MD1658Z - Method for treating seed potatoes against the nematode Ditylenchus destructor - Google Patents
Method for treating seed potatoes against the nematode Ditylenchus destructorInfo
- Publication number
- MD1658Z MD1658Z MDS20210085A MDS20210085A MD1658Z MD 1658 Z MD1658 Z MD 1658Z MD S20210085 A MDS20210085 A MD S20210085A MD S20210085 A MDS20210085 A MD S20210085A MD 1658 Z MD1658 Z MD 1658Z
- Authority
- MD
- Moldova
- Prior art keywords
- destructor
- bacillus cereus
- nematode
- potato
- fluorescens
- Prior art date
Links
- 244000061456 Solanum tuberosum Species 0.000 title claims abstract description 74
- 235000002595 Solanum tuberosum Nutrition 0.000 title claims abstract description 72
- 241000399948 Ditylenchus destructor Species 0.000 title claims abstract description 49
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- DUEPRVBVGDRKAG-UHFFFAOYSA-N carbofuran Chemical compound CNC(=O)OC1=CC=CC2=C1OC(C)(C)C2 DUEPRVBVGDRKAG-UHFFFAOYSA-N 0.000 description 2
- UQHKFADEQIVWID-UHFFFAOYSA-N cytokinin Natural products C1=NC=2C(NCC=C(CO)C)=NC=NC=2N1C1CC(O)C(CO)O1 UQHKFADEQIVWID-UHFFFAOYSA-N 0.000 description 2
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Landscapes
- Pretreatment Of Seeds And Plants (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
Description
Invenţia se referă la parazitologie, în special la un procedeu de combatere biologică a nematodului fitoparazit Ditylenchus destructor la cartofi seminceri, şi poate fi utilizat în agricultură. The invention relates to parasitology, in particular to a process for biological control of the phytoparasitic nematode Ditylenchus destructor in seed potatoes, and can be used in agriculture.
Este cunoscut faptul că una dintre bolile cartofilor, mai nocivă şi cu un impact economic major, este ditilenhoza tuberculilor - boală cauzată de nematodul Ditylenchus destructor Thorne, 1945, care se enumără printre cei mai periculoşi paraziţi, exclusiv al tuberculilor. D. destructor parazitează mai mult de 90 de specii de plante superioare, însă principala gazdă sunt tuberculii de cartof Solanum tuberosum. Pe o scară de 1:5, privind potenţialul dăunător, acesta a fost încadrat la valoarea 5, fiind considerată o specie cu capacitate de dăunare foarte mare, în special în perioada de păstrare. Principala sursă de răspândire a parazitului este materialul semincer infestat în fazele 1-2 de ditilenhoză (când simptomele de boală nu sunt clar vizibile), destinat plantării primăvara. It is known that one of the potato diseases, more harmful and with a major economic impact, is tuber dithylenosis - a disease caused by the nematode Ditylenchus destructor Thorne, 1945, which is listed among the most dangerous parasites, exclusively of tubers. D. destructor parasitizes more than 90 species of higher plants, but the main host is the potato tubers Solanum tuberosum. On a scale of 1:5, regarding the harmful potential, it was ranked at the value 5, being considered a species with very high damaging capacity, especially during the storage period. The main source of spread of the parasite is the seed material infested in phases 1-2 of dithylenosis (when the symptoms of the disease are not clearly visible), intended for planting in the spring.
Este cunoscut procedeul de tratare a materialului semincer de cartof contra nematodului Ditylenchus destructor, în care sunt utilizate următoarele preparate de origine chimică: Carbofuran 5%, Furadan 5%, Heterofos 0,05%, Carbation 0,5% [1]. The process of treating potato seed material against the nematode Ditylenchus destructor is known, in which the following preparations of chemical origin are used: Carbofuran 5%, Furadan 5%, Heterofos 0.05%, Carbation 0.5% [1].
Neajunsul acestor preparate constă în aceea că acestea sunt de origine chimică şi foarte toxice. Utilizarea lor provoacă riscuri de poluare a apei, solului şi faunei, ceea ce afectează grav sănătatea omului şi animalelor. Reieşind din cele expuse, autorii recomandă ca preparatele chimice să fie utilizate doar în cazul focarelor de fitohelmintoze, depistate la cultura cartofului. Actualmente, folosirea produselor nematocide de origine chimică a fost radical redusă în majoritatea ţărilor. Pentru combaterea nematozilor culturilor agricole, printre care Ditylenchus destructor, sunt căutate alternative mai puţin toxice. The disadvantage of these preparations is that they are of chemical origin and very toxic. Their use causes risks of water, soil and fauna pollution, which seriously affects human and animal health. Based on the above, the authors recommend that chemical preparations be used only in the case of phytohelminthiasis outbreaks detected in potato crops. Currently, the use of nematocidal products of chemical origin has been radically reduced in most countries. To combat nematodes of agricultural crops, including Ditylenchus destructor, less toxic alternatives are sought.
Mai sunt cunoscute procedee de tratare a cartofului semincer infestat, înainte de plantare în special, prin procedee cu utilizarea biopreparatelor pe bază de microorganisme saprofite antagoniste. Acestea nu exercită simptome de toxicitate asupra dezvoltării plantelor, dimpotrivă stimulează mecanismele de protecţie şi rezistenţă către diverşi agenţi patogeni, iar în final contribuie la sporirea recoltelor. Aşadar, aplicarea produselor de protecţie a plantelor pe bază de microorganisme saprofite antagoniste contra nematodelor este o soluţie reală şi de perspectivă de rezolvare a problemei în cauză. Astfel, pentru tratarea cartofului semincer contra nematodului Ditylenchus destructor se recomandă înmuierea cartofului înainte de sădire într-o soluţie ce conţine lichid de cultură al tulpini de bacterii Pseudomonas fluorescens CNM-PFB-01 cu un titru de 109 cel./ml şi apă, luate în raport de 1:(100-500) respectiv, în decurs de 20 de ore [2]. There are also known methods of treating infested seed potatoes, before planting in particular, by using biopreparations based on antagonistic saprophytic microorganisms. These do not exert symptoms of toxicity on plant development, on the contrary, they stimulate the mechanisms of protection and resistance to various pathogens, and ultimately contribute to increasing yields. Therefore, the application of plant protection products based on antagonistic saprophytic microorganisms against nematodes is a real and promising solution to solve the problem in question. Thus, for treating seed potatoes against the nematode Ditylenchus destructor, it is recommended to soak the potato before planting in a solution containing culture liquid of the bacterial strain Pseudomonas fluorescens CNM-PFB-01 with a titer of 109 cells/ml and water, taken in a ratio of 1:(100-500) respectively, within 20 hours [2].
