Wpływ długoletniego stosowania wysokich dawek herbicydów na biologię gleby. Część I. Skutki długoletniego stosowania wysokich dawek herbicydów na rośliny i mikroorganizmy glebowe

J. SOBIESZCZAŃSKI, J. ROLA, M. ŻUREK

EFFECT OF MANY YEARS APPLICATION

OF HIGH HERBICIDES DOSES ON THE SOIL BIOLOGY 1 PART I. EFFECT A F M A N Y YEARS OF HIGH HERBICIDES DOSES ON THE SUCCESSION OF SEGETAL PLAN TS AND SOIL MICROORGANISMS

Agricultural University in Wrocław, Institute of Food Science and Technology, Department of Microbiology and Bioengineering

Department of Ecology and Weed Control,

Institute of Soil Science and Plant Cultivation, Wrocław, Poland

INTRODUCTION

The great advantage of herbicides, to which they owe their wide application consists in their differentiated selectivity for plants. This property when skillfully exploited is very useful for removing weeds as rivals of plants.

Each herbicide has, however, a given spectrum of action on different plant species. This is depending on several ecologic factors, the technique and doses applied. It is known that the same herbicide in small doses, can stimulate the growth of a plant, whereas in higher doses, it exerts an inhibiting or even a killing effect.

The differentiated selectivity of herbicides is advantageous as far as the protection of environment is concerned, but one preparation, or even a whole group of derivatives is not able to solve all problems of segetal weed control. For instance, the application of the same prepa­ ration for several years is invariably followed by compensation of species resistant to the herbicide. To a certain degree this phenomenon can be prevented by differentiating doses o f the preparation, but the most efective method consists in using different herbicides.

Weed-killing preparations are known to-act selectively not only on 1 This work was supported in part by Department of Agriculture project No. 09.1.3.05.06.

166 J. S obieszczański, J. R ola, M. Ż u rek

higher plants. Still more susceptible to their action are microorganisms. To elucidate the above phenomena field experiments were undertaken in 1964 on the experimental field of the Department of Ecology and Weed Control in Wrocław.

METHODS

The experiments were carried out on degraded black soil. The size of plots was 25 m 2. From 1964 the following herbicides were used an­ nually in spring:

control — without herbicide,

Gesatop (50% simazine) 20 kg/ha,

Gesaprim (50% atrazine) 20 kg/ha,

Gesagard (50% prometryne) 20 kg/ha,

Telvar (80% monuron) 15 kg/ha,

Basfapon (85% DCP) 20 kg/ha,

Antyperz (38% TCA) 75 kg/ha.

Each year the half of the plots (belt B) was sprayed with the her-bicides and the mineral manure in the doses of N — 40 kg, P — 50 kg, К — 80 kg/ha was applied. The other half (belt A) received no manure throughout the experimental period. Each year during the vegetation season, the amount of weeds was analyzed on 3-4 occasion by estimation method to determine the rate of soil contamination by individual species. In autumn, dried weeds were cut and removed from the field. Since 1964 no agricultural procedures and mechanical control of plants were applied. Soil samples for microbiologie examination were collected from the depth 0-10 cm in autumn 1972 and in spring and summer 1973. The number of microorganisms was determined by the plate dilution method. The following media were used:

— soil extract enriched by glucose and phosphates [9] ; — cabbage extract with glucose [14];

— mineral salts and glucose [7]; — special for Pseudomonas sp. [11]; — special for fungi [13].

The above mentioned media of the parallel series were treated with the proper herbicide. The medium reaction for bacteria was established at pH of 7.2 and for fungi at pH of 6.3. After few days’ incubation at 26 °C the colonies were counted, with establishment of their character­ istics. The diagrams show the results presented as the mean arythmetic and compared with control values.

RESULTS

The experimental field was up to 1963, used to grow various vege­ tables. As the result of a fallow in 1963 a fairly exube rant growth of Agropyron repens appeared. This state persisted for several years up to 1967 (Table 1) and was followed by its systematic decline and total disappearance in 1973. Agropyron repens was replaced by other species, mostly Solidago sp., Achillea millefolium and Artemisia vulgaris, which is consistent with the lows of natural succession.

