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

Application of lilium regale germin protein gene LrGLP1

A germin and protein technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of inability to completely control fungal diseases, harm to human and animal life and health, long cycle, etc., achieve broad market application prospects, shorten the breeding cycle, Simple to use effects

Inactive Publication Date: 2014-11-05
KUNMING UNIV OF SCI & TECH
View PDF1 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional prevention and treatment of fungal diseases is mainly to screen resistant varieties, clean up diseased plants and fruits in time, and use pesticides. Health hazards, so fungal diseases cannot be completely controlled

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 lilium regale germin protein gene LrGLP1
  • Application of lilium regale germin protein gene LrGLP1
  • Application of lilium regale germin protein gene LrGLP1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: LrGLP1 Full-length cDNA cloning and sequence analysis

[0023] Lilium Minjiang was inoculated with Fusarium oxysporum, total RNA was extracted from the roots 12 hours after inoculation, the treated roots of Lily Minjiang were ground into powder with liquid nitrogen, then transferred to a centrifuge tube, and extracted by guanidine isothiocyanate method total RNA. Use reverse transcriptase M-MLV (promega) 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 50 ng oligo (dT), 2 μL dNTP Mix (2.5mM Each), the reaction volume was made up to 14.5 μL with DEPC water; after mixing, heated and denatured at 70°C for 5 min, then rapidly cooled on ice for 5 min, then added 4 μL 5×First-stand buffer, 0.5 μL RNasin ( 200U), 1 μL M-MLV (200U), mix well and centrifuge briefly, incubate at 42°C for 1.5 h, take it out and heat at 70°C for 10 min to terminate the reaction. Aft...

Embodiment 2

[0026] Embodiment 2: plant overexpression vector construction

[0027] Use the SanPrep column type plasmid DNA mini-extraction kit (Shanghai Sangong) to extract the insert LrGLP1 coli plasmid pMD18-T- LrGLP1 As well as the plant expression vector pCAMBIA2300S plasmid, 1 μL was used for agarose gel electrophoresis to detect the integrity and concentration of the extracted plasmid. restriction endonuclease Eco RI (TaKaRa) and Bam HI (TaKaRa) against plasmid pMD18-T- LrGLP1and pCAMBIA2300S for double enzyme digestion (100 μL system), the reaction system and operation process are as follows: take 20 μL pMD18-T- LrGLP1 and pCAMBIA2300S plasmid, add 10 μL 10×K buffer, 5 μL EcoRI , 5 μL Bam HI, 60 μL ddH 2 O, after mixing, centrifuge for a short time, and place at 37°C for overnight reaction. All digested products were subjected to agarose gel electrophoresis, and then SanPrep Column DNA Gel Extraction Kit (Shanghai Shenggong) was used for LrGLP1 The fragments and the la...

Embodiment 3

[0030] Example 3: Plant genetic transformation mediated by Agrobacterium and screening of transgenic plants

[0031] The transgenic recipient in this experiment was tobacco ( Nicotiana tabacum L.). Tobacco seeds were soaked in 75% alcohol for 30 s, washed with sterile water and washed with 0.1% HgCl 2 Soak for 8 minutes, then wash several times with sterile water, sow on 1 / 2 MS medium, culture in dark at 28°C for 5-8 days, transfer to light incubator after germination (25°C, 16h / d light), Subculture once a month with MS medium.

[0032] Take out the stored pCAMBIA2300S-containing pCAMBIA2300S- LrGLP1 Agrobacterium LBA4404 strain of the plasmid was inoculated in 5 mL of LB liquid medium containing 50 mg / L Km and 20 mg / L rifampin in 20 μL, and cultured at 28°C until the medium was turbid. Pipette 1 mL of turbid bacterial solution onto LB solid medium containing 50 mg / L Km, and incubate at 28°C for 48 h. Then scrape off an appropriate amount of Agrobacterium on LB solid me...

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

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention relates to an application of a lilium regale germin protein gene LrGLP1. The nucleotide sequence of the LrGLP1gene is shown as SEQ ID NO:1, a germin-like protein is encoded, and according to the invention, functional genomics related technical researches prove that the LrGLP1gene has a function of improving the fungal infection resistance of plants. An antifungal LrGLP1 gene in the invention is built into a plant expression vector, and transferred into a tobacco to be subjected to excessive expression, so that a transgenic tobacco plant has an extremely strong in-vitro antifungal activity. LrGLP1overexpressed transgenic tobaccos have an obvious inhibitory effect on the growth of alternaria alternate, sclerotinia sclerotiorum and catenuliform gibberella.

Description

technical field [0001] The invention relates to the research field of molecular biology and genetic engineering related technologies, in particular to the Minjiang lily germin protein gene with antifungal activity LrGLP1 Applications. Background technique [0002] Plant diseases refer to a series of biochemical, physiological and even morphological lesions that occur in plants under the influence of biological factors, hindering normal growth and development, and seriously endangering agricultural production. Pathogenic fungi are the main pathogens that cause plant diseases. The incidence of fungal diseases is extremely high, and when they occur on a large scale, they can cause crop yield reduction or even death. The traditional prevention and treatment of fungal diseases is mainly to screen resistant varieties, timely clean up diseased plants and fruits, and use pesticides. Health hazards, so fungal diseases cannot be completely controlled. With the establishment and ra...

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): C12N15/29C12N15/84A01H5/00
Inventor 刘迪秋张南南季博韩青葛锋陈朝银
Owner KUNMING UNIV OF SCI & TECH
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