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Application and method of using pto/atbi-i bivalent gene in promoting growth of Taxus chinensis and increasing paclitaxel content

A taxus and paclitaxel technology, applied in the biological field, can solve problems such as adverse effects on the growth of medicinal plants, no practical use value, etc., and achieve the effects of increasing the content of paclitaxel, reducing the cultivation cost and shortening the growth cycle.

Inactive Publication Date: 2016-09-28
HANGZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, many scholars have studied the effects of environmental stress on the accumulation of secondary metabolites in medicinal plants, and found that some artificial stress can increase the content of secondary metabolites in medicinal plants, but these artificial stresses often simultaneously It will have serious adverse effects on the growth of medicinal plants, so these technologies are often of no practical value in production practice

Method used

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  • Application and method of using pto/atbi-i bivalent gene in promoting growth of Taxus chinensis and increasing paclitaxel content
  • Application and method of using pto/atbi-i bivalent gene in promoting growth of Taxus chinensis and increasing paclitaxel content
  • Application and method of using pto/atbi-i bivalent gene in promoting growth of Taxus chinensis and increasing paclitaxel content

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: Transformation of Pto / AtBI-1 gene yew plant construction

[0033] (1) Target gene amplification:

[0034] The pET112-pto plasmid containing the tomato Pto cDNA sequence was used as a template to amplify and recover Pto, which was provided by Dr. Gregory Martin of Purdue University. Other plasmids containing the Pto cDNA sequence can also be used and are available from the authors of published papers on the subject. After electrophoresis of the PCR product, the amplified Pto gene (SEQ ID NO.1) was recovered for future use.

[0035] Use the pYX112-AtBI-GFP plasmid containing the tobacco AtBI-1 cDNA sequence as a template to amplify and recover AtBi-1. This plasmid is provided by Dr. Hirofumi Uchimiya of the University of Tokyo. Other plasmids containing the AtBI-1 cDNA sequence can also be used. These plasmids Available from authors who have published relevant papers. After electrophoresis of the PCR product, the amplified AtBi-1 gene (SEQ ID NO.2) was re...

Embodiment 2

[0055] Effect of Pto / AtBI-1 Gene on Growth of Taxus chinensis

[0056] (1) Using the Pto / AtBI-1 gene-transferred yew plants obtained in Example 1 as materials, perform cultivation and management according to the following transplantation cultivation points:

[0057] 1. Transplantation time: generally from the first ten days of April to the first ten days of May

[0058] 2. Soil preparation: Before transplanting, it is necessary to prepare the land. It is ideal to choose a shady or semi-shady slope or a valley area with short sunshine. The land preparation requires deep plowing and fine preparation, and the soil layer should not have water accumulation or compaction.

[0059] 3. Transplanting: The method of transplanting can be planted in holes. First, use a shovel to dig out a hole on the seedbed with a certain row spacing, and put some fertile nutrient soil in it, and spread the roots of the seedlings evenly on the surface of the small cone at the bottom of the hole. , pay a...

Embodiment 3

[0069] Effect of Pto / AtBI-1 Gene on Paclitaxel Content in Taxus Plants

[0070] (1) Obtain Pto / AtBI-1 gene yew plants and transgenic control plants respectively according to the method in implementation 1;

[0071] (2) Cultivate and manage the Pto / AtBI-1 gene-transferred yew plants, yew wild-type plants, and transgenic control plants according to the same transplanting and cultivation points as in Example 2;

[0072] (3) Sampling and measuring the paclitaxel content in the leaves and stems of Taxus chinensis at different stages of plant growth, and examining the paclitaxel content in the Pto / AtBI-1 gene-transformed Taxus chinensis plants, Taxus wild-type plants, and transgenic control plants. The results showed that the paclitaxel content in the leaves and stems of the transgenic Pto / AtBI-1 yew plants was significantly higher than that of the wild yew plants and the transgenic control yew plants. After 6 months of cultivation, the paclitaxel content in leaves and stems of tax...

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Abstract

The invention discloses an application of Pto / AtBI-I bivalent gene in promoting taxus chinensis growth and improving taxol content and method which comprises the following steps: connecting a target gene Pto and AtBI-1 to an expression carrier plasmid with 35S promoter to construct a Pto / AtBI-1 bivalent gene recombinant plasmid, introducing the bivalent gene recombinant plasmid into taxus chinensis cell through an agrobacterium-mediated method, screening a stably expressed positive clone and performing differentiation culture to obtain a transgene taxus chinensis plant with fast growth and high taxol content. By expressing the Pto / AtBI-I bivalent gene in the taxus chinensis, the growth of the taxus chinensis is promoted, the production period is shortened, the cultivation cost is lowered, and the taxol content is improved; after the transgene taxus chinensis is differentially planted for 6 months, the single weight is respectively improved by 56.7% in comparison with wild taxus chinensis, and the taxol content in taxus chinensis stem and leaf is respectively improved by 65.3% and 87.5%.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to an application and a method of using Pto / AtBI-1 bivalent gene in promoting the growth of yew and increasing the content of paclitaxel. Background technique [0002] The natural medicine industry based on medicinal plants is one of the key industries supported by the country during the 12th Five-Year Plan and one of the ten leading agricultural industries in the 12th Five-Year Plan of Zhejiang Province. Medicinal plants can not only be used as raw materials for chemicals, but also can be used as traditional Chinese medicine materials in the health care and treatment of traditional medicine. In medicinal plants, the content of medicinal ingredients is the material basis for determining their quality and development and utilization value. [0003] At present, extraction and separation from plants is still the most important way to obtain many important natural medicines. How...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00
Inventor 徐茂军阮家昭金海红范耀奇邱蓉蓉
Owner HANGZHOU NORMAL UNIVERSITY
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