CN108142442A - A kind of wintergreen extract and its application as Genes For Plant Tolerance fungus - Google Patents

A kind of wintergreen extract and its application as Genes For Plant Tolerance fungus Download PDF

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Publication number
CN108142442A
CN108142442A CN201711324938.9A CN201711324938A CN108142442A CN 108142442 A CN108142442 A CN 108142442A CN 201711324938 A CN201711324938 A CN 201711324938A CN 108142442 A CN108142442 A CN 108142442A
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China
Prior art keywords
extract
carbendazim
wintergreen
bacteria
grape
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CN201711324938.9A
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Chinese (zh)
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CN108142442B (en
Inventor
王俊丽
王忠跃
黄航君
郝娜
黄婧婧
林昱星
刘名飞
刘杰
马伟伟
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Institute of Plant Protection of Chinese Academy of Agricultural Sciences
Minzu University of China
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Institute of Plant Protection of Chinese Academy of Agricultural Sciences
Minzu University of China
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/08Magnoliopsida [dicotyledons]

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Plant Pathology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Agronomy & Crop Science (AREA)
  • Dentistry (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Environmental Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Food Preservation Except Freezing, Refrigeration, And Drying (AREA)

Abstract

The present invention relates to a kind of wintergreen extract, the wintergreen extract is alcohol extract, and wherein alcohol extract extracting method is:It is to weigh to crush sample 800g, impregnates plant drymeal with the ethyl alcohol of 5 times of amounts, stand 12h at room temperature;Ultrasonic extraction 2h;Decompression filters, and continuous to extract 3 times, merging filtrate is concentrated at 45 DEG C with Rotary Evaporators, after dry, preserved in 4 DEG C, is spare;Further include application of the wintergreen extract as Genes For Plant Tolerance fungus.Wintergreen extract used in the present invention is a kind of botanical biological pesticide, has the characteristics that antibacterial activity is high, antimicrobial spectrum is wide, is not easy to produce resistance, is nontoxic to humans and animals, can improve grape fruit foodsafety.

