGRICULTURAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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  • A method for identification of radish clubroot resistance
  • A method for identification of radish clubroot resistance
  • A method for identification of radish clubroot resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Identification of radish germplasm resistance to clubroot

[0033] The radish germplasm seeds were sterilized, rinsed and germinated, and then sown in a plug tray containing vermiculite: peat = 1:1 sterilization substrate. 6 seeds per accession, 1 per hole, were repeated 3 times, and protective rows were planted around the test material to prevent marginal effects.

[0034] After sowing, each hole was injected with 2 mL of 'XY-2' P. brassicae bacterial solution for inoculation (spore concentration 1×10 7 mL -1 ).

[0035] After 40 days of inoculation, the root disease of each plant tested was investigated, identified and analyzed, identified and graded one by one.

[0036] refer to as figure 1 The grading standard for the identification of radish clubroot disease at the seedling stage shown in: Grade 0, each root system grows normally, no gall mass; Grade 1, the main root does not swell, but there are small galls on the fibrous root; Grade 3, the main roo...

Embodiment 2

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

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

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Abstract

The invention discloses a method for identification of radish clubroot resistance. The radish clubroot resistance functional marker RsNBS135C was obtained by amplifying the radish genomic DNA by the upstream primer RsNBS135C‑F and the downstream primer RsNBS135C‑R. The nucleotide sequence amplified by the marker in the radish genomic DNA shows a specific band pattern after agarose gel electrophoresis, which can quickly and directly identify the target resistance gene locus in the radish germplasm, and is conducive to efficient and reliable utilization Target genes for clubroot disease resistance and accelerate the process of genetic improvement of clubroot disease resistance.

Description

technical field [0001] The invention belongs to the field of vegetable crop genetics and breeding, and relates to a method for identifying the resistance of radish clubroot. 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 dietary value, and 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. Pathogens infect diseased plants, resulting in abnormally enlarged roots, resulting in a decline in the quality and commercial value of vegetable products, and severely restricting the industrialization of cruciferous crops. Due to the strong vitality and high mutat...

Claims

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Application Information

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Patent Type & Authority Patents(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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