阅读说明：本技术 黄瓜CsHMGB基因在降低农药残留量以及缓解农药毒性上的应用 (Application of cucumber CsHMGB gene in reducing pesticide residue and relieving pesticide toxicity ) 是由 辛明 秦智伟 李胜男 周秀艳 王蕾 于 2020-05-20 设计创作，主要内容包括：黄瓜CsHMGB基因在降低农药残留量以及缓解农药毒性上的应用,涉及CsHMGB基因在农业领域的应用。本发明从遗传学角度出发,在植物体内代谢农药,提高黄瓜安全品质,实现减少农药残留的同时避免了环境污染等问题。沉默CsHMGB的黄瓜株系霜霉威残留量增加而过量表达CsHMGB的黄瓜株系霜霉威残留量下降。霜霉威胁迫后黄瓜受到ROS的干扰,过量表达的CsHMGB可以提高抗氧化酶SOD、POD、CAT等的活性,且提高ASA-GSH循环能力。过量表达的CsHMGB诱导解毒类基因,谷胱甘肽家族和膜转运蛋白ABC家族的部分基因上调表达。CsHMGB可通过抗氧化酶系统缓解霜霉威的毒害。(An application of a cucumber CsHMGB gene in reducing pesticide residue and relieving pesticide toxicity relates to an application of the CsHMGB gene in the agricultural field. From the genetic aspect, the invention metabolizes pesticides in plants, improves the safety quality of cucumbers, reduces pesticide residues and avoids the problems of environmental pollution and the like. The amount of propamocarb in cucumber lines silent on CsHMGB was increased and that in cucumber lines overexpressing CsHMGB was decreased. After the cucumber is stressed by the propamocarb, the cucumber is interfered by ROS, and the over-expressed CsHMGB can improve the activities of antioxidant enzymes SOD, POD, CAT and the like and improve the circulating capacity of ASA-GSH. The over-expressed CsHMGB induces the up-regulation expression of partial genes of detoxification genes, glutathione family and membrane transporter ABC family. CsHMGB can alleviate the toxicity of propamocarb through an antioxidase system.)

1. The application of the cucumber CsHMGB gene in reducing the pesticide residue and relieving the pesticide toxicity.

2. Use according to claim 1, characterized in that the pesticide is propamocarb.

Technical Field

The invention relates to application of a CsHMGB gene in the field of agriculture.

Background

Cucumber (Cucumis sativus) is one of the main cultivated crops in vegetable production of Chinese facilities, and has a wide planting area. Because of special climatic conditions and continuous planting in the greenhouse, the cucumber is very easy to be attacked by diseases and insect pests, and the cucumber downy mildew is a destructive disease.

Propamocarb is a novel, efficient and broad-spectrum carbamate bactericide, can effectively prevent downy mildew of melons, is one of common pesticides for preventing and treating downy mildew of cucumbers, but is very easy to accumulate in cucumber fruits to form pesticide residues after application due to certain volatility and systemic property, and the rest pesticide residues are deposited in soil or diffused in the atmosphere, so that on one hand, cucumber pesticide residues are caused, the quality is influenced, and the food safety problem is caused; on the other hand, the environment pollution is caused, and the human health is harmed. At present, effective ways for reducing pesticide residues in fruits and vegetables are to reduce pesticide investment and accelerate the metabolism of pesticides in plants, but exogenous substances increase production cost and even cause secondary pollution.

Disclosure of Invention

From the genetic aspect, the invention metabolizes pesticides in plants, improves the safety quality of cucumbers, reduces pesticide residues and avoids the problems of environmental pollution and the like.

The application of the cucumber CsHMGB gene in reducing the pesticide residue and relieving the pesticide toxicity.

The total length of the cucumber CsHMGB gene is 624bp, 207 amino acids are coded, and the gene has the highest homology with the melon and the balsam pear on the same evolutionary branch; the amount of propamocarb in cucumber lines silent on CsHMGB was increased and that in cucumber lines overexpressing CsHMGB was decreased. After the cucumber is stressed by the propamocarb, the cucumber is interfered by ROS, and the over-expressed CsHMGB can improve the activities of antioxidant enzymes SOD, POD, CAT and the like and improve the circulating capacity of ASA-GSH. The over-expressed CsHMGB induces the up-regulation expression of partial genes of detoxification genes, glutathione family and membrane transporter ABC family. CsHMGB can alleviate the toxicity of propamocarb through an antioxidase system.

Drawings

FIG. 1 shows the expression level of CsHMGB at different tissue sites of the D0351 cucumber line induced by the propamocarb signal in example 1.

FIG. 2 shows the expression level of CsHMGB at different tissue sites induced by propamocarb signal in the D9320 cucumber line in example 1.

FIG. 3 is a graph comparing the CsHMGB expression levels of the D0351 cucumber line in example 1 in different time periods under propamocarb stress and a control.

FIG. 4 is a graph comparing the CsHMGB expression levels of the D9320 cucumber line in example 1 stressed by propamocarb with that of a control over different periods of time.

FIG. 5 is a confocal laser scanning microscope image of Arabidopsis protoplasts transferred into a vector in example 1.

FIG. 6 is a confocal laser scanning microscope image of epidermal cells of tobacco leaves transfected with the vector in example 1.

FIG. 7 shows the whitening of leaves of cucumber plants inoculated with TRV2-PDS in example 2.

FIG. 8 is a graph comparing the CsHMGB expression levels of 3 groups of cucumber lines inoculated with TRV2-HMGB in example 2 with TRV2 empty control and wild type control.

FIG. 9 is a comparison of the propamocarb residual levels of the 3 groups of cucumber lines inoculated with TRV2-HMGB in example 2 with TRV2 empty control and wild type control.

FIG. 10 is a time chart of the residual amount of propamocarb from T1P-dais fruit sprayed with 400pm propamocarb in example 2.

FIG. 11 is a time chart of the residual amount of propamocarb from T1P-daidzein sprayed with 1mM propamocarb in example 2.

FIG. 12 is O in cucumber bodies after propamocarb stress in example 2^2-The content is compared with the figure.

FIG. 13 shows H in cucumber bodies after Pericarb stress in example 2₂O₂The content is compared with the figure.

FIG. 14 is a graph comparing the MDA content in cucumber bodies after propamocarb stress in example 2.

FIG. 15 is a graph comparing the antioxidant enzyme activity in cucumbers after propamocarb stress in example 2.

FIG. 16 is a comparative set of ASA-GSH cycles in cucumbers after propamocarb stress in example 2.

FIG. 17 is a graph showing comparison of expression levels of detoxification genes in cucumbers after the stress of propamocarb in example 2.

Detailed Description

The technical solution of the present invention is not limited to the following specific embodiments, but includes any combination of the specific embodiments.