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

Transcription factor PDD1 for regulating development of fruiting body of flammulina velutipes and coding gene and application thereof

A technology of Flammulina velutipes and transgenic, which is applied in the field of genetic engineering, can solve the problems of knocking down adenosine deaminase encoding gene and needs to be carried out, and achieve the effect of increasing the total biomass

Active Publication Date: 2018-09-21
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
View PDF1 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, fvh1 is highly expressed during the development of fruiting bodies of Flammulina velutipes, and the overexpression of the hydrophobin-encoding gene Fv-hyd1 advances the formation of primordium, histidine kinase gene and GATA Zinc finger transcription factor participate in the process of cold-induced primordium formation, Mads8, The HK535 gene was differentially expressed in primordia and hyphae. Knocking down the adenosine deaminase encoding gene Fv-ada could inhibit the mycelial growth of Flammulina velutipes. The analysis of the mating type genes of Flammulina velutipes, the laccase-encoding gene is involved in the formation of the fruiting body of Flammulina velutipes, and the overexpression of Fv-JRL1 promotes the mycelial growth and fruiting body development of Flammulina velutipes, but these studies are limited to the study of gene structure or gene function. The application in Flammulina velutipes has yet to be developed

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
  • Transcription factor PDD1 for regulating development of fruiting body of flammulina velutipes and coding gene and application thereof
  • Transcription factor PDD1 for regulating development of fruiting body of flammulina velutipes and coding gene and application thereof
  • Transcription factor PDD1 for regulating development of fruiting body of flammulina velutipes and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Example 1, the acquisition of pdd1 gene and transcription factor PDD1 and its sequence analysis

[0082] 1. Nucleotide sequence of pdd1 gene and amino acid sequence of transcription factor PDD1

[0083] Eight genes encoding both nuclear localization sequences and HMG-box structural proteins were found in the Flammulina velutipes genome. RNA-seq and quantitative PCR were used to analyze the transcription levels of these eight genes at different stages of fruiting body development, and one of the genes was found to be from the original From the base stage to the elongation stage and the mature stage, the expression is gradually up-regulated ( figure 1 ), may play an important function in the development of the fruiting bodies of Flammulina velutipes, and the gene is named pdd1 (primordium development defect1), and its nucleotide sequence is sequence 1. The pdd1 gene encodes the transcription factor PDD1, and the amino acid sequence of the transcription factor PDD1 is SE...

Embodiment 2

[0087] Example 2, construction of pdd1 overexpression mutant strain and pdd1 knockdown expression mutant strain

[0088] 1. Construction of pdd1 overexpression vector

[0089] (1) carry out PCR amplification with primer Pgpd-F and Pgpd-OE-R with the genome DNA of Flammulina velutipes wild-type strain as template, obtain Flammulina velutipes gpd promoter fragment (Pgpd-OE), this gpd promoter comprises gpd gene first Introns and exons, the target sequence size is 920bp. The primer sequences are as follows:

[0090] Pgpd-F:5'-CAGATCCCCCGAATTATTCGAGCTCGGTACAGTCGTG-3';

[0091] Pgpd-OE-R: 5'-AGAAAGAGTGGACCTGTAAAATGGTGAGCAAGAC-3'.

[0092] The PCR reaction program was: 98°C for 30s; 98°C for 10s, 55°C for 90s, 72°C for 60s (25cycles); 72°C for 10min; 4°C.

[0093] (2) PCR amplification was carried out with primers pdd1-F and pdd1-R using the genomic DNA of Flammulina velutipes wild-type strain as a template to obtain the pdd1OE fragment, the target sequence size was 1203bp, and ...

Embodiment 3

[0191] Embodiment 3, the application of pdd1 in regulating the mycelial growth of Flammulina velutipes

[0192] 1. Observation experiment of mycelial growth on plate for pdd1 mutant strain

[0193] (1) The wild type of Flammulina velutipes, pdd1 overexpression mutant strain (pdd1 OE#7 , pdd1 OE#31 and pdd1 OE#38 ) and pdd1 knockdown expression mutant strains (pdd1 RNAi#64 , pdd1 RNAi#148 and pdd1 RNAi#170 ) were respectively made into bacterial blocks (d=5mm) of the same size with a puncher, and were inoculated on drug-free CYM plates, cultured at 25°C for 7 days and photographed.

[0194] (2) During the cultivation process, the growth length of mycelium was measured every 24 hours, and recorded, and the average growth rate of each bacterial strain was calculated.

[0195] The results of the growth and growth rate analysis of different strains are as follows: On the CYM plate, the mycelia growth of the pdd1 overexpression mutant strain was similar to that of the wild typ...

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
Lengthaaaaaaaaaa
Lengthaaaaaaaaaa
Molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention discloses a transcription factor PDD1 for regulating development of fruiting body of flammulina velutipes and an encoding gene and an application thereof. The coding gene sequence of thetranscription factor PDD1 is shown in the sequence 1 in the sequence table; the amino acid sequence of the transcription factor is shown in sequence 2 in the sequence table. According to the method,the overexpression of the pdd1 and the RNAi experiment in the flammulina velutipes are carried out, and the pdd1 is a positive control factor which is necessary during the growth and the fruiting bodydevelopment of the flammulina velutipes, and influences the original basal period and the maturity of the flammulina velutipes, the number of the fruiting bodies, the height and the wet weight. The invention also discloses a flammulina velutipes mutant strain (pdd1OE#31) with short fruiting period and high yield, which not only has the characteristics of early maturity, but also can improve the yield of the flammulina velutipes, shorten the production cycle of the flammulina velutipes, reduce the energy consumption and the labor cost, and has great application prospect.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a transcription factor PDD1 that positively regulates the mycelial growth and fruiting body development of Flammulina velutipes, and a pdd1 overexpression mutant strain (pdd1) with short fruiting period and high yield. OE#31 ). Background technique [0002] Flammulina velutipes (Flammulina velutipes), also known as mushrooms, mushrooms, and money mushrooms, is a typical basidiomycete edible fungus, which is widely cultivated in the world, especially in Asian countries. Flammulina velutipes is rich in protein, dietary fiber and carbohydrates, and also rich in vitamins B1, B2, B3 and vitamins C, D, and E. As a traditional edible fungus, Flammulina velutipes not only has rich nutritional value, but also has important medicinal value. Studies in recent years have shown that Flammulina velutipes has anti-tumor, prolong life, lower cholesterol, anti-fatigue, immune regula...

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
IPC IPC(8): C07K14/375C12N15/31C12N15/80C12N1/15C12R1/645
CPCC07K14/375C12N15/80
Inventor 李少杰吴塔菊孙宪昀张振颖
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF 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