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Overexpressing brassica napus and application of MYB43 of parental species of overexpressing brassica napus in improving plant type and increasing yield

A Brassica napus overexpression technology, applied in the field of genetic engineering, can solve the problem of lack of functional research on transgenic plants, and achieve the effects of enhanced lodging resistance and increased grain yield

Active Publication Date: 2019-10-01
SOUTHWEST UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For the MYB43 gene of cruciferous plants including Arabidopsis thaliana, rape, Chinese cabbage, and cabbage, there is no report on the function of its transgenic plants

Method used

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  • Overexpressing brassica napus and application of MYB43 of parental species of overexpressing brassica napus in improving plant type and increasing yield
  • Overexpressing brassica napus and application of MYB43 of parental species of overexpressing brassica napus in improving plant type and increasing yield
  • Overexpressing brassica napus and application of MYB43 of parental species of overexpressing brassica napus in improving plant type and increasing yield

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1, cloning of Brassica napus, Chinese cabbage, Brassica oleracea MYB43 gene family

[0037] The primers used in this study are listed in Table 1.

[0038] Table 1 Primers used in this study

[0039]

[0040]

[0041] 1. Extraction of total DNA and total RNA from Brassica napus, Chinese cabbage, and Brassica oleracea

[0042] Take the young leaves of typical plants for each strain, extract the total genomic DNA by cetyltrimethylammonium bromide (CTAB) method, and evaluate the quality and concentration of nucleic acid samples by 1.0% agarose gel electrophoresis and spectrophotometry . At the same time, using the roots, stems, leaves, flowers, seeds 30 days after flowering, and pod skins 20 days after flowering of Brassica napus variety ZY821 as materials, total RNA was extracted using a column-type small amount plant total RNA extraction kit according to the instructions. DNA impurities were removed with DNase I, and the total RNA was ethanol-precipita...

Embodiment 2

[0052] Example 2, Bioinformatics analysis of Brassica napus, Chinese cabbage, Brassica oleracea MYB43 gene family

[0053] Perform sequence alignment, open reading frame (ORF) search and translation on Vector NTI Advance 11.5, perform BLAST and protein sequence CDD searches on the http: / / www.ncbi.nlm.nih.gov / website, and http: Protein structure analysis is performed on bioinformatics sites with links at http: / / bip.weizmann.ac.il / and www.expasy.org at http: / / prodes.toulouse.inra.fr / multalin / multalin.html and Multiple alignment and cluster analysis of gene and protein sequences are performed on http: / / www.ebi.ac.uk / clustalw / and other websites.

[0054] 1. Structural analysis of the MYB43 gene family in Brassica napus, Chinese cabbage and Brassica oleracea

[0055](1) Structural parameters of the MYB43 gene family in Brassica napus, Chinese cabbage and Brassica oleracea

[0056] The DNA sequences of the 8 full-length genes of Brassica napus, Chinese cabbage, and Brassica ole...

Embodiment 3

[0079] Example 3, Brassica napus, Chinese cabbage, cabbage MYB43 family organ expression characteristics

[0080] The RNA samples of 6 tissues and organs, including roots, stems, leaves, flowers, seeds 30 days after flowering, and pod skins 20 days after flowering, of Brassica napus, Chinese cabbage, and typical black seed materials of Brassica oleracea were selected as templates, and PrimeScript RT Reagent Kit with gDNA was used as templates. Eraser reverse transcribed the total cDNA, and analyzed the expression organization characteristics of MYB43 gene family by fluorescent real-time quantitative PCR (qRT-PCR). The overall expression level of MYB43 gene in each organ was detected with primer combination FBMYB43RT+RBMYB43RT. Primer combinations FBrMYB43-1RT+RBoMYB43-1RT, FBoMYB43-1RT+RBoMYB43-1RT, FBrMYB43-2RT+RBrMYB43-2RT and FBoMYB43-2RT+RBoMYB43-2RT were used to detect horizontal gene members BnMYB43-1, BnMYB43-2, BnMYB43- 3. Expression levels of BnMYB43-4, BnMYB43-1, Bn...

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Abstract

The invention discloses an overexpressing brassica napus and application of MYB43 of parental species of the overexpressing brassica napus in improving plant type and increasing yield. The gene familyof the MYB43 of brassica napus comprises BnMYB43-1, BnMYB43-2, BnMYB43-3 and BnMYB43-4 member genes; the parental specie brassica rapa has BrMYB43-1 and BrMYB43-2 member genes; the parental specie brassica oleracea has BoMYB43-1 and BoMYB43-2 member genes. After overexpression in plants, the roots of the plants become more developed and longer, the number of branch roots is more, the ratio of root to crown is increased, the plants are slightly higher, the stems become thicker, the number of effective branches per plant is increased, the number of effective pods per plant is increased, the yield of seeds per plant is increased, and the folding resistance and lodging resistance are also improved significantly. According to the overexpressing brassica napus and the application of the MYB43 of the parental species of the overexpressing brassica napus in improving the plant type and increasing the yield, a new method is provided for application of the MYB43 in improving the plant type, lodging resistance and seed yield of brassica napus, brassica rapa, brassica oleracea and other plants.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the application of overexpressing MYB43 of Brassica napus and its parent species in improving plant type and increasing yield. Background technique [0002] Plant architecture is an important trait of plants and is the result of stem development. Since Donald (1968) proposed the "ideal plant type" crop breeding model, plant type has attracted the attention of researchers. A reasonable plant type can increase the leaf area index, photosynthetic efficiency and crop fertilizer tolerance, and increase the harvest index, thereby increasing the crop yield factor and grain yield. Therefore, the genetic improvement of crop plant type is the fundamental guarantee for high crop yield. Plant height, stem diameter, effective branch number, root length, branch root number, root-to-shoot ratio and other traits are the main constituent traits of plant type, and there are extremely ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/8261
Inventor 柴友荣蒋佳怡陆奇丰谭莉廖雪莉尹能文张琳杜有杰
Owner SOUTHWEST UNIVERSITY
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