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Method for identifying sugarcane smut resistance by utilizing SCAR (Sequence Characterized Amplified Region) label

A smut and marker technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of incompatibility, low recombination probability, and high cost of breeding large populations, and reduce environmental conditions. Influence and bias of human judgment, effect of small amount of DNA and short detection time

Inactive Publication Date: 2012-07-18
FUJIAN AGRI & FORESTRY UNIV
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Problems solved by technology

[0003] At present, the identification of sugarcane resistance to smut at home and abroad still adopts artificial inoculation of sugarcane stems and planting in the field, and the disease resistance identification is carried out by observing the incidence rate throughout the growing season. 2 new plantings or 1 new planting and 1 perennial root) to make a comprehensive judgment on the identification results, and the period is as long as 2 years; secondly, this method has a large workload, occupies a large area, and is expensive, and it is difficult to identify large-scale samples. The highly heterozygous allopolyploid crops of sugarcane are also asexually propagated crops, and the hybrid F 1 Generational segregation populations are highly segregated in traits, which is the best time for selection, but the recombination probability of excellent aggregated genotypes is very low, usually the probability from seedlings to bred varieties is about 1 / 100,000, so the workload of identification and selection is huge
The selection of sugarcane smut resistance traits is also based on the large population of hybridization and segregation. The existing identification methods are not suitable for the large population requirements of breeding; The result of the interaction of factors, therefore, the environment has a great influence on the disease resistance phenotype, and the identification results of different years have relatively large differences
[0004] Therefore, the identification efficiency and accuracy of sugarcane resistance to smut are directly related to the safety of sugarcane production. However, the current identification methods for disease resistance are inefficient, long-term, expensive, and insufficient in accuracy. Accurate Disease Resistance Identification Technology

Method used

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  • Method for identifying sugarcane smut resistance by utilizing SCAR (Sequence Characterized Amplified Region) label
  • Method for identifying sugarcane smut resistance by utilizing SCAR (Sequence Characterized Amplified Region) label
  • Method for identifying sugarcane smut resistance by utilizing SCAR (Sequence Characterized Amplified Region) label

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Embodiment 1

[0019] Example 1 A method for identifying resistance to smut in sugarcane using SCAR markers

[0020] A method for identifying resistance to smut in sugarcane using SCAR markers, including the following steps:

[0021] 1. Extraction of genomic DNA

[0022] (1) Take 5 grams of young leaves of sugarcane plants to be tested, place them in a mortar, add liquid nitrogen to quickly grind them into powder, and transfer them to a 50 ml centrifuge tube before thawing;

[0023] (2) Add 15 ml of SDS extract solution preheated at 65 ℃, mix well and keep it in a 65 ℃ water bath for 40 min;

[0024] (3) Add 8 ml of 3 mol / L KAC and mix well, then ice bath for 30 min;

[0025] (4) Centrifuge at 25,000 g at 4°C for 20 min; collect the supernatant and add 12 ml of isopropanol pre-cooled at 4°C;

[0026] (5) After placing it at -20°C for 30 minutes, hook out the DNA floating on the liquid surface and place it in a clean 1.5 ml centrifuge tube. After drying, add 750 μl TE; add an equal volume of balanced sat...

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Abstract

The invention relates to a method for identifying sugarcane smut resistance by utilizing an SCAR (Sequence Characterized Amplified Region) label. The method comprises the following steps of: extracting genome DNA; establishing a detection system of an SCAR-PCR (Polymerase Chain Reaction); and detecting an SCAR-PCR product. The method for identifying sugarcane smut resistance by utilizing the SCAR label has the characteristics of high sensitivity and lower dosage of DNA, and compared with conventional artificial inoculation field resistance identification, the method has the advantages of simple and convenient operation, short detection time, high accuracy, easy judgment, good repeatability, no environment condition influence and the like. The method provides a simple, convenient and effective smut resistance identification system for the evaluation of sugarcane germplasm resources and the approval of new varieties and has significance to scientific researches and production.

Description

technical field [0001] The invention relates to a method for identifying plant disease resistance molecular markers, in particular to a method for identifying sugarcane smut resistance using SCAR markers, and belongs to the field of biotechnology. Background technique [0002] sugar cane( Saccharum officinarum ) is the most important sugar crop, sucrose accounts for more than 90% of my country's total sugar production, and sugarcane production is related to my country's sugar safety. Ustilago Ustilago scitaminean The sugarcane smut caused by it is a worldwide and the most serious sugarcane fungal disease in my country. At present, the incidence of perennial root smut in main cultivars is as high as 20%, causing serious losses. Breeding disease-resistant varieties is the most economical and effective The efficiency and accuracy of disease resistance identification directly affect the process and effect of disease resistance breeding. [0003] At present, the identification...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 阙友雄许莉萍徐景升陈如凯
Owner FUJIAN AGRI & FORESTRY UNIV
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