AsFUL gene and protein capable of accelerating plant to bloom, and application of AsFUL gene and protein
A protein and gene technology, applied in the field of protein and AsFUL genes that promote flowering of plants, can solve problems such as deficiencies, and achieve the effects of promoting flowering, improving sexual reproduction, and obvious theoretical research and economic application value
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Embodiment 1
[0048] Cloning of embodiment 1 garlic AsFUL gene
[0049]1 Experimental selection
[0050] This example uses 'Acheng Zipi' garlic as the material, provided by Harbin Changri Onion Research Institute. In view of the fact that low temperature storage before planting can regulate flowering and fertility recovery, the garlic cloves for seed were stored in a cold storage at 4°C, and after four months, they were planted in the solar greenhouse of the Institute of Flower Bioengineering, Northeast Forestry University. When the garlic grows to the three-leaf stage, use a sterile blade to collect the root, pseudostem (above the root and below the leaf), and young leaves respectively; when the plant grows 6 to 7 leaves, collect the protective leaves at the bulb formation stage, flower buds at the first stage, Two weeks later, the second phase of flower buds, the base of the flower stem wrapped by the pseudostem, and the third phase of flower buds after the garlic was completely bolted w...
Embodiment 2
[0064] Example 2 Sequence characteristics and phylogenetic analysis of garlic AsFUL gene
[0065] 1 Sequence analysis of garlic AsFUL gene
[0066] Using bioinformatics software to analyze the sequence characteristics and predict the protein function of the garlic AsFUL gene, to provide a theoretical basis for the subsequent construction of gene function mutants and genetic transformation experiments to verify gene function.
[0067] 2 Sequence analysis and structure prediction of AsFUL gene
[0068] Perform blastn (nucleotide query vs. nucleotide database) and blastp (protein query vs. protein database) analysis; use ORF finder (https: / / www.ncbi.nlm.nih.gov / orffinder / ) online analysis software to analyze the open reading frame of the gene and deduce its amino acid sequence, and submit the deduced amino acid sequence to NCBI Conserved Domains (CDD) database (http: / / www.ncbi.nlm.nih.gov / Structure / cdd / wrpsb.cgi) was used to predict the conserved domains.
[0069] Use Bioedit ...
Embodiment 3
[0073] Example 3 Expression analysis of garlic AsFUL gene
[0074] In order to analyze the spatial expression specificity of the garlic AsFUL gene in different organs of garlic, the inventors used the garlic roots, pseudostems, young leaves, early flower buds (F1), middle flower buds (F2), late flower buds (F3), bulbs, and flower stems. Total RNA was used as a template for gene-specific semi-quantitative RT-PCR and real-time fluorescent quantitative PCR detection.
[0075] 1 Experimental materials
[0076] The garlic cultivated in the greenhouse of the Institute of Flower Bioengineering, School of Life Sciences, Northeast Forestry University was used as the experimental material. The tissues and organs of the garlic were collected with RNase-free tweezers and blades, and were quickly frozen in liquid nitrogen and stored in a -80°C refrigerator for later use.
[0077] 2 Experimental Reagents and Instruments
[0078] TransZol Plant kit was purchased from TiansGen Company; chlo...
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