A protein related to plant flowering time and its application

A flowering time and plant technology, applied in the field of protein and its application, to achieve the effect of saving manpower and material resources, shortening the growth period, and increasing value

Active Publication Date: 2022-07-15
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, expansins related to plant flowering time have not been reported yet.

Method used

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  • A protein related to plant flowering time and its application
  • A protein related to plant flowering time and its application
  • A protein related to plant flowering time and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Cloning of early flowering gene PtoEXPB3

[0070] The inventors of the present invention isolated and cloned an early flowering related gene PtoEXPB3 from Populus tomentosa TC1521, as shown in Sequence 1 of the sequence listing, and named the encoded protein as PtoEXPB3 protein, as shown in Sequence 2 of the sequence listing.

[0071] The specific cloning method is as follows:

[0072] The total RNA from the leaves of Populus tomentosa TC1521 was extracted by Trizol method (Invitrogen, USA), reverse transcribed into cDNA, and the cDNA was used as a template with a primer pair (PtoEXPB3F: 5'-CCAGAAAACTGTCGTCAAAGG-3';

[0073] PtoEXPB3R: 5'-GCTTGTTTCTCTCACGTGGA-3') was used as a primer, and KODplus DNA polymerase was used to amplify the gene PtoEXPB3. The amplification program was: 94°C for 5 min; 94°C for 30 s, 55°C for 40 s, and 72°C for 1 min, a total of 35 cycles. The amplified products were separated by 1.5% agarose gel electrophoresis, and the electropho...

Embodiment 2

[0075] Example 2 Acquisition and identification of N. benthamiana transgenic PtoEXPB3 gene

[0076] 1. Construction of recombinant expression vector

[0077] Taking the recombinant vector P-Blunt-PtoEXPB3 in Example 1 as a template, using PtoEXPB3F (5'-CCC AAGCTT ATGCAGCTCTTGGGGTTAC-3', the restriction site of HindIII is underlined) and PtoEXPB3R (5'-TGC TCTAGA TTAATGGAAGAAATTGAGCCTAGAGG-3', the restriction site of XbaI is underlined) as a primer pair, and high-fidelity enzyme KOD was used to amplify and recover the PCR product. The PCR product and the PEZR(K)-LC expression vector plasmid were digested with Hind III and Xba I restriction enzymes, respectively, and the digested PEZR(K)-LC vector and the target gene fragment were recovered to obtain the recombinant vector PEZR ( K)-LC-PtoEXPB3 (schematic diagram as shown in image 3 shown), the recombinant vector PEZR(K)-LC-PtoEXPB3 is to replace the sequence between the Hind III and Xba I recognition sites of the PEZR(K)-L...

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Abstract

The present invention relates to a protein related to plant flowering time and its application. Said protein is the following protein of A1), A2) or A3): A1) the amino acid sequence is the protein of sequence 2 in the sequence table; A2) the sequence The amino acid sequence shown in Sequence 2 in the list is obtained by substitution and / or deletion and / or addition of one or several amino acid residues, which is more than 90% identical to the protein shown in A1) and is related to plant flowering time. Protein; A3) Fusion protein obtained by linking a protein tag to the N-terminus or / and C-terminus of A1) or A2). The present invention obtains an expansin gene PtoEXPB3 from Populus tomentosa. When this gene was introduced into tobacco, the flowering time of transgenic tobacco flowers was 21 days earlier than that of the wild type, indicating that PtoEXPB3 has the effect of early flowering and can be used as an important gene resource for molecular breeding of flowering plants.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protein related to plant flowering time and its application. Background technique [0002] The early flowering phenomenon (ie, early budding and flowering) of plants is related to genes, and some early flowering genes have been identified in different plants and have a positive impact on production practices. For example, Smykal et al. (2007) cloned the flowering time-related genes NtSOC1 and NtFUL from tobacco, which are related to photoperiod characteristics, and overexpression can cause early flowering in tobacco. Li Shuang et al. (2014) transferred the PsnAP1 gene of Populus nigra in tobacco, and the transgenic plants bloomed earlier. Wang (2012) found that the PtGT1 gene was overexpressed in tobacco, and the plants showed early flowering. (An et al., 2011) showed that the PtAP3 gene of Populus tomentosa plays an important role in inducing early flowering. Deng Xinjie et al....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H5/02A01H6/82
CPCC07K14/415C12N15/827
Inventor 徐吉臣张皓尹鹏刘晓徐筱
Owner BEIJING FORESTRY UNIVERSITY
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