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African agapanthus flowering gene ApFT protein and coding gene and probe thereof

A probe and gene technology, which is applied in the field of ApFT protein, the gene encoding gene and probe related to the flowering of Agacia japonica, can solve the problems such as

Inactive Publication Date: 2013-09-11
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Before the publication of the present invention, there was no patent report related to the Agapanthus FT protein sequence mentioned in this patent application

Method used

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  • African agapanthus flowering gene ApFT protein and coding gene and probe thereof
  • African agapanthus flowering gene ApFT protein and coding gene and probe thereof
  • African agapanthus flowering gene ApFT protein and coding gene and probe thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 , Cloning of the ApFT gene of Agapanthus

[0037] 1. Acquisition of plant material

[0038] The test materials were 4-year-old potted seedlings of Agapanthus praecox ssp.orientalis. Stem tips and leaves in the vegetative growth, flower bud induction, and flower bud differentiation stages were collected for RNA extraction and real-time fluorescence quantitative qRT-PCR analysis.

[0039] 2. Extraction of RNA

[0040] Total RNA was extracted with the "Plant RNA Small Extraction Kit", which was purchased from Shanghai Laifeng Biotechnology Co., Ltd. The integrity of RNA was identified by 1% agarose gel electrophoresis, and then the purity and concentration of RNA were determined on a spectrophotometer (Thermo Scientific NANODROP1000 Spectrophotometer).

[0041] 3. Full-length cloning of genes

[0042] According to the results of transcriptome sequencing, the core fragment with high homology to FT of the flowering-related gene of Agapanthus was obtained, and ...

Embodiment 2

[0056] Example 2 , Sequence information and homology analysis of the ApFT gene of Agapanthus chinensis

[0057] The full-length open reading frame sequence of the new Agapanthus abaciens ApFT gene of the present invention is 534bp, and the detailed sequence is shown in the sequence shown in SEQ ID NO.3. According to the sequence of the open reading frame, the amino acid sequence of Agapanthus ApFT was deduced, with a total of 177 amino acid residues, a molecular weight of 19490 Daltons, and an isoelectric point (pI) of 8.67. For the detailed sequence, see the sequence shown in SEQ ID NO.4;

[0058] The open reading frame sequence of Agapanthus ApFT and the amino acid sequence of its encoded protein were nucleated in Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations+PDB+SwissProt+Superdate+PIR databases using BLAST program Nucleic acid and protein homology search, it was found that it has 65.51% similarity with Satsuma citrus CuMFT1 gene (GenBank...

Embodiment 3

[0059] Example 3 、 Expression differences of ApFT gene in different tissues at different developmental stages of Agapanthus

[0060]1. Material acquisition: collect leaves and stem tips of Agapanthus vegetative growth period, flower bud induction period, and flower bud differentiation stage respectively, wrap the samples in aluminum platinum paper, put them into liquid nitrogen immediately, and then put them in -80°C Store in ultra-low temperature refrigerator until use.

[0061] 2. Extraction of RNA: Use the "Plant RNA Small Extraction Kit" (Shanghai Laifeng Biotechnology Co., Ltd.) to extract RNA from leaves and shoot tips of Agapanthus at different developmental stages.

[0062] 3. Determination of the integrity, purity and concentration of RNA: Use ordinary agarose gel electrophoresis (gel concentration 1%; 1×TAE electrophoresis buffer; 120v, 30min) to check the integrity. The maximum rRNA brightness in the electrophoresis strip should be 1.5-2.0 times the brightness of...

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Abstract

The invention relates to an African agapanthus flowering gene ApFT protein and coding gene and probe thereof, and the protein is the protein (a) or (b): (a) protein composed of the amino acid sequence shown in the SEQ ID NO.4; (b) protein derived from protein (a) which prepared by replacing, deleting or adding one or more amino acids of the amino acid sequence shown in the SEQ ID NO.4 and has the African agapanthus ApFT activity. The invention also provides a nucleic acid sequence coding the protein above and a probe for detecting the nucleic acid sequence; the spatio-temporal expression characteristics of the African agapanthus flowering related gene ApFT can be regulated by using the gene engineering technology, thereby providing a theory basis for changing the florescence regulation and control technology with high application value.

Description

technical field [0001] The present invention relates to the key gene FLOWERING LOCUS T (FT) in the developmental pathway of Agapanthus agapanthus and its encoding gene and probe, in particular to an Apania flowering related gene ApFT protein and its encoding gene and probe. Background technique [0002] Flowering is the transition of plants from vegetative to reproductive growth. The FLOWERING LOCUS T (FT) gene is the convergence point of the flower development pathway, which can integrate signals from different flower development pathways such as photoperiod pathway, vernalization pathway and autonomous pathway, and plays an important role in the process of plant flower development. The transcript levels of FT exhibited a circadian rhythm under long-day conditions. The latest research shows that the FT protein is the long-distance transport florigen signaling molecule, which moves from the leaf to the stem end through the phloem. In the shoot apical meristem, FT protein i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12Q1/68C12N15/11
Inventor 石玉波王玲卓丽环
Owner NORTHEAST FORESTRY UNIVERSITY
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