Description

A kind of wintergreen extract and its application as Genes For Plant Tolerance fungus
Technical field
The present invention relates to field of biological pesticide, and in particular to botanical biological pesticide is more particularly to a kind of wintergreen and carries Take object and its application as Genes For Plant Tolerance fungus.
Background technology
Grape grey mould, white rot, anthracnose etc. seriously affect grape quality and yield, not still in the production of China's grape The difficulty that must be solved, and be also the problem in worldwide grape production.In agricultural production, it is useization to commonly use preventing control method It learns to farm the chemical prevention method of medicine, but chemical prevention has drug resistance, again residual contamination, the wind such as wildness for causing grape pathogen Danger.Using a variety of methods, alternately cooperation Synthetical prevention is pest and disease damage prevention and control and prevents the developing direction that mycotoxin pollutes and avoid The approach of risk.Therefore other approaches and methods of non-chemical pesticide are found, are that the vital task of industry development and science are ground The important topic studied carefully.Plant source method is excavated from plant, is found first including exploitation botanical pesticide, from phytochemical The direct utilization of compound, even plant leaching liquor immersion is led, is the important content of this developing direction.
Wintergreen (Pyrola calliantha H.Andr.) is Pyrolaceae (Pyrolaceae) pyrola (Pyrola) plant, production Shaanxi, Qinghai, Gansu, Shanxi, Shandong, Hebei, Henan, Anhui, Jiangsu, Zhejiang, Fujian, Hubei, lake South, Jiangxi, Sichuan, Guizhou, Yunnan, Tibet.It is born in 700-4100 meters of needle leaf forests of height above sea level, mixed coniferous broad leaved forest or broad-leaf forest Under.This kind of herb hyoscine makees astringent, civil to be used as tonic, controls consumptive disease, cough-relieving, strengthening the muscles and bones.But it has no as life Report and research of the object pesticides application to preventing and treating grapevine diseases.
Invention content
Answering the invention mainly solves the technical problem of providing a kind of wintergreen extract and its as Genes For Plant Tolerance fungus With grape foodsafety can be improved.
In order to solve the above technical problems, the technical solution adopted by the present invention is:
A kind of wintergreen extract, the wintergreen extract are alcohol extract, and the alcohol extract extracting method is:It is It weighs and crushes sample 800g, impregnate plant drymeal with the ethyl alcohol of 5 times of amounts, stand 12h at room temperature;Ultrasonic extraction 2h;Decompression filters, Continuous to extract 3 times, merging filtrate is concentrated at 45 DEG C with Rotary Evaporators, after dry, preserved in 4 DEG C, is spare.
A kind of application of wintergreen extract as Genes For Plant Tolerance fungus is further related to, the wintergreen extract is to following disease Opportunistic pathogen of falling ill has significant bacteriostatic activity, mainly including Botrytis cinerea, fruit white rot of grape bacterium, grape anthracnose and carbon black Aspergillus germ.
Further, wherein the grape anthracnose is included to carbendazim highly resistance anthrax bacteria bacterial strain, the grey mold Germ is included to carbendazim highly resistance ash arrhizus bacteria bacterial strain.
Further, a concentration of 0.3125mg/mL-5mg/mL of the alcohol extract.
Further, as a concentration of 5mg/mL of alcohol extract, to anthrax bacteria, white rot germ, ash arrhizus bacteria and Aspergillus carbonerius Germ and inhibiting rate are respectively 45.99%, 36.75%, 24.59% and 30.63%, to carbendazim sensitivity anthrax bacteria bacterial strain The inhibiting rate of FJZZ-62, coll-9 and coll-20 are respectively 66.67%, 68.55% and 57.81%, to carbendazim highly resistance charcoal The inhibiting rate of subcutaneous ulcer germ bacterial strain FJND-9, FJND-40 and GZSD-89 are respectively 56.09%, 76.55% and 54.71%, to more The inhibiting rate of sensitive sense grape grey mould bacteria strain GXGL-7, GXNN-6 and the GXNN-7 of bacterium is respectively 42.28%, 44.79% and 55.22%, the inhibiting rate to carbendazim highly resistance grape grey mould bacteria strain HBLF-10, GXZY-4 and GXZY-7 is respectively 44.83%th, 61.00% and 57.38%.
The beneficial effects of the invention are as follows:Wintergreen extract of the present invention is one as the germ killing drugs of active ingredient Kind botanical biological pesticide, belongs to one kind of bio-pharmaceutical, meets requirement of the modern society to novel pesticide.Secondly, wintergreen carries Object is taken to have the characteristics that antibacterial activity is high, antimicrobial spectrum is wide, be not easy to produce resistance, be nontoxic to humans and animals.In addition, it is to grape fruit Real encountered pathogenic bacteria has inhibition, can improve grape fruit foodsafety.
Specific embodiment
It is described in detail with reference to preferred embodiment, so that advantages and features of the invention can be easier to by this field Technical staff's understanding, so as to make a clearer definition of the protection scope of the present invention.
