CN108770840B - Tobacco disease leaf specimen dipping preservation liquid and specimen preparation method - Google Patents

Tobacco disease leaf specimen dipping preservation liquid and specimen preparation method Download PDF

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Publication number
CN108770840B
CN108770840B CN201810907354.2A CN201810907354A CN108770840B CN 108770840 B CN108770840 B CN 108770840B CN 201810907354 A CN201810907354 A CN 201810907354A CN 108770840 B CN108770840 B CN 108770840B
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China
Prior art keywords
tobacco
specimen
leaf
disease
dipping
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Expired - Fee Related
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CN201810907354.2A
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Chinese (zh)
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CN108770840A (en
Inventor
刘旭
雷强
余伟
刘昌华
肖筠
吴斌
屈建康
余祥文
李斌
陈松
陈庆东
刘振兴
张宗锦
闫芳芳
张瑞平
张映杰
胡伟
陈倩颖
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Panzhihua Corp Of Sichuan Province Tobacco Monopoly Administration
Institute of Plant Protection Sichuan Academy of Agricultural Sciences
China National Tobacco Corp Sichuan Branch
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Panzhihua Corp Of Sichuan Province Tobacco Monopoly Administration
Institute of Plant Protection Sichuan Academy of Agricultural Sciences
China National Tobacco Corp Sichuan Branch
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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
    • A01N3/00Preservation of plants or parts thereof, e.g. inhibiting evaporation, improvement of the appearance of leaves or protection against physical influences such as UV radiation using chemical compositions; Grafting wax
    • GPHYSICS
    • G09EDUCATION; CRYPTOGRAPHY; DISPLAY; ADVERTISING; SEALS
    • G09BEDUCATIONAL OR DEMONSTRATION APPLIANCES; APPLIANCES FOR TEACHING, OR COMMUNICATING WITH, THE BLIND, DEAF OR MUTE; MODELS; PLANETARIA; GLOBES; MAPS; DIAGRAMS
    • G09B23/00Models for scientific, medical, or mathematical purposes, e.g. full-sized devices for demonstration purposes
    • G09B23/38Models for scientific, medical, or mathematical purposes, e.g. full-sized devices for demonstration purposes for botany

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Physics & Mathematics (AREA)
  • Business, Economics & Management (AREA)
  • Educational Technology (AREA)
  • Mathematical Analysis (AREA)
  • Mathematical Optimization (AREA)
  • Mathematical Physics (AREA)
  • Pure & Applied Mathematics (AREA)
  • Algebra (AREA)
  • Botany (AREA)
  • Educational Administration (AREA)
  • Computational Mathematics (AREA)
  • Theoretical Computer Science (AREA)
  • Agronomy & Crop Science (AREA)
  • Plant Pathology (AREA)
  • Toxicology (AREA)
  • Dentistry (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Environmental Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

The invention discloses tobacco disease leaf specimen dipping preservation solution and a specimen preparation method, wherein the tobacco disease leaf specimen dipping preservation solution comprises the following components in percentage by mass: 0.3-0.5% of peroxyacetic acid, 0.2-0.4% of ascorbic acid and 0.5-1.5% of citric acid. The preparation method of the tobacco diseased leaf specimen comprises the following steps: (1) washing tobacco diseased leaves with tap water, and then soaking the tobacco diseased leaves in 5% copper sulfate solution for 12-24 hours; (2) taking out the leaves treated in the step (1) and storing the leaves in the tobacco disease leaf specimen preservation solution. The three components of the peroxyacetic acid, the ascorbic acid and the citric acid are combined to stabilize the internal tissue structure and the physical properties of the tobacco leaf. The concentration percentage of the above-mentioned substances also plays a very key role in the long-term stable preservation of tobacco leaf disease specimens.

