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Method for creating cytoplasmic male sterile line by utilizing cotton transgenosis

A male sterile line, male sterile technology, applied to other methods of inserting foreign genetic materials, genetic engineering, plant genetic improvement, etc.

Active Publication Date: 2017-05-31
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In this regard, there are no reports at home and abroad

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  • Method for creating cytoplasmic male sterile line by utilizing cotton transgenosis
  • Method for creating cytoplasmic male sterile line by utilizing cotton transgenosis
  • Method for creating cytoplasmic male sterile line by utilizing cotton transgenosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Construction of Overexpression Vector PBI121-GhbZIP1-eGFP

[0067] 1. Cloning of GhbZIP1 gene cDNA

[0068] By analyzing the RNA-seq data of "cave A", and using this sequence as an intermediate fragment, specific primers were designed (primers were designed using Primer Premier 5.0 software), and the GhbZIP1 gene cDNA was cloned by combining RACE technology and hi-TAIL PCR technology full-length sequence.

[0069] Using RACE technology, synthesized reverse transcription primer AUAP (all primers are listed in Table 1), anchor primer AP, synthesized and amplified 5′ specific primers: SHP1 and SHP2, and adopted nested PCR and landing PCR technology, after two rounds of amplification to excavate The bands that meet the requirements were cloned, sequenced, and spliced ​​with the intermediate fragments to obtain a sequence with a length of 194 bp, which was repeated 81 bases with the intermediate sequence of GhbZIP1, and the length of the newly added sequence was 1...

Embodiment 2

[0079] Breeding of Example 2 Cotton Cytoplasmic Male Sterile Line

[0080] In February 2013, at the base of China Cotton Research Institute in Yacheng, Hainan, G-4 (a hybrid of "Zhongmian 16" and "King Kong King" 5 The "Zhongmian 16" was selected from the Cotton Research Institute of the Chinese Academy of Agricultural Sciences, and the "King Kong King" was purchased from the Hubei Seed Company) as the material. The recombinant expression vector PBI121-GhbZIP1-eGFP constructed in Example 1 was injected into the ovary of the test material.

[0081] In July 2013, T sown in Nanning 1 Thirty-seven male sterile strains were found in the generation population, named G-4S. Molecular identification showed that the target gene existed in G-4S plants. So far, male sterile germplasm of cotton transgenic with GhbZIP1 gene has been obtained. Then, using the male sterile germplasm resource as the female parent, the wild-type non-transgenic G-4 and its non-homologous upland cotton variet...

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Abstract

The invention provides a method for creating a cytoplasmic male sterile line by utilizing cotton transgenosis. The method comprises the following steps: taking GhbZIP1 found in upland cotton as a target gene; adopting a transgenosis method for acquiring a cytoplasmic male sterility germplasm; and performing nuclear replacement backcross with homologous or heterological maintainer line or variety (line), thereby breeding a homologous or heterological nucleation male sterile line. The cell nucleus of the male sterile line acquired according to the invention is sourced from a non-transgenosis maintainer line or variety (line), so that a transgenosis-free plasmid vector is as same as a transgenosis plant and is free from safety risk and market risk; the cytoplasm of the acquired male sterile line is sourced from cultivated variety (line), so that the composition of high-advantage cross combination is benefited; various cytoplasmic male sterile germplasms can be created, so that the potential risk in pandemic influenza of a certain disease possibly caused by the wide popularization and long-term use of single cytoplasm coeno-species in the agricultural production can be avoided; the application prospect is ultra-wide.

Description

technical field [0001] The invention relates to a breeding method, in particular to a method for creating a cytoplasmic male sterile line by transfecting cotton with GhbZIP1 gene. Background technique [0002] Cotton, Malvaceae, is one of the most important crops in the world. It is not only the most important fiber crop, but also an important oil crop, a high-protein food crop, textile, fine chemical raw material and an important strategic material. Cotton exhibits strong heterosis, and its heterosis rate is as high as 20%-30%. For a long time, the utilization of cotton heterosis has been an important content of cotton breeding at home and abroad. [0003] Cotton can use both nuclear male sterile lines and cytoplasmic male sterile lines to produce hybrid seeds. Although the use of nuclear male sterile lines is easy to find restorer lines and assemble strong dominant hybrid combinations, there are problems in the reproduction of sterile lines. Generally, male sterile stra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/89A01H1/02A01H1/04
CPCA01H1/02A01H1/04C12N15/8289C12N15/89
Inventor 周瑞阳李敏
Owner GUANGXI UNIV
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