EP0191059A1 - Method for the early determination of plant disease - Google Patents
Method for the early determination of plant diseaseInfo
- Publication number
- EP0191059A1 EP0191059A1 EP19850903976 EP85903976A EP0191059A1 EP 0191059 A1 EP0191059 A1 EP 0191059A1 EP 19850903976 EP19850903976 EP 19850903976 EP 85903976 A EP85903976 A EP 85903976A EP 0191059 A1 EP0191059 A1 EP 0191059A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- differences
- plants
- nucleic acid
- detection
- healthy
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
Definitions
- the invention is a method in which the nucleic acid extracts from healthy and diseased plants are compared with one another and the health status of the plants can be deduced from the presence or absence of certain nucleic acid molecules.
- Infectious diseases are becoming more and more serious in the field of agriculture. This is, among other things, a consequence of the cultivation of highly refined varieties in monocultures.
- Current examples are fungal diseases in cereals, viral diseases in potatoes, sugar beet and fruit trees. It is in order to carry out damage surveys for forest diseases and to obtain information about how the disease spreads necessary to check the condition of the trees at certain time intervals.
- the tree stands are examined by forest officials and divided into three damage levels. Criteria for the assessment are, for example, the needling and leafing of the crowns, discoloration of needles and leaves, so-called anxiety drives and changes in trunk and bark.
- the symptom expression is only clear in very few cases, the division into the Subjective stages take place subjectively.
- the subject of the presented invention is a gel electrophoretic method, which among other things enables a reliable distinction between sick and healthy trees to be made very early.
- a gel electrophoretic examination is shown by way of example in FIG. 1.
- 1 g each of pine trees was obtained from an apparently healthy or sick tree in a Tris-HCL buffer at pH 8.0 and an ionic strength of 100 mM in the presence of 25th SDS and 25 M mercaptoethanol together with phenol and chloroform are homogenized in an Ultra Turrax. After filtration and re-extraction with phenol / chloroform, the nucleic acid was precipitated from the plant extract by adding ethanol. The precipitate was dissolved in 200 ul electrophoresis buffer.
- the nucleic acids were separated in a two-dimensional 5% polyacrylamide gel electrophoresis, initially under denaturing conditions at 8 M urea and 50 ° C. and then in the second dimension -3- ran at 20 ° C without urea. The gel was then stained with silver nitrate.
- Fig. 1b in der_ sample of the diseased tree indicated by the arrow 'band labeled zusäzlich to the bands of the sample of the healthy tree (Fig. 1a) seen surprisingly turned out that these additional nucleic acids place independently of the Stand ⁇ and Age of the pines in all trees affected by "forest death" can be proven.
- Significant differences in the ⁇ -sand pattern after two-dimensional gel electrophoresis have now also been found in diseased spruce and beech trees.
- the method can be applied to the diagnosis of diseases of agricultural crops.
- diseases of agricultural crops examples include viral diseases that occur in cereals, potatoes, sugar beets, fruit trees and grapevines.
- the method described has hitherto been used successfully in the Rizomania disease of sugar beet, which is caused by the Beet Necrotic Yellow Vein Virus (BNYVV), and in the vein yellowing of apples, which is caused by unidentified viruses .
- BNYVV Beet Necrotic Yellow Vein Virus
- the additional nucleic acid band caused by the disease is already visible in the one-dimensional gel electrophoresis. The result is confirmed by Fig. 2; the additional band in the sample of the sick plant (left side) compared to the sample of the healthy plant (right side) is marked by an arrow.
- aqueous two-phase system With the help of the aqueous two-phase system mentioned, the extraction of the nucleic acids can be carried out with great advantage.
- an aqueous two-phase system has proven to be suitable, which consists on the one hand of an aqueous solution of a neutral salt and on the other hand of an aqueous polyethylene glycol solution, the nucleic acids accumulating in the other, ie the brine phase.
