Seed specific interference vector containing pOsOle18 promoter and application of seed specific interference vector

An interfering vector and specific technology, applied in the biological field, can solve the problems of decreased plant fertility and yield, unclear direct regulation effect, and no application research report on rice protein content of OsMADS1 gene, etc., so as to improve rice yield and improve rice. quality effect

Active Publication Date: 2021-05-11
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
View PDF5 Cites 5 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 pOsOle18 promoter and application of seed specific interference vector
  • Seed specific interference vector containing pOsOle18 promoter and application of seed specific interference vector
  • Seed specific interference vector containing pOsOle18 promoter and application of seed specific interference vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1: Construction of pOsOle18-RNAi vector

[0076]To construct the pOsOle18-RNAi vector, specifically, using the homologous recombination directional cloning method, the promoter of the rice seed-specific expression gene OsOle18 was forward cloned into the plant constitutive RNA interference binary vector pLHRNAi with a hygromycin selection marker, and The original Ubiquitin constitutive expression promoter pUbi in the pLHRNAi vector was replaced to obtain the pOsOle18-RNAi seed-specific interference vector. Wherein, the pOsOle18 is the promoter of the OsOle18 gene. The construction flowchart and carrier map of the pOsOle18-RNAi vector are as follows: figure 1 As shown, the specific construction method is as follows:

[0077] 1. OsOle18 gene expression profile analysis

[0078] 1. OsOle18 gene expression profile analysis based on gene chip database

[0079] Expression profile analysis of OsOle18 gene in Nipponbare (rice variety, subsp. japonica). First use th...

Embodiment 2

[0144] Example 2: Construction of pOsOle18::OsMADS1-RNAi vector

[0145] Schematic diagram of the structure of pOsOle18::OsMADS1-RNAi vector and the main construction process as follows Figure 7 shown. The specific operation method is as follows:

[0146] 1. OsMADS1 gene expression profile analysis

[0147] 1. Transcriptional profiling analysis of OsMADS1 gene based on gene chip database

[0148] 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

[0239] Example 3: Specific application and effect demonstration of pOsOle18::OsMADS1-RNAi seed-specific interference vector

[0240] 1. The pOsOle18::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:

[0241] 1. Callus induction and subculture of mature rice seeds

[0242] 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 pOsOle18 promoter and application of the seed specific interference vector. The invention particularly provides a seed specific interference vector which is a pOsOle18-RNAi vector, and the DNA sequence of the pOsOle18-RNAi vector is shown as SEQ ID NO 9. The pOsOle18-RNAi vector can specifically interfere with expression of a target gene in rice seeds without affecting normal expression of the target gene in other tissues and organs, thus, direct regulation and control effect of the target gene on seed development can be effectively researched, furthermore, the purpose of improving the grain shape, yield and quality of rice can be achieved, and an effective technical means and an effective solution are provided for molecular accurate design and breeding of rice yield and rice quality. Meanwhile, the invention also provides an OsMADS1 seed specific interference vector pOsOle18::OsMADS1-RNAi based on the pOsOle18-RNAi vector, and the DNA sequence of the OsMADS1 seed specific interference vector pOsOle18::OsMADS1-RNAi is shown as SEQ ID NO 10. The vector can improve the protein content in rice and increase the nutritional value of rice. The seed specific interference vector has important scientific research significance and application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a seed-specific interference vector containing a pOsOle18 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 e...

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): C12N15/82C12N15/29A01H5/10A01H6/46
CPCC12N15/8218C12N15/8234C12N15/8251C12N15/8261C07K14/415Y02A40/146
Inventor 李辉李鹏慧涂升斌黄维藻汪松虎蒋海波刘阳轩古袁扬庄勇刘志坚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 Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap