Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Application of cucumber lipid phosphatase cslppε2 in resistance to cucurbit blight

A phosphatase and cucumber lipid technology, which is applied in the fields of plant molecular biology and plant genetic engineering, can solve the problems such as the reports that cucumber lipid phosphatase is resistant to melon blight and the like, achieves wide application prospects and increases the area of ​​disease spots. Effect

Active Publication Date: 2022-05-17
INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous studies have shown that lipophosphate phosphatase plays an important role in plant abiotic stress, but there is no report of cucumber lipophosphate phosphatase CsLPPε2 in resisting melon blight

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of cucumber lipid phosphatase cslppε2 in resistance to cucurbit blight
  • Application of cucumber lipid phosphatase cslppε2 in resistance to cucurbit blight
  • Application of cucumber lipid phosphatase cslppε2 in resistance to cucurbit blight

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1 Construction of Cucumber CsLPPε2 Gene Overexpression Vector

[0093] Cucumber CsLPPε2 gene belongs to lipophosphate phosphatase protein gene. The cucumber CsLPPε2 gene CDS full-length (SEQ ID NO.2) was constructed into pGWB5 (pGWB5 plasmid map as shown in figure 1 shown), the specific position is between attR1 and attR2 behind the 35S promoter (the ccdB gene was replaced by gene recombination exchange), and an overexpression vector (pGWB5-CsLPPε2) was obtained; Fragment (CDS 1-137bp, SEQ ID NO.4) and its complementary fragment were constructed on pK7GWIW (pK7GWIW plasmid map as figure 2 shown), the specific position is between attR1 and attR2 behind the 35S promoter (the ccdB gene was replaced by gene recombination exchange), and the gene silencing vector (pK7GWIW-CsLPPε2) was obtained.

[0094] The amino acid sequence of cucumber CsLPPε2 protein is (221aa):

[0095] MRLVGWRLYSISCVWNFVDFKLGHLGSNFVSLGVAVLFGNIVDSFQSKWLVAALFAGIILLRHDAEALWTAMGSVINAVLSIVLKRILNQ...

Embodiment 2

[0102] Example 2 Constructing a Cucumber Cotyledon Model of Transient Overexpression / Gene Silencing

[0103] (1) The overexpression vector and the silencing vector in Example 1 were respectively transformed into Agrobacterium GV3101, and cultured upside down on the corresponding resistant medium for 48 hours.

[0104] (2) Pick a single clone and add it to 4 mL of LB medium containing corresponding antibiotics and rifampicin, shake the bacteria at 180 rpm for 24 hours at 28°C.

[0105] (3) Add fresh LB medium containing corresponding antibiotics and rifampicin at a ratio of 1:100, shake the bacteria at 28°C and 180 rpm until the OD600 value is about 3.0.

[0106] (4) Centrifuge at 3000rpm for 5 minutes to collect the bacteria, and use the suspension (10mM MES, 10mM MgCl 2 ) to resuspend the cells, adjust the OD600 value to about 0.4, and add 200 mM acetosyringone.

[0107] (5) Stand at room temperature for 3 hours.

[0108] (6) Prick a needle hole on both sides of the main v...

Embodiment 3

[0121] Example 3 Cucumber cotyledon resistance experiment

[0122] For the cucumber cotyledon model with transient overexpression of CsLPPε2 successfully constructed in Example 2, the cucumber cotyledon model of CsLPPε2 gene silencing, and the wild-type cucumber cotyledon, the cucumber seedlings after injection were cultured in the dark for 12 hours and then cultured in the light at 22°C for 3 days. After being isolated and inoculated with Phytophthora cucurbitus, the cotyledons of cucumber were cultured in a culture dish at 28°C for 24 hours in the dark.

[0123] The resistance function of the target gene to the disease was judged according to the size of the lesions on the isolated cucumber cotyledons of different treatments. The results are shown in the attached Figure 5 shown.

[0124] The results showed that the transient overexpression of CsLPPε2 gene (pGWB5-CsLPPε2) in cucumber cotyledon can significantly enhance the resistance of cucumber cotyledon to blight compared...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the fields of plant molecular biology and plant genetic engineering, and discloses the application of cucumber lipophosphate phosphatase CsLPPε2 in resisting melon blight. The present invention finds for the first time that cucumber lipophosphate phosphatase CsLPPε2 is resistant to melon blight: inoculating Phytophthora melonis on the cotyledons of plants overexpressing cucumber lipophosphate phosphatase CsLPPε2, the lesion area is significantly reduced, while in silent lipophosphate phosphatase Phytophthora cucurbita was inoculated on the cotyledons of plants with enzyme CsLPPε2, and the lesion area increased significantly; it can be seen that lipophosphate phosphatase CsLPPε2 plays a very important role in plant resistance to melon blight, and can be used to resist melon blight and prepare melon blight resistance The products and the cultivation of transgenic plants resistant to melon blight have broad application prospects.

Description

technical field [0001] The invention belongs to the fields of plant molecular biology and plant genetic engineering, and specifically relates to the application of cucumber lipophosphate phosphatase CsLPPε2 in resisting melon blight. Background technique [0002] Melon blight is a worldwide oomycete disease. The pathogen is Phytophthora melonis, which causes the leaves, stems, and fruits of cucumbers, pumpkins, wax gourds, and bitter melons to turn brown and cause death or dead seedlings. The disease occurs in areas where melons are grown in my country. In the north, it occurs more heavily in summer and autumn open-field cultivation and spring protection of sweet melons, and in the south in spring and summer and autumn, when the seasons are high and humid. The entire growth period and various parts of melons can be damaged. The damaged growth point of the seedlings and the base of the tender stems are water-soaked and constricted, wilting and dying. In the adult stage, the...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/16C12N15/55C12N15/84A01H5/00A01H6/34
CPCC12N9/16C12N15/8205C12N15/8218C12N15/8282C12Y301/03004
Inventor 吴廷全杜虎王瑞林毓娥徐晓美金庆敏阿奎尔·艾哈迈德杨晓珊
Owner INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com