Molecular marker primer and method for identifying common camellia oleifera and small-fruit camellia oleifera

A molecular marker, a technology for Camellia oleifera, which is applied in the field of molecular biology, can solve the problem that there is no chloroplast gene molecular marker, and achieves the effects of low cost and simple method.

Pending Publication Date: 2021-11-05
NANCHANG UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

So far, there is no research report on the application of chloroplast gene molecular markers in the identification of common Camellia oleifera and Camellia oleifera

Method used

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  • Molecular marker primer and method for identifying common camellia oleifera and small-fruit camellia oleifera
  • Molecular marker primer and method for identifying common camellia oleifera and small-fruit camellia oleifera
  • Molecular marker primer and method for identifying common camellia oleifera and small-fruit camellia oleifera

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Embodiment 1

[0032] (1) Extract the genomic DNA of common Camellia oleifera and Camellia oleifera samples from different regions:

[0033] In this embodiment, common Camellia oleifera populations in 11 regions and Camellia oleifera populations in 6 regions were selected, and 3 individuals were randomly selected from each population as test samples, see Table 1 for details.

[0034] Take 0.02 g of dry leaves of the sample to be tested, add liquid nitrogen to grind thoroughly, and use a new plant genome DNA extraction kit (TIANGEN, Shanghai) to extract the whole genome DNA of the sample.

[0035] Integrity, purity and concentration were checked by 1.0% agarose gel electrophoresis and DNA / RNA UV spectrophotometer (NanoDrop 2000, Thermo, USA).

[0036] OD 260 / OD 280 For DNA samples >1.8, dilute to 2ng / μl for subsequent amplification. DNA extracts were stored in a -20°C refrigerator for later use.

[0037] Table 1: Information on the sampling sites of Camellia oleifera and Camellia oleifer...

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Abstract

The invention discloses a molecular marker primer and method for identifying common camellia oleifera and small-fruit camellia oleifera. The molecular marker primer can amplify gene spacer regions atpH and atpI on a chloroplast genome of camellia oleifera. The method comprises the following steps of: extracting genome DNA of camellia oleifera leaves to be detected as a template, performing PCR amplification by taking the molecular marker primer as an amplification primer, and performing electrophoresis detection on the amplification product; if a strip of about 609bp appears in the electrophoresis result, determining that the camellia oleifera is common camellia oleifera; and if a strip of about 227bp appears in the electrophoresis result, determining that the camellia oleifera is small-fruit camellia oleifera. The sequence of the molecular marker primer is as follows: an upstream primer: 5'-CTCCCTCTCCTAACCAACCC-3', and a downstream primer: 5'-CCGCGGCTTATATAGGTGAA-3'. The molecular marker primer disclosed by the invention can be used for rapidly identifying common camellia oleifera and small-fruit camellia oleifera, the method is simple, rapid and accurate, and the method is a molecular means which cannot be replaced by conventional morphological characteristics for identifying common camellia oleifera and small-fruit camellia oleifera.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a molecular marker primer and a method for identifying common Camellia oleifera and Camellia oleifera. Background technique [0002] Camellia oleifera generally refers to the general term for a class of plants with high seed oil content and economic value for cultivation in the Theaceae (Theaceae) Camellia. Camellia oleifera seed oil is rich in unsaturated fatty acids mainly composed of oleic acid and linoleic acid. Its nutritional value is comparable to that of olive oil. It is a high-quality edible vegetable oil. Camellia oleifera has a long history of application in my country, with a wide distribution area and a large cultivation area. Among them, the planting area of ​​common camellia oleifera (C. oleifera) has exceeded 67 million mu, and it is the largest woody oil crop. Camellia oleifera (C. meiocarpa) next. At present, the output of camellia oleifer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 张剑戎俊陈璐陈凯赵耀
Owner NANCHANG UNIV
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