T a b l e 1

Compensation of weeds rate of s o il contamination by weeds /i n %/• Belt

Group Control Veeds ₁₉₆₄ 1965 1966 1967 1968 1969 1970 1971 1972 1973 I Agropyron repęns 67 79 70 78 35 12 23 5 5 Digitaria sanquinalis Bchinocbloa c .g a lli 1 4 n Chenopodium album 4 Galinaoga parviflora 8 4 H I Erigeron canadensis 2 Seneclo Vulgaris + Grepis biennis ₅ IT Clxsium arvense 3 3 3 2 1 2 2 1 Artemisia vulgaris + + 3 3 10 8 9 12 10 Achillea millefolium 2 2 6 5 19 15 8 Pastinaca sativa 5 2 1 1 1 Solidago sp* 4 9 38 41 44 50 56 Sonchus arvensis 1 1 Taraxacum officinale + 1 6 5 6 8 2 4

Diametrically different was the contamination on herbicide-treated plots; for instance, the plot sprayed with Gesaprim (20 kg/ha) showed, from the first day, almost total disappearance of Agropyron repens and the emergence of a hitherto nonrecorded species i.t. : Digitaria sanqui-

nalis (Table 2). On the other hand, the Gesagard-treated plot (20 kg/ha)

showed disappearence of Agropyron repens, similarly as in the control, with Taraxacum officinale and Pastinaca sativa taking its place (Table 3). On TCA- and DCP-treated plots various species of dicotyledonas weeds, with perdominance of perenniae, were highly numerous. Only Telvar- sprayed plot was almost devoid of vegetation, which indicates a con­ siderable phy to toxicity of the herbicide.

From the above findings it may be concluded that, except for Telvar, all herbicides under examination, even in such high doses, used for several years, failed to eliminate all plant species. Changed were only

168 J. S obieszczański, J. R ola, M. Ż u rek

proportions between the species, as some of them disappeared and others were replaced. This suggests a high selectivity of herbicides for certain weed species, the property which is of basic importance for their practical application.

In this connection, the question which arises is: what was the be­ haviour of soil microorganisms at that time and what changes might

T a b l e 2

Compensation of weeds rate of ecil contamination by W6eds /in %/. Belt A.

Group Weeds Gesaprim - 50 1964 1965 1966 1967 1968 1969 1970 1971 1972 1973 I Agropyron repçns Digitaria sanauineli Echinoohloa c.galli 7 1 7 + + 23 5 + 10 5 II Chenopodium album Galinsoga psxvii'Iora

III Srigeron canadensis Senecio vulgaris Crspis biennis IV Cirsium arrense Artemisia vulgaris Achillea millefolium Solidago sp. Sonchus arvensis Taraxacum officinale 10 + ł + T a b l e 3

Compensation of weeds rate of soil contamination by weeds /in %/. Belt A.

(xPOUp Weeds Gesagard; - 20 kg 1964 1965 1966 1967 1968 1969 1970 1971 1972 1973 i Agropyron repens Digitaria sanquinalis Echinochloa c.galli 64 4 56 70 53 20 24 19 + 1

i i Chenop odium album Galinsoga parviflora 2 5 1 h i Erigeron canadensis Senecio vulgaris Crepis biennis + 5 1 3 IV Cirsium arvense Artemisia vulgaris Achillea millefolium Solidago sp. Sonchus arvensis 1 4 3 4 4 3 1 4 + 1 5 Taraxacum officinale 1 1 3 5 7 4 4 26 8 Pastinaca sativa 2 7 31 9 2 13

have occurred in them after the prolonged application of high doses of the herbicide.

Many years application of high doses of the same herbicides was found to restrict the development of bacteria and fungi in the soil of manured and non-manured plots. The quantitative changes in the soil were dependent on mineral manure and the herbicide used. A number of bacterial and fungal media used made it possible to detect differences in the incidence of nutritional groups and to isolate some interesting bacterial and fungal species.

The soil extract medium enabled to establish quantitative differences in bacterial population of control and herbicide-treated plots.

Gesatop, Gesaprim, TCA and Basfapon had a stronger inhibiting effect on soil bacteria, as compared with that of Telvar and Gesagard, this being particularly conspicuous for non-manured plots.