Embodiment 1:
First, the preparation of alcohol extract
The wintergreen of the present invention is purchased from Hui nationality's medicinal material market.Lay-by material is dried, crushes, is placed at aeration-drying It saves backup.It weighs and crushes sample 800g, impregnate plant drymeal with the ethyl alcohol of 5 times of amounts, stand 12h at room temperature;Ultrasonic extraction 2h; Decompression filters, and continuous to extract 3 times, merging filtrate is concentrated at 45 DEG C with Rotary Evaporators, after dry, preserved in 4 DEG C, is spare.
2nd, the bacteriostasis of alcohol extract
Root of Dahurain angelica water extract, alcohol extract are measured to Botrytis cinerea (Botrytis using mycelial growth rate method Cinerea), fruit white rot of grape bacterium (Coniothyrium diplodiella), grape anthracnose (Glomerella ) and the bacteriostatic activity of Aspergillus carbonerius (Aspergillus carbonarius) cingulata.It is as follows:
Strain will be frozen to recover at room temperature, with a small amount of bacterium colony picking of transfer needle picking to the culture dish containing PDA culture medium Central area, sealing, no light inversion are incubated in the incubator of 25 DEG C of temperature, humidity 73%.
A certain amount of extract is weighed, alcohol extract is configured to 50mg/mL, 25mg/mL, 12.5mg/ respectively with sterile water The solution of mL, 6.25mg/mL, 3.125mg/mL.Sterile water is as blank control, and the carbendazim of 5 μ g/mL is as positive control. In aseptic workbench, by prepared liquid respectively with PDA culture medium according to 1:9 ratio mixing, makes band medicine culture A concentration of 5mg/mL, 2.5mg/mL, 1.25mg/mL, 0.625mg/mL, 0.3125mg/mL of base, pour into a diameter of 90mm's Its solidification is treated in culture dish, placement.
After actication of culture, vigorous place is grown in mycelia with aseptic card punch (Ф=5mm) and punched, bacteria cake, inoculation is made Onto band medicine culture medium, each culture dish center connects a bacteria cake, is 25-28 DEG C in temperature, humidity is in 73% incubator After being inverted culture 2-7d, colony diameter is surveyed with crossing method, calculates inhibiting rate.Each processing sets 3 repetitions.
The result shows that:Wintergreen alcohol extract shows different degrees of fungistatic effect to 4 kinds of grape disease fungus, works as alcohol extracting During a concentration of 5mg/mL of object, it is respectively to anthrax bacteria, white rot germ, ash arrhizus bacteria and Aspergillus carbonerius germ and inhibiting rate 45.99%th, 36.75%, 24.59% and 30.63%.
3rd, alcohol extract is to the inhibiting effect of carbendazim sensitivity anthrax bacteria bacterial strain
As a concentration of 5mg/mL of wintergreen alcohol extract, to carbendazim sensitivity anthrax bacteria bacterial strain FJZZ-62, coll-9 and The inhibiting rate of coll-20 is respectively 66.67%, 68.55% and 57.81%.
4th, alcohol extract is to the inhibiting effect of carbendazim highly resistance anthrax bacteria bacterial strain
100 μ g/mL carbendazim are respectively 9.80% to the inhibiting rate of bacterial strain FJND-9, FJND-40 and GZSD-89, 21.50% and 10.01%.Wintergreen alcohol extract has stronger inhibition to carbendazim highly resistance anthrax bacteria bacterial strain.When wintergreen alcohol During a concentration of 5mg/mL of extract, to the inhibiting rate point of carbendazim highly resistance anthrax bacteria bacterial strain FJND-9, FJND-40 and GZSD-89 Not Wei 56.09%, 76.55% and 54.71% (table 3), effect be substantially better than 100 μ g/mL carbendazim.0.625mg/mL alcohol extractings Object is suitable with the effect of 100 μ g/mL carbendazim to the inhibiting effect of carbendazim highly resistance anthrax bacteria bacterial strain.
5th, alcohol extract is to the inhibiting effect of carbendazim sensitivity ash arrhizus bacteria bacterial strain
As a concentration of 5mg/mL of wintergreen alcohol extract, to carbendazim sensitivity grape grey mould bacteria strain GXGL-7, GXNN- The inhibiting rate of 6 and GXNN-7 is respectively 42.28%, 44.79% and 55.22%.
6th, alcohol extract is to the inhibiting effect of carbendazim highly resistance ash arrhizus bacteria bacterial strain
100 μ g/mL carbendazim are respectively 53.45% to the inhibiting rate of bacterial strain HBLF-10, GXZY-4 and GXZY-7, 55.11% and 14.76%.As a concentration of 5mg/mL of wintergreen alcohol extract, to carbendazim highly resistance grape grey mould bacteria strain The inhibiting rate of HBLF-10, GXZY-4 and GXZY-7 are respectively 44.83%, 61.00% and 57.38%.Wintergreen alcohol extract (5mg/mL) is less than 100 μ g/mL carbendazim to the inhibiting effect of bacterial strain HBLF-10, and the inhibition of GXZY-4 and GXZY-7 is made With being better than 100 μ g/mL carbendazim.
The foregoing is merely the embodiment of the present invention, are not intended to limit the scope of the invention, every to utilize this hair The equivalent structure or equivalent flow shift that bright description is made directly or indirectly is used in other relevant technology necks Domain is included within the scope of the present invention.

