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AdMsrB1 gene for increasing ACC content and application thereof

A plant and content technology, applied in the field of plant molecular biotechnology and genetic engineering, can solve problems such as difficulty in ensuring long-term market supply, difficulty in controlling fruit sales period, and affecting fruit flavor and quality.

Active Publication Date: 2020-01-31
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the post-ripening process of kiwi fruit in the shelf life is too fast or too slow, it will affect the flavor and quality of the fruit, and it is difficult to control the fruit sales period and ensure long-term market supply

Method used

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  • AdMsrB1 gene for increasing ACC content and application thereof
  • AdMsrB1 gene for increasing ACC content and application thereof
  • AdMsrB1 gene for increasing ACC content and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] The expression pattern of embodiment 1.AdMsrB1 gene under air (control) and exogenous ethylene treatment

[0012] Research methods:

[0013] The mature fruit of ‘Hayward’ kiwi fruit was harvested in a commercial orchard in Shaanxi Province in 2015. The selection criteria for the experimental fruit was that the fruit shape was uniform in size and free from mechanical damage, disease and insect pests. At 20°C, the fruit was sealed in a 20L bucket, treated with 100μl / L ethylene and air (as a control) for 24 hours, and then taken out and stored at 20°C. The sampling points were ethylene treatment groups 0, 0.5, 1, 2, 4, 8d, the control group was 0, 1, 2, 4, 8d. When collecting fruit samples, only the pulp was taken, and the peel, seeded part and central column were removed. After freezing in liquid nitrogen, store at -80°C.

[0014] Use the online software Primer 3.0 (v.0.4.0; http: / / bioinfo.ut.ee / primer3-0.4.0 / ) to design real-time fluorescent quantitative PCR primer pa...

Embodiment 2

[0018] Example 2. Determination of ACC content in tobacco leaves transiently overexpressing 35S::AdMsrB1

[0019] Research methods:

[0020] 1. Transient overexpression in tobacco leaves

[0021] The full-length coding region of AdMsrB1 was loaded into pGreen II 002962-SK binary expression vector, and the primer sequences used were SEQ: NO.5 and SEQ: NO.6. The recombinant vector was transformed into GV3101 strain by electric shock method, and Glycerol bacterium was preserved. Agrobacteria containing 35S::AdMsrB1 and empty SK vector were activated and resuspended in permeate (10 mM MES, 150 mM acetosyringone, 10 mM MgCl 2 , pH5.6) and adjust OD 600 to 0.75. The target gene (35S::AdMsrB1 recombinant vector) and the negative control (empty SK vector) were injected on both sides of the main vein of the fifth true leaf of common tobacco, and repeated at least 5 times. The injected tobacco was cultured in an artificial climate chamber for 5 days, and samples were taken for the ...

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Abstract

The invention provides an AdMsrB1 gene for increasing the ACC content and the application thereof. The AdMsrB1 gene comes from kiwifruit. The cDNA sequence, obtained through clone; of the AdMsrB1 geneis utilized for constructing a plant expression vector, overexpression is carried out in common tobacco leaves, the content of 1-amino cyclopropane carboxylic acid (ACC), which is an intermediate product of the biosynthetic pathway of ethylene, is obviously increased from 2.52 nmolg-1 to 5.41 nmolg-1, and the increasing rate is about 200%. The content of ACC can be increased by carrying out overexpression on the methionine sulfoxide reductase gene, and then ethylene biosynthesis in plants can be influenced.

Description

technical field [0001] The invention belongs to the fields of plant molecular biotechnology and genetic engineering, and relates to methionine sulfoxide reductase AdMsrB1. It involves a gene of methionine sulfoxide reductase from kiwi fruit, named AdMsrB1. The invention also relates to the amino acid sequence coded by the gene, the carrier containing the gene, and the application of the gene in genetic engineering for increasing the content of 1-aminocyclopropanecarboxylic acid (ACC) product in the ethylene biosynthesis pathway. Background technique [0002] Kiwi fruit belongs to Actinidiaceae (Actinidiaceae) and is a perennial deciduous vine. The fruit is sweet and sour, rich in amino acids, vitamins, mineral elements and other nutrients. Globally, the planting area of ​​kiwifruit is expanding year by year. China is the origin center of kiwifruit. With the improvement of kiwifruit commercial cultivation technology and the rapid development of the industry, my country's pl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N9/02C12N15/82A01H5/00A01H6/82
CPCC12N9/0051C12N15/8249C12Y108/04012
Inventor 殷学仁傅蓓凌张爱迪
Owner ZHEJIANG UNIV
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