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

Seed-specific interference vector containing pOsGlb-1 promoter and application thereof

A technology of posglb-1 and posglb-1-rnai, which is applied in the field of seed-specific interference vectors, can solve the problems of no OsMADS1 gene application research reports on rice protein content, decreased plant fertility and yield, and unclear direct regulatory effects, etc. , to achieve the effect of improving the nutritional quality of rice and increasing the protein content of rice

Active Publication Date: 2021-05-28
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at the same time, the fertility and yield of interfering plants are extremely significantly reduced, making it difficult for practical application (Liao PF, Ouyang JX, Zhang JJ, et al. OsDCL3b affects grain yield and quality inferior[J]. Plant Molecular Biology, 2019, 99( 3): 193-204.)
[0007] At the same time, although the OsMADS1 gene has an important regulatory function on rice flower development, its direct regulation on rice seed development and rice quality development is still unclear, and there are no research reports on the application of OsMADS1 gene in improving rice protein content. and invention patents

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
  • Seed-specific interference vector containing pOsGlb-1 promoter and application thereof
  • Seed-specific interference vector containing pOsGlb-1 promoter and application thereof
  • Seed-specific interference vector containing pOsGlb-1 promoter and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1: Construction of pOsGlb-1-RNAi vector

[0075]Construct the pOsGlb-1-RNAi vector, specifically using the homologous recombination directional cloning method to forward clone the promoter of the rice seed-specific expression gene OsGlb-1 into a plant constitutive RNA interference binary vector with a hygromycin selection marker pLHRNAi, and replace the original Ubiquitin constitutive expression promoter pUbi in the pLHRNAi vector, so as to obtain the pOsGlb-1-RNAi seed-specific interference vector. Wherein, the pOsGlb-1 is the promoter of the OsGlb-1 gene. The construction flowchart and carrier map of the pOsGlb-1-RNAi vector are as follows: figure 1 As shown, the specific construction method is as follows:

[0076] 1. OsGlb-1 gene expression profile analysis

[0077] 1. OsGlb-1 gene expression profile analysis based on gene chip database

[0078] Expression profile analysis of OsGlb-1 gene in Nipponbare (Nipponbare, rice variety, Japonica subspecies). Fir...

Embodiment 2

[0143] Example 2: Construction of pOsGlb-1::OsMADS1-RNAi vector

[0144] Schematic diagram of the structure of pOsGlb-1::OsMADS1-RNAi vector and the main construction process as follows Figure 7 shown. The specific operation method is as follows:

[0145] 1. OsMADS1 gene expression profile analysis

[0146] 1. Transcriptional profiling analysis of OsMADS1 gene based on gene chip database

[0147] First, the expression profile of the OsMADS1 (gene number: Os03g0215400) gene in Nipponbare was analyzed using the RiceXPro rice gene chip expression database, as shown in Figure 8 As shown (according to the picture, you need to enter the URL http: / / ricexpro.dna.affrc.go.jp / category-select.php, enter Os03g0215400 in the Search options on the web page, then click search, and then click FeatureNum in the new web page After downloading the serial number, check the Spatio-temporal profile on the new webpage, you can find this graph, and click Downloadselected graph on the webpage to...

Embodiment 3

[0238] Example 3: Specific application and effect demonstration of pOsGlb-1::OsMADS1-RNAi seed-specific interference vector

[0239] 1. The pOsGlb-1::OsMADS1-RNAi and pUbi::OsMADS1-RNAi plasmids constructed in Example 2 and Comparative Example were sent to Hangzhou Baige Biotechnology Co., Ltd. to transform the rice variety Nipponbare respectively. The conversion process is as follows:

[0240] 1. Callus induction and subculture of mature rice seeds

[0241] Select mature rice seeds, peel off the glumes, pour them into a 50ml centrifuge tube, add 75% ethanol to sterilize for 1 minute, pour off the ethanol, rinse with sterile water once, pour it out, then add 30% sodium hypochlorite to disinfect for 20 minutes, pour out the sodium hypochlorite, and use Rinse with sterile water 5-6 times. Use a pipette to absorb excess water, and transfer the seeds to the induction medium, 20-25 seeds per dish. After the callus grows, the original embryo can be directly used for transformatio...

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 technical field of biology, and particularly relates to a seed-specific interference vector containing a pOsGlb-1 promoter and application of the seed-specific interference vector. The invention specifically provides the seed-specific interference vector which is a pOsGlb-1-RNAi vector and has a DNA sequence as shown in SEQ ID No. 9. By utilizing the pOsGlb-1-RNAi vector, the expression of a target gene in rice seeds can be specifically interfered without influencing the normal expression of the target gene in other tissue and organs, so not only can the direct regulation effect of the target gene on the seed development be effectively researched, but also the purposes of improving the grain shape of rice, rice yield and rice quality can be achieved; and the invention provides an effective technical means and solution for molecular precise design and breeding of rice with high yield and high quality. Meanwhile, the invention also provides an OsMADS1 seed-specific interference vector pOsGlb-1: OsMADS1-RNAi based on the pOsGlb-1-RNAi vector, wherein the DNA sequence of the OsMADS1 seed-specific interference vector is as shown in SEQ ID NO 10; By utilizing the OsMADS1 seed-specific interference vector, the protein content of rice can be increased, and the nutritional value of the rice is increased. The seed-specific interference vector of the invention has important scientific research significance and application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a seed-specific interference vector containing a pOsGlb-1 promoter and application thereof. Background technique [0002] Rice is the main ration of more than half of the world's population, and it is also the basic source of human starch and protein. The growth and development of rice seeds not only controls the thousand-grain weight, which is one of the three elements of rice yield, but also is an important factor that directly determines the nutritional quality of rice such as rice protein content. The morphological development of rice seeds and rice protein content are regulated by a series of important genes. Therefore, studying the functions of these important regulatory genes and their regulatory mechanisms on seed morphological development and rice protein content is the basis for using these important genes to carry out precise seed design breeding and genetic en...

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): C12N15/82C12N15/113C12N15/66A01H5/10A01H6/46
CPCC12N15/8218C12N15/8261Y02A40/146
Inventor 李辉李鹏慧涂升斌黄维藻汪松虎蒋海波庄勇刘阳轩刘志坚古袁扬魏明唐钰莹乐露路龙祥胡永志喻兰
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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