A fter addition of the herbicide to the medium, the quantitative changes in the bacterial population from the control plots were less than for herbicide-treated ones. In this case, the reaction of micro­ organisms to Basfapon was the strongest (Fig. 1). The enriched medium confirmed a considerable reduction in the number of bacterial population in herbicide-treated plots, but it showed the appearance of single colonies in the presence of Basfapon in the herbicide-treated plot (Fig. 2).

Fig. 1. The number of bac­ teria growing in soil extract medium per 1 g of soil from

the field experiment Ь — unfertilized ; 2 — control without herbicide, 2 — G esatop,

3 — G esaprim , 4 — Gesagard,

5 — T elvar, e — T C A , 7 — B a s­

fapon

The size and appearance of colonies from the herbicide containing medium did not differ from those found in the medium without the herbicide.

170 J. Sobieszczański, J. Rola, M. Żurek miß1 12 Ю

n п

h

111

Q'Zl

t

Fig. 2. The number of bacteria growing in soil extract medium with 1000 ppm suitable herbicide; explanation as in Fig. 1

After a few weeks incubation some colonies were surrounded by clear zones due to the herbicide transformations in the medium. A slight­ ly different pattern was seen in the cabbage extract medium (Fig. 3). The recorded number of bacteria was much higher than in the previous case. This effect was particularly evident in Gesaprim, TCA, Basfapon, as well as in Gesatop and Gesagard treated plots. Highly pronounced quantitative differences recorded for manured plots. As in the previous case, the highest number of bacteria was found in control plots.

Fig. 3. The number of bac­ teria growing in cabbage- - extract medium; explanation

The presence of herbicide added to the cabbage extract reduced still more the number of bacteria (Fig. 4). A clear-cut reduction in the growth of bacterial population was found for the soil suspension of control plots.

In the experimental plots the number of bacteria varied: Gesatop, Gesagard and TCA-treated plots showed no reducing effect but Telvar and Basfapon in the doses of 1000 ppm were toxic for bacteria (Fig. 4). As for control plots, Telvar was found to reduce the number of bacteria, where-as Basfapon did so both in control and experimental plots. The mineral medium with glucose (Fig. 5) enabled the detection of a greater number of bacteria in most manured plots; they were less numerous in non-manured ones.

mi

Fig. 4. The number of bacteria growing in cab­ bage extract with 1000 ppm suitable herbicide; explanation as in Fig. 1

2 3 4 5 6 7

CZJ-

min 20

18 16 14

Fig. 5. The number of bac­ teria growing in mineral Kaufman-Kerney medium [7]; explanation as in Fig. 1 12 6 ab

I

/71 bpi

172 J. Sobieszczański, J. R ola, M. Ż u rek

The treatment of the soil with Gesagard and Telvar caused a pro­ nounced reduction in the bacterial growth, while Basfapon failed to show any toxic action and consequently the number of the growing bacterial colonies was comparatively high (Fig. 5). The addition of the herbicide to the medium resulted in strong selection of bacteria parti­ cularly by Telvar and weaker by Gesatop and Gesaprim (Fig. 6).

mln w\ u 0$

0,8

1

П

Plots with herbicides

Fig. 6. The number of bac­ teria growing in mineral Kaufman-Kerney medium [7] With 1000 ppm suitable her­ bicide; explanation as in

with 1000 ppm suitable her­ bicide; explanation as in

Fig. 1

The use of selective media made it possible to establish more pre­ cisely the reaction of certain groups or genera of microorganisms to long-termed application of herbicides in the field. The special medium for Pseudomonas sp. revealed that the application of TCA, Basfapon, and Gesagard led to increased incidence of Pseudomonas sp. in the soil. Gesatop. Gesaprim and Telvar exhibited a reducing effect on the num­ ber of the bacteria, as compared with the controls. Analysed were only non-manured plots (Fig. 7). The colony of UV fluorizing bacteria were not present in the soil treated with Basfapon, TCA and Telvar only few o f them being found in the soil after treatment with Simazine, Ge­ saprim and Gesagard.

