Plant parthenocarpy regulation gene and use thereof
A parthenocarpic and gene technology, applied in the direction of plant gene improvement, application, plant products, etc., can solve the problem of molecular regulation of eggplant parthenocarp that has not been seen
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Embodiment 1
[0016] Embodiment 1: Cloning and separation of ARFs genes in eggplant
[0017] 1.1 Test materials and reagents
[0018] Materials: Hangzhou tomato plants were provided by Vegetable Institute of Zhejiang Academy of Agricultural Sciences. The plants are planted at room temperature, and the roots, stems, leaves, flower buds, flowers, and fruits are collected during the vigorous growth period, and stored in a -72°C ultra-low temperature refrigerator after liquid freezing. Reagents: Total RNA extraction kit (RNeasy Plant Mini Kit) was purchased from QIAGEN. MMLV reverse transcriptase was purchased from Promega, and Taq DNA polymerase and pMD-19T vector were purchased from Treasure Biological Company (Dalian) Co., Ltd. (TaKaRa). SMARTTMRACE cDNA Amplification Kit (PT3269-1) was purchased from Clontech, and the primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. Gel recovery kit was purchased from Hangzhou Vgene Company. Sequencing was performe...
Embodiment 2
[0046] Example 2 Full-length cDNA sequence and structural domain analysis
[0047] The newly cloned eggplant SmARF8 sequence has a full length of 3750bp, an open reading frame of 2676bp, and encodes 891 amino acids. S1ARF8 agreement reached 54.4%.
[0048] The amino acid sequence analysis of SmARF8 protein showed that, like Arabidopsis ARF8 gene, it had the typical characteristics of this family gene sequence. We used the clustW 1.83 program to analyze the amino acid sequences of rice OsARF8, Arabidopsis ARF8, cucumber CsARF3, tomato S1ARF8 and eggplant SmARF8 genes, and then analyzed them with Gendoc. ARFs family genes contain a conserved domain of about 300aa at the N-terminal, which is verified to have the function of a DNA binding domain in the ARF1 gene (Ulmasov T, Hagen G, Guilfoyle TJ.1997a.ARF1, a transcription factor that binds to auxin response elements. Science.1997 Jun 20; 276(5320):1865-1868), known as the DNA-binding domain (DBD), can specifically bind to the c...
Embodiment 3
[0049] Phylogenetic tree analysis of different species ARF family genes of embodiment 3
[0050] In order to compare the genetic relationship between the cloned eggplant SmARF8 gene and ARFs family genes in other species, the published ARFs family gene sequences in Arabidopsis, rice, cucumber and tomato were respectively downloaded. The full-length amino acid sequence of each gene was aligned, and a phylogenetic tree was constructed with phylip software. The result is as Image 6 As shown, it can be seen that the SmARF8 gene is clustered with ARF8, OsARF8, S1ARF8, CsARF3, CsARF4, OsARF6a, OsARF6b on one branch, and other genes are not clustered with them. indicating that they are very closely related. Among them, it has a very close relationship with the gene ARF8, which regulates parthenocarpy in Arabidopsis, indicating that the gene has the function of regulating parthenocarpy.
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