- the Zu ⁇ by administration of ethanol precipitated nucleic acids in 8 parts of H.-0 and treated with 21 parts of 30% (w / w) potassium phosphate, and ' 11 parts 30% (w / w) polyethylene glycol 6000 mixed.
- the solution is briefly centrifuged to separate the phases.
- the nucleic acids are quantitatively in the lower, the salt phase, from which they are precipitated on ice with 0.1% cetyltrimethylammonium bromide (CTAB) for 30 min.
- CTAB cetyltrimethylammonium bromid
- Cetyltrimethylammonium bromide precipitate is washed twice with 70% ethanol, dried and taken up in H_0.
- the cleaning of the nucleic acids from the interfering plant constituents is so effective that differences in the nucleic acid pattern are already visible in the one-dimensional gel electrophoresis (see FIG. 2).
- the use of extraction with the aid of the aqueous two-phase system saves additional cleaning procedures in all subsequent steps.
- RNA or nucleic acid sequence it is entirely possible to carry out the tests for a specific RNA or nucleic acid sequence not only by gel electrophoresis, but also with the aid of modern molecular hybridization techniques. If such a nucleic acid is found in leaf or needle samples, such as the additional RNA in pine tissue in the example presented, it can be assumed that the tree is diseased, even if there has not yet been any visible symptom manifestation.
- the test thus enables objective early detection and mapping of the damage. Trees can be felled in good time before the wood quality has decreased due to the disease. Resistant or more resistant trees can be recognized early.
- the test is also very easy and quick to carry out and is suitable for serial examinations of a few hundred trees per person per examiner.
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Botany (AREA)
- Mycology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
On peut montrer que des modifications typiques existent dans des échantillons d'acide nucléique des plantes atteintes de maladie. Indépendamment du lieu d'origine et de l'âge des plantes, des bandes supplémentaires apparaissent lors de la séparation gel-électrophorétique à deux dimensions des acides nucléiques. Ces acides nucléiques supplémentaires peuvent être testés, par exemple avec des techniques d'hybridation moléculaire. Des diagnostics objectifs précoces peuvent ainsi être obtenus, qui permettent de prendre à temps les mesures de combat nécessaires ainsi que d'établir la cartographie de grands espaces des dégâts causés aux plantes. Le test est très simple et s'adapte à des investigations en série.It can be shown that typical modifications exist in nucleic acid samples from plants with disease. Regardless of the place of origin and the age of the plants, additional bands appear during the two-dimensional gel-electrophoretic separation of nucleic acids. These additional nucleic acids can be tested, for example with molecular hybridization techniques. Early objective diagnoses can thus be obtained, which make it possible to take the necessary combat measures in time as well as to establish the mapping of large areas of damage caused to plants. The test is very simple and adapts to serial investigations.
Description
Claims
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE3428386 | 1984-08-01 | ||
DE3428386 | 1984-08-01 | ||
DE3503978 | 1985-02-06 | ||
DE19853503978 DE3503978A1 (en) | 1984-08-01 | 1985-02-06 | METHOD FOR THE EARLY DETECTION OF PLANT DISEASES |
Publications (1)
Publication Number | Publication Date |
---|---|
EP0191059A1 true EP0191059A1 (en) | 1986-08-20 |
Family
ID=25823503
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP19850903976 Withdrawn EP0191059A1 (en) | 1984-08-01 | 1985-07-25 | Method for the early determination of plant disease |
Country Status (4)
Country | Link |
---|---|
EP (1) | EP0191059A1 (en) |
AU (1) | AU4723185A (en) |
DE (1) | DE3503978A1 (en) |
