Mechanized hybrid rice seed production using inducible female sterility

An induced and hybrid rice technology, applied in the field of agricultural biology, can solve problems such as large-scale commercial application, and achieve the effects of reducing seed production procedures, improving efficiency, and reducing seed production costs

Inactive Publication Date: 2016-02-10
HUNAN HYBRID RICE RES CENT
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned methods have many defects and cannot fundamentally realize the mechanized seed production of hybrid rice, and there is no large-scale commercial application in production at present.

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
  • Mechanized hybrid rice seed production using inducible female sterility
  • Mechanized hybrid rice seed production using inducible female sterility
  • Mechanized hybrid rice seed production using inducible female sterility

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Construction of rice expression vectors carrying gene expression cassettes of rice female fertility genes driven by inducible promoters

[0029] An expression cassette was inserted into the plant expression vector pHB: inducible promoter (SEQ ID NO.4), wild-type PTB1 (rice female fertility gene) coding region (SEQ ID NO.2), rbcSPolyA terminator (SEQ ID NO.5). The complete expression cassette sequence is shown in SEQ ID NO.6. See attached for complete construction vector map figure 1 .

[0030] The constructed vector plasmid was introduced into Agrobacterium strain EHA105 for genetic transformation of rice. The introduction method adopts the electric shock transformation method, mainly referring to the instruction manual of the electric shock instrument of bio-rad company, and the specific steps are as follows:

[0031] (1) Take out the DH10B competent cells stored at -80°C and freeze and thaw them on ice, place the 1mm electric shock cup on ice for pre-coo...

Embodiment 2

[0036] Embodiment 2: Transgenic rice obtains

[0037] 2.1 Plant material selection

[0038] The rice (Oryzasativa L.) variety used for genetic transformation was selected as the corresponding rice female sterile line carrying the PTB1 gene (SEQ ID NO.1).

[0039] 2.2 Induction of rice embryogenic callus

[0040] Take the immature rice seeds 10-15 days after pollination, peel off the seed coat, soak in 70% alcohol for 3 minutes, then soak in 0.1% mercury chloride for 15 minutes (or transfer to 20% sodium hypochlorite solution and sterilize on a shaking table for 25 minutes ), wash 3-5 times with sterile water in the ultra-clean workbench, squeeze out the sterilized immature embryos with tweezers, inoculate them on the induction medium, put about 35 seeds in each dish, and culture them in dark at 28°C for 4-5 days. Cut off the radicle, continue to culture for 12-15 days, and subculture after the callus grows up, once every two weeks, for a total of 2-3 times.

[0041]The matu...

Embodiment 3

[0053] Example 3: Molecular detection of transgenic plants

[0054] 3.1 DNA large-scale extraction

[0055] T 0 Transgenic rice DNA was extracted using the CTAB method (Murry and Thompson, 1980). Weigh 2-4 grams of fresh leaves, cut them into pieces, put them into a pre-cooled mortar at -20°C, grind them into powder with liquid nitrogen, transfer them to a pre-cooled grinding bottle and store them at -20°C for later use. When extracting, add 10ml of 1.5XCTAB preheated to 100°C, stir quickly, shake in a water bath at 56°C for 20 minutes, and then extract with an equal volume of chloroform:isoamyl alcohol (24:1). After centrifugation, the supernatant was extracted once with 1 / 10 volume of 10% CTAB buffer preheated to 56°C and an equal volume of chloroform:isoamyl alcohol (24:1), and then 1% CTAB was added to the supernatant to precipitate DNA After centrifugation, add 1mol / L NaCl added with RNaseA to dissolve the DNA, completely dissolve the alcohol precipitate at 56°C overni...

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 present invention discloses a female sterile rice breeding method. According to the method, an inducible promoter driven rice female fertile gene expression cassette is transformed into female sterile rice; during female sterile line breeding, the transgene rice uses a corresponding induction agent to induce recombinant fertile gene expression so as to achieve fertile selfing breeding; and during hybrid rice seed production, the transgene rice presents completely female sterility when the induction agent is not applied, such that the transgene rice can be adopted as a male parent so as to be used for seed production, and is subjected to hybridization with a female parent to produce hybrid seeds. According to the present invention, during the harvesting of the seed production process, the male parent is female sterility so as not to bear fruits, such that the male parent and the female parent can be subjected to box type sowing or mixed sowing, and the mixed harvesting can be achieved during the harvesting; and the difficult problem of the female sterile rice breeding is solved, the mechanized hybrid rice seed production is achieved, and especially the mechanized hybrid rice seed production in the mixed sowing and mixed harvesting manner is achieved.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology and discloses a hybrid rice mechanized seed production method using induced female sterility. Background technique [0002] my country's current hybrid rice seed production technology adopts the parental planting in stages, mostly the male parental planting first, and the female parental planting later; the parental parental plants are planted separately in box-type, the female parental lines are multiple rows, and the male parental lines are double-rowed; the parental parental plants are harvested separately , or cut off the male parent after pollination and before maturity. Therefore, hybrid rice seed production is a technology with high labor intensity, complicated procedures and high production costs. With the gradual implementation of urbanization, industrialization, agricultural industrialization and rural land transfer policies, the young and middle-aged labor force left behind in ...

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): A01H1/02C12N15/82
Inventor 曹孟良夏玉梅袁隆平沈春修方真
Owner HUNAN HYBRID RICE RES CENT
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