Hibiscus cannabinus L. expression sequence tag SSR (Simple Sequence Repeat) DNA markers

A technique for expressing sequence tags and DNA markers, which can be applied in recombinant DNA technology, DNA/RNA fragments, microbial determination/inspection, etc., and can solve problems such as late start of research

Inactive Publication Date: 2015-02-04
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on molecular markers of kenaf started relatively late at home and abroad, and only a small number of laboratories have started related work. At present, the microsatellite markers of kenaf can be found to be part of the genomic SSR (public number CN1302111 C)

Method used

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  • Hibiscus cannabinus L. expression sequence tag SSR (Simple Sequence Repeat) DNA markers
  • Hibiscus cannabinus L. expression sequence tag SSR (Simple Sequence Repeat) DNA markers
  • Hibiscus cannabinus L. expression sequence tag SSR (Simple Sequence Repeat) DNA markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Based on the transcriptome data of kenaf, 24 different types of transcription factor expression sequence tags were determined, and 24 new microsatellite markers of kenaf were developed, as shown in Tables 1 and 2.

Embodiment 2

[0029] Embodiment 2: Utilize the kenaf germplasm resources of different sources, through PCR amplification and electrophoresis detection, determine 24 microsatellite markers with abundant polymorphism, as table 3 and figure 1 shown.

[0030] Table 3 Polymorphism and amplification efficiency of kenaf EST-SSR

[0031]

[0032] ABCD indicates the level of clarity of the electrophoretic band pattern.

Embodiment 3

[0033] Example 3: Analysis of genetic diversity of kenaf germplasm resources using the microsatellite markers developed above. The test materials come from the Research Office of Hemp Genetics and Breeding of Fujian Agriculture and Forestry University, namely Fuhong 992, Fuhong 2, Fuhonghang 1B, Fuhong B3, Fuhong R-1, KG121, Zanyin 1 and Fuhong 952B . Among them, Zanyin No. 1 is an excellent strain introduced and bred from Zambia, with red stems, round leaves, and tall plants; while other materials are high-generation strains bred by the research group, with green stems, split leaves, and plants higher. Using NTsys software to calculate the genetic similarity coefficient of 8 kenaf materials. The calculation method is unweighted cluster analysis (UPGMA), and the reading band adopts the method of 0, 1, 0 means no band, and 1 means band. The results of cluster analysis showed that these 24 microsatellite markers could be well used in the genetic diversity analysis of kenaf. ...

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Abstract

The invention relates to a molecular marker technology and specifically relates to hibiscus cannabinus L. expression sequence tag SSR (Simple Sequence Repeat) DNA markers. The following steps are included: new SSR sites are developed based on a hibiscus cannabinus L. transcriptome sequencing sequence, and the primer sequences of the SSR sites are amplified and polymorphic analysis is performed on the SSR sites. The hibiscus cannabinus L. expression sequence tag SSR DNA markers are characterized in that the DNA sequences of 24 SSR markers are separated and identified and the primer sequences of the 24 SSR sites are amplified based on the transcription factor expression sequence tags of hibiscus cannabinus L. transcriptome sequencing. The 24 new hibiscus cannabinus L. SSR DNA markers can be applied to researches such as genetic diversity analysis of the hibiscus cannabinus L. breeds, genetic linkage map establishment and molecular marker assisted selection, and are good in repeatability; as a result, the 24 new hibiscus cannabinus L. SSR DNA markers are reliable and effective molecular markers.

Description

technical field [0001] The invention relates to a molecular marker technology, in particular to a kenaf expression sequence tag microsatellite DNA molecular marker. Background technique [0002] Kenaf ( Hibiscus cannabinus L.) is an annual bast fiber crop belonging to the Malvaceae Hibiscus genus. It has the characteristics of wide adaptability, salt-alkali tolerance, waterlogging tolerance, barren tolerance, drought resistance, high fiber yield, good quality and fast growth. Its fibers can be used as textile raw materials, industrial raw materials and biomass energy, etc., and are highly valued by the industry in the development of new products and materials. [0003] Microsatellite (also known as simple repeat sequence, SSR) is a piece of DNA composed of a short sequence of 1-6 nucleotides repeated in tandem multiple times. The length of the amplified microsatellite sequence is polymorphic because the number of repetitions of the repeating unit in the microsatellite...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 张立武祁建民万雪贝
Owner FUJIAN AGRI & FORESTRY UNIV
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