Description

Tobacco disease leaf specimen dipping preservation liquid and specimen preparation method
Technical Field
The invention belongs to the technical field of plant specimen preparation, and relates to tobacco disease leaf specimen dipping preservation solution and a specimen preparation method.
Background
Tobacco disease specimens teaching and scientific research are important tools for field disease identification. The existing tobacco disease leaf specimen preservation methods are many, and a dipping method and a drying method are commonly used. However, the tobacco leaves are wide and contain much moisture, and drying methods such as sun drying or baking are easy to change color, poor in color retention effect, unobvious in scab, short in storage time and the like. Whereas the impregnation method generally: (1) boiling the diseased leaves with water by adopting a mixed diluent of copper acetate crystals and acetic acid, and then preserving the diseased leaves in formalin; (2) the disease leaves are boiled in water by adopting a dilution liquid of copper sulfate crystals and then are put into a mixed liquid of sulfurous acid, alcohol and the like for storage. Both methods for dip preservation of samples present the following problems: (a) the concentration and the treatment time of the treatment solution can be determined only by a period of time; (b) the treated leaves are withered and changed in color when being fresh, so that some disease spots are not clear and disease characteristics are not obvious after treatment. (c) When the heat treatment liquid is heated in the treatment process of the disease specimen, the heat treatment liquid can volatilize and emit pungent smell, and the health of people is seriously affected. (d) The time for preserving the diseased leaf specimen is not long and the color is easy to fade.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: the method solves the problems that the existing tobacco diseased leaf impregnated specimen fades after being stored for a long time, the storage time is short, the specimen changes yellow and black in weeks or months, the specimen cannot be distinguished, and pungent smell exists in the treatment process.
The technical scheme of the invention is as follows: a tobacco disease leaf specimen dipping preservation solution comprises the following components in percentage by mass: 0.3 to 0.5 percent of peroxyacetic acid, 0.2 to 0.4 percent of ascorbic acid, 0.5 to 1.5 percent of citric acid and the balance of water.
On the other hand, the invention also provides a manufacturing method of the tobacco disease leaf specimen, which comprises the following steps:
(1) washing tobacco diseased leaves with tap water, and then soaking the tobacco diseased leaves in 5% copper sulfate solution for 12-24 hours;
(2) taking out the leaves treated in the step (1) and storing the leaves in the tobacco disease leaf specimen dipping and storing solution.
Compared with the prior art, the invention has the following beneficial effects:
the tobacco disease leaf specimen is soaked in 5% copper sulfate diluent for sterilization, and then is stored in a mixed solution consisting of peroxyacetic acid, ascorbic acid and citric acid. The three components of the peroxyacetic acid, the ascorbic acid and the citric acid are combined to stabilize the internal tissue structure and the physical properties of the tobacco leaf. The concentration percentage of the above-mentioned substances also plays a very key role in the long-term stable preservation of tobacco leaf disease specimens.
Detailed Description
The tobacco leaf disease specimens were stored by dipping in the samples of examples 1 to 5 and comparative examples 1 to 2, respectively, in the same storage apparatus and under the same storage conditions. Each group was set to 5 replicates for a total of three replicates. The condition of the specimen is observed and recorded every day, whether the specimen fades or not, whether the preservation solution is turbid or not and whether the lesion mark is obvious or not. And finally, the fading and greening time of the tobacco leaf disease specimen, turbid preservation solution and the number of stored months are counted, and the fading, turbid preservation solution and storage time in the storage of the tobacco leaf disease specimen are used for comprehensively evaluating the effects of the preservation solutions and the preparation method.
Example 1:
a tobacco leaf disease specimen preserving fluid and a manufacturing method thereof comprise the following steps:
1. mixing A, B solutions according to the specification before using, placing at 30 deg.C, sealing, standing for 48 hr to obtain stock solution, diluting with purified water to obtain 0.3% diluted solution, and adding ascorbic acid and citric acid to obtain soaking and preserving solution. The dipping preservation solution consists of 0.3 percent of peroxyacetic acid, 0.4 percent of ascorbic acid and 0.5 percent of citric acid.
2. Cleaning a tobacco leaf disease specimen with tap water, placing the tobacco leaf disease specimen into a 5% copper sulfate solution, soaking the old and tender degree of the visual leaf for 12-24 hours, and taking out the visual leaf disease specimen.
3. Taking out the tobacco disease specimen and storing the tobacco disease specimen in the dipping preservation solution.
Example 2:
a tobacco leaf disease specimen preserving fluid and a manufacturing method thereof comprise the following steps:
1. mixing A, B solutions according to the specification before using, placing at 30 deg.C, sealing, standing for 48 hr to obtain stock solution, diluting with purified water to obtain 0.4% diluted solution, and adding ascorbic acid and citric acid to obtain soaking and preserving solution. The dipping preservation solution consists of 0.4 percent of peroxyacetic acid, 0.3 percent of ascorbic acid and 1.0 percent of citric acid.
2. Cleaning a tobacco leaf disease specimen with tap water, placing the tobacco leaf disease specimen into a 5% copper sulfate solution, soaking the old and tender degree of the visual leaf for 12-24 hours, and taking out the visual leaf disease specimen.
