Method for identifying pennisetum forage grass by utilizing ISSR (Inter Simple Sequence Repeat) molecular marker technology

A molecular marker and technical identification technology, which is applied in the field of forage identification using molecular marker technology to achieve the effect of ensuring vigorous promotion, reducing workload and ensuring authenticity

Inactive Publication Date: 2012-07-18
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is no report on the application of ISSR molecular markers to distinguish and identify Pennisetum forage grasses at home and abroad

Method used

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  • Method for identifying pennisetum forage grass by utilizing ISSR (Inter Simple Sequence Repeat) molecular marker technology
  • Method for identifying pennisetum forage grass by utilizing ISSR (Inter Simple Sequence Repeat) molecular marker technology
  • Method for identifying pennisetum forage grass by utilizing ISSR (Inter Simple Sequence Repeat) molecular marker technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Screening of ISSR molecular markers of Pennisetum

[0037] (1) Based on the collected materials of 2 main species of Pennisetum and their hybrids, they are: Hybrid Pennisetum No. 1, Hybrid Pennisetum No. 2, and Hybrid Pennisetum No. 3 , Elephant Grass, Hybrid Pennisetum, Purple Pennisetum, Broadleaf Pennisetum;

[0038] (2) Seedling cultivation: select 100 uniform seeds of each variety, and cultivate them for 20 days under greenhouse conditions to form normal seedlings as test materials;

[0039] (3) A new type of plant genome DNA extraction kit (Tiangen Reagent Company) was used to extract the total DNA of a single seedling, and three single plants were randomly selected for each material, and 0.8% agarose gel electrophoresis was used for detection and DNA was determined with a nucleic acid protein analyzer. The concentration of the sample is shown in Table 1, and the DNA pool of each material was prepared so that the final concentration of DNA was 10 ng / μL;...

Embodiment 2

[0055] Example 2 Identification method 1

[0056] (1) Extract Pennisetum DNA and construct a DNA pool;

[0057] Select the seven Pennisetum materials that need to be identified: Hybrid Pennisetum No. 1, Hybrid Pennisetum No. 3, Hybrid Pennisetum No. 3, Elephant Grass, Hybrid Pennisetum, Ziguang Pennisetum, and Broad-leaved Pennisetum For one or more of them, select 100 uniform seeds from each material to be identified, and cultivate them under greenhouse conditions for 20 days to form normal seedlings. Take the leaves of a single seedling and use Tiangen’s new plant genome DNA extraction reagent Genomic DNA of Pennisetum genus was extracted from each box, and three individual plants were extracted from each material, detected by 0.8% agarose gel electrophoresis, and the DNA sample concentration was determined by a nucleic acid and protein analyzer. Three individual plants of each material were used The DNA was mixed and diluted in equal amounts to prepare DNA pools for the DN...

Embodiment 3

[0064] Example 3 Identification method 2

[0065] (1) Extract Pennisetum DNA and construct a DNA pool;

[0066] Select the seven Pennisetum materials that need to be identified: Hybrid Pennisetum No. 1, Hybrid Pennisetum No. 3, Hybrid Pennisetum No. 3, Elephant Grass, Hybrid Pennisetum, Ziguang Pennisetum, and Broad-leaved Pennisetum For one or more of them, select 100 uniform seeds from each material to be identified, and cultivate them under greenhouse conditions for 20 days to form normal seedlings. Take the leaves of a single seedling and use Tiangen’s new plant genome DNA extraction reagent Genomic DNA of Pennisetum genus was extracted from each box, and three individual plants were extracted from each material, detected by 0.8% agarose gel electrophoresis, and the DNA sample concentration was determined by a nucleic acid and protein analyzer. Three individual plants of each material were used The DNA was mixed and diluted in equal amounts to prepare DNA pools for the DN...

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Abstract

The invention belongs to the field of forage grass germplasm resource identification, in particular to a method for identifying pennisetum forage grass by utilizing an ISSR (Inter Simple Sequence Repeat) molecular marker technology, wherein primers used by ISSR-PCR (Polymerase Chain Reaction) are UBC815 and UBC835, and the nucleotide sequences of the primers are respectively UBC815-CTCTCTCTCTCTCTCTG and UBC835-AGAGAGAGAGAGAGYC. The method fills the blank of distinguishing pennisetum forage grass categories (varieties) by applying the ISSR molecular marker method, and an electrophoresis ideograph finally drawn by the method for distinguishing seven pennisetum forage grass materials for tests can be used as a basis for distinguishing and identifying the seven materials. The method gets rid of unreliable factors of traditional morphology identification and can quickly and accurately distinguish the seven pennisetum forage grass materials, and an identification result can be used as a reliable basis for approving categories, varieties and strains so as to guarantee the great popularization of germplasm on production.

Description

technical field [0001] The invention belongs to the identification field of forage germplasm resources, and in particular relates to a method for identifying forage grass using molecular marker technology. Background technique [0002] Pennisetum Rich. is an annual or perennial grass forage, which is a high-quality forage resource. Among them, elephant grass (P.purpureum) has the characteristics of high yield, rich nutritional value, and high temperature and rainy characteristics, and American pennisetum (P.americanum) has high quality and good palatability. American pennisetum (male parent) and elephant grass (female parent) The interspecific triploid hybrid hybrid pennisetum has the advantages of both male and female parents, and has the characteristics of strong adaptability and long grass supply period. As an excellent green fodder for herbivores, it plays an important role in breeding production. role. With the deepening of research, the introduction and selection of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张蕴薇王乐杨富裕毛培胜于晓丹刘斯佳李洪超
Owner CHINA AGRI UNIV
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