Soybean biological clock gene GmFKF2, encoding protein, and applications thereof

A gene and soybean technology, applied in the regulation of plant photoperiod and flowering time, in the field of soybean circadian clock genes, can solve the problems of changing the sensitivity of plants to photoperiod and flowering time, so as to promote plant flowering and prolong plant growth period effect

Inactive Publication Date: 2014-01-08
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Changing plant photoperiod sensitivity and flowering time by modulating the soybean circadian clock regulator gene FKF1 has not been reported so far

Method used

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  • Soybean biological clock gene GmFKF2, encoding protein, and applications thereof
  • Soybean biological clock gene GmFKF2, encoding protein, and applications thereof
  • Soybean biological clock gene GmFKF2, encoding protein, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Cloning of soybean circadian clock genes GmFKF1 and GmFKF2

[0044] The GmFKF1 gene was amplified from soybean Kennong 18 (Glycine max L. Kennong18) by RT-PCR using forward primer 5'-ATGGCTGTGACAAAAGAGAAAGAGGAT-3' and reverse primer 5'-TCACAGGTCAGAGTCCTGTCGAC-3'.

[0045] The GmFKF2 gene was amplified from soybean Kennong 18 (Glycine max L. Kennong18) by RT-PCR using forward primer 5'-ATGGCCATGCCAAAAGAGAAA-3' and reverse primer 5'-TTCACAGGTCAGAGTCATGTCGA-3'.

[0046] The PCR reaction program was: pre-denaturation at 95°C for 5min, denaturation at 95°C for 30S, annealing at 50°C for 35S, extension at 72°C for 1min and 50S, 30 cycles, and extension at 72°C for 10min.

[0047] Obtaining of GmFKF1 and GmFKF2 cloning vectors: clone the amplified PCR products directly into such as figure 2 on the indicated pGWCm vector. Firstly, the pGWCm vector was hydrolyzed with Ahd I endonuclease, and then the digested product was recovered with a gel recovery kit to obtain t...

Embodiment 2

[0049] Example 2 Amino Acid Sequence Analysis of Proteins Encoded by Soybean Circadian Clock Genes GmFKF1 and GmFKF2

[0050] The similarity between GmFKF1 and GmFKF2 protein sequences encoded by soybean circadian clock genes GmFKF1 and GmFKF2 and Arabidopsis thaliana FKF1 (AtFKF1) protein is high: the similarity between GmFKF1 and GmFKF2 and AtFKF1 is 83.5% and 81.4%, and the similarity between GmFKF1 and GmFKF2 The origin is 95.5%, and they are highly similar in the conserved functional domain (MYBDAN-binding domain), such as figure 1 shown. Therefore, it is speculated that GmFKF1 and GmFKF2 have similar functions to Arabidopsis FKF1 and have the activity of promoting plant flowering.

Embodiment 3

[0051] Example 3 Expression levels of soybean circadian clock genes GmFKF1 and GmFKF2 in different tissues and organs and different developmental stages in soybean

[0052] The expression of soybean circadian clock genes GmFKF1 and GmFKF2 in soybean was determined by quantitative real time RT-PCR. Real-time fluorescent quantitative PCR was carried out using ABI StepOne, and SYBR Green I was used to detect the fluorescent signal.

[0053] FKF1 upstream primer: 5'-GCATAAAACCAGACCCTCTTCTCTC-3';

[0054] FKF1 downstream primer: 5'-CAATCATGTAATAGTGGCTCATATAC-3';

[0055] FKF2 upstream primer: 5'-AAAGATTTTGGGTGGGTTTCC-3';

[0056] FKF2 downstream primer: 5'-GTGTAATAGACGGTGGAGGAG-3'.

[0057] The reaction system is:

[0058]

[0059] The reaction parameters were two-step method: 95°C 10S, hot start; 95°C 5S, 60°C 1min, 40 cycles. Gene expression was normalized and plotted using gene chip data analysis software Genesis.

[0060] Soybean circadian clock genes GmFKF1 and GmFKF2...

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Abstract

The invention provides a soybean biological clock gene GmFKF2 protein, (1) the protein is composed of amino acid sequences represented in the SEQ ID NO.1; (2) or the protein is derived from the protein mentioned in (1) through substituting, losing or adding one or several amino acids in the amino acid sequence represented in the SEQ ID No.1, and has the same activity as that of the protein mentioned in (1). The invention at the same time provides a gene GmFKF2, which encodes the protein mentioned above, through over-expressing the gene, plant blossom can be prominently promoted, and the growth period can be prolonged. The soybean biological clock gene GmFKF2 and encoding protein thereof can be used to solve the problems of flowering asynchronism in the crossbreeding process, growth period control of various plants, vegetables, fruits, and flowers, photoperiod sensitivity, and species introduction.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a soybean circadian clock gene, its encoded protein, and their application in the regulation of plant photoperiod and flowering time. Background technique [0002] Soybean is one of the important crops, and it is an important source of plant protein, edible oil, biodiesel, and secondary metabolites such as isoflavones and lecithin. Soybean is a short-day plant, and flowering is strictly controlled by photoperiod, so excellent varieties from different regions cannot be introduced to each other, and the growth period is also restricted by environmental photoperiod. If the sensitivity of soybean to photoperiod can be reduced, and the limitation of photoperiod on soybean flowering can be broken, the problem of soybean introduction can be solved, so as to realize the exchange of high-quality varieties between regions, enrich excellent germplasm resources in various regions, and regu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N1/15C12N1/21C12N1/19C12N15/11A01H5/00
CPCC07K14/415C12N15/827
Inventor 傅永福张晓玫李芳
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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