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Precocious trifoliate orange flowering gene spliced variant PtFT2 and application of precocious trifoliate orange flowering gene spliced variant PtFT2 to regulating flowering of plants

A gene and plant technology, which is applied to the flowering gene PtFT2 of Citrus aurantium and its application field in regulating plant flowering, can solve the problems such as the application report on the regulation of plant flowering time of the flowering gene PtFT2 of the isolated and cloned Citrus aurantium, and the like. The effect of shortening flowering time and shortening childhood

Inactive Publication Date: 2017-09-26
HUAZHONG AGRI UNIV
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  • Description
  • Claims
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Problems solved by technology

[0004] However, so far, there have been no reports on the isolation and clone of the flowering gene PtFT2 and its application in regulating plant flowering time.

Method used

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  • Precocious trifoliate orange flowering gene spliced variant PtFT2 and application of precocious trifoliate orange flowering gene spliced variant PtFT2 to regulating flowering of plants
  • Precocious trifoliate orange flowering gene spliced variant PtFT2 and application of precocious trifoliate orange flowering gene spliced variant PtFT2 to regulating flowering of plants
  • Precocious trifoliate orange flowering gene spliced variant PtFT2 and application of precocious trifoliate orange flowering gene spliced variant PtFT2 to regulating flowering of plants

Examples

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Effect test

Embodiment 1

[0026] Example 1 Isolation of PtFT2 of Trifoliate Fructus

[0027]Precocious trifoliate orange (Precocious trifoliate orange) is a variant plant of trifoliate orange (Liang Suiquan et al., 1999). Compared with common trifoliate orange, the childhood period is significantly shortened. Trifoliate trifoliate and common trifoliate were used as materials to explore the flowering mechanism of citrus (Ai et al.2012; Li et al.2010; Zhang et al.2011). In order to clone the homologous gene of FT in Trifoliate trifoliate Fructus, search for the FT gene of Arabidopsis thaliana in NCBI, find the CDS sequence of the AtFT gene of Arabidopsis thaliana, and put it in the Clementine database (http: / / phytozome.jgi. doe.gov / pz / portal.html#) to find out the FT gene (geneID=Ciclev10012905m) in Clementine, and use the Primer Primer 5.0 software to design primers to amplify the FT homologous gene in Trifoliate Fructus. increase.

[0028] Primer sequences for amplifying FT homologous genes:

[0029...

Embodiment 2

[0036] Example 2 Bioinformatics analysis of PtFT2 gene

[0037] 1) Evolutionary analysis of PtFT2 gene: The FT protein sequences of some common species of dicotyledonous and monocotyledonous plants were searched in NCBI, and the comprehensive analysis of NCBI Blast found that this gene has a high degree of homology with most similar FT genes. The homology between PtFT and LcFT similar gene of litchi is as high as 94%, and it also has great similarity with Populus, apple and so on. Some representative plants of monocotyledons and dicotyledons were selected, and MEGA4.0 was used to construct the evolutionary tree, and the Bootstrap value was 1000, from figure 1 It can be seen from the figure that the position of PtFT protein in plant evolution is high, and it has a high homology with dicotyledonous plants, so it can fully explain that the PtFT gene does belong to the FT gene family.

[0038] 2) Sequence conservation analysis of PtFT amino acids: FT has obvious early flowering ...

Embodiment 3

[0039] Example 3 Genetic Transformation of Tobacco with PtFT2 Gene

[0040] 1. Vector construction: in order to verify the function of the PtFT2 gene, according to the multiple cloning site and the PtFT2 sequence of the pCAMBIA1301 carrier (purchased from the CAMBIA laboratory in Australia), the amplification design containing The primer sequence of the restriction site: forward primer: 5-ccatggATGTCTAGCAGGGACAGAGA-3 reverse primer e: 5-ggtaaccTCATCGTGTCATAGTCCTTCC-3 (note: the lowercase letters are the added restriction site sequence). The obtained correct PtFT2 was connected to the pMD18-T bacteria liquid as a template for PCR amplification to obtain a PtFT2 fragment with a restriction site, which was convenient for the construction of subsequent vectors. The annealing temperature for PCR amplification was 60°C. The PCR reaction conditions are: 95°C, 5min; 95°C, 30s; 56°C, 30s; 72°C, 90s; 72°C, 10min. 35 cycles. Then the amplified fragments were separated by 1% agarose ge...

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Abstract

The invention belongs to the technical field of plant genetic engineering, and in particular relates to the flowering gene PtFT2 of trifoliate trifoliate fruit and its application in regulating plant flowering. The present invention newly discovered a flowering regulatory gene PtFT2 from Trifoliate Fructus Trifoliate Fructus. Through gene isolation, structural analysis, cloning and functional verification, the PtFT2 gene was obtained from the RNA level for the first time. The gene PtFT2 is the spliced ​​version of the PtFT1 gene. Compared with the genome, the excised version lacks the first intron in gene structure, and its translation terminates early after the second exon. The nucleotide sequence of the gene is shown in SEQ ID NO:1. Transforming tobacco with PtFT2 gene can obviously promote the flowering time of tobacco earlier, and the gene is expected to play an important role in shortening childhood and earlier flowering of perennial woody fruit trees.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to the flowering gene PtFT2 of trifoliate trifoliate fruit and its application in regulating plant flowering. The present invention has newly discovered a flowering regulatory gene PtFT2 from Citrus citrifolia, through gene isolation, structural analysis, cloning and functional verification, for the first time obtained PtFT2 from citrus at the RNA level, and its nucleotide sequence is shown in SEQ ID NO: 1 Show. Genetic transformation of PtFT2 into tobacco, the PtFT2 gene can significantly promote the flowering of tobacco, advance the flowering time, significantly shorten the childhood of tobacco, shorten the breeding process for perennial woody fruit trees, and play an important role in the genetic improvement of plants. Background technique [0002] Flowering and fruiting is an important biological process in the life cycle of flowering plants, the c...

Claims

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Application Information

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IPC IPC(8): C12N15/29A01H5/00
CPCC07K14/415C12N15/827
Inventor 张金智侯小进胡春根靖艳玲
Owner HUAZHONG AGRI UNIV
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