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

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

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[0012] Example 1: VIGS treatment of tomato

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

Example Embodiment

[0027] Example 2: Low temperature stress test of SlSLD gene silenced plants (treatment group) and control group

[0028] The plants of the SlSLD gene silenced (treatment group) and the control group were subjected to stress treatment at 4°C, and photographed to record the phenotypic changes of the plants in a low temperature environment. From figure 1 It can be seen that when the SlSLD gene silenced and control plants were treated at low temperature for 6h, the leaf edges appeared curled, and the changes in the treatment group were more obvious; at 24h, the control group still maintained the state at 6h, and the leaves appeared Slight curl, but the whole plants in the treatment group appeared wilting and could no longer grow normally.

Example Embodiment

[0029] Example 3: Determination of plant physiological indicators after low temperature stress test

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

[0031] To prepare the supernatant, weigh 0.1g tomato leaves, add 2.5mL of pre-cooled 50mM phosphate buffered saline solution (pH7.0), grind on ice, centrifuge the homogenate at 4℃, 4,000×g for 10min, and collect The supernatant is placed in a refrigerator at 4°C and used for the determination of indicators 2-5 below.

[0032] 1) Determination of relative electrolytic leakage (REL)

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

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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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