Dezavantajul acestui procedeu constă în nivelul scăzut de creştere a productivităţii plantelor. The disadvantage of this process is the low level of plant productivity growth.
În calitate de cea mai apropiată soluţie pentru tratarea cartofului semincer contra nematodului Ditylenchus destructor este procedeul de înmuiere a cartofului înainte de sădire într-o soluţie ce conţine lichid de cultură al tulpinii de bacterii Rizobium japonicum CNMN- RB-06 cu titrul de 109 cel./ml şi apă, luate în raport de 1:200 respectiv, în decurs de 72 ore [3]. As the closest solution for treating seed potatoes against the Ditylenchus destructor nematode, the procedure of soaking the potato before planting in a solution containing culture liquid of the bacterial strain Rhizobium japonicum CNMN-RB-06 with a titer of 109 cells/ml and water, taken in a ratio of 1:200 respectively, within 72 hours [3].
Dezavantajul acestui procedeu constă în nivelul scăzut de creştere a productivităţii plantelor. The disadvantage of this process is the low level of plant productivity growth.
Actualmente, în majoritatea ţărilor dezvoltate (SUA, India, Germania, Franţa, Marea Britanie, Brazilia, Federaţia Rusă etc.), pentru combaterea speciilor parazite de nematozi sunt elaborate şi utilizate biopreparate nematocide pe bază de fungi prădători şi bacterii parazite din genurile Arthrobotrys, Catenaria, Paecilomyces, Pasteuria, Pseudomonas, Rhizobium, Bacillus etc. În sursele literare nu este descrisă acţiunea nematocidă a speciei de bacterii saprofite Bacillus cereus asupra nematodului tuberculilor de cartofi Ditylenchus destructor. Însă, conform standardelor internaţionale, cartoful semincer trebuie să fie liber de speciile parazite de nematozi nocivi pentru această cultură, printre care Meloidogyne chitwoodi, Meloidogyne fallax şi Ditylenchus destructor (standardul pentru cartofii de sămânţă EAK OOH S-1). Currently, in most developed countries (USA, India, Germany, France, Great Britain, Brazil, Russian Federation, etc.), to combat parasitic nematode species, nematocidal biopreparations based on predatory fungi and parasitic bacteria from the genera Arthrobotrys, Catenaria, Paecilomyces, Pasteuria, Pseudomonas, Rhizobium, Bacillus, etc. are developed and used. The nematocidal action of the saprophytic bacterial species Bacillus cereus on the potato tuber nematode Ditylenchus destructor is not described in literary sources. However, according to international standards, seed potatoes must be free from parasitic nematode species harmful to this crop, including Meloidogyne chitwoodi, Meloidogyne fallax and Ditylenchus destructor (EAK OOH S-1 seed potato standard).
Problema pe care o soluţionează prezenta invenţie este combaterea eficientă a nematodului Ditylenchus destructor, prezent la cartoful semincer, cu utilizarea unui biopreparat, care manifestă suplimentar acţiune stimulatoare asupra dezvoltării şi productivităţii acestora. The problem solved by the present invention is the effective control of the nematode Ditylenchus destructor, present in seed potatoes, with the use of a biopreparation, which additionally exhibits a stimulating action on their development and productivity.
Speciile de bacterii aerobe, formatoare de spori din genul Bacillus, se includ în grupa microorganismelor de mare perspectivă în biotehnologia actuală, ca producătoare ale substanţelor biologic active. The species of aerobic, spore-forming bacteria of the genus Bacillus are included in the group of microorganisms with great potential in current biotechnology, as producers of biologically active substances.
Esenţa invenţiei constă în faptul, că se propune un procedeu de tratare a cartofului semincer contra nematodului Ditylenchus destructor, care include înmuierea cartofului semincer infestat înainte de plantare într-o soluţie ce conţine lichid cultural (LC) al tulpinii de bacterii Bacillus cereus var. fluorescens CNMN-BB-07 şi apă, luate în raport de 1:400, în decurs de 16 ore. Lichidul cultural se obţine prin cultivarea bacteriilor pe un mediu agarizat până la titrul de 109 cel./ml şi centrifugare. The essence of the invention lies in the fact that a method is proposed for treating seed potatoes against the nematode Ditylenchus destructor, which includes soaking the infested seed potatoes before planting in a solution containing culture liquid (CL) of the Bacillus cereus var. fluorescens CNMN-BB-07 bacterial strain and water, taken in a ratio of 1:400, within 16 hours. The culture liquid is obtained by cultivating the bacteria on an agarized medium to a titer of 109 cells/ml and centrifugation.
Rezultatul tehnic al invenţiei constă combaterea nematodului fitoparazit al tuberculilor Ditylenchus destructor în proporţie de 99%, şi suplimentar stimularea încolţirii cartofului semincer, sporirea creşterii, dezvoltării şi productivităţii plantelor. The technical result of the invention consists in combating the phytoparasitic tuber nematode Ditylenchus destructor in a proportion of 99%, and additionally stimulating the germination of seed potatoes, increasing the growth, development and productivity of plants.
Conform surselor bibliografice, tulpina Bacillus cereus var. fluorescens posedă capacitatea de a reduce bolile provocate de diverşi agenţi patogeni ai plantelor - fungi, bacterii, viruşi, determinând paralel, acţiune stimulatoare asupra dezvoltării plantelor prin capacitatea sa de a produce substanţe de tipul auxinelor, citochininelor, giberelinelor. According to bibliographic sources, the Bacillus cereus var. fluorescens strain has the ability to reduce diseases caused by various plant pathogens - fungi, bacteria, viruses, while also having a stimulating effect on plant development through its ability to produce substances such as auxins, cytokinins, and gibberellins.
Bacteriile Bacillus cereus sintetizează un pigment galben-verzui şi substanţe antibiotice de natură polipeptidică, care manifestă proprietăţi antimicrobiene pronunţate asupra speciilor de fungi fitopatogeni, printre care din genul Fusarium - F. oxysporum, F. solani, care provoacă diferite boli grave tuberculilor de cartofi. Bacillus cereus bacteria synthesize a yellow-green pigment and antibiotic substances of a polypeptide nature, which exhibit pronounced antimicrobial properties on phytopathogenic fungal species, including those from the genus Fusarium - F. oxysporum, F. solani, which cause various serious diseases of potato tubers.
