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Soy biological clock gene GmLCL1, encoding protein and application thereof

A soybean and protein technology, applied in the field of genetic engineering, can solve problems such as changing plants' sensitivity to photoperiod and flowering time

Active Publication Date: 2014-09-17
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

[0005] In order to solve the problem of changing the sensitivity and flowering time of plants to the photoperiod by adjusting the soybean circadian clock, the object of the invention is to provide a soybean GmLCL1 protein

Method used

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  • Soy biological clock gene GmLCL1, encoding protein and application thereof
  • Soy biological clock gene GmLCL1, encoding protein and application thereof
  • Soy biological clock gene GmLCL1, encoding protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Cloning of soybean circadian clock gene GmLCL1

[0030] Genes were amplified from soybean Kennong 18 (Glycine max L. Kennong 18) by RT-PCR using forward primer 5'-ATGGACGCATACTCCTCCGGC-3' and reverse primer 5'-TCAAGTCGAAGTCTCCCCTTCCA-3' respectively.

[0031] The PCR reaction program was: 95°C 5min pre-denaturation, 95°C 30S, 50°C 35S, 72°C 2min 30S, 30 cycles, 72°C 10min extension.

[0032] Acquisition of GmLCL1 cloning vector: clone the amplified PCR product directly into such as TA cloning method 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 the T vector. Then the PCR product and the T vector were ligated at 16° C. for 12 hours, the ligated product was transformed into Escherichia coli DH5α, amplified therein, positive clones were screened, and sequenced.

[0033] The gene sequence of the obtained GmLCL1 gene is shown...

Embodiment 2

[0034] Example 2 Amino acid sequence analysis of protein encoded by soybean circadian clock gene GmLCL1

[0035] The similarity between the GmLCL1 protein sequence of the soybean circadian clock gene and the Arabidopsis LHY protein is 55.8%, and it is highly similar in the conserved functional domain (MYB DAN-binding domain), such as figure 1 shown. Therefore, it is speculated that GmLCL1 has a similar function to Arabidopsis LHY, and has the activity of inhibiting plant flowering.

Embodiment 3

[0036] Example 3 Expression levels of soybean circadian clock gene GmLCL1 in different tissues and organs and different developmental stages in soybean

[0037] The expression of soybean circadian clock gene GmLCL1 in soybean was determined by quantitative real time RT-PCR. Real-time fluorescent quantitative PCR was carried out by ABIStepOne, and SYBR Green I was used to detect the fluorescent signal.

[0038] Upstream primer: 5'-TATCATTTCTTTTATCCCTCCAACGG-3'

[0039] Downstream primer: 5'-ACACACATCTACACGACAAGTTTC-3'

[0040] The reaction system is:

[0041]

[0042] 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.

[0043] The soybean circadian clock gene GmLCL1 is expressed in soybean cotyledons, stems, leaves, flowers, and petioles, and the highest expression level is in the second compound leaf at the flowering stage, as shown in ...

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Abstract

The present invention provides a soy GmLCL1 protein, which is: 1) a protein comprising an amino acid sequence represented by SEQ ID NO.1; or 2) a protein, wherein an amino acid sequence represented by SEQ ID NO.1 is processed by substituting, deleting, or adding one or a plurality of amino acids to form the amino acid sequence of the protein, and the protein is derived from the protein 1) and has the same activity as the protein 1). The present invention further provides a gene GmLCL1 encoding the protein. With over-expression of the gene, plant flowering can be significantly inhibited, and a growing period can be prolonged. The gene and the encoding protein can be used for solutions of problems of asynchronous flowering period in cross breeding, and growing period controls, photoperiod sensitivity and introduction of various crops, vegetables, fruits and flowers.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a soybean biological clock gene, its encoded protein and its 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 (Zhang et al., 2008). 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 variou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N15/82C12N1/21C12N5/10A01H5/00
Inventor 傅永福吴卓晶张晓玫
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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