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Method for quickly identifying genetic purity of glutinous corn hybrid

An identification method, the technology of waxy corn, applied in the biological field, can solve the problems of low accuracy, heavy workload, and long time consumption, and achieve the effects of stable labeling, high accuracy, and improved efficiency

Inactive Publication Date: 2013-08-07
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the genetic purity detection of fine corn seeds mainly includes field planting identification and isoenzyme identification, but field identification requires sowing seedlings, field planting management, and observation of morphological characteristics, which is time-consuming, heavy workload, and susceptible to seasonal changes. Restriction and influence of environmental factors (Kong Guangchao et al. Study on the method of testing the purity of corn seeds [J]. Seeds, 2000, (3): 26-28)
Because the morphological difference between the maize pseudo-hybrids and the true hybrids is not obvious before harvest, it is difficult to distinguish between the seedling stage and the adult plant stage, especially when the genetic basis between the parents is very close, it is even more difficult to distinguish
The analysis of isozymes is easily affected by the environment and the parts of the materials taken, and for some hybrid varieties with small genetic differences between the parents, it is sometimes difficult to detect and identify the isozymes for purity, and the accuracy is not high (Liu Liwang et al. Molecular marker technology in Application in Variety Identification and Purity Detection of Vegetable Crops [J]. Molecular Plant Breeding, 2004, 2(4): 563-568)

Method used

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  • Method for quickly identifying genetic purity of glutinous corn hybrid
  • Method for quickly identifying genetic purity of glutinous corn hybrid
  • Method for quickly identifying genetic purity of glutinous corn hybrid

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Embodiment 1

[0033] Rapid identification of genetic purity of waxy maize hybrid seeds.

[0034] The main operating procedures for the rapid identification of the genetic purity of waxy maize variety ‘Su Yunuo 2’ using SRAP and RAPD molecular markers are as follows:

[0035] 1 DNA extraction

[0036] (1) Take about 0.1 g of the young leaves of waxy corn variety ‘Su Yunuo 2’, grind them with liquid nitrogen, add 500 μl of CTAB lysate (2% CTAB, 2mol L -1 NaCl, 20mmol·L -1 EDTA, 100mmol·L -1 Transfer Tris-HCl (pH 8.0) and 0.2% β-mercaptoethanol (V / V) into a 1.5ml centrifuge tube, and place in a water bath at 65°C for 30-50min;

[0037](2) Take out and centrifuge at 16°C and 12,000rpm for 10min, transfer the supernatant into a newly sterilized 1.5ml centrifuge tube, add an equal volume of chloroform:isoamyl alcohol (24:1, volume ratio) mixture, and then Shake gently, and centrifuge at room temperature at 12,000rmp for 10min;

[0038] (3) Aspirate the supernatant into a new centrifuge tu...

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Abstract

The invention belongs to the technical field of biology, and relates to a method for quickly identifying the genetic purity of a glutinous corn hybrid. The method comprises the following steps of: extracting genomic deoxyribonucleic acid (DNA) from glutinous corn, performing polymerase chain reaction (PCR) amplification by using the screened sequence-related amplified polymorphism (SRAP) effective primer combination NAUSRem7 / NAUSRpm1 and random amplified polymorphic DNA (RAPD) effective primers NAUSR709 and NAUSR712, respectively performing non-denaturing polyacrylamide gel electrophoresis and agarose gel electrophoresis on a PCR product obtained through amplification, and shooting a DNA electrophoresis pattern; and comparing and analyzing the size and position difference of polymorphism amplified fragments formed due to the difference of DNA fragment sequences in the electrophoresis pattern, and identifying the genetic purity of the glutinous corn F1 hybrid, namely a Suyunuo 2 seed. The detection method has the advantages of marking stability, high accuracy, no influence of the growth stage and environment of a sample to be detected, low cost, capability of being performed in thewhole growth season, and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for quickly identifying the genetic purity of waxy corn hybrids, which uses SRAP and RAPD molecular marker techniques to detect the genetic purity of seeds of the waxy corn variety 'Suyunuo 2'. Background technique [0002] Seeds are extremely important means of production in agriculture, and the quality of seeds is directly related to the economic interests of seed producers, operators and users. The important basis for judging the quality of seeds is the four indicators of genetic purity, germination rate, clarity and moisture. The latter three index tests can be carried out indoors, but genetic purity identification is relatively difficult. Accurate, rapid and convenient identification of seed genetic purity is an important part of modern agricultural production and seed work. Maize belongs to Poaceae Zea (Zea) maize species (mays) (Gai Junyi. Monographs on Crop Breeding ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 柳李旺刘君龚义勤徐良陈莹陆虎华
Owner NANJING AGRICULTURAL UNIVERSITY
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