Tulpina de bacterii Bacillus cereus var. fluorescens a fost izolată din solul de ciornoziom carbonatic gras, mediu lutos, humus 3,5, în Republica Moldova, de către cercetătorul ştiinţific Valerina Slanina din cadrul Institutului de Microbiologie şi Biotehnologie al MEC în perioada anului 2005, şi este depozitată în Colecţia Naţională de Microorganisme Nepatogene (CNMN) al aceluiaşi Institut. The bacterial strain Bacillus cereus var. fluorescens was isolated from the rich carbonate chernozem soil, loamy medium, humus 3.5, in the Republic of Moldova, by the scientific researcher Valerina Slanina from the Institute of Microbiology and Biotechnology of the Ministry of Education and Culture in 2005, and is stored in the National Collection of Non-pathogenic Microorganisms (CNMN) of the same Institute.
Pentru cultivarea şi multiplicarea tulpinii Bacillus cereus var. fluorescens sunt utilizate mediile cu agar de cartofi, cât şi mediile Petrenco cu următoarea componenţă: peptonă-15,0%; glicerină-2,0%; MgCl2-0,7%; K2SO4-2,0%; agar-2,0% şi 1 litru de apă distilată (pH = 7,0-7,2). Cultivarea se realizează în termostat la temperatura de +28°-+30° C, timp de 24-48 ore. Apoi prin centrifugare este obţinut Lichidul cultural (LC) sau supernatantul. For the cultivation and multiplication of the Bacillus cereus var. fluorescens strain, potato agar media and Petrenco media with the following composition are used: peptone-15.0%; glycerin-2.0%; MgCl2-0.7%; K2SO4-2.0%; agar-2.0% and 1 liter of distilled water (pH = 7.0-7.2). Cultivation is carried out in a thermostat at a temperature of +28°-+30° C, for 24-48 hours. Then, by centrifugation, the culture liquid (LC) or supernatant is obtained.
Lichidul cultural al tulpinii de bacterii Bacillus cereus var. fluorescens obţinut a fost utilizat la testarea contactului direct (in vitro) cu nematodul D. destructor. The culture liquid of the Bacillus cereus var. fluorescens bacterial strain obtained was used to test direct contact (in vitro) with the nematode D. destructor.
Obţinerea rezultatului menţionat a fost posibilă datorită utilizării lichidului cultural al tulpinii de bacterii Bacillus cereus var. fluorescens. Obtaining the aforementioned result was possible due to the use of the culture liquid of the Bacillus cereus var. fluorescens bacterial strain.
Rezultatul invenţiei este ilustrat în figurile 1- 6, care reprezintă: The result of the invention is illustrated in figures 1-6, which represent:
- fig. 1, eficacitatea letală (%) a LC al tulpinii de bacterii Bacillus cereus var. fluorescens CNMN-BB-07, luat în diferite concentraţii, la diferite intervale de timp, în contact cu D. destructor; - Fig. 1, lethal efficacy (%) of the LC of the bacterial strain Bacillus cereus var. fluorescens CNMN-BB-07, taken in different concentrations, at different time intervals, in contact with D. destructor;
- fig. 2, acţiunea destructivă a LC al tulpinii de bacterii Bacillus cereus asupra nematozilor Ditylenchus: a - înainte de contact; b, c, d, e - la diferite intervale de timp; - Fig. 2, the destructive action of LC of the Bacillus cereus bacterial strain on Ditylenchus nematodes: a - before contact; b, c, d, e - at different time intervals;
- fig. 3, a - cartofi seminceri inoculaţi cu D. destructor; b - după incubare, înainte de plantare; - fig. 3, a - seed potatoes inoculated with D. destructor; b - after incubation, before planting;
- fig. 4, cartofi experimentali: a - în proces de plantare; b - fenofaza înflorire-maturizare; - fig. 4, experimental potatoes: a - in the planting process; b - flowering-maturation phenophase;
- fig. 5, recoltele cartofilor experimentali (tuberculi/parcelă), obţinute din: a- parcela Variantei V2 - cartofi seminceri inoculaţi, trataţi înainte de plantare cu LC al tulpinii de bacterii Bacillus cereus (1:400), la un interval de timp de 16 ore; b- parcela Martorului M1- inoculaţi cu D. destructor, nontrataţi; - fig. 5, experimental potato yields (tubers/plot), obtained from: a- Variant V2 plot - inoculated seed potatoes, treated before planting with LC of the Bacillus cereus bacteria strain (1:400), at a time interval of 16 hours; b- Control M1 plot - inoculated with D. destructor, untreated;
- fig. 6, cartofi experimentali Roko, recoltă, tuberculi/cuib: a - materialul semincer inoculat, prelucrat cu LC al tulpinii de bacterii Bacillus cereus - V2; b - martorul M1, materialul semincer inoculat, nontratat (original). - fig. 6, experimental Roko potatoes, harvest, tubers/nest: a - inoculated seed material, processed with LC of the Bacillus cereus - V2 bacterial strain; b - control M1, inoculated, untreated seed material (original).
Analizele de laborator au fost efectuate la 14 soiuri de tuberculi de cartofi seminceri colectaţi din depozite în perioada de toamnă-iarnă-primăvară din diverse gospodării ale zonelor de Nord, Centru şi Sud ale Republicii Moldova. Rezultatul investigaţiilor contra fitohelmintozelor la soiurile de cartofi seminceri au demonstrat că sunt infestate de D. destructor în proporţii variate. Prezenţa parazitului a fost depistată în majoritatea cazurilor la cartoful semincer din gospodăriile individuale (60% din totalul gospodăriilor cercetate). Laboratory analyses were performed on 14 varieties of seed potato tubers collected from storage during the autumn-winter-spring period from various households in the North, Center and South of the Republic of Moldova. The results of investigations against phytohelminthiasis on seed potato varieties showed that they are infested with D. destructor in varying proportions. The presence of the parasite was detected in most cases on seed potatoes from individual households (60% of all households surveyed).
Pentru aprecierea gradului de atac al nematodului Ditylenchus destructor a fost utilizată scara de 5 grade (după Ivaniuc, Iliaşenco, 2010): gradul I - soiuri de cartofi rezistente la invazie (0-1,0%); gradul II - soiuri slab infestate (1,0-10,0%); gradul III - moderat infestate (11,0-25,0%); gradul IV - serios infestate (26,0-50%); gradul V - puternic infestate, atacate mai mult de 50%. To assess the degree of attack of the nematode Ditylenchus destructor, a 5-grade scale was used (after Ivaniuc, Iliaşenco, 2010): grade I - potato varieties resistant to invasion (0-1.0%); grade II - lightly infested varieties (1.0-10.0%); grade III - moderately infested (11.0-25.0%); grade IV - seriously infested (26.0-50%); grade V - heavily infested, more than 50% attacked.
