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

Plant expression vector for targeted silence of phytophthora capsici cellulose synthase 3 and application of plant expression vector

A plant expression vector and cellulose synthase technology, applied in the field of agricultural biology, can solve problems such as undiscovered plant diseases, and achieve the effects of delaying the generation of drug resistance, reducing the amount of use, and being beneficial to environmental protection

Pending Publication Date: 2022-08-05
CHINA AGRI UNIV
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After searching, there is currently no relevant literature on the control of plant diseases caused by Phytophthora capsici based on RNA silencing technology

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
  • Plant expression vector for targeted silence of phytophthora capsici cellulose synthase 3 and application of plant expression vector
  • Plant expression vector for targeted silence of phytophthora capsici cellulose synthase 3 and application of plant expression vector
  • Plant expression vector for targeted silence of phytophthora capsici cellulose synthase 3 and application of plant expression vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Sequence source for inhibiting the expression of Phytophthora capsicum oxysterol protein 1 gene of the present invention

[0039] The sequences in the two plant expression vectors of the present invention are derived from different regions of the cDNA sequence of PcCes3, and the sequence is shown in SEQ ID No. Therefore, it can be used as a plant expression vector for enhancing the resistance of plants to Phytophthora capsicum. The cDNA sequence of PcCes3 was derived from the gene numbered JX905357 in the database (GenBank) of Phytophthora capsici.

[0040] The 1801-2602 cDNA sequence of PcCes3, a total of 802 nucleotides (sequence 2 in the sequence table), was selected as the expression sequence of the plant expression vector with the long loop hairpin structure, and the 3005-3559 555 nucleotide sequence (the sequence in the sequence table). 1) As a plant expression vector with a short loop hairpin structure, the required amplification primers are shown in T...

Embodiment 2

[0044] Example 2 Construction of plant expression vector of the present invention

[0045] The primers involved in amplifying the sequences of the present invention are entrusted to the Synthesis Biosynthesis, and the sequences are obtained by amplification using Q5 High Fidelity DNA Polymerases (NEB), which are recovered by gel electrophoresis for vector construction.

[0046] For the short loop hairpin structure vector, the fragment described in sequence 1 (with Phytophthora capsici as a template, and the fragments obtained by amplification with pENsenseC5F and pENsenseC5R in Table 2 as primers were obtained by double digestion with Cla I and EcoR I) To insert between the pENTR1a plasmid restriction site Cla I and EcoR I, after PCR, sequencing, restriction enzyme digestion, electrophoresis verification, the fragment described in sequence 1 (with Phytophthora capsici For template, take pENantisenseC5F and pENantisenseC5R in table 2 as primer amplification obtained fragment ob...

Embodiment 3

[0051] Example 3 Using the plant expression vector involved in the present invention to construct a transgenic plant

[0052] Test plants and bacterial liquid: aseptically cultured tobacco seedlings, Agrobacterium tumefaciens strain GV3101 containing the plant expression vector involved in the present invention.

[0053] Test medium:

[0054] (1) 1 / 2 MS solid medium: MS (Murashige & Skoog medium) 2.2g, sucrose 30g, stir and dissolve, adjust pH to 5.6-5.8 with NaOH, add 8g agar to make up to 1L, sterilize at high temperature and autoclave.

[0055](2) MS solid (liquid) pre-medium: MS 4.44g, MES 2.1235g, sucrose 30g, after stirring and dissolving, adjust pH to 5.8 with NaOH, add 2.4g Phytogel, 1mL 6-BA (1mg / mL) After autoclaving, 1 mL of acetosyringone (AS, 100 μM), 1 mL of naphthalene acetic acid (NAA, 1 mg / mL) were added, and no phytogel was added to the liquid medium.

[0056] (3) Screening medium (budding): MS 4.44g, sucrose 30g, stir and dissolve, adjust pH to 5.8 with N...

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 discloses a plant expression vector capable of effectively interfering a phytophthora capsici cellulose synthase 3 gene through host-induced gene silencing and application of the plant expression vector. The plant expression vector disclosed by the invention can be transcribed to form double-stranded RNA with a reverse complementary structure, and the double-stranded RNA is formed by respectively inserting a nucleotide sequence shown as SEQ ID No.1 or SEQ ID No.2 into the vector twice in the forward direction and the reverse direction. The invention also discloses application of the expression vectors in preventing and treating plant diseases caused by phytophthora capsici. The plant expressing the vector provided by the invention has obviously enhanced disease resistance to phytophthora capsici, the expression of the cellulose synthase 3 gene of the phytophthora capsici infecting the plants is significantly inhibited, and the cellulose synthase 3 gene is necessary for the development and pathopoiesis of the phytophthora capsici. Therefore, the plant expression vector can effectively inhibit plant infection caused by phytophthora capsici, and an effective way is provided for preventing and treating related diseases caused by phytophthora capsici.

Description

technical field [0001] The invention relates to the technical field of agricultural biotechnology, in particular to two plant expression vectors with different secondary structure transcripts for inhibiting the expression of Phytophthora capsici cellulose synthase 3 gene, and the use of these plant expression vectors in the prevention and treatment of plant diseases caused by Phytophthora capsici application on. Background technique [0002] Phytophthora capsici is an important phytopathogenic oomycete that is distributed all over the world and can infect more than 70 species of Solanaceae, legumes and most cucurbits, causing damping, wilting and root, Stem and fruit rot, from seedling stage to fruit stage, can cause serious economic losses. The pepper blight caused by Phytophthora capsicum is a devastating disease that can be spread by rain, soil and air currents, causing symptoms such as leaf withering, fruit rot, and even the death of the whole plant, which has a great i...

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 Applications(China)
IPC IPC(8): C12N15/113C12N15/84A01H5/00A01H6/82
CPCC12N15/1137C12N9/1059C12N15/8218C12N15/8282C12N2310/14
Inventor 刘西莉王治文钟珊高翔李瑜张博瑞张思聪李腾蛟
Owner CHINA AGRI UNIV
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