Primer combination for genetic integrity analysis of common wild rice and application of primer combination

A primer combination, wild rice technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, recombinant DNA technology, etc., can solve problems such as distribution area degradation

Inactive Publication Date: 2016-05-04
江西省农业科学院水稻研究所 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Common wild rice is the most widely distributed wild rice species in my country. However, due to environmental damage and other reasons, its distribution area has been greatly degraded in recent decades.

Method used

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  • Primer combination for genetic integrity analysis of common wild rice and application of primer combination
  • Primer combination for genetic integrity analysis of common wild rice and application of primer combination
  • Primer combination for genetic integrity analysis of common wild rice and application of primer combination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] The genetic integrity analysis of embodiment 1 common wild rice germplasm

[0059] 1. Experimental materials

[0060] The common wild rice in Dongxiang, Jiangxi Province was used as the test material, and a certain amount of seeds were transplanted to the field for routine management. Randomly select 160 individual plants, take their young leaves, and freeze them at -20°C for later use.

[0061] 2. Genomic DNA extraction: Genomic DNA of 160 individual leaves were extracted by CTAB method.

[0062] 3. Primer synthesis and screening

[0063] (1) Synthetic primers: With reference to the rice SSR primer sequence, 550 pairs of primers were randomly selected, and the primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd.

[0064] (2) Primer screening: using 160 single-plant DNA templates, using all 550 pairs of primers to amplify, the amplified products were subjected to polyacrylamide gel electrophoresis and gel silver staining, and the non-polymorphic one...

Embodiment 2

[0068] Example 2 Analysis of Genetic Integrity of Common Wild Oryza Germplasm in Different Vitality Populations

[0069] 1. Experimental materials

[0070] The common wild rice in Dongxiang, Jiangxi was used as the test material. Common wild rice seeds were collected, and high-temperature and high-humidity methods were used to age the seeds to obtain populations with different germination rates (see Table 2 for details). Each population took a certain amount of seeds to grow seedlings and transplanted them to the field for routine management. 160 individual plants were randomly selected from each group, and the young leaves were taken and frozen at -20°C for future use.

[0071] 2. Genomic DNA extraction: Genomic DNA of 160 individual leaves were extracted by CTAB method.

[0072] 3. Comparison of genetic integrity of different vitality populations: Using the above 25 pairs of core primers, the genomic DNA of 5 different vitality populations of common wild rice was amplified...

Embodiment 3

[0077] Example 3 is used for the primer combination of common wild rice genetic integrity analysis

[0078] SSR01 forward primer SEQ ID NO.1caatttccataggctgcatg

[0079] Reverse primer SEQ ID NO.2gcttgggttagcgacgac

[0080] SSR02 forward primer SEQ ID NO.3tctttggaggcgagctgag

[0081] Reverse primer SEQ ID NO.4cgaagggtacatctgcttag

[0082] SSR03 forward primer SEQ ID NO.5gtacggaaaacatggtaggaag

[0083] Reverse primer SEQ ID NO.6tcgagggaaggatctggtc

[0084] SSR04 Forward Primer SEQ ID NO.7ggagcgggagagagagcccacg

[0085] Reverse primer SEQ ID NO.8 tgccgatgaaggactgcgacgc

[0086] SSR05 Forward Primer SEQ ID NO.9ctgatcgagagcgttaaggg

[0087] Reverse primer SEQ ID NO.10gggatcaaaccacgtttctg

[0088] SSR06 Forward Primer SEQ ID NO.11ctactcctctgtccctcctctc

[0089] Reverse primer SEQ ID NO.12 ccagctagacacaatcgagc

[0090] SSR07 forward primer SEQ ID NO.13agcgacgccaagacaagtcggg

[0091] Reverse primer SEQ ID NO.14tccacgtcgatcgacacgacgg

[0092] SSR08 Forward Primer SEQ ID NO....

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PUM

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Abstract

The invention relates to genetic integrity analysis primers, and particularly discloses SSR molecular marker core primers for genetic integrity analysis of common wild rice and application of the core primers. A batch of the SSR core primers used for the genetic integrity analysis of the common wild rice are screened out by taking the common wild rice in Jiangxi province as a material through a large number of experimental studies, a primer combination for the genetic integrity analysis of the common wild rice is built, and an analyzing method is built on the basis of the primer combination. The analyzing method is suitable for analyzing the genetic integrity analysis of the common wild rice. Through the group quantity of the screened primer combination and at least 90 single plants, a standard method is supplied to accurate analysis of genetic integrity detection in the germplasm preservation and reproduction updating process of the common wild rice.

Description

technical field [0001] The invention relates to genetic integrity analysis primers, in particular to SSR molecular marker core primers for common wild rice genetic integrity analysis. Background technique [0002] Common wild rice is the close ancestor species of cultivated rice, which contains a large number of excellent genetic resources. It has good compatibility and high hybrid seed setting rate, and is a precious genetic resource for cultivated rice breeding. Common wild rice is the most widely distributed wild rice species in my country. However, due to environmental damage and other reasons, its distribution area has been greatly degraded in recent decades. The main ways to protect common wild rice germplasm resources include: in situ conservation, germplasm nursery protection and gene bank preservation of seeds. The goal of its conservation and preservation is very clear: to maintain its genetic integrity and diversity. [0003] The genetic integrity of the germpla...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 黎毛毛辛霞吴锦文尹广鹍余丽琴张金梅李慧陈晓玲熊玉珍卢新雄
Owner 江西省农业科学院水稻研究所
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