Interesting results were obtained with the soil suspension applied to the medium enriched by 10 ppm of the proper herbicide. In the case, the appearence of Pseudomonas sp. in the herbicide treated plots was higher than in non treated ones with the soil suspension (Fig. 8).

big. 7. The number of

Pseudomonas sp. growing

in special Sands and Ro- vira medium [11]; expla­

nation as in Fig. 1 mln 12 10 CD 4 0 M

igjAifflfc]

П

Lfe

C .Zl-Plot without herbicide

I 1

J A «

5

Fig. 8. The number of Pseudomonas sp. growing in special Sands and Rovira medium [11] with 10 ppm suitable herbicide; explanation as

in Fig. 1 mln W 0,9 0,8 0,7 0,6 0,5 § 0,3 0,2 0J 0 2 3

□

174 J. Sobieszczański, J. R ola, M. Ż u rek mirta 9 0 8 0 - 706 0 5 0 - \40-

30-20

-Fig. 9. The number of fungi growing in special Williams and Schmitthenner medium [13]; explanation as in Fig. 1

n

4 5 6 7

Piots with herbicides

min 70 60 50 40 %30 10 \

П

bi

n

U .~ 2 -P tot without herbicide

1— \- Plots with herbicides

Fig. 10. The number of fungi growing in special Williams and Schmitthenner medium with 1000 ppm suitable herbi­ cide; explanation as in Fig. 1

The special medium for fungi revealed that the treatment of the soil with the examined herbicides resulted in appreciable reduction of fungi and changes in their qualitative composition. A strong reduction in the number of soil fungi was caused by: Basfapon, TCA, Telvar, a weaker action, in this respect, being exterted by Gesatop, Gesaprim and Gesa- gard (Fig. 9).

After addition of the proper herbicide to the medium, the number of fungi from the control soil was found to be slightly lower (Fig. 10),

whereas that from the herbicide treated plots was several times lower. Apparently, the additional effect of the herbicide in the medium sup­ pressed other fungal species or restricted their growth, which could not be observed with single application of the herbicide (Fig. 10).

The appearance of clear zones around the colonies growing on media containing herbicides could be seen after a few weeks’ incubation of bacteria. In the soil of the experimental plots — Bacterium sp., Arthro-

bacter sp., Pseudomonas sp., were predominant, Bacillus sp. being less

frequent, particularly in TCA and Basfapon treated plots. The prevalent fungal species were: Penicillum, Aspergillus, Mucor and Trichoderma sp.

DISCUSSION

The soil microorganisms react to the presence of high doses of the herbicides, as do a number of weeds. High doses of the herbicides were found to cause a pronounced reduction in the number of bacterial po­ pulation of the soil. No such phenomena were seen in other experiments with normal herbicide doses, when used for several years [2, 3]. Other studies have, however, pointed to the possibility of qualitative and qua- titative transformations in the bacteria under the influence of herbicides [1, 6, 12]. This effect is dependent on the chemical structure and doses of the herbicide used [10, 12, 16]. Nonetheless, the same herbicides can exert a stimulating or toxic effect on various groups and genera of micro­ organisms [1, 10].

Mineral manure was found to be of benfit in the present studies. Bacterial population in the manured soil were more numerous, with varying effect o f certain components on the development o f microflora by attenuating the toxic action of herbicides [5, 6]. Better development and higher incidence of bacteria such as Pseudomonas, as observed in some TCA, Basfapon, Telvar and Gesagard treated plots, imply a great possibility of their adaptation the medium with the ability of meta­ bolizing some of herbicides, which has been pointed out by other in­ vestigations [4, 8].

REFERENCES

[1] B a k a ł i w a n o w D . : Biological activity of certain herbicides on microscopic soil fungi. Symp. Biol. Hung., 1972, 373-377.

[2] B a l i c k a N., S o b i e s z c z a ń s k i J., N i e w i a d o m a T.: The effect of herbicides on soil microflora. III. The action of herbicides on soil micro­ organisms. Acta microb. poi. Ser. B, 18, 1969, 7-10.

[3] B a l i c k a N., S o b i e s z c z a ń s k i J.: The effect of herbicides on soil microflora. I. Effect of herbicides on the number of soil microorganisms in field experiments. Acta microb. poi. Ser. B, 18, 1969, 1-6.