Claims (5)

1. a kind of wintergreen extract, it is characterised in that:The wintergreen extract is alcohol extract, and the alcohol extract extracts Method is:It weighs and crushes sample 800g, impregnate plant drymeal with the ethyl alcohol of 5 times of amounts, stand 12h at room temperature;Ultrasonic extraction 2h;Subtract Pressure filters, and continuous to extract 3 times, merging filtrate is concentrated at 45 DEG C with Rotary Evaporators, after dry, preserved in 4 DEG C, is spare.
2. application of a kind of wintergreen extract as described in claim 1 as Genes For Plant Tolerance fungus, which is characterized in that described Wintergreen extract has significant bacteriostatic activity to following disease pathogen, mainly including Botrytis cinerea, fruit white rot of grape Bacterium, grape anthracnose and Aspergillus carbonerius germ.
3. application as claimed in claim 2, which is characterized in that the wherein described grape anthracnose is included to carbendazim highly resistance Anthrax bacteria bacterial strain, the ash arrhizus bacteria are included to carbendazim highly resistance ash arrhizus bacteria bacterial strain.
4. application as claimed in claim 2 or claim 3, which is characterized in that a concentration of 0.3125mg/mL-5mg/ of the alcohol extract mL。
5. application as claimed in claim 4, which is characterized in that as a concentration of 5mg/mL of alcohol extract, to anthrax bacteria, white rot Germ, ash arrhizus bacteria and Aspergillus carbonerius germ and inhibiting rate are respectively 45.99%, 36.75%, 24.59% and 30.63%, right The inhibiting rate of carbendazim sensitivity anthrax bacteria bacterial strain FJZZ-62, coll-9 and coll-20 are respectively 66.67%, 68.55% and 57.81%, the inhibiting rate to carbendazim highly resistance anthrax bacteria bacterial strain FJND-9, FJND-40 and GZSD-89 is respectively 56.09%, 76.55% and 54.71%, to the inhibiting rate point of carbendazim sensitivity grape grey mould bacteria strain GXGL-7, GXNN-6 and GXNN-7 Not Wei 42.28%, 44.79% and 55.22%, to carbendazim highly resistance grape grey mould bacteria strain HBLF-10, GXZY-4 and The inhibiting rate of GXZY-7 is respectively 44.83%, 61.00% and 57.38%.
CN201711324938.9A 2017-12-13 2017-12-13 Wintergreen extract and application thereof as plant antibacterial material Expired - Fee Related CN108142442B (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113773970A (en) * 2021-10-14 2021-12-10 四川农业大学 Aspergillus niger BZZJ-17 with plant growth promoting effect

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101401591A (en) * 2008-11-24 2009-04-08 王占斌 Method for extracting bactericide from wintergreen
CN101862028A (en) * 2010-06-09 2010-10-20 广东神农烟科技术有限公司 Wintergreen extraction method, cigarette comprising same and preparation method thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101401591A (en) * 2008-11-24 2009-04-08 王占斌 Method for extracting bactericide from wintergreen
CN101862028A (en) * 2010-06-09 2010-10-20 广东神农烟科技术有限公司 Wintergreen extraction method, cigarette comprising same and preparation method thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
王亚飞 等: "鹿蹄草等4种中草药提取液对桃褐腐菌抑制作用研究", 《安徽农业科学》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113773970A (en) * 2021-10-14 2021-12-10 四川农业大学 Aspergillus niger BZZJ-17 with plant growth promoting effect
CN113773970B (en) * 2021-10-14 2023-05-23 四川农业大学 Aspergillus niger BZZJ-17 with plant growth promoting effect

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