Specific molecular markers for detection of Aegilops comosa 2M, 3M, 6M and 7M chromosomes in wheat, kit and method

A molecular marker and chromosome technology, applied in the field of molecular biology, can solve the problem of lack of awn goat grass and other problems

Active Publication Date: 2016-10-12
CROP RES INST SHANDONG ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problem that there is no PCR marker for the Chromosome of Goatwort in the prior art, the present invention provides a method that can quickly, effectively and simply identify whether the sample to be tested contains Chromosomes 2M, 3M, 6M and 7M in Goatwort. specific molecular markers

Method used

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  • Specific molecular markers for detection of Aegilops comosa 2M, 3M, 6M and 7M chromosomes in wheat, kit and method
  • Specific molecular markers for detection of Aegilops comosa 2M, 3M, 6M and 7M chromosomes in wheat, kit and method
  • Specific molecular markers for detection of Aegilops comosa 2M, 3M, 6M and 7M chromosomes in wheat, kit and method

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Experimental program
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Effect test

Embodiment 1

[0052] Designing and screening PLUG primers for homology groups 2, 3, 6, and 7 of chromosomes 41 and 2, 3, 6, and 7 using A. chinensis, Chinese spring-A. For pairs, 84 pairs, 53 pairs and 73 pairs, the amplified products were digested with restriction endonuclease HaeIII and TaqI respectively, and the PCR amplification products from the same primer (undigested, HaeIII digested and TaqI digested three case) agarose gel electrophoresis, it was found that the CD913720 of the second homologous group of chromosomes ( figure 1 A) and CK171581 ( figure 1 B), CD491237 of the third homology group of chromosome ( figure 1 C) and CK207576 ( figure 1 D), CK168221 of the sixth homologous group of chromosome ( figure 1 E) and CK206466 ( figure 1 F), CK214770 of the seventh homologous group of chromosome ( figure 1 G) and CK203272 ( figure 1 H) Additional polymorphic bands can be amplified in A. chinensis and Chinese spring-A. 650bp, 1200bp and 770bp ( figure 1 Arrows in A-H), howeve...

Embodiment 2

[0056] Up to now, since there is no report on the creation of the 1M addition line of the Chinese spring-A. g Chromosome donors may have undergone partial recombination or variation of chromosomes during the polyploidization and evolution of the species, resulting in slightly different chromosomes from A. chinensis and the M chromosome of A. , with M g Indicated to show the difference. Therefore, in the absence of the Chinese spring-A. g The additional system is used for marker positioning.

[0057] In order to locate the above polymorphic bands from A. chinensis on one or several chromosomes of A. chinensis 1M-7M, the 8 pairs of PLUG primers screened above were used to pair Chinese spring-A. Grass Diploid, Chinese Spring, Chinese Spring - Goat Grass 1M g Addition line, Chinese Spring-Dingmang goat grass 2M addition line, Chinese spring-Dingmang goat grass 3M addition line, Chinese spring-Dingmang goat grass 4M addition line, Chinese spring-Dingmang goat grass 5M addition ...

Embodiment 3

[0063] In order to verify the practicability of the above 8 markers, the 8 pairs of primers screened above were used to amplify the 4 populations respectively to verify the practicability of the markers.

[0064] Among them, CD913720 and CK171581 of the second chromosome homology group were used to amplify P1-P42 progeny materials of the Chinese Spring-Dingmis chinensis 2M addition line / Chinese Spring hybrid F2 progeny, and the results showed that both CD913720 and CK171581 could be used in wheat- Amplified polymorphisms in the amphidiploid, wheat-A. acanthus 2M addition line, P2-P6, P16-P20, P25, P26, P28, P31, P32, P36, P39, P40 and P42 Marked CD913720-PLUG and CK171581-PLUG, but no corresponding polymorphic bands were amplified in Chinese Chunhe P1, P7-P15, P21-P24, P27, P29, P30, P33-P35, P37, P38 and P41 (Table 2). Among them, the amplification results of CD913720 on P1-P21 are as follows image 3 As shown in A, the amplification result of CK171581 on P22-P42 is as foll...

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Abstract

The invention relates to the field of molecular biology, in particular to specific molecular markers for detection of Aegilops comosa 2M, 3M, 6M and 7M chromosomes in wheat, and the specific molecular markers are DNA base sequences respectively shown as SEQ ID No.1 and -18 in the sequence table. A to-be-detected chromosome sample and a specific molecular marker composition are put into a kit to conduct amplification. The specific molecular markers and the kit can be used for detection or assisted detection of the existence of Aegilops comosa 2M, 3M, 6M and 7M chromosomes in wheat. The invention provides an effective detection means for screening and identification of wheat-Aegilops comosa distant hybridization new germplasms involving Aegilops comosa 2M, 3M, 6M and 7M chromosomes, provides a detection method for transfer of excellent genes on Aegilops comosa 2M, 3M, 6M and 7M chromosomes to wheat by means of chromosome segments, and has positive industrialization value.

Description

technical field [0001] The present invention relates to the field of molecular biology, in particular to specific molecular markers for detection of 2M, 3M, 6M and 7M chromosomes in wheat, and to a kit containing the specific molecular markers, and for the detection of 2M in wheat. , 3M, 6M and 7M chromosomes. Background technique [0002] Aegilops comosa (Aegilops comosa, 2n=2x=14, genome MM), is highly resistant to various diseases such as wheat stripe rust, leaf rust, stem rust and powdery mildew (Friebe B, Jiang JM, Raupp WJ, et al.Characterization of wheat-alien translocations conferring resistance to diseases and pests: current status. Euphytica, 1996, 91:59-87; Gill BS, Sharma HC, Raupp WJ, et al. Evaluation of Aegilops species for resistance to wheat powdery mildew, wheat leaf rust, Hessian fly and greenbug. Plant Dis, 1985, 69:314-316), is an excellent gene source for wheat breeding. Since the 1960s, Riley's research group in the UK, Miller's research group and Do...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 刘成宫文萍韩冉刘建军李豪圣宋健民刘爱峰曹新有程敦公李根英楚秀生黄承彦赵振东
Owner CROP RES INST SHANDONG ACAD OF AGRI SCI
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