Conform acestei scări, cartofii cercetaţi au o infestare de 11-30%, în dependenţă de zonă şi corespund soiurilor moderat afectate. According to this scale, the potatoes studied have an infestation of 11-30%, depending on the area, and correspond to moderately affected varieties.
Cercetările au fost axate pe unul dintre soiurile cultivate pe larg în Republica Moldova - Roko. Conform Catalogului Soiurilor de Plante cultivate în Republica Moldova (2016), elaborat de Ministerul Agriculturii şi Industriei Alimentare, soiul Roko a fost înregistrat în anul 2011, iar ulterior reînregistrat. De asemenea, cartoful Roko este inclus în Catalogul Comun al Soiurilor de Plante Agricole, elaborat de Uniunea Europeană (2019). Cartoful alimentar de soiul Roko posedă aspect calitativ ca marfă, indicele calităţii fiind de 95%, este preferat de cumpărători datorită gustului. Cantitatea de amidon constituie 12-16%, fiind apreciaţi ca cartofi alimentari, iar recolta poate atinge la 600 q/ha. The research was focused on one of the varieties widely cultivated in the Republic of Moldova - Roko. According to the Catalogue of Plant Varieties Cultivated in the Republic of Moldova (2016), developed by the Ministry of Agriculture and Food Industry, the Roko variety was registered in 2011, and later re-registered. The Roko potato is also included in the Common Catalogue of Agricultural Plant Varieties, developed by the European Union (2019). The Roko food potato has a qualitative aspect as a commodity, the quality index being 95%, it is preferred by buyers due to its taste. The amount of starch is 12-16%, being appreciated as food potatoes, and the harvest can reach 600 q/ha.
Este cunoscut faptul că principala sursă de răspândire a nematodului Ditylenchus destructor la cultivarea cartofului sunt tuberculii infestaţi, care se plantează primăvara ca material semincer. O altă sursă de infestare a cartofului o constituie solul, unde nematozii pot rămâne activi pe o perioadă de aproximativ 3-4 ani. It is known that the main source of spread of the nematode Ditylenchus destructor in potato cultivation are infested tubers, which are planted in spring as seed material. Another source of potato infestation is the soil, where the nematodes can remain active for a period of approximately 3-4 years.
Conform celor 5 faze de dezvoltare a ditilenhozei, evaluate în cercetările precedente efectuate la soiul Roko, s-a observat, că cei mai periculoşi sunt cartofii de sămânţă în primele faze (1, 2), deoarece simptomele exterioare, care apar pe tuberculi în aceste faze, încă nu sunt vizibile nici la o examinare minuţioasă, de aceea asemenea cartofi, din neatenţie ar putea fi plantaţi. According to the 5 stages of dithylinosis development, evaluated in previous research conducted on the Roko variety, it was observed that the most dangerous are seed potatoes in the first stages (1, 2), because the external symptoms that appear on the tubers in these stages are not yet visible even upon careful examination, therefore such potatoes could be inadvertently planted.
În faza a 2-a simptomele de deteriorare la suprafaţa tuberculilor sunt observate doar la înlăturarea cojii. Conform recalculelor efectuate, în asemenea porţiuni are loc popularea monotipică primară a ţesutului vegetal (doar cu populaţia speciei parazite de nematozi- Ditylenchus destructor), cele mai numeroase (50% din totalul de unităţi/gram) fiind formele juvenile ale nematodului (J2, J3, J4) cu dominanţa J2 (n=10,1x103 indivizi/gram ţesut infestat). Numărul excesiv de larve, ce se hrănesc intens cu ţesutul sănătos al pulpei cartofului, în decursul a 18-20 de zile ajunge la stadiul matur. O singură femelă depune circa 200-250 de ouă. În condiţiile republicii, nematodul D. destructor formează 4-5 generaţii, iar reproducerea intensă a parazitului provoacă majorarea densităţii indivizilor şi, în consecinţă, reducerea recoltei. In the 2nd phase, the symptoms of damage on the surface of the tubers are observed only when the peel is removed. According to the recalculations performed, in such portions the primary monotypic population of the plant tissue takes place (only with the population of the parasitic nematode species - Ditylenchus destructor), the most numerous (50% of the total units/gram) being the juvenile forms of the nematode (J2, J3, J4) with the dominance of J2 (n=10.1x103 individuals/gram of infested tissue). The excessive number of larvae, which feed intensively on the healthy tissue of the potato pulp, reaches the mature stage within 18-20 days. A single female lays about 200-250 eggs. Under the conditions of the republic, the nematode D. destructor forms 4-5 generations, and the intense reproduction of the parasite causes an increase in the density of individuals and, consequently, a reduction in the harvest.
Exemple de realizare a invenţiei Examples of embodiments of the invention
Deoarece nu dispunem de informaţii despre capacitatea nematocidă a tulpinii de bacterii cercetate Bacillus cereus var. fluorescens CNMN-BB-07 în contact cu nematozii fitoparaziţi ai tuberculilor de cartofi D. destructor, precum şi despre acţiunea acesteia asupra creşterii şi dezvoltării plantelor de cartofi, a fost necesar de efectuat testarea Bacillus cereus în două direcţii, conform exemplelor nr. 1 şi nr. 2. Since we do not have information about the nematocidal capacity of the investigated bacterial strain Bacillus cereus var. fluorescens CNMN-BB-07 in contact with the phytoparasitic nematodes of potato tubers D. destructor, as well as about its action on the growth and development of potato plants, it was necessary to test Bacillus cereus in two directions, according to examples no. 1 and no. 2.
Exemplul 1- Testarea contactului direct (in vitro) a LC al tulpinii de bacterii Bacillus cereus var. fluorescens CNMN-BB-07, cu nematodul fitoparazit al tuberculilor de cartofi - Ditylenchus destructor. Example 1- Testing the direct contact (in vitro) of the LC of the bacteria strain Bacillus cereus var. fluorescens CNMN-BB-07, with the phytoparasitic nematode of potato tubers - Ditylenchus destructor.
