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Crested wheatgrass P genome specific sequence

A specific sequence, genome technology, applied in genetic engineering, plant genetic improvement, biochemical equipment and methods, etc., can solve problems that have not yet been reported in research.

Inactive Publication Date: 2007-03-07
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, there has not been a special report on the specific sequence of the Genus P genome and the specific molecular markers that identify the P chromosome.

Method used

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  • Crested wheatgrass P genome specific sequence
  • Crested wheatgrass P genome specific sequence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1, the acquisition of P genome-specific sequence

[0019] Using RAPD primer Opx-7: 5'-GAGCGAGGCT-3' (SEQ ID NO: 1), the P genome DNA of Agrogenus genus and Triticea ABD genome DNA were screened, and the specific amplified fragment of P genome was isolated and cloned.

[0020] Among them, the 20ul system of PCR amplification: 10×PCRbuffer (containing 20mMol / L Mg 2+ ) 2ul

[0021] dNTP(10mM) 0.3ul

[0022] Primer (8uM) 0.7ul

[0023] Taq enzyme (2u / ul) 0.5ul

[0024] Template DNA (50ng / ul) 1ul

[0025] wxya 2 O 15.5ul

[0026] PCR program: first fully denatured at 94°C for 4min; then denatured at 94°C for 30S, annealed at 37°C for 45S, extended at 72°C for 1min, 45 cycles; finally extended at 72°C for 10min. Store at 4°C.

[0027] 1.2% agarose gel, 100V constant voltage electrophoresis, UV gel imaging.

[0028] The results of PCR amplification are shown in Figure 1. A specific band not found in the Triticea ABD genome was amplified in the diploid, tetr...

Embodiment 2

[0030] Embodiment 2, wheatgrass P genome-specific sequence verification

[0031] According to the sequencing results of the specific amplified fragments of the P genome, a pair of SCAROPX-7 primers were designed by DNASTAR design software optimization: SCAROPX-7F: 5'-GCTACCATTCCAACTTCTACG-3' (sequence 3), corresponding to 75-95nt of sequence 2 ; SCAROPX-7R: 5'-GCGAGGCTATGTTGGTTTTGA-3' (SEQ ID NO: 4), reverse complementary to 1014-1034nt of SEQ ID NO:2.

[0032] SCAROPX-7 primers were used to amplify different genomic DNAs of the Triticum family (annealing temperature 62° C.), and the amplification and electrophoresis conditions were the same as in Example 1. The results showed that in diploid wheatgrass Z804, tetraploid wheatgrass Z559 and hexaploid wheatgrass Z1750, a specific fragment (SCAR marker) with a length of 960 bp was amplified, while in wheat ABD genome and most wheat The specific fragment of 960bp was not obtained in other genomes of the family (C, D, E, AG, I, R....

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Abstract

the invention discloses an astopic sequence of crested wheatgrass P gene group, which is characterized by the following: adopting gene group DNA as mould to do PCR augmentation for RAPD primer; obtaining 1036bp nucleotide sequence; or using SCAR primer to do PCR augmentation to obtain 960bp nucleotide sequence; possessing a couple of oligonucleotide with sequence 3 and sequence 4 for SCAR primer; providing new astopic probe and molecular mark for P chromatin rapidly and exactly.

Description

technical field [0001] The present invention relates to the specific sequence of wheatgrass P genome. Background technique [0002] Wheat is the main food crop of human beings and plays an important role in agricultural production. At present, the narrow genetic base of cultivated wheat has caused serious obstacles to wheat improvement, and wheat wild relatives are a huge gene pool for wheat genetic improvement. [0003] There are 25 genera and nearly 400 species in the Triticea family, occupying a pivotal position in the grass family. The basic genome of plants of the genus Agropyron is Group P (Dong Yuchen, Zheng Diansheng. China Wheat Genetic Resources. China Agricultural Press, 2000, 174-177), distributed in the cold temperate plateau and sandy land of Eurasia, not only adapting to drought for a long time , cold climate and poor soil caused by abiotic adversity, and has strong resistance to major diseases such as powdery mildew, yellow dwarf and rust of wheat. In addit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29
Inventor 刘伟华吴嫚李立会杨欣明高爱农李秀全李洪杰
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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