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Cold-injury-resistance gene of lycopersicon esculentum mill and application

A tomato and gene technology, applied in the field of plant biology, to achieve the effects of improving adaptability, promoting stable and high yield, and expanding the planting range

Active Publication Date: 2017-05-31
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no researchers have explored △ 8 -The role of sphingomyelin dehydrogenase in cold adaptation of tomato

Method used

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  • Cold-injury-resistance gene of lycopersicon esculentum mill and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Example 1: VIGS processing of tomato

[0013] The formula of LB solid medium used below is: 10g / L tryptone (Tryptone), 5g / L yeast extract (Yeast extract), 10g / L sodium chloride (NaCl), 15-20g / L agar powder, use Adjust the pH value to 7.4 with 10M NaOH, and the balance is water; sterilize at 121°C for 20 minutes, and when the temperature drops to about 55°C (not hot), add Kanamycin (Kanamycin, Kan) at a final concentration of 30mg / L, or Add final concentration of 30mg / L Kanamycin (Kanamycin, Kan), 50mg / L Rifampicin (Rifampicin, Rif), 50mg / L Gentamicin (Gentamicin, Gen), pour plate, save for later use; LB liquid The formula of the medium is: 10g / L tryptone (Tryptone), 5g / L yeast extract (Yeast extract), 10g / L sodium chloride (NaCl), adjust the pH value to 7.4 with 10M NaOH, and the balance is water; Sterilize at 121°C for 20 minutes, and after the temperature drops to about 55°C (not hot to the hands) or after cooling, add Kanamycin (Kanamycin, Kan) at a final concentrat...

Embodiment 2

[0027] Embodiment 2: the low temperature stress test of the plants (treatment group) and control group of SlSLD gene silencing

[0028] The plants of the SlSLD gene silenced (treatment group) and the control group were subjected to stress treatment at 4°C, and the phenotypic changes of the plants in the low temperature environment were recorded by taking pictures. From figure 1 It can be seen that, when the plants of the SlSLD gene silenced and the control group were treated at low temperature for 6 hours, the edges of the leaves curled, and the changes in the treatment group were more obvious; at 24 hours, the control group still maintained the state at 6 hours, and the leaves appeared Slight curling, while the whole plants in the treatment group wilted and could no longer grow normally.

Embodiment 3

[0029] Embodiment 3: Determination of physiological indexes of plants after low temperature stress test

[0030] The physiological indexes of tomato leaves treated with low temperature for 6 hours mainly included electrical conductivity, malondialdehyde content, soluble polysaccharide content, superoxide dismutase activity, peroxidase activity and chlorophyll content. The specific operation is as follows:

[0031] Preparation of supernatant: Weigh 0.1 g of tomato leaves, add 2.5 mL of pre-cooled 50 mM phosphate buffered saline (pH 7.0), grind on ice, centrifuge the homogenate at 4°C, 4,000×g for 10 min, collect The supernatant was placed in a 4°C refrigerator for the determination of the following items 2-5.

[0032] 1) Determination of conductivity (Relative electrolytic leakage, REL)

[0033] Take 0.05 g of low-temperature-treated tomato leaves of the control group and the treatment group, and wash the electrolyte on the surface of the leaves with double distilled water. ...

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Abstract

The invention discloses a cold-injury-resistance gene of lycopersicon esculentum mill and an application. On the basis of virus induced gene silencing (VIGS) technology, the novel chilling-injury-resistance related gene SISLD of the lycopersicon esculentum mill is discovered, and the gene, through silencing, can result in obvious decrease in low temperature tolerance ability of a tomato plant. Based upon analysis on an amino acid sequence encoded by the gene, protein encoded by the gene is [delta]8-sphingomyelin dehydrogenase, which is related to lipid metabolism of neurolemma, and the enzyme (the [delta]8-sphingomyelin dehydrogenase) plays an important role in a reaction that a plant resists to low-temperature stress. According to the research, a nucleotide sequence and an amino acid sequence of the gene are provided; and meanwhile, the invention also relates the application of the gene in cold-resistance breeding of tomato. The SISLD gene, when applied to anti-freezing breeding of the tomato, can improve the adaptability of tomato's high-quality varieties, extend a planting range of the tomato's high-quality varieties and promote stable production and high-yield production of the tomato's high-quality varieties.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and in particular relates to a tomato chilling damage resistance gene and its application. Background technique [0002] Sphingolipids is a general term for a class of complex lipids containing fatty acids and long-chain sphingosine skeletons. It is de novo synthesized on the endoplasmic reticulum by catalyzing serine and palmitoyl-CoA by serine palmitoyl-CoA transferase (SPT). This type of lipid is an important part of the plant cell plasma membrane, tonoplast membrane and inner membrane system, accounting for about 40% of the lipids on the membrane, and mainly exists in the outer leaflet of the cell membrane. The outer leaves of the cell membrane affect the integrity of the membrane and the permeability of ions, and the physiological state of the cell membrane under low temperature conditions is an important indicator of the plant's resistance to chilling damage. In addition, sphingolipids ...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/53C12N15/82A01H5/00
CPCC12N9/0071C12N15/8273C12Y114/19
Inventor 周瀛杨子银曾兰亭蒋跃明段学武
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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