A kind of plant gene and its application
A gene and plant technology, applied in the application field of regulating plant disease resistance, can solve problems such as separation, and achieve the effect of improving plant disease resistance
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Embodiment 1
[0018] Example 1: High-throughput transcriptome sequencing of marigold
[0019] Since the marigold genome has not been determined, in order to obtain the sequence of functional gene transcripts, a total of 9 samples were subjected to high-throughput DNA extraction by extracting RNA from leaves, immature flowers, and mature flowers of three single plants using tissue samples of marigold cultivars. Transcriptome sequence determination and assembly annotation.
[0020] 1. Reagents
[0021] The plant RNA extraction reagent Trizol was purchased from Invitrogen Company, DNase I (Dnase I) was purchased from TaKaRa Company, the RNA library preparation kit (RNA Library Prep Kit) was from Beijing Biomike Biotechnology Co., Ltd., and the rest of the reagents were imported. Or domestic analytically pure products.
[0022] 2. Plant material
[0023] Marigold (Tagetes erecta L.) cultivar Juwang was purchased from Chifeng Xinhui Gardening Company.
[0024] 3. Method
[0025] 3.1 RNA ext...
Embodiment 2
[0058] Embodiment 2: the cloning of TeLTF4 gene of marigold
[0059] According to the data assembly of Marigold TeLTF4 gene in Example 1, use the primer design software PrimerPremier 5.0 to design PCR-specific primers, extract total RNA from mature flowers of Marigold, and reverse transcription PCR (reversetranscription-PCR, RT-PCR) clone to obtain TeLTF4 Gene.
[0060] 1. Reagents
[0061] Plant RNA extraction reagent Trizol was purchased from Invitrogen Company, DNase I (Dnase I) was purchased from TaKaRa Company, reverse transcriptase, dNTP, and high-fidelity DNA polymerase were purchased from Beijing Quanshijin Biotechnology Co., Ltd., and primers were purchased from Sangon Biotechnology Co., Ltd. Synthesized by Engineering (Shanghai) Co., Ltd., and the rest of the reagents were imported or domestically produced analytically pure products.
[0062] 2. Vector and strain
[0063] The cloning vector pEASY-Blunt Simple Cloning Vector was purchased from Beijing Quanshijin Bi...
Embodiment 3
[0111] Embodiment 3: TeLTF4 gene expression analysis
[0112] The full-length sequence of the TeLTF4 gene was cloned according to Example 2, and quantitative PCR primers were designed using the primer design software Primer Premier 5.0, and total RNA was extracted from marigold roots, leaves, and flowers respectively, and cDNA was obtained by reverse transcription, and the expression level of the TeLTF4 gene was determined. quantitative analysis.
[0113] 1. Reagents
[0114] The RNA extraction and reverse transcription reagents were as described in Example 2; the real-time quantitative PCR reagent TransStart Green qPCRSuperMix was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; the primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. Imported repackaged or domestically produced analytically pure products.
[0115] 2. Strains
[0116] Pseudomonas syringae pathogenic strain Pst DC3000 (Pseudomonas syringae pv.tomato DC3000) was obtained from the ...
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