Method for developing genic-SSR molecular markers of Dongxiang wide rice

A molecular marker, wild rice technology, applied in the field of molecular biology, can solve limited problems, achieve the effect of overcoming the difficulty of acquisition, simple method and easy operation

Pending Publication Date: 2016-10-12
JIANGXI NORMAL UNIV
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Problems solved by technology

However, the use of Dongxiang wild rice transcriptome data to develop genetic-SSR molecular markers has not been reported. At the same time, the available genetic-SSR molecular markers of Dongxiang wild rice are also very limited.

Method used

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  • Method for developing genic-SSR molecular markers of Dongxiang wide rice
  • Method for developing genic-SSR molecular markers of Dongxiang wide rice
  • Method for developing genic-SSR molecular markers of Dongxiang wide rice

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Embodiment 1

[0043] Using the above method, the high-throughput sequencing data of the above-ground and underground tissue transcriptomes of Dongxiang wild rice were downloaded from the network database of the National Center for Biotechnology Information (NCBI) in the United States. Then, 76,258 Dongxiang wild rice non-redundant Unigene-EST sequences were assembled (Table 1). A total of 21,226 SSR loci were discovered by using MISA software. The frequency distribution of SSR variant types is shown in Figure 1. Primer 3.0 software was used to develop SSR primers 3681 pairs.

[0044] 25 pairs were randomly selected from 3681 pairs of designed SSR primers, and the genomic DNA of Dongxiang wild rice and 15 modern cultivated rice (Table 2) were used to verify the universality of the primers. The specific material information is shown in Table 2. The results showed that 18 of the 25 pairs of primers selected could detect clear target amplification bands in Dongxiang wild rice (Table 3), and the...

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Abstract

A method for developing genic-SSR molecular markers of Dongxiang wide rice comprises the following steps: acquiring the Unigene-EST sequence of Dongxiang wild rice; identifying the SSR sequence of Dongxiang wild rice; designing genic-SSR primers of Dongxiang wild rice; determining the validity and the generality of the genic-SSR primers of Dongxiang wild rice. According to the method provided by the invention, an SSR locus is excavated from the Unigene-EST sequence of Dongxiang wild rice based on a bioinformatics method, development of the genic-SSR molecular markers is carried out, and the problem that the genic-SSR molecular markers of Dongxiang wide rice are difficult to acquire in the prior art is solved; when Dongxiang wild rice and modern cultivated rice of different varieties are adopted for PCR identification of the developed genic-SSR primers, the obtained results are true and reliable. The method provided by the invention is simple, and simple and convenient to operate; the number of the acquired molecular markers is huge, so that a foundation is laid for genetic research, molecular breeding and the like of Dongxiang wild rice.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a method for developing a Dongxiang wild rice genic-SSR molecular marker. Background technique [0002] For a long time, due to breeders' one-sided pursuit of high yield and simplification of variety promotion, the genetic diversity of modern cultivated rice has been significantly reduced. Wild rice resources are in a state of natural growth, containing rich genetic resources and genetic diversity, and are extremely valuable genetic resources for rice improvement. Dongxiang wild rice originated in Dongxiang County, Jiangxi Province (28 o 14'), is the northernmost common wild rice discovered so far in China and even the world, rich in many excellent characteristics (such as high yield, cytoplasmic sterility, fertility recovery, strong tillering, etc.) , drought resistance, disease resistance, insect resistance, etc.), using the excellent genetic resources of Dongxiang wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/156
Inventor 张帆涛张朦谢建坤孙佳
Owner JIANGXI NORMAL UNIV
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