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Nucleic acid molecule, vector, cell, application of nucleic acid molecule, vector and cell, and screening method of plant apomixis clone seeds based on paternal imprinting gene

A nucleic acid molecule and transgenic plant technology, applied in the field of nucleic acid molecules, vectors and cells and their applications and the screening of plant apomictic cloned seeds based on paternally imprinted genes, can solve the problem of inability to sort seeds, low purity requirements, inability to The realization of the application of hybrid apomixis system, etc.

Pending Publication Date: 2021-05-11
HUNAN HYBRID RICE RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the ratio of cloned seeds in this apomictic system is only 29%, far below the 96% purity requirement in production, and cloned seeds and seeds formed by normal pollination and fertilization cannot be sorted
So that it is still impossible to realize the application of hybrid apomixis system

Method used

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  • Nucleic acid molecule, vector, cell, application of nucleic acid molecule, vector and cell, and screening method of plant apomixis clone seeds based on paternal imprinting gene

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Embodiment approach

[0038] According to a preferred embodiment of the present invention, the nucleic acid molecule further comprises an expression cassette E2 carrying an embryo autonomous gene.

[0039] More preferably, the expression cassette E2 includes from upstream to downstream: a second promoter, an embryo autonomous gene and a second terminator, and more preferably, in order to further improve the purity of cloned seeds, the expression cassette E2 includes an egg cell-specific promoter Son, embryo autonomous gene and second terminator.

[0040] 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 ZmEAl promoter; more preferably AtDD45, the sequence of AtDD45 is preferably shown in SEQ ID NO.7.

[0041] According to the present invention, the e...

Embodiment 1

[0094] This example is used to illustrate the acquisition of transgenic plants

[0095] 1. Vector construction of apomictic vector p76C

[0096] 1. Synthetic expression cassettes E1 and E2

[0097] Synthetic expression cassette E1, the sequence is shown in SEQ ID NO.1: including the fusion sequence of the paternal imprinted gene functional regions ICR, DMR2', DMR1 (SEQ ID NO.2), the embryo-specific expression promoter OsESP1 (SEQ ID NO.3 ), red fluorescent protein gene DsRed2 (SEQ ID NO.4), NOS terminator (SEQ ID NO.5).

[0098] The synthetic expression cassette E2, whose sequence is shown in SEQ ID NO.6, includes egg cell-specific promoter AtDD45 (SEQ ID NO.7), embryo autonomous gene selection from BBM1 (SEQ ID NO.8), NOS terminator (SEQ ID NO. .5).

[0099] E1 and E2 were constructed as a linked expression cassette, named 76E.

[0100] 2. Construction of expression vector p76C

[0101] Two targets were designed in the coding regions of PAIR1, REC8, and OSD1 genes (PAI...

Embodiment 2

[0119] This example is used to illustrate the acquisition of transgenic plants

[0120] The transgenic plants were prepared according to the method of Example 1, except that the functional region of the paternally imprinted gene in the expression cassette E1 did not include ICR, DMR2', and DMR1.

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Abstract

The invention belongs to the technical field of plant molecular biology and agricultural biology, and particularly relates to a nucleic acid molecule, a vector, a cell, application of the nucleic acid molecule, the vector and the cell, and a screening method of plant apomixis clone seeds based on a paternal imprinting gene. The nucleic acid molecule comprises an expression cassette E1 based on the imprinting gene, wherein the expression cassette E1 comprises a paternal imprinting gene functional region, a first promoter, a screening marker gene and a first terminator from upstream to downstream. When the nucleic acid molecule provided by the invention is used for screening the apomixis clone seeds, obtained positive transformed plants are self-crossed to generate zygotic embryo seeds with fluorescence and asexual embryo seeds without fluorescence. Through photoelectric sorting, the asexual embryo seeds with fixed heterosis can be efficiently sorted, so that a method for applying a hybrid apomixis system is realized.

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, a cultivation A method for transgenic plants, a method for identifying said transgenic plants, a method for screening seeds of high-purity clones of plants based on paternally imprinted genes, and a method for screening apomictic lines based on paternally imprinted genes. Background technique [0002] In recent years, plant biotechnology has developed rapidly, and the research and improvement of crops by means of molecular biology and genetic engineering have achieved good application prospects in agricultural production. Recently, important progress has been made in the field of apomixis research by means of molecular biology and genetic engineering. At the end of 2018, the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/65C12N15/29A01H5/10A01H6/46A01H1/02A01H1/04
CPCC12N15/8212C12N15/8234C12N15/8233C12N15/8287C07K14/415A01H1/02A01H1/04
Inventor 曹孟良
Owner HUNAN HYBRID RICE RES CENT
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