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SCAR marker linked closely with sorghum aphid-resistant gene and application thereof

A kind of aphid-resistant gene and marker technology, which is applied in the fields of application, plant gene improvement, DNA/RNA fragments, etc., can solve the problems of labor-intensive, time-consuming, and drug resistance of sorghum aphids, so as to save the workload of field breeding, save manpower, The effect of broad promotion and application prospects

Inactive Publication Date: 2010-04-07
SHENYANG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to solve the technical problems of labor-intensive, time-consuming and cost-intensive use of chemical agents to prevent and control sorghum aphids, and produce drug resistance and pollute the environment by selecting aphid-resistant varieties in the most effective way against the above-mentioned deficiencies in the prior art.

Method used

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  • SCAR marker linked closely with sorghum aphid-resistant gene and application thereof
  • SCAR marker linked closely with sorghum aphid-resistant gene and application thereof
  • SCAR marker linked closely with sorghum aphid-resistant gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Acquisition of RAPD polymorphic marker linked to aphid resistance gene

[0066] 500 random primers were screened by RAPD technology, and a total of 1614 DNA bands were amplified, among which the amplified products of primers OPN-07, OPN-08 and OPY-14 had linked polymorphic fragments. primers for further linkage analysis, co-analyzed F 2 There were 132 isolated populations, of which 98 were resistant to aphids and 34 were susceptible to aphids. 0PN-07 727 , OPN-08 373 and OPY-14 600 The recombination rates were 3.0%, 6.1% and 9.1%, respectively. Using the Mapmaker 3.0 software package, the map distance units are 3.2cM, 6.5cM and 11.7cM respectively.

[0067] OPN-07 727 , OPN-08 373 the F 2 See the results of single plant amplification figure 1 , 2 . From figure 1 It can be seen that OPN-07 was amplified in anti-parent (1), anti-pool (3) and anti-aphid individual plants (6, 9, 11, 13, 15, 16, 18) 727 This polymorphic fragment was not amplified in Ganqin (2), ...

Embodiment 2

[0070] Recovery, Cloning and Sequencing of RAPD Markers Linked to Aphid Resistance Genes

[0071] Will OPN-07 727 and OPN-08 373 Two polymorphic fragments were recovered, cloned and sequenced. Sequencing results The polymorphic fragment amplified by OPN-08 is 373bp, and the nucleotide sequence of the polymorphic fragment amplified by OPN-07 is shown in Table 2 and Table 3. According to the sequencing results, the two polymorphic fragments were named OPN-07 727 and OPN-08 373 . The bold underlined bases in the figure are random primer sequences.

[0072] Table 2 RAPD marker OPN-07 727 The sequencing results of

[0073] Table2 Sequenced results of polymorphism fragment of OPN-07

[0074] 1AAACGACGGC CAGTGCCAAG CTTGCATGCC TGCAGGTCGA CGATT CAGCC

[0075] 51 CAGAG GAGAC ATGTTTCCAC CTATTCTTTG AATGTATCTT CAGCCAAGCC

[0076] 101TGCTGGAATC TACTAAGCAT TAACTGGGAC AAATCCCTCC AACCTTTTGG

[0077] 151 TATGCTGATC AAAGTTAGAA CAGATTTTGG AATGCACATC TTTAGAGAAA

[0078] 201TTTTC...

Embodiment 3

[0108] RAPD markers converted to SCAR markers

[0109] According to the specific fragment OPN-07 727 , OPN-08 373 Based on the principles of both-end sequence and primer design (avoid hairpin structure, appropriate G+C content), two pairs of specific primers were designed at both ends of the sequence.

[0110] Use this pair of primers to detect BTAM428 (anti-parent), ICS-12B (sense relative) and F after hybridization between the two 2 The results showed that the SCAR markers and the RAPD markers of the aphid-resistant gene were completely consistent in the resistant ponds, susceptible ponds and individual plants, and the OPN-08 373 a and OPN-08 373 b is the primer for PCR amplification (i.e. SCAR analysis), the results are shown in image 3 . From image 3 It can be seen that OPN-08 was amplified in anti-parent (1), anti-chi (3) and anti-aphid single plants (5-13) 373 One band, but no fragments were amplified in Ganqin (2), Ganchi (4) and Ganyi single plant (14-22).

...

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Abstract

The invention relates to an SCAR marker linked closely with sorghum aphid-resistant gene and an application thereof. The BTAM428*ICS-12B hybrid progeny is used as raw material to perform molecular tagging to sorghum aphid-resistant gene by using BSA method and adopting RAPD and SCAR screening techniques to obtain two RAPD markers OPN-07727 and OPN-07373 linked closely with sorghum aphid-resistant gene. The markers OPN-07727 and OPN-07373 of the gene is closely linked with sorghum aphid-resistant gene and the nucleotide sequences of the markers can be obtained by sequencing. Meanwhile, the invention changes the markers OPN-07727 and OPN-07373 to stable SCAR markers, the markers can be used to perform detection to the F2 generation and partial sorghum resistant varieties to find molecular marker specific bands from resistant varieties so as to provide reference for molecular marker assistant breeding, overcome the difficulty that the workload is very great by adopting the common method for assistant selection in phenotype analysis and provide technical support for realizing molecular marker assistant breeding and reducing the breeding cycle.

Description

Technical field: [0001] The invention relates to a closely linked SCAR marker of sorghum aphid-resistant gene and its application, in particular to the RAPD marker of sorghum aphid-resistant gene and the SCAR marker transformed from the marker, and the application of the marker in identification of aphid-resistant varieties. Background technique: [0002] Sorghum (Sorghum bicolor (L.) Moench) is one of the most important cereal crops in the world. It is mainly distributed in tropical arid and semi-arid regions, and is also planted in temperate and cold regions. From a global perspective, it is second only to wheat, rice, corn, and barley, ranking fifth. [0003] Sorghum aphid [Melanaphis sacchari (Guenée)] is one of the main pests that frequently occur in sorghum production areas in my country, generally causing a yield reduction of about 15%, and a serious yield reduction of more than 30%. In the past, the use of chemical agents to prevent and control was time-consuming, l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11A01H1/04
Inventor 李玥莹邹剑秋李雪梅林凤刘天华马纯艳彭霞陶思源徐昕陈琳
Owner SHENYANG NORMAL UNIV
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