Construction method of activation tag carrier and apocynum venetum seed mutant library

A construction method and technology of apocynum, applied in the field of plant genetics and breeding, to achieve the effects of avoiding lethal effects, overcoming the influence of seed setting rate, and high conversion rate

Active Publication Date: 2019-02-15
INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the continuous improvement of plant transformation technology, activating mutation libraries have been successfully established in many important annual economic crops such as cotton, tobacco, and salt mustard, but there are still no relevant reports in perennial herbaceous crops such as apocynum

Method used

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  • Construction method of activation tag carrier and apocynum venetum seed mutant library
  • Construction method of activation tag carrier and apocynum venetum seed mutant library

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0041]1. Construction of recombinant Agrobacterium strains containing the activation tag vector pSKI074-GUS:

[0042] A GUS transient transfection reporter gene (nucleotide sequence from Genebank, Gene ID: 946149) was introduced into the pSKI074 vector containing four tandem 35S enhancers using a multi-step restriction enzyme ligation reaction. The activated tag in the transformed pSK074-GUS activation tag vector is four tandem 35S enhancers and contains a GUS reporter gene for transient transfer detection. The constructed plasmid vector map is as follows: figure 1 .

[0043] 2. Preparation of recombinant Agrobacterium-infected bacterial suspension:

[0044] Agrobacterium LB4404 was transfected with the plasmid vector constructed above, and the transfection method was electric shock transformation method to obtain the recombinant strain of Agrobacterium;

[0045] Pick the recombinant strain and inoculate it in 5ml of LB liquid medium containing 50mg / L kanamycin. Shake cultu...

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Abstract

The invention relate to the technical field of plant genetic breeding, in particular to a construction method of an activation tag carrier and apocynum venetum seed mutant library. Compared with the prior art, a large-scale apocynum venetum mutant seed group can be rapidly obtained through selection of explants, genetic transformation and construction of a regeneration system; an activation tag sequence in the mutant group comprises four series-connected 35s enhancers, activation tags are randomly distributed in the overall apocynum venetum genome, expression of adjacent genes is enhanced, mutants are formed by induction, the produced mutants are in dominant mutation, and lethal effect produced by loss-of-function mutation is avoided.

Description

technical field [0001] The invention relates to the technical field of plant genetics and breeding, in particular to a method for constructing an activation tag carrier and a mutant bank of apocynum seeds. Background technique [0002] Apocynum venetum L. is a plant of the genus Apocynum venetum. Also known as kenaf, tea flower, red willow and so on. Our country has a long history of resource utilization of apocynum plant: the whole plant of apocynum apocynum can be used as medicine, and the young leaves are steamed, fried and kneaded and then drunk as tea, which has the functions of cooling and removing internal heat, preventing dizziness and strengthening the heart. The fiber of apocynum has health functions such as anti-ultraviolet, antibacterial, antistatic, etc., and is known as the "king of wild fibers". Raw materials such as high-grade clothing materials, fishing net threads, leather threads, and high-grade paper are used in defense industry, aviation, navigation, t...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/66C12N15/65A01H5/10A01H6/00
CPCC12N15/65C12N15/66C12N15/8271
Inventor 高钢朱爱国喻春明陈平陈继康陈坤梅
Owner INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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