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A drought-resistant and salt-tolerant gene of alfalfa

A drought-resistant and salt-tolerant gene, alfalfa technology, applied in the field of genetic engineering, can solve the problems of alfalfa, such as lack of stress-resistant genetic resources, and difficulty in realizing stress-resistant varieties.

Active Publication Date: 2016-05-04
杨凌金栗农业科技有限公司
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, alfalfa lacks stress-resistant genetic resources, and it is not easy to select stress-resistant varieties through conventional cross-breeding

Method used

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  • A drought-resistant and salt-tolerant gene of alfalfa
  • A drought-resistant and salt-tolerant gene of alfalfa
  • A drought-resistant and salt-tolerant gene of alfalfa

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Embodiment Construction

[0017] The method of the present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0018] The combination of these two methods is expected to obtain a full-length cDNA gene.

[0019] The outline of the 5′RACE steps is as follows: figure 1 Shown: first-strand primer GSP1 annealed to mRNA; mRNA was transcribed into cDNA using SuperScriptIII (invitrogen); RNA was degraded using RNaseH (MBI); cDNA was purified using glass milk; purified cDNA was tailed using dCTP and TdT; The combined anchor primer and nested GSP2 PCR amplified dC-tailed cDNA; the primary PCR product was reamplified using AUAP, UAP and nested GSP.

[0020] The outline of the 3′RACE steps is as follows: figure 2 Shown: 3′RACE utilizes the naturally occurring Poly(A) tail at the 3′-terminus of mRNA as a universal priming site for PCR amplification, and the mRNAs are first reverse-transcribed with reverse transcriptase and Oligo-dT linker ...

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Abstract

The invention discloses a medicago sativaL drought resistance and salt tolerance gene, which has an amino acid nucleotide sequence represented by SEQ ID NO.1 in the sequence table. After the gene is transformed into medicago sativaL, drought resistance and salt tolerance of the medicago sativaL can be increased, and the strong stress resistance medicago sativaL variety can be rapidly bred with a genetic engineering manner. The strong stress resistance medicago sativaL variety can be obtained by adopting the following process: 1, linking the gene and a plant expression vector pCAMBIA1301 by using T4 ligase to construct a plant expression vector; 2, transforming vector expression plasmid into Agrobacterium tumefaciens GV3101, adopting a leaf disc method to infect tobacco leaves, and adopting hygromycin to screen resistant plants; and 3, transplanting the obtained plants into soil, and observing growth states and phenotypes of the plants.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering and relates to a drought-resistant and salt-tolerant gene of alfalfa. Background technique [0002] Alfalfa (Medicagosativa L.) is the most widely distributed perennial leguminous herbage in the world, with a planting area of ​​more than 30 million hectares worldwide 2 , my country is one of the countries that cultivate and utilize more alfalfa in the world. Because of its rich nutrition, strong palatability, wide cultivation area, and high protein content, it has the reputation of "the king of forage grass" and plays an important role in animal husbandry production. In recent years, due to the increasingly severe climate drought in our country, over-exploitation and extensive management of pastures have greatly restricted the production of alfalfa. Therefore, it is of great practical significance to carry out technical research on improving alfalfa's stress resistance at this stage. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00
Inventor 韩博杨培志呼天明余成群张攀王卫栋张志强曹玉曼杨唯梓
Owner 杨凌金栗农业科技有限公司
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