Exemplul 2 - Testarea (in vivo) a LC al tulpinii de bacterii Bacillus cereus var. fluorescens CNMN-BB-07, în cazul contactului cu cartofii seminceri de soiul Roko, infestaţi cu nematodul tuberculilor de cartofi Ditylenchus destructor. Example 2 - Testing (in vivo) of the LC of the bacterial strain Bacillus cereus var. fluorescens CNMN-BB-07, in case of contact with seed potatoes of the Roko variety, infested with the potato tuber nematode Ditylenchus destructor.
Exemplul 1 Example 1
Testarea a avut loc în condiţii de laborator prin metoda de contact direct (Kurt, 1973) al LC al tulpinii de bacterii Bacillus cereus var. fluorescens CNMN-BB-07 (6x109 cel./ml), cu nematodul fitoparazit al tuberculilor de cartofi Ditylenchus destructor. Contactul a avut loc în cutiile Petri cu diametrul de 5 cm (fiecare variantă fiind luată în 3 reprize) unde în fiecare cutie s-au adăugat, cu acul entomologic steril, câte 50 de indivizi maturi (femele + masculi) de D. destructor. Apoi în cutiile Petri, peste nematozii selectaţi, s-a adăugat LC al tulpinii de bacterii Bacillus cereus. Testarea a avut loc în 4 variante - V1, V2, V3, V4 şi un martor - M, în raportul următor (fig.1): The testing took place under laboratory conditions by the direct contact method (Kurt, 1973) of the LC of the Bacillus cereus var. fluorescens CNMN-BB-07 bacterial strain (6x109 cells/ml) with the potato tuber nematode Ditylenchus destructor. The contact took place in Petri dishes with a diameter of 5 cm (each variant being taken in 3 batches) where 50 mature individuals (females + males) of D. destructor were added to each dish, using a sterile entomological needle. Then, in the Petri dishes, over the selected nematodes, LC of the Bacillus cereus bacterial strain was added. The testing took place in 4 variants - V1, V2, V3, V4 and a control - M, in the following ratio (fig.1):
1. V1 - D. destructor + LC al tulpinii Bacillus cereus var. fluorescens CNMN-BB-07, nediluat; 1. V1 - D. destructor + LC of the Bacillus cereus var. fluorescens strain CNMN-BB-07, undiluted;
2. V2 - D. destructor + LC al tulpinii Bacillus cereus var. fluorescens CNMN-BB-07, în diluţie de 1:50 ; 2. V2 - D. destructor + LC of the Bacillus cereus var. fluorescens strain CNMN-BB-07, in a dilution of 1:50;
3. V3 - D. destructor + LC al tulpinii Bacillus cereus var. fluorescens CNMN-BB-07, în diluţie de 1:100 ; 3. V3 - D. destructor + LC of the Bacillus cereus var. fluorescens strain CNMN-BB-07, in a dilution of 1:100;
4. V4 - D. destructor + LC al tulpinii Bacillus cereus var. fluorescens CNMN-BB-07, în diluţie de 1:200 ; 4. V4 - D. destructor + LC of the Bacillus cereus var. fluorescens strain CNMN-BB-07, in a dilution of 1:200;
5. M - D. destructor + apă distilată (martorul). 5. M - D. destructor + distilled water (control).
Experienţele au avut loc la temperatura cuprinsă între 25-27°C, la diverse intervale de timp: 18, 24, 48 ore şi 5-7-14 zile. În calitate de martor (M) au fost luaţi nematozii D. destructor în contact cu apă distilată. The experiments took place at a temperature between 25-27°C, at various time intervals: 18, 24, 48 hours and 5-7-14 days. As a control (M), D. destructor nematodes in contact with distilled water were taken.
Conform rezultatelor obţinute, testarea in vitro a demonstrat eficacitate nematocidă sporită, în varianta V1 (LC, luat în concentraţie iniţială) (fig. 1). According to the results obtained, in vitro testing demonstrated increased nematocidal efficacy in variant V1 (LC, taken in initial concentration) (fig. 1).
Rezultatele experienţelor au demonstrat, că are loc o reacţie momentană a nematozilor, care capătă formă de spirală, chiar în primele 20-60 minute de contact. Eficacitatea letală a LC Bacillus cereus var. fluorescens CNMN-BB-07 în concentraţie iniţială (V1) a constituit 98,5-100% la o expunere de 24 ore. LC al tulpinii testate fiind diluat în proporţii de 1:50 (V2); 1:100 (V3) şi 1:200 (V4), de asemenea, posedă activitate nematocidă. Astfel, la o expunere de 48 de ore efectul letal a constituit: 50%; 40% şi 27,3%, respectiv (fig. 1). The results of the experiments demonstrated that a momentary reaction of nematodes occurs, which takes the form of a spiral, even in the first 20-60 minutes of contact. The lethal efficacy of LC Bacillus cereus var. fluorescens CNMN-BB-07 in the initial concentration (V1) was 98.5-100% at a 24-hour exposure. LC of the tested strain being diluted in proportions of 1:50 (V2); 1:100 (V3) and 1:200 (V4), also possesses nematocidal activity. Thus, at a 48-hour exposure the lethal effect was: 50%; 40% and 27.3%, respectively (fig. 1).
Modul de acţiune asupra nematodului: distrugerea completă a organelor interne, deformarea corpului (fig. 2: b; c; d; e). În majoritatea cazurilor, la femele, simptomele pronunţate de contact apar în porţiunile unde este situat orificiul vulvar al acestora, precum şi gonada cu ouă din interiorul corpului. A fost evidenţiat faptul că masa omogenă descompusă a organelor interne, formată în rezultatul contactului cu bacteriile, este eliminată în afara corpului prin rupturile cuticulei. Paralel s-a observat eliminarea unei substanţe gazoase. Este cunoscut faptul că speciile de bacterii antagoniste aerobe din genul Bacillus produc un şir de enzime: transferaze, hidrolaze, lipaze, care dezagreghează amidonul, pectina, celuloza, grăsimile, proteinele. Din figura 2 (c, d, e) rezultă că bacteriile studiate, în contact cu nematodul D. destructor exercită o acţiune a enzimelor produse de aceste bacterii, datorită cărora organele interne ale nematozilor sunt supuse proceselor de hidroliză. Mode of action on the nematode: complete destruction of internal organs, deformation of the body (Fig. 2: b; c; d; e). In most cases, in females, pronounced symptoms of contact appear in the portions where their vulvar orifice is located, as well as the gonad with eggs inside the body. It has been shown that the homogeneous decomposed mass of internal organs, formed as a result of contact with bacteria, is eliminated outside the body through the ruptures of the cuticle. At the same time, the elimination of a gaseous substance was observed. It is known that species of aerobic antagonistic bacteria of the genus Bacillus produce a number of enzymes: transferases, hydrolases, lipases, which disaggregate starch, pectin, cellulose, fats, proteins. From figure 2 (c, d, e) it results that the studied bacteria, in contact with the nematode D. destructor, exert an action of enzymes produced by these bacteria, due to which the internal organs of the nematodes are subjected to hydrolysis processes.
