Cloning of inducible promoter Rd29A in japanese lawngrass and functional identification

A technology of promoter and zoysia, applied in the field of DNA extraction of zoysia, can solve the problems of plants being injured due to lack of water, plant death, etc.

Inactive Publication Date: 2012-07-18
BEIJING FORESTRY UNIVERSITY +2
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  • Summary
  • Abstract
  • Description
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Problems solved by technology

[0002] Drought, high salinity and low temperature stress will cause water shortage and damage to plants, and cause plant death in severe cases

Method used

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  • Cloning of inducible promoter Rd29A in japanese lawngrass and functional identification
  • Cloning of inducible promoter Rd29A in japanese lawngrass and functional identification
  • Cloning of inducible promoter Rd29A in japanese lawngrass and functional identification

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0007] 1 Materials and methods

[0008] 1.1 Plant material

[0009] Zoysia species were collected and preserved by the Turfgrass Research Institute of Beijing Forestry University in Yazigou, Liaoyang County, Liaoning Province.

[0010] 1.2 Experimental method

[0011] 1.2.1 Extraction of Zoysias Genomic DNA and Escherichia coli Plasmid DNA

[0012] Genomic DNA of Zoysia officinalis was extracted by SDS method (attached figure 1 ), the extraction of Escherichia coli plasmid DNA and related operations were carried out by alkaline lysis method (refer to the molecular cloning experiment manual for specific experimental operation steps).

[0013] 1.2.2 Rd29A promoter target gene cloning

[0014] According to the reported Rd29A promoter gene sequence and analyzed by the molecular biology software Oligo, the following two pairs of primers were designed, and the corresponding enzyme cutting sites BamHI and HindIII were introduced to facilitate directional ligation during vector co...

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Abstract

The invention relates to clone and functional identification of a stress-induced promoter Rd29A in zoysia. The invention comprises the following main steps: the zoysia is taken as a plant material, and total DNA of the plant is extracted; a pair of special primers is designed by biological software, and the sequence of the Rd29A promoter is obtained by the PCR amplification method; two plant eukaryon expression vectors, namely pBIG(35S-GFP) and pBIRG(Rd29A-GFP) are respectively established on the basis of pBI121; epidermic cells of an onion are transformed by the particle gun method; and the activity of the Rd29A promoter in adjusting and controlling GFP gene expression in receptor cells is detected. The invention can screen out the stress-induced promoter Rd29A which is suitable for genetransfer of turfgrass; and the Rd29A promoter is a special promoter which is induced to express by adverse circumstances such as drought, saline and alkaline, low-temperature and so on, is also an ideal induced promoter in the plant resistance genetic engineering, and has important significance on the growth and development of the turfgrass under the condition of adverse circumstances.

Description

technical field [0001] The invention belongs to the field of plant biotechnology or the field of crop breeding. Specifically, it involves related technologies of Zoysias DNA extraction, primer design, gene cloning and vector construction, and biolistic transformation. Thus, the stress-inducible promoter Rd29A was obtained from Zoysia pratense, and its function was identified. Background technique [0002] Drought, high salinity and low temperature stress will all cause water shortage and damage to plants, and cause plant death in severe cases. In order to adapt to or resist these unfavorable environmental conditions, plants will make various responses to maintain their own water status, so as to avoid or reduce the harm of water shortage to cells. Molecular responses and signaling. According to reports in recent years, the expression of many plant genes is induced by the above-mentioned drought, high-salt and low-temperature stress. Rd29A gene encodes a highly hydrophili...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10C12N15/29C12N15/82C12Q1/68
Inventor 韩烈保王莹曾会明刘君
Owner BEIJING FORESTRY UNIVERSITY
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