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Upland cotton GhLFY protein and coding gene and application thereof

A protein and gene technology, applied in the field of upland cotton GhLFY protein and its coding gene and application, can solve the problems of insufficient prematurity of short-season cotton

Active Publication Date: 2012-10-10
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current short-season cotton has the problem of insufficient early maturity

Method used

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  • Upland cotton GhLFY protein and coding gene and application thereof
  • Upland cotton GhLFY protein and coding gene and application thereof
  • Upland cotton GhLFY protein and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1, the cloning of cotton gene GhLFY

[0054] 1. Cloning of cotton gene GHLFY

[0055] 1. Extraction of RNA

[0056] According to the CTAB method, RNA extraction steps:

[0057] (1) Place a 50ml centrifuge tube on the centrifuge tube rack, add 15ml extract solution to the centrifuge tube, add 300ul β-mercaptoethanol, and preheat the water bath at 65°C for 10 minutes;

[0058] (2) Clean and dry the mortar and pestle, pour a little alcohol into it to burn and sterilize, add liquid nitrogen to pre-cool after cooling, put 2-3g of different tissues of the early-maturing cotton variety Zhongmian 36 into the cooled mortar , quickly grind, and after grinding into a fine powder, scrape it into the preheated extract at 65°C with a pre-cooled spoon, shake vigorously, and put it in a warm bath at 65°C for 10 minutes, shaking 3-4 times during the period;

[0059] (3) Add an equal volume of chloroform: isoamyl alcohol (24:1), shake vigorously for 10 minutes, 13000 rpm, a...

Embodiment 2

[0120] Embodiment 2, the application of cotton gene GhLFY in promoting plant flowering

[0121] 1. Construction of expression vector

[0122]Using the cDNA of Zhongmian Institute 36 terminal bud as a template (or artificially synthesized sequence 1 as a template), the infusion primer (F: 5'-CACTGTTGATACATATGCTTCAAAAATGGACCCTGAGAC-3' (sequence 3); R: 5'-TGTTGATTCAGAATTCTTCAGTTCCAACTTAAACCATACA-3' ( Sequence 4), amplified to obtain a PCR product containing the complete open reading frame of the GhLFY gene, after sequencing, it is the 3rd-1300 nucleotides from the 5' end of Sequence 1 in the sequence listing;

[0123] The plant expression vector pRI101 (Takara Code: D3262) was digested with NdeI and EcoRI to obtain the vector backbone;

[0124] The above vector backbone was ligated with the above PCR product using Clotech's Infusion ligation kit.

[0125] The enzyme digestion system of pRI101 is as follows:

[0126]

[0127] The connection system of the Infusion Ligation Ki...

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Abstract

The invention discloses upland cotton GhLFY protein and a coding gene and application thereof. The protein is protein in (a) or (b): (a) protein consisting of an amino acid sequence shown as Sequence 2 in a sequence table; and (b) plant flower-related protein derived from Sequence 2 via substitution and / or deletion and / or addition of one or a plurality of amino acid residue(s) of the amino acid sequence shown as Sequence 2 in the sequence table. Experiments show that a new gene GhLFY is discovered, the transgenic plant modified genetically by the new gene can flower in advance, seeds can also be obtained in advance, the number of rosette leaves of the plant can be reduced, and the number of cauline leaves can be increased; and thus the gene can be applied in cultivation of cotton with a short growth period, and a foundation is laid for cultivation of genetically modified plants.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a GhLFY protein of upland cotton and its coding gene and application. Background technique [0002] Cotton is one of the most important economic crops in our country and plays an important role in national production. However, the development of cotton is limited to a certain extent due to the land competition between grain and cotton. Breeding and promotion of short-season cotton varieties can alleviate the conflict between grain and cotton land, and is an effective way to achieve double harvests of grain and cotton. However, the current short-season cotton has the problem of insufficient prematurity. [0003] As one of the important traits of cotton, early maturity has become an important breeding goal of upland cotton. Studies have shown that flowering sooner or later is closely related to the early maturity of cotton. Therefore, the flowering-related genes were cloned, and the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11A01H5/00
Inventor 喻树迅李洁范术丽宋美珍庞朝友魏恒玲
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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