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Codominant marking method and its uses in plant breeding

A co-dominant marker, plant technology, applied in the fields of application, plant genetic improvement, botanical equipment and methods, etc., can solve the problems of complex steps, difficult to popularize, high cost, and achieve good repeatability, large amount of information, and low cost. Effect

Inactive Publication Date: 2009-06-10
北京市中农良种有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, RFLP, COS and other markers have disadvantages such as high cost, complicated steps, long cycle, and high-quality DNA requirements, which are difficult to promote in assisted breeding, and RFLP markers need to use radioactive isotopes

Method used

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  • Codominant marking method and its uses in plant breeding
  • Codominant marking method and its uses in plant breeding
  • Codominant marking method and its uses in plant breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Establishment of Codominant-SCAR, a co-dominant marker technique

[0036] Tomato materials TA1527 and E6203 were sown in the greenhouse. When the seedlings grew to 6-8 true leaves, the DNA in the tender leaves was extracted for analysis. DNA extraction was carried out according to the routine CTAB (Williamson V M. et al., 1994) mini-extraction method, and the DNA was stored in a -20°C refrigerator for future use.

[0037] Taking the COS marker T0974 as the research object, it was first converted into a CAPS marker, and obtained the PCR products of the CAPS markers of the parent materials TA1527 and E6203 (Wang Xiaoxuan. Dissertation of the Chinese Academy of Agricultural Sciences, 2004). The PCR products were sequenced (sequenced by Sanbo Polygala Company), and the obtained sequencing results were named T0974-2F and T0974-3R, respectively.

[0038] The T0974-2F and T0974-3R sequences were homologously compared with DNAMAN, and then Primer Pmemier5.0 was used...

Embodiment 2

[0046] Example 2: Genetic identification of 12 near-isogenic lines of TA517

[0047] Tomato materials TA517, TA1527, E6203, and 12 near-isogenic lines of TA517 were applied: 02P68, 02P69, 02P70, 02P71, 02P72, 02P73, 02P74, 02P75, 02P76, 02P77, 02P78, 02P79. All the test materials were sown in the greenhouse, and when the seedlings grew to 6-8 true leaves, the DNA in the tender leaves was extracted for analysis. DNA extraction, primer design and PCR conditions were carried out according to Example 1.

[0048] The results show that (attached figure 2 ): The polymorphisms of 67, 75, 68, 69, 71 and 72 were consistent, and a 270bp band was obtained; the polymorphisms of 1A, 70, 73, 74, 76, 78 and 79 were consistent, and a 200bp band was obtained The bands are consistent with the expected results (Wang Xiaoxuan. Dissertation of Chinese Academy of Agricultural Sciences, 2004, 26-30).

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Abstract

This invention discloses a Codominant SCAR method, which comprises the following steps: making the same source comprarison on the male and female parent DNA list of the RFLP, AFLP, CAPS, RAPD, COS maker, then using Primer5 or other software to design eduction to obtain the apparent SCAR eduction that is accordant with the male and female parent material gene types, and then adding the parent eduction into the PCR reaction system to obtain the special bands of a spectrum which can simultaneously indentify the male and female parent and its mixed gene type.

Description

technical field [0001] The invention relates to a plant co-dominant-SCAR (Codominant-SCAR) method. Specifically, it is a method established by combining the principle of SCAR labeling with multi-primer PCR technology. Further, the present invention utilizes this method to carry out genetic identification on 12 tomato near-isogenic lines. Background technique [0002] In 1992, Tanksley et al. published a high-density tomato linkage map dominated by RFLP markers, with an average of one marker per 1.2 cM (centimeter) (Tanksley et al., 1992). Since then, the tomato map has been continuously saturated with various markers ( Http: / / www.sgn.cornell.edu / maps / tomato_arabidopsis ). The completion of Arabidopsis genome sequencing has accelerated the saturation of molecular markers on the tomato map. For example, the COS (Conserved Ortholog Set) marker technology developed by using the existing tomato EST sequence and Arabidopsis genome sequence information has increased the number o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68A01H1/00
Inventor 杜永臣国艳梅王孝宣高建昌朱德蔚戴善书
Owner 北京市中农良种有限责任公司