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

Nucleic acid molecule, vector and cell, application of nucleic acid molecule, vector and cell and application of nucleic acid molecule, vector and cell and method for efficiently screening plant apomixis cloned seeds through asexual embryos and

A technology of nucleic acid molecules and recombinant vectors, which is applied in the fields of plant molecular biology and agricultural biotechnology, and can solve problems such as low purity requirements, the inability to realize the application of hybrid apomictic systems, and the inability to sort zygotic embryo seeds.

Pending Publication Date: 2022-05-17
HUNAN HYBRID RICE RES CENT
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the highest rate of cloned seeds in the reported apomictic system is only 29%, which is far below the purity requirements for production, and cloned seeds and zygotic embryo seeds formed by normal pollination and fertilization cannot be sorted, so that Realize the application of hybrid apomixis system

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
  • Nucleic acid molecule, vector and cell, application of nucleic acid molecule, vector and cell and application of nucleic acid molecule, vector and cell and method for efficiently screening plant apomixis cloned seeds through asexual embryos and
  • Nucleic acid molecule, vector and cell, application of nucleic acid molecule, vector and cell and application of nucleic acid molecule, vector and cell and method for efficiently screening plant apomixis cloned seeds through asexual embryos and

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0059] According to a preferred embodiment of the present invention, the nucleic acid molecule further comprises an expression cassette E2 carrying an embryo autonomous gene.

[0060] More preferably, the expression cassette E2 includes the following elements from upstream to downstream: a first promoter, an embryo autonomous gene and a first terminator, more preferably an egg cell-specific promoter, an embryo autonomous gene and a first terminator.

[0061] According to the present invention, the egg cell-specific promoter can be a variety of conventional promoters capable of specifically promoting the expression of embryo autonomous gene expression, preferably selected from but not limited to AtDD45, Os03g0296600pro, ECA1-like1 pro, DCL2, AT1G74480.1 and ZmEAlpromoter; more preferably AtDD45, the sequence of AtDD45 is preferably shown in SEQ ID NO.2 (SEQ ID NO.1 sequence 1bp-1002bp).

[0062] According to the present invention, the embryo autonomous gene can be a conventiona...

Embodiment 1

[0116] This example is used to illustrate the vector construction of apomictic vectors p72C and p73C

[0117] 1. Construction of MiMe and BABY BOOM-like gene editing vector p72MB

[0118]Design 2 targets in PAIR1, REC8, OSD1 gene coding regions respectively (design the 2 targets shown in SEQ ID NO.10 and SEQ ID NO.11 in the PAIR1 gene coding region, design SEQ ID NO.12 and SEQ ID NO. 2 targets shown in ID NO.13, 2 targets shown in SEQ ID NO.8 and SEQ ID NO.9 designed in OSD1 gene coding region); 2 targets were designed in BBM1 intron region (SEQ ID NO.14 and shown in SEQ ID NO.15); respectively design a target in the coding region of BBM2 and BBM3 gene (design a target shown in SEQ ID NO.16 in the coding region of BBM2 gene, design SEQ ID in the coding region of BBM3 gene 1 target shown in NO.17), and then constructed into the gene editing vector Cas9 vector, named p72MB.

[0119] 2. Construction of MiMe and BABY BOOM-like gene editing vector p73MB

[0120] Design 2 targets...

Embodiment 2

[0138] This example is used to illustrate the acquisition of transgenic plants

[0139] Select a single colony of Agrobacterium containing rice expression vectors p72C and p73C and inoculate them on LB medium containing 50 mg / L kanamycin, culture in dark at 26°C for 2 days, wash the Agrobacterium cells with NB-AS liquid medium, shake at 28°C at 180rpm Cultivate for 90-120min. Adjust the concentration of colony to OD600 of 0.8-1.0, respectively for the transformation of hybrid rice.

[0140] Disinfect the hybrid rice seeds, select the seeds with full grains, soak them in 75% alcohol for 30s, pour off the alcohol, rinse them with sterile water, and wash them with HgCl 2 Disinfect for 8 minutes, wash 2 times with sterile water, soak for 1 minute each time, and soak for 1 hour with sterile water. The sterilized seeds were inoculated on the induction medium and grown under light for 7 days.

[0141] Pool the sterile callus together. Put them into the Agrobacterium suspension co...

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 plant molecular biology and agricultural biology, and particularly relates to a nuclear nucleic acid molecule, a vector, a cell and application thereof and efficient screening of plant apomixis cloned seeds through asexual embryos and embryo-free seeds, the nucleic acid molecule comprises an expression cassette E1, and the expression cassette E1 carries an element for generating MiMe mutants and an element for silencing BABY BOOM-like genes. The screening method comprises the following steps: introducing nucleic acid molecules containing expression cassettes E1, E2 and E3 or recombinant vectors containing the nucleic acid molecules into plants; selfing the obtained transgenic plant to obtain an asexual embryo seed with a selection marker and an embryo-free seed without the selection marker; and the asexual embryo seeds and the non-embryo seeds are sorted through the selection markers, and the asexual embryo seeds serving as clone seeds are obtained. According to the method, the asexual embryo seeds and the non-embryo seeds can be obtained during selfing of the transgenic plant, and the separation of the asexual embryo seeds is facilitated, so that the cloned seeds can be obtained at a high ratio.

Description

technical field [0001] The invention belongs to the fields of plant molecular biology and agricultural biotechnology, and specifically relates to a nucleic acid molecule, a recombinant vector, a transgenic cell, the application of the nucleic acid molecule, the recombinant vector or the transgenic cell, and a cultivation method. A method of transgenic plants, a method of identifying said transgenic plants, an efficient screening method of plant apomictic clone seeds, and a method of screening apomictic lines. Background technique [0002] Crops can fix heterosis through apomixis, which has been called "the holy grail of agriculture". Apomixis is the reproductive mode in which plants produce embryos and seeds without fusion of sperm and egg cells during the process of producing sporophytes from gametophytes. Including haploid apomixis and diploid apomictic two basic types. Because diploid apomixis can produce offspring with exactly the same genotype as the maternal plant an...

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): C12N15/29C12N15/82A01H5/10A01H6/46C12Q1/6895
CPCC07K14/415C12N15/8267C12Q1/6895C12Q2600/158
Inventor 曹孟良夏玉梅余木兰唐宁詹祎捷淡俊豪卜小兰王耀石跃斌
Owner HUNAN HYBRID RICE RES CENT
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