WO (1) | WO1986000934A1 (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
NL8801147A (en) * | 1988-05-02 | 1989-12-01 | Tno | METHOD FOR DETECTING GENETIC VARIATION, AND SUITABLE KIT. |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4405720A (en) * | 1981-03-04 | 1983-09-20 | The United States Of America As Represented By The Department Of Health And Human Services | Silver stains for protein in gels |
US4480040A (en) * | 1981-12-03 | 1984-10-30 | The United States Of America As Represented By The Secretary Of Agriculture | Sensitive and rapid diagnosis of viroid diseases and viruses |
-
1985
- 1985-02-06 DE DE19853503978 patent/DE3503978A1/en not_active Withdrawn
- 1985-07-25 WO PCT/DE1985/000251 patent/WO1986000934A1/en not_active Application Discontinuation
- 1985-07-25 AU AU47231/85A patent/AU4723185A/en not_active Abandoned
- 1985-07-25 EP EP19850903976 patent/EP0191059A1/en not_active Withdrawn
Non-Patent Citations (1)
Title |
---|
See references of WO8600934A1 * |
Also Published As
Publication number | Publication date |
---|---|
DE3503978A1 (en) | 1986-02-06 |
WO1986000934A1 (en) | 1986-02-13 |
AU4723185A (en) | 1986-02-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
DE1698180B2 (en) | METHOD OF MANUFACTURING A BLOOD SERUM REFERENCE STANDARD | |
DE3402647C2 (en) | Process for the recovery of colony stimulating factor and kallikrein from human urine | |
DE2602997C2 (en) | Lyophilized collagen preparation and process for their manufacture | |
DE102005013261A1 (en) | Reagent and method for preventing time-dependent expression in biological cells | |
DE2819574A1 (en) | METHOD AND DEVICE FOR PROCESSING A BODY FLUID SAMPLE | |
DE4327429C2 (en) | Method and device for brain wave analysis | |
McCormick | Notes on the anatomy of the young tuber of Ipomoea batatas Lam | |
EP0191059A1 (en) | Method for the early determination of plant disease | |
DE3424640C2 (en) | ||
DD239672A5 (en) | PROCESS FOR THE FRONTAL IDENTIFICATION OF PLANT DISEASES | |
DE1076888B (en) | Process for the production of active substances that promote breathing for therapeutic purposes | |
EP0547465B1 (en) | Process of preparation of extracts of Petasites rhizomas | |
DE60003723T2 (en) | INCREASING THE BIOLOGICAL ACTIVITY OF PLANTS | |
EP1984740B1 (en) | Sample collection method | |
DE675605C (en) | Process for the separation and extraction of the valuable components of bee and snake venom | |
DE4221537A1 (en) | Dry witch hazel extract, process for its preparation and its use as a medicine | |
DE2657926B2 (en) | Method for determining the antistreptolysin content in the blood | |
DE2205897C3 (en) | Process for the preparation of antithymocyte serum | |
DD202600A5 (en) | METHOD FOR THE INVESTIGATION OF CELLS BY ELECTROPHORESIS | |
AT352292B (en) | PROCEDURES FOR DETERMINING INFECTIOUS AND IMMUNOLOGICAL PROCEDURES AND MEANS OF BLOOD TESTS | |
DE202022104675U1 (en) | A system for the botanical and physico-chemical standardization of Murraya koengii | |
Häusler et al. | Zur Wirkung von Allylchlorid bei chronischer gewerblicher Exposition | |
DE19530332C2 (en) | Extraction of fungal cell DNA and methods of detecting fungal cells in clinical material | |
Wuensch et al. | Verteilung der Molybdatoarsensäure und Molybdänsäure zwischen wäßriger Lösung und einigen organischen Lösungsmitteln | |
DE3115761A1 (en) | METHOD FOR PRODUCING UNCOUTLABLE BLOOD USING PROTEOLYTIC ENZYMES AND PROTEIN CONCENTRATE OBTAINED therefrom |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AT BE CH DE FR GB IT LI LU NL SE |
|
17P | Request for examination filed |
Effective date: 19860813 |
|
RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: SCHUMACHER, JUERGEN Owner name: RIESNER, DETLEV |
|
17Q | First examination report despatched |
Effective date: 19870812 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
18D | Application deemed to be withdrawn |
Effective date: 19871223 |