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
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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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