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PCR primers, methods and kits for analyzing grass genetic diversity

A technology of genetic diversity and gramineous plants, applied in methods and kits, and in the field of PCR primers for analyzing the genetic diversity of gramineous plants, to achieve high detection efficiency

Active Publication Date: 2019-12-06
TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI
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Problems solved by technology

[0004] At present, no SRAP molecular marker primers have been reported for the study of obtuse leaf grass germplasm resources

Method used

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  • PCR primers, methods and kits for analyzing grass genetic diversity
  • PCR primers, methods and kits for analyzing grass genetic diversity
  • PCR primers, methods and kits for analyzing grass genetic diversity

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Embodiment Construction

[0054] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Mg used in the present invention 2+ , dNTPs, Taq DNA polymerase and standard molecular weight DNA Marker for gel detection were purchased from Beijing Tiangen Co., Ltd.

[0055] In the embodiment of the present invention, 13 obtuse leaf grasses collected by the Tropical Crops Variety Resources Research Institute of the Chinese Academy of Tropical Agricultural Sciences were used as test materials (see Table 1).

[0056] Table 1 Experimental materials of blunt leaf grass

[0057]

[0058] 1. Genomic DNA extraction

[0059] Using the modified CTAB method, the steps are as follows:

[0060] (1) Take fresh young leaves, add liquid nitrogen to quickly grind them into powder, transfer them to a 2mL centrifuge tube, add 1mL pre-cooled CTAB-free buffer, and ice-bath for 10min. Centrifuge at 7 000r / min for 5min, discard the supernatant;

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Abstract

The invention provides PCR (polymerase chain reaction) primers, method and kit for analyzing genetic diversity in gramineous plants. According to the technical scheme provided herein, 3 DNA templates different in character are picked from 13 DNA parts, 21 pairs of primers with clear polymorphic bands are screened out of 100 pairs of primers, the 21 pairs of primers are used to mark the 13 DNA parts, electrophoresis, imaging, photographing and data statistics are performed, UPGMA (unweighted pair-group method with arithmetic means) cluster analysis is performed on genetic similarity and genetic distances of the plants, and diversity analysis and interspecific relation research are performed. The research indicates that gramineous plant germplasm resources show rich genetic diversity; the genetic groups of these materials have no great relationship with geographic origin and geographic environment; SRAP (sequence-related amplified molecular polymorphism) marker is a molecular marker suitable for the research on gramineous plant germplasm resources.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a PCR primer, method and kit for analyzing the genetic diversity of gramineous plants. Background technique [0002] Genetic diversity is an inherent characteristic of every organism and is the product of long-term adaptation and evolution. The genetic diversity of obtuse leaf grass is very important in its own right. When the genetic diversity of a species disappears, it will not recover, and when the genetic diversity decreases to a certain level, inbreeding depression will occur and even lead to species extinction. my country is one of the origins of plants of the genus Oxyphyllum, and the distribution range is also very wide. At present, some scholars have studied Oxyphyllum, such as Huangjuan and Laomao. But in-depth research at the molecular level is still rare. [0003] The invention and application of SRAP (Sequence-related Amplified Polymorphism) marker, a new...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143
Inventor 黄春琼刘国道白昌军
Owner TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI
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