Conform rezultatelor obţinute menţionăm, că lichidul de cultură al bacteriei Bacillus cereus în diverse concentraţii (nediluat şi în diluţii de 1:50; 1:100; 1:200), la o expunere de 24-48 de ore de contact direct cu nematozii D. destructor, exercită eficacitate sporită, mortalitatea fiind de 98-50-40 şi 27,3%, respectiv. Menţionăm faptul că bacteriile Bacillus cereus posedă paralel activitate sporită asupra speciilor de fungi patogeni pentru cultura cartofului din genul Fusarium. Însă, pentru testare in vivo, în scopul evitării acţiunii fitotoxice asupra dezvoltării plantelor de cartof în perioada de vegetaţie, se recomandă de a utiliza acest preparat în diluţii mai mari. According to the results obtained, we note that the culture liquid of the bacterium Bacillus cereus in various concentrations (undiluted and in dilutions of 1:50; 1:100; 1:200), at a 24-48 hour exposure of direct contact with the nematodes D. destructor, exerts increased efficacy, the mortality being 98-50-40 and 27.3%, respectively. We note that the bacteria Bacillus cereus possess parallel increased activity on the species of fungi pathogenic for potato culture of the genus Fusarium. However, for in vivo testing, in order to avoid the phytotoxic action on the development of potato plants during the vegetation period, it is recommended to use this preparation in higher dilutions.
Exemplul 2 Example 2
Testarea (în condiţii in vivo) a capacităţii nematocide a LC al bacteriilor Bacillus cereus var. fluorescens CNMN-BB-07, în cazul contactului cu cartofii seminceri de soi Roko, inoculaţi cu nematodul tuberculilor de cartofi Ditylenchus destructor. În experienţe de câmp (anii 2015-2019) a fost testat contactul LC al bacteriilor Bacillus cereus var. fluorescens CNMN-BB-07 cu cartofii seminceri inoculaţi cu nematodul tuberculilor de cartofi Ditylenchus destructor, faza a 2-a de ditilenhoză. Testing (in vivo conditions) of the nematocidal capacity of the LC of the bacteria Bacillus cereus var. fluorescens CNMN-BB-07, in case of contact with seed potatoes of the Roko variety, inoculated with the potato tuber nematode Ditylenchus destructor. In field experiments (2015-2019), the contact of the LC of the bacteria Bacillus cereus var. fluorescens CNMN-BB-07 with seed potatoes inoculated with the potato tuber nematode Ditylenchus destructor, the 2nd phase of dithylenchosis, was tested.
În experienţe, în calitate de material semincer, au fost selectaţi tuberculi de cartofi de soiul Roko sănătoşi, care au fost calibraţi, greutatea medie a unui tubercul selectat a variat între 50,5-55,2 grame, iar diametrul între 5,0-6,73x3,8-5,1 cm. Tuberculii de cartofi de soiul Roko infestaţi în faza a 2-a de ditilenhoză, din experiment, au fost obţinuţi prin inocularea preventivă a fiecărui tubercul, cu un anumit număr de indivizi D. destructor (15 femele+15 masculi/tubercul). Tuberculii experimentali au fost expuşi incubării pe o perioadă de 35-40 de zile. In the experiments, as seed material, healthy potato tubers of the Roko variety were selected, which were calibrated, the average weight of a selected tuber varied between 50.5-55.2 grams, and the diameter between 5.0-6.73x3.8-5.1 cm. Potato tubers of the Roko variety infested in the 2nd phase of dithylinosis, in the experiment, were obtained by preventive inoculation of each tuber with a certain number of D. destructor individuals (15 females + 15 males / tuber). The experimental tubers were exposed to incubation for a period of 35-40 days.
Scopul incubării: în acest timp nematozii D. destructor au pătruns în ţesutul vegetal al pulpei tuberculilor inoculaţi, unde a avut loc ciclul evolutiv al nematozilor, paralel toţi cartofii au încolţit (mărimea colţilor fiind de 0,8-1,2 mm) (fig. 3 a,b). Rezultatele inoculării cu nematodul D. destructor au arătat că extensivitatea invaziei acestora constituie 100%, iar intensivitatea - 200-280 indivizi/tubercul, ceea ce corespunde fazei a 2-a de ditilenhoză, când în tuberculi are loc popularea primară monotopică doar cu specia D. destructor. The purpose of incubation: during this time, the D. destructor nematodes penetrated the plant tissue of the pulp of the inoculated tubers, where the evolutionary cycle of the nematodes took place, in parallel all the potatoes sprouted (the size of the fangs being 0.8-1.2 mm) (Fig. 3 a,b). The results of inoculation with the D. destructor nematode showed that the extensiveness of their invasion is 100%, and the intensity - 200-280 individuals/tuber, which corresponds to the 2nd phase of dithylinosis, when the tubers are primarily monotopic populated only with the D. destructor species.
În cercetările precedente multianuale, efectuate în experienţe vegetative, s-a constatat, că LC al tulpinii de bacterii Bacillus cereus, utilizat la tratarea cartofilor seminceri infestaţi, fiind luat în concentraţii mai mari - Varianta V1 - nediluat şi variantele V2, V3 - diluat în proporţii mici, de 1:50, 1:100, corespuzător, provoacă o eficacitate letală înaltă a nematozilor D. destructor, diminuând infestarea recoltelor obţinute din cartofii trataţi până la 0%, însă, paralel, exercită acţiune fitotoxică asupra dezvoltării tuberculilor, precum şi asupra recoltelor. In previous multi-year research, carried out in vegetative experiments, it was found that the LC of the Bacillus cereus bacterial strain, used to treat infested seed potatoes, being taken in higher concentrations - Variant V1 - undiluted and variants V2, V3 - diluted in small proportions, 1:50, 1:100, respectively, causes a high lethal efficacy of the D. destructor nematodes, reducing the infestation of the crops obtained from the treated potatoes to 0%, but, in parallel, exerts a phytotoxic action on the development of tubers, as well as on the crops.
