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Red skinned pear polygalacturonase-inhibiting protein gene (PpPGIP) and application

A technology of polygalactose and protein inhibition, which is applied in the field of molecular biology and genetic engineering, can solve problems such as fungal diseases that cannot be fundamentally solved, reduce the use of disease-resistant agents, reduce environmental pollution, and broaden market application prospects Effect

Inactive Publication Date: 2010-06-16
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Each of the above three methods has certain disadvantages and limitations, and cannot fundamentally solve the problem of fungal diseases

Method used

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  • Red skinned pear polygalacturonase-inhibiting protein gene (PpPGIP) and application
  • Red skinned pear polygalacturonase-inhibiting protein gene (PpPGIP) and application
  • Red skinned pear polygalacturonase-inhibiting protein gene (PpPGIP) and application

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Embodiment

[0024] 1. PpPGIP full-length gene cloning and sequence analysis

[0025] Grind the red peel of the torched pear into powder with liquid nitrogen, transfer it to a centrifuge tube, and extract total RNA by the guanidine isothiocyanate method. Use reverse transcriptase M-MLV (Tiangen) to synthesize the first strand of cDNA using total RNA as a template. The reaction system and operation process are as follows: Take 5 μg Total RNA, add 50ng oligo(dT)15, 2μL dNTP (2.5mM each) in sequence , ddH 2 O (RNase-free) to a reaction volume of 13.5 μL; after mixing, heat denaturation at 70°C for 5 min, then rapidly cool on ice for 5 min, then add 4 μL 5×First-stand buffer, 0.5 μL RNasin (200 U), 1 μL M- MLV (200U), 1 μL DTT (0.1M), mix well and centrifuge for a short time, incubate at 42°C for 1.5h, take it out and heat at 95°C for 5min to terminate the reaction. After the first strand of cDNA was synthesized, it was stored at -20°C for future use.

[0026] Using the synthesized first-st...

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Abstract

This invention relates to a fungal resistance gene of red skinned pear, namely a polygalacturonase-inhibiting protein gene (PpPGIP), and application. The PpPGIP gene has a base sequence which is stated in a sequence-list SEQ ID; the length of the PpPGIP gene is 1,032bp, wherein 990bp is an open reading frame; and the PpPGIP gene codes polygalacturonase-inhibiting proteins containing 330 amino acids. Functional genomics related technology prove that the PpPGIP gene has the function of improving antifungal function of plants. The antifungal PpPGIP gene is constructed on a plant expression vector and is excessively expressed after being transferred into tobacco, the transgenic tobacco for expressing PpPGIP has strong in vitro antifungal activity and the obvious inhibition effect on a plurality of funguses like Aspergillus niger, Phomopsis funguses, Alternaria alternata, penicillium, and the like.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and genetic engineering, in particular to a polygalacturonase inhibitory protein gene PpPGIP with antifungal activity of torched pear and its application. Background technique [0002] In agricultural production, plant diseases are a very difficult problem, especially fungal diseases, which account for more than 80% of the total plant diseases, and several or even dozens of fungal diseases can be found on a crop. For nearly a century, breeders and plant pathologists have made great efforts to control fungal diseases of plants. So far, there are three main methods for controlling plant fungal diseases. One is to adopt several different cultivation management methods, such as crop rotation to avoid the use of soil containing pathogenic fungi; pesticide. Each of the above three methods has certain disadvantages and limitations, and cannot fundamentally solve the problem of fungal diseas...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00
Inventor 刘迪秋李文娴王继磊葛锋方松刚王光勇田荣欢
Owner KUNMING UNIV OF SCI & TECH
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