176 J. Sobieszczański, J. Rola, M. Żurek

[4] J o h n s o n H. W. , B r i g g s G. G., A l e x a n d e r M.: Metabolism of 3-chloro- benzoicacid by Pseudomonas. Soil Biol. Biochem. 4, 1972, 187-190.

[5] K a u f m a n D. D., W i l l i a m s L. E.: Effect of mineral fertilization and soil reaction on soil fungi. Phytopathology 54, 1964, 2, 134-139.

[6] K a u f m a n D. D.: Effects of s-triazine and phenylurea herbicides on soil fungi in corn and soybean-croppde soil. Phytopathology 54, 1964, 897. [7] K a u f m a n D. D., K e a r n e y P. C.: Microbial degradation of isopropal-

-N-3-chlorophenylcarbamate and 2-chloroethyl-N-3-chlorophenyl carbamate. Applied Microbiology 13, 1965, 3.

[8] K e a r n e y P. C., K a u f m a n D. D., Von E n d t D. W. , G a r d i a F. S.: TCA metabolism by soil microorganisms. Agricultural and Food Chemistry 17,

1969, 3, 581-584.

[9] P o c h o n J.: Manuel technique d’analyse microbiologique du sol. Massou et Cie, Paris 1954.

[10] R o d r i g u e z - K a b a n a R., C u r l E, A.. F u n d e r b u r k K. H., Jr: Effect of Atrazine on growth response of Sclerotium Rolfsi and Trichoderma viride. Canadian Jour of Microbiology 13, 1967, 1343-1349.

[11] S a n d s D. C., R о v i r a D. A .: Isolation of fluorescent Pseudomonas with a selective medium. Applied Microbiology 20, 1970, 513-514.

[12] S m i t h N. R., D a v i s o n V. T., W n z e l M. E.: The effect of certain herbicides on soil microorganism. Proc. Soil. Sei. Soc. Amer. 10, 1945, 197-201. [13] W i l l i a m s L. E., S c h m i t t h e n n e r : Genera of fungi in Ohio soils.

Ohio Agr. Expt. Sta. Res. Circ. 38, 1956, 7.

[14] W o z n a k o w s k a j a J. M., Ż i l c o w a О. G.: Pitatielnyje sriedy dlja izu- czenija korniewych mikroorganizmow. Trudy Wsiessojuzn. Nauczn. Issled., Instituta Siel. Choz., Microbiol. 15, 156-163, Leningrad 1958.

J. S O B IE S Z C Z A Ń S K I, J. R O LA , M. ŻU R EK

W P Ł Y W DŁUGOLETNIEGO STO SO W A N IA W YSO K IC H D A W E K HERBICYDÓW NA BIOLOGIĘ GLEBY

CZĘSC I. S K U T K I D Ł U G O LET N IEG O S T O S O W A N IA W Y S O K IC H D A W E K H E R B IC Y D Ó W N A R O ŚL IN Y I M IK R O O R G A N IZ M Y G LEBOW E

Instytut Technologii Żywności Akademii Rolniczej we Wrocławiu oraz Instytut Gleboznawstwa i Uprawy Roślin

S t r e s z c z e n i e

Celem pracy było bliższe poznanie skutków wieloletniego stosowania na tym samym poletku wysokich dawek tego samego herbicydu. Do badania wzięto: Gesatop, Gesaprim, Gesagard, Basfapon w dawkach po 20 kg/ha, Telvar 15 kg/ha i Antyperz 75 kg/ha.

Poletka były podzielone na dwa pasy: A — nie nawożone i В — nawożone solami mineralnymi (N4o, P50, K 80, kg/ha). Przez cały czas trwania doświadczeń, od roku 1964, poletka są nie uprawiane, a chwasty jesienią po wyschnięciu są usuwane z poletek.

