Paeonia ostii PoCCoAOMT gene cDNA full-length sequence and application thereof in drought resistance of plants

A technology of sequence and phoenix dan, applied in the field of plant genetic engineering, can solve the problems of no genome report, weak genetic background of phoenix dan, no description of specific sequence, etc., to achieve the effect of enhancing drought tolerance.

Active Publication Date: 2020-07-24
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The genetic background of Fengdan is relatively weak, and there is no report on the genome
As for the research on the CCoAOMT gene of Fengdan, Li Guo (Peony 'Fengdan' DFR gene cloning and functional verification and lignin synthesis gene discovery, Northwest Agriculture and Forestry University master's thesis, 2017) used Fengdan seed shell and kernel as materials Transcriptome sequencing was carried out and a total of 13 CCoAOMT gene fragments were identified, but the specific sequence was not described, and the full-length cDNA sequence of the CCoAOMT gene was not obtained through cloning, and there was no report on the application of Fengdan CCoAOMT gene to plant drought resistance

Method used

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  • Paeonia ostii PoCCoAOMT gene cDNA full-length sequence and application thereof in drought resistance of plants
  • Paeonia ostii PoCCoAOMT gene cDNA full-length sequence and application thereof in drought resistance of plants
  • Paeonia ostii PoCCoAOMT gene cDNA full-length sequence and application thereof in drought resistance of plants

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1 Cloning of the full-length cDNA sequence of PoCCoAOMT gene

[0024] The 3'end cDNA sequence of PoCCoAOMT gene was obtained: Fengdan leaves under drought stress were selected as materials, and the total RNA was extracted with MiniBEST Plant RNA Extraction Kit (TaKaRa). Use 3'full RACE CoreSet Ver.2.0 (TaKaRa) reverse transcription to produce the first strand of cDNA. The reverse transcription system is: 1μL RNA, 1μL 3'-RACEAdaptor, 1μL dNTP Mixture (10mM each), 2μL 5×M -MLV Buffer, 0.25μL RNaseInhibitor, 0.25μL Reverse Transcriptase M-MLV(RNase H - ), 4.5μL RNase Free ddH 2 O; Reverse transcription program: reaction at 42°C for 60 minutes, extension at 70°C for 15 minutes. On this basis, 3'-RACE performs two rounds of PCR amplification respectively. The first round of PCR amplification system is: 2μL cDNA, 8μL 1×cDNA Dilution BufferⅡ, 2μL 3'-RACEOuter Primer, 2μL Gene specific Outer Primer(10μM)(5'-CGTGAAGTAACCGCAAAA-3'(SEQ ID NO.1)) 、5μL 10×LA PCR BufferⅡ(Mg + ...

Embodiment 2

[0026] Example 2 Homology comparison analysis of the amino acid sequence of Fengdan and other 8 plant CCoAOMT genes

[0027] The amino acid sequences inferred from Fengdan and other 8 plant CCoAOMT genes are expressed in FASTA format, saved as TXT file, and then loaded into DNAMAN5.2.2 software for homology comparison, and the most homologous amino acid can be observed. Sequence, see results figure 2 .

Embodiment 3

[0028] Example 3 Analysis of amino acid conservative motifs inferred from Fengdan and other 8 plant CCoAOMT genes

[0029] The amino acid sequences inferred from Fengdan and other 8 plant CCoAOMT genes are expressed in FASTA format, saved as a FASTA file, and then loaded into the MEME online tool to analyze the conservative motifs of the CCoAOMT protein. Set according to the default conditions, you can observe To 8 conservative motifs, see the results image 3 .

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Abstract

The invention discloses a paeonia ostii PoCCoAOMT gene cDNA full-length sequence and application thereof in drought resistance of plants. The paeonia ostii PoCCoAOMT gene cDNA full-length sequence isdisclosed as SEQ ID NO. 6. The new paeonia ostii PoCCoAOMT gene cDNA full-length sequence is cloned, and the nucleotide sequence of the new paeonia ostii PoCCoAOMT gene cDNA full-length sequence and the obtained amino acid sequence are determined. Besides, the eukaryotic expression vector of the PoCCoAOMT gene is transformed into the plant, especially tobacco, and the plant has obviously enhanceddrought resistance.

Description

Technical field [0001] The invention belongs to the technical field of plant genetic engineering, and specifically relates to a full-length cDNA sequence of Fengdan PoCCoAOMT gene and its application in plant drought resistance. Background technique [0002] In recent years, due to the greenhouse effect, global warming has been increasing, and extreme drought disasters have appeared in many places, which have seriously threatened the agricultural and forestry production in the region. Fengdan (Paeonia ostii) belongs to the perennial woody plant of the genus Paeonia in the family Paeoniaceae. It is a variety of Yangshan peony and has high ornamental value, medicinal value and oil value. Fengdan was approved by the Ministry of Health as an oil-use peony resource on March 22, 2011. Because of its advantages of easy survival, large seed yield, and high content of unsaturated fatty acids, it has been popularized in more than 20 provinces and regions across the country, especially in m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/54C12N15/11C12N15/84A01H5/00A01H6/82
CPCC12N9/1007C12N15/8273C12Y201/01104
Inventor 赵大球陶俊张夏燕孙静孟家松张克亮
Owner YANGZHOU UNIV
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