Method for increasing content of vitamins of rice grains

A technology of rice grains and vitamins, applied in the field of molecular biological plant genetic engineering, to achieve the effects of promoting human health, increasing vitamin E content, and improving quality

Inactive Publication Date: 2018-07-03
袁米农业科技有限公司
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, except for the research by Sookwong et al. on increasing the content of tocotrienol in the rice bran layer by hybridization, there are no other reports on improving the vitamin E content of rice or changing the composition of vitamin E isomers.

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
  • Method for increasing content of vitamins of rice grains
  • Method for increasing content of vitamins of rice grains
  • Method for increasing content of vitamins of rice grains

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Embodiment 1: the construction of recombinant vector and the establishment of transformed Agrobacterium:

[0076] 1. According to figure 2 According to the technical route, the plasmid pUC18-Glb with the glutenin GluB-1 promoter (Glb) (according to the EMBL sequence X5433314 was PCR amplified and cloned into the pUC18 vector (Takara product), the method refers to Lee et al., 2001). Digest with EcoR I and HindIII, separate and recover the target fragment, connect with pCAMBIA1300 (Cambia company product) after double digestion with EcoR I and HindIII by T4 ligase to form an intermediate vector, and then isolate the clone from Arabidopsis The cDNA fragment of the γ-tocopherol methyltransferase (γ-TMT) gene (sequence see figure 1 ) was digested with BamHI and SacI, separated and recovered the target fragment, and formed a recombinant vector with the intermediate vector digested with BamHI and SacI by T4 ligase, and the above restriction enzymes (EcoR I, HindIII, Bam H I...

Embodiment 2

[0079] Example 2: Genetic Transformation of Agrobacterium:

[0080] 1. Induction:

[0081] The seeds of the mature rice variety (Wuyujing No. 7) were shelled, then treated with 70% volume ratio of absolute ethanol for 1 min, and 60.15% concentration of sodium hypochlorite (NaClO) seeds were shaken and sterilized for 20 min, and then rinsed with sterile water for 4 -5 times, and then put the seeds on japonica rice induction medium (N6D2) and culture them in a dark place for 4 weeks at a temperature of 25±1°C.

[0082] 2. Succession:

[0083] Select bright yellow, compact and relatively dry embryogenic calli, place them on the japonica rice subculture medium, and culture them in the dark for 2-3 weeks at a temperature of 25±1°C.

[0084] 3. Pre-culture:

[0085] Select compact and relatively dry embryogenic calli, put them on the japonica rice pre-medium and culture them in the dark for 4-5 days at a temperature of 25±1°C.

[0086] 4. Agrobacterium culture:

[0087] In the ...

Embodiment 3

[0100] DNA was extracted from leaves of transgenic plants, and polymerase chain reaction (PCR) was performed. PCR program: 94°C pre-denaturation for 5 minutes; 35 cycles (94°C denaturation for 1 minute; 55°C annealing for 1 minute; 72°C extension for 2 minutes), 72°C extension 7min. Detect positive transformed plants, and the single plant that can amplify the 1064bp size band is the positive transformed plant ( image 3 ). The RNA of the grains of the positive individual plants was extracted for Northern hybridization to detect the expression of the transferred fragments in the grains, and all the transferred fragments of the positive individual plants were expressed ( Figure 4 ). The large samples of DNA extracted from positive individual plants were digested with SacI and XbaI (products of Takara Company) respectively, and Southern hybridization was performed to identify the fragment integration copy number of positive transformed individual plants, and six independent tr...

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 method for increasing the content of vitamins of rice grains. The method comprises the following steps: A, constructing a recombinant vector; B, obtaining a transgenic plant;C, harvesting selfed seeds of single plant; D, selfing and seeding for the T1 generation; E, selfing and seeding for the T2 generation; F, selecting phenotype; and further comprises the steps of constructing the recombinant vector, namely separating the cDNA fragment of a gamma-tocopherol transmethylase gene from arabidopsis thaliana, and digesting and linking the cDNA fragment and specific gluten GluB-1 promoter of rice grains onto the expression vector pCAMBIA1300 to form the recombinant vector. According to the method, the content of vitamin E of rice grains is improved, the rice grain quality is improved, and a novel rice material with improved quality can be created.

Description

technical field [0001] The invention relates to the technical field of molecular biology plant genetic engineering, in particular to a method for increasing the vitamin content of rice grains. Background technique [0002] Vitamin E is a fat-soluble antioxidant, which is divided into two categories: tocopherols (α, β, γ, δ) and tocotrienols (α, β, γ, δ), which play an important role in the regulation of plant growth and development. effect. In addition, it is also involved in signal transduction in plant cells (Packer et al., 2001; Munne-Bosch S et al., 2007), regulating gene expression, etc. (Landes N et al., 2003; Abate A et al., 2000 ). [0003] Natural vitamin E is mainly synthesized in plants. Humans cannot synthesize it by themselves. They must be obtained from diet, especially from plants. Natural vitamin E is superior to artificially synthesized vitamins in terms of physiological activity, absorption rate, retention time in the body and safety. E, so it is of grea...

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): C12N15/82A01H1/02A01H1/04C12N15/66A01H5/10A01H6/46A23L7/10A23L33/00
CPCA01H1/02A01H1/04A23L7/10A23L33/00A23V2002/00C12N15/66C12N15/8243A23V2200/30
Inventor 张国栋刘佳音王晶邹丹丹吴洁芳张佩修旺珊米铁柱
Owner 袁米农业科技有限公司
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