Plant stress tolerance associated protein GmNF-YB6 as well as coding gene and application thereof

A technology of plant stress tolerance and stress tolerance, applied in the direction of plant genetic improvement, botany equipment and methods, applications, etc., can solve the problems of comprehensive improvement of plant stress resistance that are difficult to achieve, and achieve the effect of improving drought tolerance

Active Publication Date: 2014-01-15
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

[0008] Since the stress tolerance of plants is a complex trait regulated by multiple genes, it is difficult to achieve comprehensive improvement of plant stress resistance by introducing a single functional protein gene

Method used

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  • Plant stress tolerance associated protein GmNF-YB6 as well as coding gene and application thereof
  • Plant stress tolerance associated protein GmNF-YB6 as well as coding gene and application thereof
  • Plant stress tolerance associated protein GmNF-YB6 as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1, the cloning of GmNF-YB6

[0060] 1. Isolation of mRNA

[0061] Soybean (Glycine max L.) variety Tiefeng No. 8 grown in hydroponics for about 10 days (references: Sun Xiao, Dong Jianhui, Chen Ming, Xu Zhaoshi, Ye Xingguo, Li Liancheng, Qu Yanying, Ma Youzhi. 2008. Cloning and regulatory segment analysis of soybean stress resistance gene GmDREB3 promoter. Acta Crops Sinica, 34(8):14751479) Four-leaf stage seedlings were drought-treated for 2 hours, quick-frozen in liquid nitrogen, and stored at -80°C for later use. mRNA was isolated using Quikprep Micro mRNA Purification Kit (Pharmacia). First-strand cDNA synthesis was performed with Reverse Transcriptase XL (AMV). The ds cDNA was synthesized by SMART method, and the PCR products were detected by 1.0% agarose gel electrophoresis.

[0062] 2. Acquisition of the full-length sequence of the GmNF-YB6 gene

[0063] The full-length sequence of soybean CCAAT-box nuclear transcription factor B family gene was ob...

Embodiment 2

[0065] Example 2, real-time fluorescent quantitative PCR analysis of the expression characteristics of GmNF-YB6

[0066] 1. Coercion treatment

[0067] Seedling age is No. 8 seedling of soybean variety Tiefeng of 10 days, carries out following treatment:

[0068] (1) Drought treatment: Take out the potted soybean seedlings to absorb the moisture from the roots, place them on dry filter paper, and cultivate them in a dry environment for 30 minutes, 1 hour, 2 hours, 5 hours, 12 hours, and 24 hours. Quick-frozen in nitrogen and stored at -80°C for future use.

[0069] (2) Low-temperature treatment: Place the soybean seedlings in a 4°C incubator, incubate under light for 30 minutes, 1 hour, 2 hours, 5 hours, 12 hours, and 24 hours, take them out and freeze them with liquid nitrogen, and store them at -80°C for later use.

[0070] (3) High temperature treatment: Soybean seedlings were placed at 42°C, light-cultured for 30 minutes, 1 hour, 2 hours, 5 hours, 12 hours, and 24 hours,...

Embodiment 3

[0081] Embodiment 3, the activation characteristic of GmNF-YB6

[0082] YEP-GAP Vectors: Chinese Academy of Agricultural Sciences Crop Science Research Guaranteed to be Available to the Public; References Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi-Shinozaki K, Shinozaki K. Two transcription factors, DREB 1 and DREB2, with an EREBP / AP2DNA binding domain separate two cellular signal transduction pathways in drought-and low-temperature-responsive gene expression, respectively, in Arabidopsis, Plant Cell 1998 Aug;10(8):1391-1406.

[0083] YPD liquid medium: 10g / L of yeast extract (Bacto-Yeast Extract) for bacterial culture, 20g / L of tryptone (Bacto-Peptone) for bacterial culture, adjust the pH to 5.8, sterilize at 121°C / 15min, drop After reaching 60°C, add 40% Glucose to make the final concentration 20g / L.

[0084] SD / His - / Ura - / Trp - Selective medium: Yeast nitrogen base without amino acids (Yeast nitrogen base) 6.7g / L, auxotrophic mixture (drop-out media without H...

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Abstract

The invention discloses a plant stress tolerance associated protein GmNF-YB6 as well as a coding gene and an application thereof. The protein disclosed by the invention is the following (a) or (b): (a) a protein consisting of amino acid sequences shown in the sequence 1 in a sequence table; and (b) a protein which is formed through substitution and / or deletion and / or addition of one or more amino acid residues of the amino acid sequences in the sequence 1, is associated with plant stress tolerance and is derived from the sequence 1. The GmNF-YB6 disclosed by the invention can be expressed under the inductivity of low temperature, drought and high temperature, can specifically regulate transcriptional expression of genes containing CCAAT-box cis elements (core element CCAAT), can improve the drought tolerance, freezing resistance and high temperature resistance of plants, provides a basis for artificially controlling expression of stress resistance and stress tolerance associated genes, plays an important role in stress resistance and stress tolerance enhanced plant breeding so as to provide a basis for artificially controlling expression of stress tolerance associated genes, and plays an important role in stress tolerance enhanced plant breeding.

Description

technical field [0001] The invention relates to a plant stress tolerance-related protein GmNF-YB6 and its coding gene and application. Background technique [0002] Adversity stresses such as drought, high salinity and low temperature severely restrict the growth and development of soybean. Therefore, understanding the response and signal transduction mechanism of soybean to stress conditions and improving the stress resistance of soybean varieties have become one of the important tasks of soybean genetic research and variety improvement. [0003] Under adversity stress, plants will produce a series of responses, accompanied by many physiological, biochemical and developmental changes. Clarifying the response mechanism of plants to stress will provide scientific evidence for the research and application of stress-resistant genetic engineering. At present, the research on plant stress resistance has gradually penetrated into the cellular and molecular levels, combined with ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
CPCC07K14/415C12N15/8271C12N15/8273
Inventor 徐兆师马有志郑炜君李连城陈明
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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