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Alfalfa cold-resistant gene MsF-box as well as encoding protein and application thereof

An msf-box, alfalfa technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problems of low overwintering rate, freezing injury, death and other problems of alfalfa, and achieve the effect of enriching cold-resistant genetic resources and improving plant cold resistance.

Active Publication Date: 2016-01-06
HARBIN NORMAL UNIVERSITY
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Problems solved by technology

In high-latitude and high-altitude areas in northern my country, alfalfa generally has a low overwintering rate and is prone to freezing damage and death.

Method used

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  • Alfalfa cold-resistant gene MsF-box as well as encoding protein and application thereof
  • Alfalfa cold-resistant gene MsF-box as well as encoding protein and application thereof
  • Alfalfa cold-resistant gene MsF-box as well as encoding protein and application thereof

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preparation example Construction

[0030] The cloning of described MsF-box gene and the preparation method of plant expression vector comprise the following steps:

[0031] 1) Primer design

[0032] MsF-boxF:5'TGGGTTAGGTTAGGATTA3'

[0033] MsF-boxR: 5'TTTCTAGCCTTCTTACAGT3'

[0034] 2) Cloning of MsF-box gene

[0035] The cDNA of alfalfa (Medicagosativa L.) was used as a template, the above primers were used to amplify the MsF-box gene, and ligated into the pMD18T vector to construct the pMD18T-MsF-box vector.

[0036] 1) Construction of plant expression vector pCBM-MsF-box

[0037] Recombinant plasmid pMD18T-MsF-box and pCBM expression vector plasmid were double digested with BamHI and PstI respectively, and ligated to obtain plant expression vector pCBM-MsF-box.

[0038] The obtaining of the transgenic tobacco comprises the following steps:

[0039] 1) Using the Agrobacterium-mediated genetic transformation method to introduce pCBM-MsF-box into Agrobacterium tumefaciens EHA105, and obtain Agrobacterium-po...

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Abstract

The invention discloses an alfalfa cold-resistant gene MsF-box as well as encoding protein and application thereof, and belongs to the technical field of plant genetic engineering. An open reading frame of the MsF-box disclosed by the invention is 1422bp in total length, and 473 amino acids are coded. The gene plant expression vector pCMB-MsF-box is transformed into tobacco by virtue of an agrobacterium-mediated genetic transformation method. On the basis of analysis on cold resistance of the transgenic tobacco, the MsF-box gene is capable of improving the cold resistance of the tobacco; and researches on the alfalfa cold-resistant gene MsF-box lay a foundation for future genetic improvement of alfalfa.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to an alfalfa cold-resistant gene MsF-box, its encoded protein and its application. Background technique [0002] Alfalfa (Medicagosativa L.) has high protein content, comprehensive nutritional value and long useful life. Adult alfalfa plants can survive winter smoothly in Altay and Tacheng. In high-latitude and high-altitude areas in northern my country, alfalfa generally has a low overwintering rate and is prone to freezing damage and death. Overwintering has gradually become a key issue restricting the successful establishment of alfalfa grassland in northern my country. Therefore, it is of great theoretical and practical significance to study the cold resistance of alfalfa. Zhaodong City, Heilongjiang Province is the hometown of alfalfa. The average temperature in winter in Heilongjiang Province is about -20℃, and Zhaodong alfalfa can survive th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00
Inventor 郭长虹丛庆宋丽莉张琪崔慧萍
Owner HARBIN NORMAL UNIVERSITY
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