Application of hexose transporter gene CsSWEET1a of camellia sinensis

A transporter and transgene technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of induction and function that have not been reported, and achieve the effect of enhancing low temperature resistance and good application value

Active Publication Date: 2020-02-07
TEA RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The CsSWEET1a gene sequence involved in the present invention has been published in NCBI (accession number: KP025702). Studies have shown that the expression of CsSWEET1a is significantly induced by low temperature stress, but its function has not been reported

Method used

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  • Application of hexose transporter gene CsSWEET1a of camellia sinensis
  • Application of hexose transporter gene CsSWEET1a of camellia sinensis
  • Application of hexose transporter gene CsSWEET1a of camellia sinensis

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Experimental program
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Effect test

Embodiment 1

[0024] Embodiment 1: the overexpression vector construction of tea tree CsSWEET1a gene

[0025] According to the CsSWEET1a sequence disclosed in the NCBI database, its nucleotide sequence is shown in SEQ ID NO: 1, and the amino acid sequence of its encoded protein is shown in SEQ ID NO: 2, primers F and R were designed, and the tea tree leaf cDNA was used as a template , the gene CsSWEET1a was amplified by PCR. The amplified sequence was connected to the pEASY bluntzero vector, and finally connected to the Super promoter-pCambia1300 plant expression vector through a double enzyme digestion ligation reaction.

[0026] The primers used were F: 5'- CCCAAGCTT ATGGGTAATACTGCGCATTTCG-3', shown in SEQ ID NO: 3;

[0027] R: 5'- GCGTCGAC CTACTTGCTCGATCGCTTCTCT-3', shown in SEQ ID NO: 4.

Embodiment 2

[0028] Embodiment 2: Construction and screening of CsSWEET1a gene overexpression plant

[0029] The sequence-correct vector constructed in Example 1 was transformed into the Agrobacterium strain GV3101, and then transformed into the wild-type Arabidopsis plant to obtain the seeds of the transgenic contemporary seedling of Arabidopsis thaliana. The screening resistance gene carried by the Super promoter-pCambia1300 vector is hygromycin, and the transgenic seedlings of Arabidopsis thaliana were screened with hygromycin. The T2 generation seeds were screened for hygromycin resistance, and a line with a segregation ratio of about 3:1 between resistant seedlings and non-resistant seedlings was selected, indicating that in this line, the carrier sequence with the target gene is a single copy form inserted. The plants with hygromycin resistance in these lines were removed, harvested from individual plants, and then screened for hygromycin resistance. If there was no separation, it i...

Embodiment 3

[0031] Example 3: Sugar transport ability of plants overexpressing CsSWEET1a

[0032] After sterilizing Arabidopsis seeds, sow them on 1 / 2MS, 1 / 2MS+1.5% sucrose and 1 / 2MS+1.5% glucose medium, after 18 days of growth, take pictures to record the phenotype, see figure 2 A-D, see the underground part figure 2 E-H. After sterilizing Arabidopsis seeds, sow them on 1 / 2MS, 1 / 2MS+3% sucrose and 1 / 2MS+3% glucose medium, after 24 days of growth, take pictures to record the phenotype, see figure 2 I-L. The results showed that compared with the wild type, the overexpressed plants could absorb more sucrose or glucose, and therefore exhibited glycotoxic phenotypes such as leaves turning purple and roots turning brown.

[0033] After sterilizing the Arabidopsis seeds, they were sown in 1 / 2MS+1.5% sucrose medium, and after 18 days of growth, they were treated at room temperature and low temperature respectively. The low-temperature treatment operation is as follows, 4°C for 2 days, th...

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Abstract

The invention discloses application of a hexose transporter gene CsSWEET1a of camellia sinensis in improving low temperature resistance of plants. Biological function verification by overexpression ofthe gene in arabidopsis proves that the CsSWEET1a gene provides a gene resource for breeding of new varieties of low-temperature-resistant plants by increasing the content of sucrose and glucose in plants and improving the low temperature resistance of the arabidopsis, and has better potential application value.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and in particular relates to the application of the tea tree hexose transporter gene CsSWEET1a. Background technique [0002] During the growth and development of plants, they will inevitably encounter various adverse environments from the outside world. In the context of global climate change, extreme temperatures appear more and more frequently, and low temperature stress has seriously affected the quality and yield of commercial crops. The ability of plants to tolerate low temperature determines their geographical distribution, which is related to whether they can survive the winter safely and whether they can resist the late spring cold in early spring. Tea tree is an important leaf-based economic crop. It originated in tropical or subtropical regions and is sensitive to low temperature. The low temperature adaptability of tea tree is closely related to tea quality and product...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/29A01H5/00A01H6/20
CPCC07K14/415C12N15/8273
Inventor 王璐姚利娜王新超杨亚军
Owner TEA RES INST CHINESE ACAD OF AGRI SCI
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