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Breeding method of brassica napus radish cytoplasm restorer line

A technology of Brassica napus and restorer lines, applied in the fields of botany equipment and methods, application, plant gene improvement, etc., can solve the problems of backward chromosome bridges, poor self-seed setting rate, etc., achieve short cycle, strong recovery ability, good affinity effect

Active Publication Date: 2021-06-25
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chen Weijiang et al. used the special material radish-blue (AACCRR, 2n=58) and Brassica napus through grafting to breed a homozygous restorer line CLR650. Because the imported radish chromosomes are large, the restorer line still suffers from chromosomal lag. Meiotic abnormalities such as chromosome bridges and chromosome bridges, and poor self-seed setting rate (Chen Weijiang et al., 2012)

Method used

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  • Breeding method of brassica napus radish cytoplasm restorer line
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  • Breeding method of brassica napus radish cytoplasm restorer line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025]Example 1 Using distant hybridization combined with backcrossing to transfer the radish cytoplasmic restoration gene from Brassica napus into Brassica napus

[0026] (1) October 2013-May 2014: The radish cytoplasmic restorer line RLM198 (referred to as RLM198 in mustard type rapeseed) is a radish cytoplasmic restorer line improved by Indian scientist Professor Prakash, and has been published (Kirti et al. .,1995), using it as the female parent. Brassica napus Yu7-120 is derived from a recurrent improved population of Brassica napus, which has also been published (Zhao et al., 2016), and it is used as the male parent. The Brassica napus RLM198 and Brassica napus Yu7-120 are preserved in the Rapeseed Research Laboratory of Huazhong Agricultural University, and promise to release the biological materials to the public for experimental research within 20 years from the date of application. The male parent (Brassica napus Yu7-120) and the female parent (Brassica napus RLM198...

Embodiment 2

[0032] Example 2 Restoration of Gene Linkage Marker Validation

[0033] Brassica napus radish sterile cytoplasmic restorer line R2000 is an excellent restorer line improved by the French Academy of Agricultural Sciences through radiation mutagenesis. At the same time, a series of linked genetic markers (US008766040B2) were developed for scanning the radish restoration gene segment. The linkage markers of the restorer lines were divided into 5 groups, and the loci controlling the glucosinolate content were in the 4th group. Therefore, using this set of markers, the size of the radish segments among the restorer lines could be scanned and compared.

[0034] Compare the restorer line CLR650 created by Mr. Chen Weijiang of Hunan Academy of Agricultural Sciences, the restorer line R2000 created by French Academy of Agricultural Sciences, the restorer line 206R of Brassica napus and the restorer line BJO of mustard type rape created in the present invention, and Huayouza 62 ( HYZ62...

Embodiment 3

[0041] Example 3 constructs F 2 Population, Genetic Mapping of Restoration Genes Using Rapeseed 60K SNP Chip and Mixed Pool Resequencing

[0042] A F 2 For the fertility segregation population, appropriate amount of young leaves were taken from fertile and sterile individual plants, and DNA was extracted by the improved CTAB method. Precipitate DNA with 2.5 times of ice ethanol. After half an hour in the refrigerator, use a 100ul pipette tip to pick out the precipitated DNA into 1ml of 70% alcohol to ensure the purity of the DNA. After RNase digestion and quality inspection, the extracted DNA was handed over to Shanghai Passino Biotechnology Co., Ltd. for Illumina second-generation library construction, and sequenced on a Hiseq X-TEN sequencer. An F hybrid constructed from Brassica napus 206R and Brassica napus Zhongshuang11 2 The population adopts the same method of library construction and sequencing, and is sequenced on the Hiseq2500 sequencer.

[0043] to two F's 2 Th...

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Abstract

The invention discloses a breeding method of a brassica napus radish cytoplasm restoring line. The breeding method comprises the following steps: by taking a mustard type rape radish cytoplasm restorer line RLM198 as a female parent and brassica napus Yu7-120 as a male parent, introducing a restoring gene of mustard type rape cytoplasm into a brassica napus genome through continuous backcross, and conducting selfing to obtain a homozygous stable restorer line. With adoption of genetic linkage and whole genome re-sequencing analysis method, it is proved that the radish section of the obtained restorer line is short and located at the tail end of the A10 chromosome, and the obtained novel restorer line is sufficient in pollen amount, good in restoring rate, normal in seed setting and free of linkage burdens and can be directly applied to production of rape hybrids in China.

Description

technical field [0001] The invention belongs to the field of rapeseed breeding, and in particular relates to a method for breeding a new type of rapeseed radish cytoplasm restorer line. Background technique [0002] Cytoplasmic male sterility is the main way to utilize crop heterosis, and it is used in the hybrid production of various crops. In terms of molecular mechanism, cytoplasmic male sterility is the result of the joint action of nucleus and cytoplasm, which is manifested as: some specific open reading frames (Open reading frames, ORFs) on the mitochondrial genome are expressed in large quantities, and are reversed during pollen development. The regulatory pathway leads to pollen abortion, while the corresponding restorer gene (Restoreof fertility, Rf) on the nucleus genome acts positively on the ORFs, causing them to degrade or fail to transcribe or translate normally, thereby restoring fertility (Chen and Liu, 2014). [0003] Rapeseed is one of the most important ...

Claims

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Application Information

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IPC IPC(8): A01H1/02A01H1/04
CPCA01H1/02A01H1/04
Inventor 涂金星吴增祥
Owner HUAZHONG AGRI UNIV
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