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Method using sequence characterize amplified region (SCAR) signs to identify sugarcane brown rust disease resistance

A SCAR-B and labeling technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of insufficient accuracy of identification results, difficult large-scale sample identification, high cost of management and investigation, and achieve simplicity Effective brown rust resistance identification system, high accuracy and short detection time

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

The main problems of this method are: first, it takes a long time, each crop season lasts for one year, and the identification results of at least two crop seasons are required for comprehensive judgment; second, the workload is large, the area is large, and the management and investigation costs High, it is difficult to identify large-scale samples, which is contradictory to sugarcane crossbreeding, which is based on the selection of large populations of segregated generations (i.e. seedlings)
For now, 1 million seedlings are planted in my country every year. Obviously, the current brown rust resistance identification technology can only be carried out on a few breeding materials after many years of identification and selection of traits such as yield and quality. Come on, at most dozens of breeding materials can be used to identify the resistance to brown rust. Therefore, many opportunities for selection must be lost; the third is in the system of interaction of pathogen, sugarcane genotype and environment , the disease can only occur if all three factors are present, that is to say, even if there is a pathogen in the field, the sugarcane genotype is also susceptible to the disease, and suitable environmental conditions are required for the disease to occur. The three are indispensable. Therefore, the field is based on Induced disease resistance identification method, identification results are unpredictable
There are large differences in the identification results in different places and different years, and the accuracy of the identification results is not enough

Method used

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  • Method using sequence characterize amplified region (SCAR) signs to identify sugarcane brown rust disease resistance
  • Method using sequence characterize amplified region (SCAR) signs to identify sugarcane brown rust disease resistance

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Embodiment 1

[0020] Example 1 A method for identifying sugarcane brown rust resistance using SCAR markers

[0021] A method utilizing SCAR markers to identify sugarcane brown rust resistance, comprising the following steps:

[0022] 1. Genomic DNA extraction

[0023] (1) Take 5 grams of young leaves of the sugarcane plant to be tested, put them in a mortar, add liquid nitrogen to grind them into powder, and transfer them to a 50 ml centrifuge tube before thawing;

[0024] (2) Add 15 ml of SDS extract preheated at 65 °C, mix well and keep warm in a water bath at 65 °C for 40 min;

[0025] (3) Add 8 ml of 3 mol / L KAC and mix well, then place in ice bath for 30 min;

[0026] (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;

[0027] (5) After standing at -20 ℃ for 30 minutes, hook out the DNA floating on the liquid surface, put it in a clean 1.5 ml centrifuge tube, add 750 μl TE solution after drying; add an equal volume o...

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Abstract

The invention relates to a method using sequence characterize amplified region (SCAR) signs to identify sugarcane brown rust disease resistance, which comprises extraction of genome deoxyribonucleic acid (DNA), detection system establishment of SCAR- polymerase chain reaction (PCR) signs and detection of SCAR-PCR products. The method using SCAR signs to identify sugarcane brown rust disease resistance has the advantages of being high in flexibility and small in using amount of DNA. Compared with general resistance identification, the method has the advantages of being convenient in operation,short in detection time, high in accuracy, easy in judgment, good in repeatability, free of effect of environmental conditions, and the like. The method provides a convenient and effective brown rustdisease resistance identification system for assessment of sugarcane germplasm resources and accreditation of new products and has significance on enhancing brown rust resistance sugarcane breading and protection on brown rust resistance sugarcane germplasm resources.

Description

technical field [0001] The invention relates to a method for identifying plant disease resistance by using molecular marker technology, in particular to a method for identifying sugarcane brown rust resistance by using SCAR markers, and belongs to the field of biotechnology. Background technique [0002] sugar cane( Saccharum officinarum ) is a plant of the Gramineae Saccharum genus, native to tropical and subtropical regions, with the advantages of high light efficiency, high light saturation point, low carbon dioxide compensation point, low photorespiration rate and high photosynthetic intensity. Sugarcane is the most important sugar crop in my country and the green energy crop with the highest biomass. Its production is not only related to the safety of sugar in my country, but also provides sufficient raw materials for the development of energy alcohol. [0003] by Puccinia nigricans ( Puccinia melanocephala ) caused by sugarcane brown rust is one of the most importan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 阙友雄许莉萍黄宁陈如凯
Owner FUJIAN AGRI & FORESTRY UNIV
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