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Establishment and application of new maintainer lines and sterile lines of plants

A sterile line, plant cell technology, applied in the fertility recovery of homozygous recessive GM male sterile plants, the field of transgenic plants, can solve the problems of high cost and complex technology, achieve stable fertility, eliminate concerns, and ensure The effect of purity

Active Publication Date: 2017-06-23
SHENZHEN INST OF MOLECULAR CROP DESIGN +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Focusing on how to improve the stability of the sterile line and break the limitations of the resource utilization of hybrid varieties, and further solve the technical bottleneck problems of complex and high-cost existing seed production technology, we have established a new hybrid breeding technology

Method used

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  • Establishment and application of new maintainer lines and sterile lines of plants
  • Establishment and application of new maintainer lines and sterile lines of plants
  • Establishment and application of new maintainer lines and sterile lines of plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Embodiment 1: Construction of rapeseed expression vector pBnSI:

[0065] Before constructing the plant expression vector of rapeseed, the inventors first transformed rapeseed separately for each expression cassette in the expression vector, and further verified the function of each expression cassette. The results show that each expression cassette can work well when transforming rape alone, and achieve the expected design effect. Further inventors constructed the following expression vectors. For the herbicide resistance gene, refer to Chinese patent 201310116228.2, and the present invention is incorporated into this patent by reference.

[0066] By assembling the individual elements of the BrSZ1, Pr2, BrMS2 and Pr4 expression cassettes, constructs such as figure 1 The indicated pBnSI vectors. The vector contains 4 expression cassettes, wherein the first expression cassette is the expression cassette of barnase, which is composed of the pollen special expression pro...

Embodiment 2

[0068] Embodiment 2: Rapeseed Transformation

[0069] The plasmid pBnSI was transferred into Agrobacterium EHA105 strain by electric shock method, and rapeseed containing homozygous brms2 and brms1 recessive sterility loci was genetically transformed by Agrobacterium-mediated dipping method, and 22 single-copy transgenic plants were obtained Material. The rape transformation recipient material containing homozygous brms2 and brms1 recessive sterility sites is Brassica napus.

Embodiment 3

[0070] Example 3: Detection of Pollen Fertility of Transgenic Rapeseed Plants

[0071] Analysis of 22 single-copy transgenic rapeseed plants (containing homozygous brms2 and brms1 recessive sterility loci) plants obtained in Example 2 found that there was no obvious morphological difference between the transgenic plants and the non-transgenic control plants, but the pollen Fertility is markedly different.

[0072] The transgenic plant material obtained by transforming rapeseed with pBnSI construct was tested for pollen dyeability rate, and wild type rapeseed was tested for pollen dyeability rate ( figure 2 ).

[0073] The method adopted is: during the flowering stage of rapeseed, a single plant is randomly selected from the transgenic rapeseed plant and its wild-type control plant, one flower is taken from each plant, and one anther is taken from each flower, placed in the center of the glass slide, dripped Add a drop of 1% Alexander solution, release the pollen with tweeze...

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Abstract

The present invention provides a plant maintainer line, a sterile line and the use thereof. The fertility restoration gene, the pollen lethal gene and the herbicide resistance gene are simultaneously transferred into a homozygous recessive male sterile plant. Two types of seeds are produced after mating: sterile line seeds without the transgene, without herbicide resistance, can be screened out by herbicides; Line pollination crosses are used for the reproduction of sterile lines. The sterile line created by the invention has stable fertility and is not affected by the environment, and the plant recessive sterility gene is applicable to most plant varieties.

Description

technical field [0001] The invention relates to the fields of plant molecular biology and breeding. In particular, embodiments of the invention relate to transgenic plants comprising seed production techniques. More specifically, the present invention relates to the restoration of the fertility of homozygous recessive male sterile plants and its use. Further, the present invention relates to a method for constructing a plant maintainer and propagating a male sterile line, more specifically, the present invention relates to a construct, a plant cell, tissue or organ, a method for propagating a plant male sterile line, a A method for restoring the male fertility of plant sterile plants, a method for preparing plant seeds, a method for preparing hybrid plants, and the use of plant male sterile lines in preparing hybrid rice. Background technique [0002] Hybrid breeding is the main way of breeding new varieties and the most important method of modern breeding. The creation, u...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H1/02C12N15/65
CPCC12N15/8287A01H1/023
Inventor 唐晓艳周君莉
Owner SHENZHEN INST OF MOLECULAR CROP DESIGN
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