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Apple fruit hexose transporter gene MdHT2.2 and application thereof

A transporter and transgenic plant technology, applied in the application field of apple fruit hexose transporter gene MdHT2.2, can solve the problem of unknown sugar metabolism content regulation and achieve the effect of increasing sugar content

Active Publication Date: 2019-05-17
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the mechanism of phloem unloading has been studied by many people, its regulation of glucose metabolism and content is still unknown

Method used

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  • Apple fruit hexose transporter gene MdHT2.2 and application thereof
  • Apple fruit hexose transporter gene MdHT2.2 and application thereof
  • Apple fruit hexose transporter gene MdHT2.2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1, analyzing the gene expression pattern of the apple fruit hexose transporter gene MdHT2.2 of the present invention:

[0041] The materials were selected from "Gala" apples in the North Campus North Campus of Northwest Agriculture and Forestry University, and the rootstock was Xinjiang wild apples (M.sieversii). The planting density is 3×4m, running north-south. Throughout the growth process, regularly spray fungicides and insecticides to control diseases and insect pests. Flowers, young fruits, mature fruits, pericarp, pulp, young leaves, mature leaves, old leaves, roots, fruit stalks, petioles, phloem, and xylem were collected as samples, and all samples were quickly frozen in liquid nitrogen immediately after harvesting, and the total The RNA was reverse transcribed, and the resulting first-strand cDNA was used to amplify the apple fruit hexose transporter gene MdHT2.2.

[0042] Wherein, utilize CTAB method when extracting total RNA, (CTAB extraction buffe...

Embodiment 2

[0046] Example 2, the gene expression of apple fruit hexose transporter gene MdHT2.2 of the present invention and its relationship with fruit sugar accumulation

[0047] 1. qPCR analysis of apple fruit hexose transporter gene MdHT2.2 during apple fruit development

[0048] The materials were taken from the Northwest A&F University North Campus Garden "Gala" apple fruit samples were taken 16, 34, 55, 75, 98, 122 days after flowering, and between 3:00 and 4:00 in the afternoon. When sampling, in all 15 trees, 6 fruits on 3 trees were used as a repetition, a total of 5 repetitions. Weigh quickly after sampling, cut into pieces after removing the nucleus, and freeze immediately. In order to compare the expression patterns of related genes in source and sink tissues, shoot tips and mature leaves were collected at 34 days after flowering. All samples were stored in ultra-low temperature freezers.

[0049] Table 1 primers used in the present invention

[0050]

[0051] 2. Chan...

Embodiment 3

[0057] Embodiment 3, the cloning of apple fruit MdHT2.2 gene

[0058] Using "Gala" fruit as the test material, the total RNA of the pulp was extracted and reverse-transcribed, and the obtained first-strand cDNA was used to amplify the MdHT2.2 gene. Extraction of total RNA from pulp, cDNA synthesis and primer design were the same as

[0059] Example 1.

[0060] Amplification primers are the same as in Table 1. The 50 μl reaction system includes: 10 μl 5×Phusion HF for GC Buffer (NEB Company), 1 μl 10 mM dNTPs, 2.5 μl forward primer, 2.5 μl reverse primer, 0.5 μl Phusion high-fidelity DNA polymerase (NEB company), 1 μl cDNA. The PCR reaction program was: pre-denaturation at 98°C for 30s; denaturation at 98°C for 10s, annealing at 65°C for 30s, extension at 72°C for 30s, 35 cycles; and extension at 72°C for 10min after the cycle was completed. After the PCR product was electrophoresed on a 1% agarose gel, the Figure 6 The indicated bands of interest were recovered with a gel...

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Abstract

The invention discloses an apple fruit hexose transporter gene MdHT2.2, the nucleotide sequence of which is shown in a sequence table 1. The invention also discloses a recombinant expression vector, an expression box, a transgenic cell line, a transgenic plant tissue or recombinant bacteria which contain the apple fruit hexose transporter gene MdHT2.2, and also discloses a protein coded by the apple fruit hexose transporter gene MdHT2.2, wherein the amino acid sequence is shown in a sequence 2, and the application of the apple fruit hexose transporter gene MdHT2.2. The apple fruit hexose transporter gene MdHT2.2 can synthesize apple fruit hexose transporter; the apple fruit hexose transporter can promote the transport of glucose, fructose and the like into cytoplasm and improve the fruit quality; the application of the apple fruit hexose transporter in transgenic plants with early flowering has obvious effects in the fields of improving the hexose content of the plant fruits, improvingthe size of the plant fruits, dwarfing the plants and the like.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to an apple fruit hexose transporter gene MdHT2.2, and the invention also relates to the protein of the apple fruit hexose transporter gene MdHT2.2, and at the same time, the invention also relates to apple fruit hexose transporter Application of protein gene MdHT2.2. Background technique [0002] In plant cells, sugar is not only a basic molecule that provides energy for plant growth and development, but also a substance for osmotic regulation, nutritional components, and signaling molecules. In fruit crops, the soluble sugar content determines the fruit quality, especially the sweetness, while the sugar content and components are regulated by the transportation, synthesis and metabolism of sugar. The accumulation of sugar is the main reason for the formation of fruit quality, which is mainly affected by Mediated by sugar transporter (Sugar transporter). [0003] In mos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/82
Inventor 李明军王正阳祝令成杨静静魏晓钰马锋旺
Owner NORTHWEST A & F UNIV
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