Soybean nuclear factor protein and encoding genes of protein and applications of protein and encoding genes

A technology for encoding genes and proteins, which can be used in applications, genetic engineering, plant genetic improvement, etc., and can solve the problem of little research on NFYB transcription factors.

Inactive Publication Date: 2013-03-13
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, compared with other plants such as Arabidopsis, rice, maize, and

Method used

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  • Soybean nuclear factor protein and encoding genes of protein and applications of protein and encoding genes
  • Soybean nuclear factor protein and encoding genes of protein and applications of protein and encoding genes
  • Soybean nuclear factor protein and encoding genes of protein and applications of protein and encoding genes

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Experimental program
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Effect test

Embodiment 1

[0057] Embodiment 1, the acquisition of GmNFYB1

[0058] Soybean (Glycine max) extracted with Trizol reagent (Dongnong 50, Wu Tianlong, Yang Qingkai, Ma Zhanfeng, Wu Zongpu, Zhao Shuwen, Gao Fenglan, Li Wenbin, Zhang Guodong, Yang Qi, Meng Qingxi, Wang Jinling. Dongnong Xiaodoudou No. 1 new soybean variety selection Report, [J]. Journal of Northeast Agricultural University, 1994, (25) 1:104; the public can obtain it from Northeast Agricultural University, hereinafter referred to as wild soybean). Leaves total RNA, the first strand of cDNA was synthesized by reverse transcription as a template, with a sense primer: GG TCTAGA CAAAGGTGCATTGGTGGT (the underlined part is the XbaI restriction site, sequence 3), antisense primer: AT GAGCTC CGTACAAGCATTCAAGGGA (the underlined part is the Sac Ⅰ restriction site, sequence 4), perform PCR reaction, the PCR condition is 94°C for 5min; 35 cycles: 94°C for 30s, 60°C for 30s, 72°C for 2.5min; 72°C for 5min. The PCR product was detected b...

Embodiment 2

[0059] Example 2, Obtaining and Functional Research of Transgenic GmNFYB1 Arabidopsis

[0060] 1. Construction of plant expression vector pCAMBIA-GmNFYB1

[0061] The vector pCAMBIA3301 vector (sold by CAMBIA Company of Australia) and pBI121 (purchased from CLONETECH Company) were double-digested with restriction endonucleases HindIII and EcoRI respectively, and the large fragment of pCAMBIA-D3 vector of 11257bp after digestion was recovered and digested The 3032bp small fragment of the pBI121 vector was connected to construct the expression vector pCAMBIA-pBI121.

[0062] The PCR product obtained in Example 1 was digested with XbaI and SacI, and the obtained fragment was connected with the pCAMBIA-pBI121 vector fragment that had undergone the same digestion, and the ligated product was transformed into Escherichia coli to obtain a transformant, and the plasmid of the transformant was extracted and sent to Sequencing showed that the plasmid was a recombinant vector obtained b...

Embodiment 3

[0168] Example 3, Obtaining and Functional Research of Transgenic GmNFYB1 Soybean Plants

[0169] 1. Genetic transformation of soybean cotyledon nodes mediated by Agrobacterium

[0170] 1. Preparation and transformation of soybean cotyledon nodes

[0171] (1) Bacterial solution preparation

[0172] Pick a single colony of Agrobacterium and inoculate it in 3mLYEP liquid medium containing rifampicin 50mg / L, streptomycin 25mg / L and ampicillin 100mg / L, cultivate overnight at 28°C and 200rpm, and then transfer the bacterial solution to 100ml Shake the above-mentioned fresh YEP liquid medium to A600=0.8, centrifuge at 5000rpm for 10min, and resuspend in the infection solution (liquid CCM medium), adjust the OD600 to 0.6.

[0173] (2) Preparation of soybean cotyledon nodes

[0174] Take the mature soybean Dongnong 50 seeds with smooth surface and no disease spots, put them in NaClO / HCl (24:1), and sterilize them in a filter with slightly excess HCl for 8 hours, then inoculate them...

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Abstract

The invention discloses a soybean nuclear factor protein and encoding genes of the protein and applications of the protein and the encoding genes. The protein provided by the invention is named GmNFYB1 and derived from soybean (Glycine Max (L.) Merrill.) Dongnong 50, and the protein can be: 1) a protein consisting of amino acid residues shown in a sequence 2 in a sequence table; or 2) a protein which is related to the stress tolerance of plants, is derived from the protein (1) and is obtained in a way that one or more than one amino acid residue in an amino acid sequence in the sequence 2 is substituted and/or deleted and/or added. Experimental results show that under the drought stress, transgenic plants into which the soybean GmNFYB1 (soybean nuclear factor binding protein) is transferred have longer roots, higher proline content and are higher in yield.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a soybean nuclear factor protein and its coding gene and application. Background technique [0002] Abiotic stresses such as drought and salinity seriously affect the yield and quality of crops. It is an effective way to improve the stress resistance of crops by means of genetic engineering. The elucidation of the molecular mechanism of resistance to abiotic stress is the prerequisite for the application of genetic engineering improvement. Transcription factors play a very important role in the signal transduction of abiotic stress. Soybean, as an important food crop, is widely planted all over the world. However, compared with other plants such as Arabidopsis, rice, maize and wheat, there is little information about NFYB transcription factors in soybean. Therefore, it has become an effective breeding method to use bioengineering technology to change the stress resistance respons...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/21C12N15/11C12N15/84A01H5/00
Inventor 李文滨王志坤李永光李胜畅
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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