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Construction method for plant root related functional gene research model

A technology related to function and plant root system, which is applied in the field of construction of gene research models related to plant root system, can solve the problems of high labor intensity, inability to apply on a large scale, low frequency, etc., reduce experimental procedures, and overcome low regeneration efficiency , the effect of quick analysis

Inactive Publication Date: 2013-01-23
HANGZHOU NORMAL UNIVERSITY
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Problems solved by technology

Although most plants, including cucumbers, have established transgenic methods such as Agrobacterium-mediated method, gene gun method, pollen tube passage method, electric shock method, and injection method; however, these plant transgenic methods have low technical efficiency (the frequency of transgenic regenerated plants Low), experimental cycle (need to spend a lot of time), labor intensity (requires a large amount of experimental operations), etc., so it cannot be applied on a large scale (Zhang Bing, Wei Zhiming. "Problems and Countermeasures in Plant Genetic Transformation ".2000, 36(3): 245-251

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  • Construction method for plant root related functional gene research model
  • Construction method for plant root related functional gene research model
  • Construction method for plant root related functional gene research model

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Embodiment 1

[0027] 1. Experimental materials

[0028] The cucumber variety Zhongnong No. 8 was purchased from the Vegetable and Flower Research Institute of the Chinese Academy of Agricultural Sciences; the wild-type Agrobacterium rhizogenes K599 was preserved in this experiment (the National Collection of Plant Pathogenic Bacteria in U.K. purchased and obtained); the plant transgene expression vector pRI101-AN was purchased from TaKaRa Company, The plasmid pGFP containing the gfp gene was purchased from Clontech; other reagents: plant ash, vermiculite, 1 / 4MS liquid medium, LB solid medium, LB liquid medium, streptomycin sulfate (Str), kanamycin (Km ), acetosyringone (As), sterile water.

[0029] Equipment: autoclave, plastic or clay flowerpot (diameter 15cm), sterile syringe (needle diameter 0.36mm), artificial climate chamber (temperature 20-25°C, 12h / 12h alternating light and dark, light intensity 1200-2400Lux ), PCR amplification instrument (PE Company 9700 type), electrophoresis ins...

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Abstract

The invention provides a construction method for a plant root related functional gene research model. The method comprises the following steps of: connecting a plant root related functional gene to a transgenic expression vector pRI101-AN to obtain recombinant plasmid containing a target gene and transferring the recombinant plasmid into agrobacterium rhizogene K599; infecting a part which is 0.2-0.5cm below a cotyledon of a living cucumber seedling germinated for 4 days, and inducing to form a transgenic hairy adventitious root; shearing off a main root below the part where the hairy root isformed when the hairy adventitious root grows to be 5-10cm long; burying the formed hairy adventitious root into soil; and continuously growing for 1-2 months to obtain the plant root related functional gene research model. In the method, by using a cucumber plant with the transgenic hairy adventitious root, a cucumber root related functional candidate gene can be quickly identified, and the identification can be performed without obtaining the transgenic plant, so that an experimental procedure can be greatly reduced, the defect of low transgenic regeneration efficiency is overcome, and quick analysis of functions of the root related gene can be performed on a large scale.

Description

(1) Technical field [0001] The invention relates to a pretreatment method for rapidly analyzing root-related functions and genes, that is, a method for constructing a research model of plant root-related functional genes. (2) Background technology [0002] Cucumber is not only an important dual-purpose vegetable crop, but also a model plant for studying plant sex differentiation determination, plant vascular biology (including xylem and phloem), and plant long-distance signal transduction (Huang Sanwen et al. The genome of the cucumber, Cucumis sativus L., Nature Genetics, 2009, 41(12): 1275-1281). [0003] On the other hand, the root system plays an important role in the growth and biological yield of the aboveground parts of plants. The root system has the functions of fixation, absorption, synthesis and secretion. Contaminated soil etc. has become an increasingly active research field (Kijne JW et al. Root lectins and rhizobia. Plant Physics, 1997, 115(3): 315-320). [...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82
Inventor 向太和孟莎莎曹庆丰杜苏瑞
Owner HANGZHOU NORMAL UNIVERSITY
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