În experienţele de faţă, o parte dintre tuberculii infestaţi la 100% în faza a 2-a de ditilenhoză au fost supuşi tratamentului prin înmuiere în LC al tulpinii Bacillus cereus, luat în diluţii mai mari - lotul variantelor V1 (1:200) şi V2 (1:400), iar o altă parte de cartofi infestaţi nu au fost trataţi, formând astfel lotul martor M1. Pe lotul martorului M2 au fost sădiţi cartofi sănătoşi fără prelucrare. In the present experiments, a part of the tubers infested at 100% in the 2nd phase of dithylenosis were subjected to treatment by soaking in LC of the Bacillus cereus strain, taken in higher dilutions - the batch of variants V1 (1:200) and V2 (1:400), and another part of the infested potatoes were not treated, thus forming the control batch M1. Healthy potatoes were planted on the control batch M2 without processing.
Schema testării (în condiţii in vivo) a contactului cartofilor seminceri de soiul Roko (inoculaţi cu D. destructor ) cu LC al tulpinii de bacterii Bacillus cereus var. fluorescens, înainte de plantare a avut loc în raportul următor: The testing scheme (under in vivo conditions) of the contact of seed potatoes of the Roko variety (inoculated with D. destructor) with LC of the bacterial strain Bacillus cereus var. fluorescens, before planting took place in the following report:
1. V1 - cartofi inoculaţi + LC al bacteriilor Bacillus cereus var. fluorescens CNMN-BB-07, în diluţie de 1:200, timpul expunerii - 16 ore; 1. V1 - inoculated potatoes + LC of the bacteria Bacillus cereus var. fluorescens CNMN-BB-07, in a dilution of 1:200, exposure time - 16 hours;
2. V2 - cartofi inoculaţi + LC al bacteriilor Bacillus cereus var. fluorescens CNMN-BB-07, în diluţie de 1:400, timpul expunerii - 16 ore; 2. V2 - inoculated potatoes + LC of the bacteria Bacillus cereus var. fluorescens CNMN-BB-07, in a dilution of 1:400, exposure time - 16 hours;
3. M1 - cartofi infestaţi, netrataţi; 3. M1 - infested, untreated potatoes;
4. M2 - cartofi sănătoşi, liberi de nematode, netrataţi. 4. M2 - healthy potatoes, free of nematodes, untreated.
LC al culturii Bacillus cereus a fost diluat cu apă luat în raport de 1:200 şi 1:400. Tratarea s-a efectuat la temperatura camerei (15-20°C), prin metoda de înmuiere, la o expunere de 16 ore (tab.1). The LC of the Bacillus cereus culture was diluted with water in a ratio of 1:200 and 1:400. The treatment was carried out at room temperature (15-20°C), by the soaking method, with an exposure of 16 hours (tab. 1).
Cartofii experimentali au fost plantaţi în câmp deschis, în sol liber de nematodul D. destructor. Pe toată perioada de vegetaţie s-au efectuat cercetări privind dezvoltarea plantelor, scopul final fiind evaluarea eficacităţii de sporire/activare a diverselor raporturi de diluţii ale LC Bacillus cereus asupra plantelor. The experimental potatoes were planted in open field, in soil free from the nematode D. destructor. Throughout the vegetation period, research was carried out on plant development, the final goal being to evaluate the enhancement/activation efficacy of various dilution ratios of LC Bacillus cereus on plants.
Datele obţinute (tab. 1) denotă că LC al tulpinii de bacterii Bacillus cereus în diferite concentraţii exercită diverse acţiuni asupra dezvoltării şi productivităţii tuberculilor. În loturile V1 (1:200) preparatul este puţin toxic, iar plantele s-au dezvoltat mai slab, având un număr de lăstari mediu, o înălţime medie, precum şi un diametru al acestora mai mic comparativ cu loturile V2 (1:400), unde cartofii au fost trataţi cu LC Bacillus cereus în concentraţie mai mică. The data obtained (tab. 1) show that LC of the Bacillus cereus strain in different concentrations exerts various actions on the development and productivity of tubers. In batches V1 (1:200) the preparation is slightly toxic, and the plants developed weaker, having an average number of shoots, an average height, as well as a smaller diameter compared to batches V2 (1:400), where the potatoes were treated with LC Bacillus cereus in a lower concentration.
De asemenea, LC al bacteriilor Bacillus cereus diluat în proporţii de 1:400 în lotul variantei V2, posedă o eficacitate biologică majoră împotriva agenţilor patogeni ai cartofului, manifestând o înaltă capacitate nematocidă. În acest lot s-a evidenţiat o sporire a dezvoltării plantelor, comparativ cu cele din alte loturi experimentale (V1, M1, M2). Also, the LC of Bacillus cereus bacteria diluted in proportions of 1:400 in the V2 variant batch, possesses a major biological efficacy against potato pathogens, manifesting a high nematocidal capacity. In this batch, an increase in plant development was highlighted, compared to those in other experimental batches (V1, M1, M2).
Bacteriile din genul Bacillus sunt capabile de a sintetiza auxine şi gibereline, care stimulează dezvoltarea plantelor şi participă la sporirea imunităţii acestora la diferiţi patogeni. Din datele expuse în tabel observăm că plantele variantei V2 au o înălţime medie cu aproximativ 12 cm mai mare şi un număr de lăstari/plantă de 2 ori mai mare, în raport cu plantele martorului M1. Bacteria of the genus Bacillus are capable of synthesizing auxins and gibberellins, which stimulate plant development and participate in increasing their immunity to various pathogens. From the data presented in the table, we note that the plants of the V2 variant have an average height of approximately 12 cm higher and a number of shoots/plant 2 times higher, compared to the M1 control plants.
