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A kind of recombinant expression vector including Phytophthora inducible gene promoter and its application

An expression vector and an inducible technology are applied in the application field of the Phytophthora inducible gene promoter and recombinant expression vector, and can solve the problems of lack of a pathogenic inducible promoter, complicated control methods of Phytophthora sojae, and insignificant control effect.

Inactive Publication Date: 2017-01-18
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to solve the problems in the prior art that the control methods for Phytophthora soybean are complicated, the control effect is not obvious, and the novel disease-resistant genetic engineering technology lacks available pathogen-inducible promoters, and a Phytophthora inducible promoter is provided. son

Method used

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  • A kind of recombinant expression vector including Phytophthora inducible gene promoter and its application
  • A kind of recombinant expression vector including Phytophthora inducible gene promoter and its application
  • A kind of recombinant expression vector including Phytophthora inducible gene promoter and its application

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Embodiment 1

[0038] The soybean GmaKSTI36 gene is a gene that is rapidly up-regulated in the early stage of Phytophthora infection

[0039] 1. Screening of microarray data

[0040] According to the microarray data of three different soybean varieties: V710370, Williams, and Sloan (the three soybean varieties are preserved by Phytophthora sojae and Plant Interaction Laboratory of Nanjing Agricultural University) during the infection process of Phytophthora sojae ( Zhou L, Mideros SX, Bao L, et al. Infection and genotype remodel the entire soybean transcriptome [J]. BMC Genomics, 2009, 10(1): 49-59.), this literature analyzed the different soybean varieties in Phytophthora infestation The results showed that the expression of almost all soybean genes were regulated to varying degrees by Phytophthora infection. Based on the gene chip data in this literature, we first screened the chip genes with differentially up-regulated expression. The data of the three soybean varieties were analyzed se...

Embodiment 2

[0049] The promoter of embodiment 2 soybean GmaKSTI36 gene is a pathogen-inducible promoter

[0050] 1. Cloning of the promoter and construction of the vector

[0051] According to Chai C, Lin Y, Shen D, et al.Identification and FunctionalCharacteri-zation of the Soybean GmaPPO12 Promoter Conferring Phytophthorasojae In-duced Expression[J].Plos One,2013,8(6):1-6. Constructed in the article The plant expression vector method of promoter fusion GUS reporter gene was used to obtain the 35S:GUS recombinant plasmids fused with the GUS reporter gene by the constitutive promoter CaMV35S respectively, and the promoter region 1,500 bp upstream of the ATG transcription initiation site of soybean GmaKSTI36 gene was the vaccine. The GmaKSTI36:GUS recombinant plasmid with a mold-inducible promoter fused with the GUS reporter gene, and the 35S min:GUS recombinant plasmid with the GUS reporter gene fused to the 35S small promoter 35S min in the -46~+8 region of the constitutive promoter CaMV...

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Abstract

The invention belongs to the biotechnical field and provides a recombinant expression vector including a phytophthora-induced gene promoter and application of the phytophthora-induced gene promoter and the recombinant expression vector. A promoter of soybean GmaKSTI36 gene is fused with a GUS reporter gene to construct in a plant expression vector pMDC162, and pathogenic-bacteria-induced expression characteristics of the promoter are analyzed by virtue of an agrobacterium-mediate tobacco transient expression and soybean root hair stable expression system. According to the results, the promoter of the GmaKSTI36 gene can be used for quickly and efficiently driving the up-regulation inducible expression of the GUS reporter gene under the induction of the phytophthora in two different expression plant systems. And thus, the promoter of the GmaKSTI36 gene of the soybean is a phytophthora-induced promoter, and the valuable phytophthora-induced promoter is provided for plant anti-phytophthora genetic engineering.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a recombinant expression carrier including a Phytophthora inducible gene promoter and the application of the Phytophthora inducible gene promoter and the recombinant expression carrier. Background technique [0002] Effective control of plant diseases has always been a core issue in phytopathology research. For a long time, the use of chemical pesticides has been the main technology for disease control, but the problems faced in practical application are: on the one hand, there is a lack of effective pesticides, on the other hand, there are problems such as phytotoxicity and environmental pollution during the process of using pesticides. Later, a disease-resistant breeding strategy was proposed, but it was difficult to popularize and apply due to a series of problems such as long cycle, high cost, large scale and low precision. With the development of molecular biology and biotechnolo...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N5/10A01H5/00
Inventor 窦道龙柴春月曾文韬
Owner NANJING AGRICULTURAL UNIVERSITY
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