Analiza ' wkładu gatunkowego chwastów wykazała, że stosowane herbicydy wykazują dużą selektywność i ograniczają wzrost niektórych chwastów; wyniki

tych badań przedstawione są w tabelach 1-3. Efektem wieloletniego stosowania herbicydów jest zjawisko kompensacji chwastów. Podobne zjawisko zanotowano, badając w jesieni 1972 i wiosną 1973 stan mikrobiologiczny gleby wymienionych wyżej poletek. Badania wykazały, że wieloletnie stosowanie wysokich dawek her­ bicydów ograniczyło liczbę bakterii i grzybów w glebie poletek nawożonych i nie nawożonych. Zastosowanie różnych pożywek pozwoliło na wykrycie różnic w iloś­ ciowym występowaniu grup pokarmowych i wyodrębnienie interesujących ga­ tunków. Wyniki przedstawione zostały w odpowiednich diagramach.

Wzbogacenie pożywek odpowiednim herbicydem wzmogło selekcję drobno­ ustrojów. Po parutygodniowej inkubacji wokół niektórych kolonii notowano pojawienie się jasnych stref, co świadczyłoby o zdolności tych bakterii względnie grzybów do przemiany określonego herbicydu. Zmiany ilościowe i jakościowe w pewnym stopniu mogły być związane ze zjawiskiem kompensacji chwastów.

W poletkach traktowanych herbicydami dominowały następujące rodzaje bak­ terii: Bacterium sp., Arthrobacter sp., Pseudomonas sp. sporadycznie występowały

Bacillus sp., a z grzybów Penicillum sp., Aspergillus sp., Mucor sp., Trichoderma sp.

E. С О Б Е Щ А Н Ь С К И , Ю . РОЛЯ, М. Ж У Р Е К ВЛИЯНИЕ ДОЛГОЛЕТНЕГО ПРИМЕНЕНИЯ ВЫ СОКИХ ДОЗ ГЕРБИЦИДОВ Н А БИОЛОГИЮ ПОЧВЫ Ч А С Т Ь I. В Л И Я Н И Е ДОЛГОЛЕТНЕГО П РИ М Е Н Е Н И Я В Ы С О К И Х ДОЗ ГЕРБИЦИ ДОВ Н А ПО Ч В ЕН Н Ы Е Р А С Т Е Н И Я И М И К Р О О Р Г А Н И ЗМ Ы Институт технологии промышленности Сельскохозяйственной академии во Вроцлаве и Институт почвоведения и растениеводства Р е з ю м е Целью работы было более подробное изучение последствий долго­ летнего применения на одной и той же делянке высоких доз одного и того же гербицида. В опытах применяли препараты: Гесатоп, Гесаприм. Гесагард и Басфапон в дозе по 20 кг/га, Тельвар в дозе 15 кг/га и Антыпэж в дозе 75 кг/га. Делянки были разделены на две полосы: А — неудобряемые и В — удо­ бряемые минеральными солями (N4o, Р5 0, К 80 кг/га). На протяжении всей продолжительности опытов от 1964 года делянки оставались без обработки, а засохшие сорняки удалялись с делянок. Анализ видового состава сорняков показал, что применяемые гербициды характеризуются высокой селектив­ ностью, ограничивая рост некоторых сорняков. Результаты опытов представ­ лены в таблицах 1-3. Эффектом долголетнего применения гербицидов есть явление компенсации сорняков. Подобное явление было отмечено в прове­ денных в 1972 и 1973 гг. исследованиях микробиологического состояния почвы указанных делянок. Результаты исследований показали, что долголетнее при­ менение высоких доз гербицидов ограничивало число бактерий и грибов в по­ чве удобряемых и неудобряемых делянок. Применение разных питательных сред позволило выявить различия в количестве питательных групп и вы ­

178 J. Sobieszczański, J. R ola, M. Ż urek делить интересующие нас виды. Результаты иллюстрируются на соответству­ ющих чертежах. Обогащение питательных сред соответствующим гербицидом способствовали повышению селекции микроорганизмов. Через несколько недель инкубации вокруг некоторых колоний появлялись светлые зоны, что может свидетель­ ствовать о способности данных бактерий или грибов изменять определенный гербицид. Количественные и качественные изменения могут быть в некоторой степени связанными с явлениями компенсации сорняков. На делянках обработанных гербицидами преобладали следующие бактерии:

Bacterium sp., Arthrobacter sp., Pseudomonas sp. Спорадически появлялись

Bacillus sp., а из грибов Pénicillium sp., Aspergillus sp., Mucor sp., Tirchoderma sp.