Tabel Table
Evaluarea eficacităţii tratării cartofilor seminceri experimental infestaţi cu LC al bacteriilor Bacillus cereus var. fluorescens CNMN-BB-07 Evaluation of the efficacy of treating seed potatoes experimentally infested with LC of the bacteria Bacillus cereus var. fluorescens CNMN-BB-07
Var. Diluţia/ timpul expunerii, ore Faza înflorire - maturizare R e c o l t a r e n. lăs tari lor înăl- ţimea plant., cm n. tu- ber- culi/ cuib d. mediu /tuber- cul, cm m. tuberculi, gr m. tuber-culi/ cuib, gr D. destructor, % Putre-gaiuri, % Coro- pişni-ţe, % 1. V1 1:200/ 16 ore 2,3 47,0 5,0 3,23/ 4,46 43,4 217,0 0 0 0 2. V2 1:400/ 16 ore 6,0 67,38 9,6 5,3/ 4,51 65,3 626,8 1 - 4 0 10,4 3. M1 inoculat, netratat 3,5 55,6 7,0 3,72/ 4,63 48,4 377,52 40,42 22,5 40,3 4 M2 sănătos, netratat 4,4 62,36 8,7 5,0/3,8 58,3 508,2 0 7,9 12,3Var. Dilution/ exposure time, hours Flowering - maturation phase Harvesting n. shoots plant height, cm n. tubers/ nest medium d. /tuber- cul, cm m. tubers, gr m. tubers/nest, gr D. destructor, % Rot-jaws, % Carpenter-worms, % 1. V1 1:200/ 16 hours 2.3 47.0 5.0 3.23/ 4.46 43.4 217.0 0 0 0 2. V2 1:400/ 16 hours 6.0 67.38 9.6 5.3/ 4.51 65.3 626.8 1 - 4 0 10.4 3. M1 inoculated, untreated 3.5 55.6 7.0 3.72/ 4.63 48.4 377.52 40.42 22.5 40.3 4 M2 healthy, untreated 4.4 62.36 8.7 5.0/3.8 58.3 508.2 0 7.9 12.3
La recoltare numărul tuberculilor/cuib, precum şi greutatea tuberculilor/cuib a fost în mediu de 1,4 ori mai mare pe lotul experimental V2 (1:400), în raport cu lotul martor M1 (infestat, netratat) (Tabelul 1). Extensivitatea invaziei cu D. destructor la cartofii recoltaţi de pe lotul experimental V2, trataţi cu LC Bacillus cereus a diminuat de la 100%, înainte de plantare, până la 1-4%, la recoltare, pe când de pe lotul M1, aceasta a fost cu mult mai mare, atingând cota de 40,42%. Metoda de tratare cu LC Bacillus cereus (V2) a condus paralel la o reducere semnificativă (0%) a manifestării putregaiurilor atât la recoltare, cât şi în timpul depozitării cartofilor, de asemenea, a redus cu mult (până la 10,4%) atacul de coropişniţe. Menţionăm faptul că recolta M1, în raport cu recolta V2, este foarte atacată de putregaiuri (22,5%), precum şi de coropişniţe (40,3%). Din datele tabelului 1 se observă că 7,9% din recolta obţinută de pe lotul M2 (cartofi sănătoşi, netrataţi) este atacată de diferite infecţii bacteriene şi fungice, iar 12,3% din tuberculii obţinuţi sunt traumaţi de coropişniţe. At harvest, the number of tubers/nest, as well as the weight of tubers/nest, was on average 1.4 times higher in the experimental group V2 (1:400), compared to the control group M1 (infested, untreated) (Table 1). The extent of invasion by D. destructor in potatoes harvested from the experimental group V2, treated with LC Bacillus cereus, decreased from 100%, before planting, to 1-4%, at harvest, while in the M1 group, it was much higher, reaching 40.42%. The treatment method with LC Bacillus cereus (V2) led in parallel to a significant reduction (0%) in the manifestation of rots both at harvest and during potato storage, and also greatly reduced (up to 10.4%) the attack of rootworms. We mention that the M1 harvest, in relation to the V2 harvest, is highly attacked by rots (22.5%), as well as by mealybugs (40.3%). From the data in Table 1, it is observed that 7.9% of the harvest obtained from the M2 batch (healthy, untreated potatoes) is attacked by various bacterial and fungal infections, and 12.3% of the tubers obtained are traumatized by mealybugs.
Luând în calcul atât datele obţinute de noi, cât şi rezultatele cercetărilor efectuate, observăm că LC al tulpinii de bacterii Bacillus cereus var. fluorescens CNMN-BB-07 manifestă suplimentar un şir de capacităţi: stimulează încolţirea cartofilor seminceri, sporeşte creşterea, dezvoltarea şi productivitatea cartofilor. Tulpina cercetată exercită atât activitate nematicidă, cât şi bactericidă şi fungicidă, fiind şi stimulatoare a dezvoltării plantelor, ceea ce este de perspectivă şi de importanţă majoră în controlul dezvoltării fitohelmintozelor. Conform datelor obţinute de noi, precum şi celor expuse în sursele bibliografice, tulpina Bacillus cereus var. fluorescens, posedă capacitatea de a reduce bolile provocate de fungii fitopatogeni din genul Fusarium (F.oxysporum, F.solani), care provoacă diferite boli cu o răspândire majoră la cultura Solanum tuberosum. Paralel această tulpină exercită acţiune stimulatoare asupra dezvoltării plantelor şi sporirii recoltelor, prin capacitatea sa de a produce auxine, citochinine, gibereline. Taking into account both the data obtained by us and the results of the research carried out, we note that the LC of the Bacillus cereus var. fluorescens CNMN-BB-07 bacterial strain additionally manifests a number of capacities: it stimulates the germination of seed potatoes, increases the growth, development and productivity of potatoes. The investigated strain exerts both nematicide, bactericide and fungicidal activity, being also a stimulant of plant development, which is of perspective and of major importance in controlling the development of phytohelminthiasis. According to the data obtained by us, as well as those presented in the bibliographic sources, the Bacillus cereus var. fluorescens strain possesses the capacity to reduce diseases caused by phytopathogenic fungi of the genus Fusarium (F.oxysporum, F.solani), which cause various diseases with a major spread in the Solanum tuberosum crop. In parallel, this strain exerts a stimulating action on plant development and crop growth, through its capacity to produce auxins, cytokinins, gibberellins.
Procedeul dat reprezintă o metodă biologică eficientă în combaterea nematozilor fitoparaziţi cu un impact stimulator înalt asupra culturilor agricole. This process represents an effective biological method in combating phytoparasitic nematodes with a high stimulating impact on agricultural crops.
1. Чуканцева Н.К. Рекомендации по защите картофеля от стеблевой нематоды в Центральном Черноземном Районе. Воронеж, ВНИИЗР, 1980, 15 р. 1. Chukantseva N.K. Recommendations for the protection of potatoes from the stem nematode in the Central Chernozem District. Voronezh, ВНИИЗР, 1980, 15 р.
2. MD 719 Y 2014.01.31 2. MD 719 Y 2014.01.31
3. MD 1297 F1 1999.08.31 3. MD 1297 F1 1999.08.31
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