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Breeding method of male sterile cytoplasm NAU2A of rapeseed

A technology of male sterility and male sterility line, applied in the fields of genetic engineering and plant breeding, can solve the problems of unstable fertility, single sterile cytoplasm, cytoplasmic male sterile cytoplasm, etc., and achieve a wide range of applications.

Inactive Publication Date: 2010-02-17
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the utilization of heterosis in rapeseed and Brassicaceae, there are problems such as single sterile cytoplasm and unstable fertility. The cytoplasmic male sterility of the present invention provides a new means for solving the problem of single cytoplasm, and can be used for heterosis in rapeseed and Brassicaceae. use

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Protoplasts were extracted from leaves of Descurainian sophia and cotyledons of Brassica napus, PEG-high pH, ​​high calcium plus DMSO fusion method, and cultured in a shallow liquid layer to study the effects of Descurainian sophia and Brassica napus 19 regenerated plants were obtained by protoplast fusion factors of Brassica napus. The results showed that: 4% cellulase + 0.5% isolated enzyme + 5 mmol / L MES enzymatic hydrolysis for 10 hours and 1% cellulase + 0.2% isolated enzyme + 3 mmol / L MES enzymatic hydrolysis for 14 hours were suitable for obtaining high-yield Artemisia annua and rape respectively Protoplasts; 30% PEG+0.3mol / L glucose+50mmol / L CaCl2 2H2O+15%DMSO obtained a 10% fusion rate; the protoplast culture effect of the improved B5 medium was better; different fresh cultures were added each time The addition of MES has a good effect on preventing browning; the suitable differentiation medium is MS+4.0mg / L 6-BA+0.3mg / L NAA+5.0mg / L AgNO3; the suitable rooting ...

Embodiment 2

[0043] Sterile plants appeared in fusion hybrids obtained by using protoplast fusion technology. The sterile line was used to cross with NAU2B, and the completely sterile line was further selected from cross F1 to backcross with NAU2A. After 5 generations of backcross, the stable sterile line NAU2A was obtained. The sterility of NAU2A showed stable sterility between years. Through the test cross, the restorer line has been found, and the three-line matching has been realized, which provides a new pollination control system for the utilization of heterosis. The study pioneered the cytoplasmic method, and through this method, cytoplasmic recombination was found from the fusion hybrid to the first generation of backcrossing; the study of gene cloning showed that the cytoplasmic mtDNA remained stable after the first generation of backcrossing, and no recombination occurred.

[0044] NAU2A is one of a series of recombinant cytoplasms obtained by protoplast fusion. In the present ...

Embodiment 3

[0046] Using the NAU2A sterile cytoplasm to cross the Chinese cabbage (Brassica napus) variety "Shanghaiqing", the F1 generation is still sterile, but after the NAP-type cytoplasmic Brassica napus is crossed with Shanghaiqing, the pollen amount of the F1 is basically normal, indicating that NAU2A sterile cytoplasm can be transferred to close relatives of rapeseed, providing a new method for the utilization of heterosis.

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Abstract

The invention discloses a creating method and characteristics of male sterile cytoplasm NAU2A of Brassica napus, belonging to the fields of genetic engineering and plant breeding. The protoplast technology of Brassica napus is employed to obtain a blending progeny, and a sterile plant is found in the blending progeny. An NAU2B maintainer line is crossed and backcrossed with the sterile plant to obtain a stable sterile line NAU2A. Mitochondrial DNA of the sterile line NAU2A is extracted and the PCR technology is employed to clone and identify the sterile cytoplasm to obtain a characteristic sequence SEQ ID NO.1 and an amino acid sequence SEQ ID NO.2 of the mitochondrial DNA of the sterile line NAU2A. The cytoplasm and the characteristic gene sequence of the invention are reported for the first time. The NAU2A cytoplasm can be widely applied to heterosis utilization of cruciferae and has good application prospect.

Description

technical field [0001] The invention discloses a Brassica napus cytoplasm and its characteristic mitochondrial DNA sequence, belonging to the fields of genetic engineering and plant breeding. Specifically, it involves the utilization of heterosis in cruciferous plants, and provides a new type of sterile cytoplasmic cytoplasm for the breeding of cruciferous plant hybrids. technical background [0002] Cytoplasmic male sterility can be applied to the production of hybrids, which has huge benefits and has been generally recognized by people. The cytoplasmic male sterile line is used to configure hybrids, which can increase the yield of rapeseed by more than 15%, and can also significantly increase the yield of cruciferous vegetables. [0003] At present, in rapeseed and even cruciferous plants, the main applications of cytoplasmic male sterility are: polimaCMS, Shaanxi 2A, Ogu CMS (see: Liu Pingwu et al., Molecular markers of pol CMS fertility restoration genes in Brassica nap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H5/00C12N15/05A01H1/02C12N15/29C07K14/415
Inventor 管荣展姜淑慧田琳琳张红生杜同庆陈健美郑秀张丽君张凤启
Owner NANJING AGRICULTURAL UNIVERSITY
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