3. Taking out the tobacco disease specimen and storing the tobacco disease specimen in the dipping preservation solution.
Example 3:
a tobacco leaf disease specimen preserving fluid and a manufacturing method thereof comprise the following steps:
1. mixing A, B solutions according to the specification before using, placing at 30 deg.C, sealing, standing for 48 hr to obtain stock solution, diluting with purified water to obtain 0.5% diluted solution, and adding ascorbic acid and citric acid to obtain soaking and preserving solution. The dipping preservation solution consists of 0.5 percent of peroxyacetic acid, 0.2 percent of ascorbic acid and 1.5 percent of citric acid.
2. Cleaning a tobacco leaf disease specimen with tap water, placing the tobacco leaf disease specimen into a 5% copper sulfate solution, soaking the old and tender degree of the visual leaf for 12-24 hours, and taking out the visual leaf disease specimen.
3. Taking out the tobacco disease specimen and storing the tobacco disease specimen in the dipping preservation solution.
Example 4:
a tobacco leaf disease specimen preserving fluid and a manufacturing method thereof comprise the following steps:
1. mixing A, B solutions according to the specification before using, placing at 30 deg.C, sealing, standing for 48 hr to obtain stock solution, diluting with purified water to obtain 0.5% diluted solution, and adding ascorbic acid and citric acid to obtain soaking and preserving solution. The dipping preservation solution consists of 0.5 percent of peroxyacetic acid, 15 percent of alcohol and 0.5 percent of citric acid.
2. Cleaning a tobacco leaf disease specimen with tap water, placing the tobacco leaf disease specimen into a 5% copper sulfate solution, soaking the old and tender degree of the visual leaf for 12-24 hours, and taking out the visual leaf disease specimen.
3. Taking out the tobacco disease specimen and storing the tobacco disease specimen in the dipping preservation solution.
Example 5:
a tobacco leaf disease specimen preserving fluid and a manufacturing method thereof comprise the following steps:
1. mixing A, B solutions according to the specification before using, placing at 30 deg.C, sealing, standing for 48 hr to obtain stock solution, diluting with purified water to obtain 0.5% diluted solution, and adding ascorbic acid and citric acid to obtain soaking and preserving solution. The immersion preservation solution consists of 0.4 percent of peroxyacetic acid, 0.1 percent of ascorbic acid and 0.5 percent of citric acid.
2. Cleaning a tobacco leaf disease specimen with tap water, placing the tobacco leaf disease specimen into a 5% copper sulfate solution, soaking the old and tender degree of the visual leaf for 12-24 hours, and taking out the visual leaf disease specimen.
3. Taking out the tobacco disease specimen and storing the tobacco disease specimen in the dipping preservation solution.
Comparative example 1:
a plant leaf disease specimen preserving fluid and a manufacturing method thereof comprise the following steps:
1. cleaning a tobacco leaf disease specimen with tap water, placing the tobacco leaf disease specimen into a 5% copper sulfate solution, soaking the old and tender degree of the visual leaf for 12-24 hours, and taking out the visual leaf disease specimen.
2. Storing in a mixture of formaldehyde 2.5% and ethanol 15%.
Comparative example 2:
a plant leaf disease specimen preserving fluid and a manufacturing method thereof comprise the following steps:
1. cleaning a tobacco leaf disease specimen with tap water, placing the tobacco leaf disease specimen into a 5% copper sulfate solution, soaking the old and tender degree of the visual leaf for 12-24 hours, and taking out the visual leaf disease specimen.
2. Stored in 1.5% sulfurous acid.
The effects of the embodiment are as follows:
comparison of preservation effects of tobacco leaf disease specimen preservation solutions
Preservation method Condition of specimen Colour of preservation solution Characteristic of lesion Storage time (moon)
Example 1 No fading and greenness loss No turbidity is seen Is obvious 24
Example 2 No fading and greenness loss No turbidity is seen Is obvious 24
Example 3 No fading and greenness loss No turbidity is seen Is obvious 24
Example 4 Fading and greening at the beginning of 6 months Onset of turbidity at 7 months Unclear vision 7
Example 5 Fading and greening at 13 months Onset of turbidity at 14 months Unclear vision 14
Comparative example 1 Fading and green loss at 12 months Onset of turbidity at 13 months Is not too clear 13
Comparative example 2 The color of the solution begins to fade and lose green after 11 months Turbidity started at 12 months Is not too clear 12
The tobacco leaf disease specimens prepared in examples 1 to 3 and the tobacco specimens prepared in comparative examples were stored and preserved under the same environmental conditions. The leaf specimens in the comparative examples gradually fade after being stored for 11-12 months, and the scabs gradually become light and unclear. However, the color of the tobacco leaf disease specimen in the preservation solution of the embodiment 1-3 of the invention is not changed, and the disease spots are still obvious. Example 4 the solution started to fade and turn green in 6 months, and became cloudy in 7 months, with a storage time of 7 months, without adding ascorbic acid. Example 5 started to fade, lose green and the lesions were less clear after 13 months.
Compared with the defects of the existing plant leaf specimen preservation solution, the invention has the following beneficial effects: the method for manufacturing the tobacco leaf disease specimen is simple to operate, and the manufactured tobacco leaf disease specimen is long in storage time. The problem of leaf sample discolour can not appear in the preservation in-process to green has reduced the pollution to the environment and to the influence of operating personnel health.

