Method for constructing boehmeria nivea core idioplasm by using EST-SSR (Expressed Sequence Tag-Simple Sequence Repeat) molecular marker

A core germplasm and molecular marker technology, which is applied in the field of plant germplasm resource evaluation, can solve the problem that the core germplasm of ramie has not yet been constructed, and achieve the effects of reliable core germplasm, short time consumption, and overcoming various shortcomings.

Inactive Publication Date: 2014-03-19
INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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Problems solved by technology

[0004] At present, there is no method for constructing ramie core collection using SSR molecular markers

Method used

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  • Method for constructing boehmeria nivea core idioplasm by using EST-SSR (Expressed Sequence Tag-Simple Sequence Repeat) molecular marker
  • Method for constructing boehmeria nivea core idioplasm by using EST-SSR (Expressed Sequence Tag-Simple Sequence Repeat) molecular marker
  • Method for constructing boehmeria nivea core idioplasm by using EST-SSR (Expressed Sequence Tag-Simple Sequence Repeat) molecular marker

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Experimental program
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Effect test

Embodiment 1

[0022] 1 Materials and methods

[0023] 1.1 Materials

[0024] 108 ramie germplasm resources. The above materials were all planted in the National Changsha Ramie Germplasm Garden.

[0025]

[0026]

[0027]

[0028] 1.2 Method

[0029] 1.2.1 Genomic DNA extraction

[0030] Take 108 newly issued shoots of ramie germplasm (Table 1) plants in the National Germplasm Changsha Ramie Garden, and use the Tiangen kit to extract DNA. After the concentration of the extracted DNA is detected by electrophoresis, the concentration of the sample DNA is calculated and diluted to the required concentration.

[0031] 1.2.2 Primer design and synthesis

[0032] Use SSRhunter combined with manual search to search for ramie EST sequences containing SSRs. The search criteria are: the number of repeats of single nucleotides ≥ 16, the number of repeats of dinucleotides ≥ 6, the number of repeats of trinucleotides ≥ 4, four cores The repeating times of nucleotides≥3, and the repeating ti...

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Abstract

The invention discloses a method for constructing boehmeria nivea core idioplasm by using an EST-SSR (Expressed Sequence Tag-Simple Sequence Repeat) molecular marker. The method comprises the steps of firstly extracting DNA (Deoxyribonucleic Acid) of boehmeria nivea idioplasm resources, then, carrying out PCR (Polymerase Chain Reaction) amplification through EST-SSR primers, and recording the banding pattern of each idioplasm resource according to amplification bands; analyzing the obtained banding patterns, and constructing the boehmeria nivea core idioplasm by using heuristic search and the principle of the DNA amplified banding pattern maximization of the idioplasm resources. According to the method, the forming of the EST-SSR primers and the experimental operating processes are simple, the operation is easy, the amplified banding patterns are clear and stable, and the repeatability is good. Compared with field-property core idioplasm construction, the molecular marker based boehmeria nivea core idioplasm construction has the characteristics of time saving, labor saving, good stability and the like. According to the core idioplasm constructed based on the molecular marker, the maximization variety of the DNA amplified banding patterns is guaranteed, the representativeness is very good, and the core idioplasm has important significance in preservation, evaluation and utilization of the idioplasm of boehmeria nivea.

Description

technical field [0001] The invention belongs to the technical field of plant germplasm resources evaluation, and in particular relates to a method for constructing ramie core germplasm by using EST-SSR molecular markers. Background technique [0002] Ramie (Boehmeria nivea L.Gaud), also known as "Chinese grass", originated in China, is a perennial herb of the genus Ramie in the family Urticaceae, and is a natural textile raw material with Chinese characteristics [1]. In addition to the traditional fiber value, ramie's medicinal value, feed use, water and soil conservation, industrial raw materials, bio-energy and other multi-functional uses have begun to attract attention [2-5] . So far, my country is the country that collects and preserves the most germplasm resources of the genus Ramie. The national germplasm Changsha Ramie Garden alone has more than 2,000 materials, and the cultivation resources have been widely used in breeding and production. [6] . Plant genetic reso...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q2531/113
Inventor 栾明宝陈建华王晓飞许英孙志民邹自征
Owner INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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