Tea tree hexose transporter gene CsSWETE17 and application thereof in regulating and controlling plant vegetative growth and seed size
A technology of vegetative growth and transporter, applied in the field of biological genetic engineering, can solve problems such as unreported functions
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Embodiment 1
[0026] Cloning of CsSWEET17 Gene of Tea Tree and Construction of Overexpression Vector
[0027] According to the CsSWEET17 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 CsSWEET17 was amplified by PCR. The amplified sequence was connected to the pENTRTM / D-TOPO vector, and finally connected to the pH7FWG2 plant expression vector through LR reaction.
[0028] The primers used were F: 5'-CACCATGGCTAGCTTGAGCTTCATC-3'; R: 5'-AGGGTGATCCTTGGTGCTTCC-3'.
Embodiment 2
[0030] Construction and Screening of CsSWEET17 Gene Overexpression Plants
[0031] 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 pH7FWG2 vector is hygromycin, and the transgenic seedlings of Arabidopsis thaliana were screened with hygromycin. Perform hygromycin resistance screening, and select a strain with a segregation ratio of about 3:1 between resistant seedlings and non-resistant seedlings, indicating that in this strain, the vector sequence with the target gene is inserted in a single copy form. Remove the hygromycin-resistant plants in these lines, harvest the seeds from each plant, and then screen for hygromycin resistance. If there is no separation, it means that the transgenic line is homozygous, and the homozygous can be ...
Embodiment 3
[0034] Vegetative growth phenotypes of plants overexpressing CsSWEET17
[0035] After the Arabidopsis seeds were sterilized, they were sown on 1 / 2 MS medium, and after 7 days of growth, they were moved into seedling blocks for growth. When cultivating to seedling age is 13 days and 18 days, take pictures and record respectively, see figure 2 and image 3. The 18-day-old seedlings were also weighed for aboveground fresh weight (the wild type and OE-1, 15 seedlings of OE-9 were counted respectively), see Figure 4 . The results showed that OE-1 and OE-9 overexpression plants both showed increased leaf area and increased fresh weight of aerial parts compared with the wild type.
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