Claims (2)

1. The tobacco disease leaf specimen dipping preservation solution is characterized by comprising the following components in percentage by mass: 0.3 to 0.5 percent of peroxyacetic acid, 0.2 to 0.4 percent of ascorbic acid, 0.5 to 1.5 percent of citric acid and the balance of water.
2. A manufacturing method of tobacco disease leaf specimen is characterized by comprising the following steps:
(1) washing tobacco diseased leaves with tap water, and then soaking the tobacco diseased leaves in 5% copper sulfate solution for 12-24 hours;
(2) taking out the leaf treated in the step (1) and storing the leaf specimen in the tobacco disease leaf specimen immersion storage solution according to claim 1.
CN201810907354.2A 2018-08-09 2018-08-09 Tobacco disease leaf specimen dipping preservation liquid and specimen preparation method Expired - Fee Related CN108770840B (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102379280B (en) * 2011-08-05 2013-07-31 云南省烟草农业科学研究院 Tobacco leaf specimen treatment liquid and flue-cured tobacco leaf specimen batch production and preservation method
CN103283718A (en) * 2013-07-04 2013-09-11 云南农业大学 Method for manufacturing tobacco disease specimen
CN103404510A (en) * 2013-08-22 2013-11-27 湖北省烟草公司恩施州公司 Method for making tobacco rhizome disease specimens
CN103548818A (en) * 2013-11-25 2014-02-05 中国热带农业科学院香料饮料研究所 Preparation method for orchidaceae succulent plant dipping specimen
CN103766328A (en) * 2014-01-14 2014-05-07 云南省烟草农业科学研究院 Manufacturing method for plant leaf dipping specimen

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102379280B (en) * 2011-08-05 2013-07-31 云南省烟草农业科学研究院 Tobacco leaf specimen treatment liquid and flue-cured tobacco leaf specimen batch production and preservation method
CN103283718A (en) * 2013-07-04 2013-09-11 云南农业大学 Method for manufacturing tobacco disease specimen
CN103404510A (en) * 2013-08-22 2013-11-27 湖北省烟草公司恩施州公司 Method for making tobacco rhizome disease specimens
CN103548818A (en) * 2013-11-25 2014-02-05 中国热带农业科学院香料饮料研究所 Preparation method for orchidaceae succulent plant dipping specimen
CN103766328A (en) * 2014-01-14 2014-05-07 云南省烟草农业科学研究院 Manufacturing method for plant leaf dipping specimen

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
棕榈植物病害浸渍标本保绿技术研究;朱辉 等;《中国热带农业》;20090630;参见摘要 *
植物病害标本固绿保绿技术研究;黄艳花 等;《安徽农业科学》;20071231;第35卷(第29期);参见摘要 *
烟草病害标本制作方法;张永富 等;《中国烟草》;19861225;参见摘要 *

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