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Application of potato vacuolar monosaccharide transport protein StTMT2 gene

A transporter, potato technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems that have not yet been reported on the study of monosaccharide transporter

Active Publication Date: 2020-10-23
宁夏农林科学院农业生物技术研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that there are 54 sugar transporters in the potato genome, of which 20 are monosaccharide transporters, but there is no research report on the analysis of the function of monosaccharide transporters, especially those located in the tonoplast membrane

Method used

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  • Application of potato vacuolar monosaccharide transport protein StTMT2 gene
  • Application of potato vacuolar monosaccharide transport protein StTMT2 gene
  • Application of potato vacuolar monosaccharide transport protein StTMT2 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Cloning of Potato Tonoplast Membrane Monosaccharide Transporter StTMT2 Gene and Construction of Expression Vector

[0049] The present invention uses RT-PCR technology to clone the cDNA sequence of the potato tonoplast monosaccharide transporter StTMT2 gene from the leaves of the potato cultivar 'Atlantic', and constructs the pCAMBIA1302-35S-StTMT2 recombinant plant expression vector for genetic transformation.

[0050] The amplification method is as follows: firstly, the total RNA of potato leaves is extracted using the Plant Total RNA Isolation Kit (purchased from Novizyme Company), and secondly reverse-transcribed into The total cDNA (see the instructions for the specific process), and then use this cDNA as a template to carry out PCR amplification with TMTF and TMTR (SEQ ID NO.3 and SEQ ID NO.4) as primers, and the amplified products are subjected to 1% agarose gel After a single band was detected by electrophoresis, the target band was recovered with a gel recovery...

Embodiment 2

[0055] Genetic Transformation of Potato Tonoplast Monosaccharide Transporter StTMT2 Gene

[0056] Inoculate 50 mL of LB liquid medium (containing 50 mg·L -1 In Kan), in a shaker at 28°C, shake culture at 180rpm for 24 hours, take it out, centrifuge and discard the supernatant, collect the bacteria, resuspend the bacteria with MS liquid medium, measure the OD value of the bacteria at about 0.6-0.8, and use it for the invasion dye transformation. Take out the test-tube potato in the ultra-clean workbench, cut it into thin slices of about 1-2 mm, put it into a sterile conical flask, pour the above-mentioned resuspended Agrobacterium bacteria solution, and dip it for 12 minutes (shaking the conical flask constantly during the period), pour out the bacteria liquid and use sterile filter paper to absorb the bacterial solution remaining on the surface of potato chips, and transfer the potato chips to co-cultivation medium (MS 4.42g·L) with 2 layers of filter paper -1 , sucrose 30g...

Embodiment 3

[0064] Genetic Transformation of the Potato Tonoplast Monosaccharide Transporter StTMT2 Gene in Arabidopsis thaliana

[0065] Planting of Arabidopsis: put an appropriate amount of Arabidopsis seeds in a centrifuge tube, add 75% alcohol to the ultra-clean workbench for disinfection for 2 minutes, then add 1 mL (10% NaCLO+0.01% Triton) for disinfection for 10 minutes, and sterilize with Wash with distilled water 4 times, 1 min each time. Finally, sow the seeds evenly on the MS solid medium, place the plate at 4°C for vernalization for 2 to 3 days, place it in an artificial incubator with 16h light / 8h dark, and cultivate it at 22°C for about 10 days. Transplant when four true leaves have grown. Mix nutrient soil, vermiculite and perlite in a ratio of 6:3:1, put it into a nutrient bowl after high-temperature sterilization, and pour the nutrient solution thoroughly. Select healthy young seedlings and transplant them into nutrient pots, cover them with plastic wrap to keep them mo...

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Abstract

The invention relates to application of a potato vacuolar monosaccharide transport protein StTMT2 gene, and belongs to the technical field of plant genetic engineering. The invention provides application of a potato vacuolar monosaccharide transport protein StTMT2 gene or a protein or a recombinant vector containing the gene, an expression cassette or recombinant bacteria in the aspects of improving plant yield, sugar content, photosynthetic rate, chlorophyll content, glycometabolism related gene expression and the like. The application disclosed by the invention has important production significance for improving the sugar content and yield of crops, screening germplasm materials, directionally breeding and the like.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to the application of the potato tonoplast membrane monosaccharide transporter StTMT2 gene. Background technique [0002] Potato (Solanum tuberosum L.) is the third largest food crop in the world. It can be used as both grain and vegetable, with high nutritional value and long industrial chain. China is the country with the largest potato planting area in the world, but there is a certain gap between the yield per unit area and that of the United States, the Netherlands, Japan and other countries. Tubers are the only form of potato production. The starch in tubers comes from the monosaccharides and sucrose synthesized by photosynthesis of leaves, which are transported from source organs to tubers for long-distance storage, or partially decomposed into monosaccharides to provide energy for tuber development and so on. [0003] The transport of sugar from source org...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/82A01H6/20
CPCC07K14/415C12N15/8245
Inventor 巩檑聂峰杰陈虞超张海雯宋玉霞甘晓燕张丽刘璇杨文静张国辉
Owner 宁夏农林科学院农业生物技术研究中心
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