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Sucrose transporter and its application in regulating male sterility in plants
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A male sterility and plant technology, applied in the fields of application, plant peptides, plant products, etc., can solve the problems of complex metabolic transport mechanism and achieve the effect of improving breeding efficiency
Active Publication Date: 2020-01-07
CHINA AGRI UNIV
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After sugar reaches the sink organ, it will be unloaded in a variety of ways. The process of decomposition or retransport requires the participation of various transport proteins and enzymes, and its metabolic transport mechanism is quite complicated.
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Embodiment 1
[0054] Embodiment 1, the acquisition of CsSUT1 protein and its coding gene
[0055] Sequence analysis, section interception and functional verification were carried out on the genomes of various cucumber varieties, and a sucrose transporter protein was found from Xintaimici cucumber, which was named CsSUT1 protein, as shown in sequence 1 of the sequence list. The gene encoding CsSUT1 protein is named CsSUT1 gene, as shown in sequence 2 of the sequence listing.
Embodiment 2
[0056] Example 2, analysis of CsSUT1 gene expression in different tissue parts of cucumber
[0057] The root (R), stem (S), young leaf (YL), mature leaf (ML), male flower (MF), female flower (FF) and fruit (F) of Xintaimithorn cucumber grown to the early melon stage were collected respectively .
[0058] The total RNA of the above materials was extracted, and the first-strand cDNA was synthesized, and the expression of CsSUT1 gene was detected by qRT-PCR (Tubllin gene was used as an internal reference gene), and the primer pair composed of primer YSUT1-F and primer YSUT1-R was used to detect For the expression of CsSUT1 gene, a primer pair consisting of primer TUB-F and primer TUB-R was used to detect the expression of Tubllin gene.
[0059] YSUT1-F: 5'-CGTGGTTACAAAGGTTGCTGAG-3';
[0060] YSUT1-R: 5'-GCGGATACGATGAACTGTGGA-3';
[0061] TUB-F: 5'-ACGCTGTTGGTGGTGGTAC-3';
[0062] TUB-R: 5'-AGAGGGGTAAACAGTGAATC-3'.
[0063] The result is as figure 1 shown. The results show...
Embodiment 3
[0064] Embodiment 3, the acquisition of transgenic plants
[0065] 1. Construction of interference carrier
[0066] 1. Extract the total RNA of Xintaimi thorn cucumber, and reverse transcribe it into cDNA, use the cDNA as a template, use the primer pair composed of CsSUT1-RNAi-1F and CsSUT1-RNAi-1R to perform PCR amplification, and recover the PCR amplification product .
[0069] In CsSUT1-RNAi-1F and CsSUT1-RNAi-1R, the underline marks the restriction sites of AscI and SwaI, respectively.
[0070] 2. Digest the PCR amplified product of step 1 with restriction endonucleases AscI and SwaI, and recover the digested product.
[0071] 3. The vector pFGC1008 was double-digested with restriction enzymes AscI and SwaI, and the vector backbone of about 10907 bp was recovered.
[0072] 4. Ligate the digested product of step 2 with the vector backbone ...
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Abstract
The invention discloses a sucrose transporter and application thereof in controlling plant male sterility. The invention provides a protein which is obtained from cucumber and named CsSUT1 protein and is the following (a1) or (a2): (a1) a protein consisting of an amino acid sequence shown by the sequence 1 in a sequence table; or (a2) a protein derived from the sequence 1, related to the plant male fertility and obtained by substituting and / or deleting and / or adding one or more amino acid residues of the amino acid sequence shown by the sequence 1 in the sequence table. A coding gene (CsSUT1 gene) of the protein is also protected by the invention. Experiments prove that down regulation of the expression quantity of the CsSUT1 gene may lead to male sterility of a cucumber plant. The sucrose transporter may be applied to cucumber breeding and plays a positive role in increasing the breeding efficiency.
Description
technical field [0001] The invention relates to a sucrose transporter and its application in regulating male sterility of plants. Background technique [0002] Cucumber belongs to Cucurbitaceae Melon, is an important horticultural crop, widely cultivated all over the world, and is one of the main vegetable crops in my country. Sugar is the main product of plant photosynthesis, the main form of long-distance transportation and storage of plants, and an important signal molecule that coordinates the relationship between plant source and sink. The sugar produced by the leaves is transported through the phloem for a long distance, and then reaches and unloaded in sink organs such as flowers, fruits, and seeds. For most plants, sucrose is the main transport form of their photosynthetic products, and a few crops use the raffinose family series of oligosaccharides as the main transport form. After sugar reaches the sink organ, it will be unloaded in a variety of ways. The process...
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