Drobnoustroje jako czynnik zmieniający działanie herbicydów na rośliny

R O C ZN IK I G L E B O Z N A W C Z E T. X X V I , Z. 2, W A R S Z A W A 1975

J. SO BIESZCZANSKI

MICROORGANISMS AS A FACTOR CHANGING

THE ACTION OF HERBICIDES ON PLANTS 1

Agricultural University of Warsaw, Institute of Food Science and Technology, Department of Microbiology and Bioengineering, Poland

The observations made during laboratory experiments with culti­

vation o f young plants point to the fact that the herbicide alone added

to the medium acts on a plant in a different way than the same her­

bicide in the medium on which strains of bacteria or fungi had been

cultured. The present paper is concerned with this phenomenon.

M ATERIAL AND METHODS

An active Pseudomonas strain sp. No. 161 was used for the ex­

periments. The bacteria were incubated in liquid meat peptone broth

for 14 days at 27°C. Then the bacterial culture was acidified to pH 2,5

and extracted with ethyl acetate [10]. The extract was condensed 10-fold,

with ethyl acetate evaporation at 40°C. Thus ” a crude preparation

161 S” , was obtained; its plant growth regulating activity was checked

in short term micropot experiments using "Królowa M ajowych” lettuce

seedlings as a plant test [14, 15]. Nantejska carrot, Miejscowa seradella

and Kleszczewska lucerne were used fo r longterm experiments. Seeds

were soaked in the crude preparation for this purpose and placed on

Petri dishes with paper discs lying on wet cotton. After germination,

seeds with root lenght of 0,3-0,5 cm were planted in cuvettes (50 in

each), filled with 2 kg of sterile river sand. The sand had been pre­

viously mixed with minerals salts (for Papillionacea in the amount of

25 mg and the rest of plants of 100 mg of nitrogen in NH4N 0 3; 73 mg

1 Supported financially by the USA Department of Agriculture, Agricultural Research Service Grant FG Po 249.

8 0 J. Sobieszczańsk i

of K 20 in K H P 04 and 19 mg of K 20 in KCl; 50 mg of M gS 04. 7 H20

and 70 mg of P20 5 in K2H P 0 4. The medium was pH 7.

The herbicide used was a technically pure CIPC (40% a.s.) — iso­

propyl N-(3 chlorphenyl carbamate) 2 in the doses of 0,05 ppm.

The control experiments were made with:

— herbicide free sand-water soaked seeds,

— sand with herbicide-water soaked seeds.

The plants were growing in glasshouse. The experiment lasted from

a few weeks to some months. After its termination the plants were

removed to measure the lenght of roots number and lenght of stalks

and leaves and the number of nodules on the roots of lucerne. The dry

weight of plants was determined.

RESULTS

L u c e r n e A/I — the experiment from : Nov. 12.71 to May 5.72.

The presence of CIPC added into the medium in the doses of 0,05 ppm

has a distinct, inhibitory effect on the plant growth, as indicated by

limited formation of lateral roots and the nodules (Fig. 1 a, b). Satu­

ration of lucerne seeds with the crude 161 S preparation, protected

in some ways the plants; it caused apart from reduction of CIPC phyto­

toxicity, an increase in the grown and development of the plants (Fig.

1, 1-4). The 161 S preparation promoted an increase in the number of

nodules on the roots and stimulated propagation of the plants. The

effect tended to become more intense with decreasing concentration

of the preparation (Fig. 1 — 1, 2, 3, 4 and Fig. 2). The same results

were found for the dry mass (Fig. 3).

L u c e r n e — B/II — experiment from June 15 to July 11.72. In

this period lucerne reacted to the presence of the crude preparation

ealier than in the previous case (Fig. 1 -B and Fig. 3-B).

L u c e r n e — C/III — experiment from July 20.72 to January 8.73

and D/IV from October 19.72 to June 18.73 (Fig. 4). Both experiments

with lucerne were similar, in spite of seasonal differences (Fig. 4-C, D,

and Fig. 5-C. D).

S e r a d e l l a — A /I — experiment from November 19.71 to June

21.72. The herbicide in the medium inhibited the growth of the plants

reduced the number of root nodules and pods formation (Fig. 6-b).

Saturation of the seeds with the crude preparation 161 S decreased the

negative effect of CIPC; moreover there was a stimulating effect on

2 Obtained through the courtesy of the Fabrik van Chemische Produkten Vondenligenplaat N.V-Rotterdam — Holland.

I.'icroorganisms as a factor changing the action., ₈₁

Fig. 1. The effect of crude sub­ stances from liquid culture of

Pseudomonas sp. 161 on the

growth of lucerne cultivated with CIPC (0,05 ppm)

con trol: a — water with CIPC, b — w ater without CIPC; dilution of crude preparation : 1 — 1 :10, 2 —

1:100, 3 — 1:1000, 4 — 1:10 000

0,05jijm C3PC O.Oöjijim CJPC

Fig. 2. Effect of crude substances from liquid culture of Pseudomonas sp. 161 on the growth and development of lucerne cultivated with CIPC (0,05 ppm) — July 20.72 — January 8.73

82 J. S obieszczańsk i

the formation of nodules and, in some cases, of pods (Pig. 6, 1-4). The

weight of the dry mass was also higher (Fig. 7).

In two short experiments from : B/II September 20.72 to January 27.73

and C/III May 25.73 to August 15.73 — the results were similar (Fig.

8, 9).

Differences in the growth and appearence of the plants are illu­

strated by Fig. 10.

C a r r o t — three experiments were made at the following dates :

A/I from November 19.71 to June 29.72, B/II from October 5.72 to

February 26.73 and C/III from January 20.73 to May 23.73.

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Fig. 3. The effect of crude substance from liquid culture of Pseudomonas sp. 161 on the dry mass of lucerne cultivated

with CIPC (0,05 ppm)

explanations а, Ъ — as in Fig. 1; dilution o f crude preparation: 1 — 1:100, 2 — 1:1000, 3 —

1:10 000, 4 — 1:100 000

аЪ 1 2 3 4 аЪ 1 2 3 4 0,05jijun CJPC 0,05 jijim CJPC

The results from the experiments confirmed the observations on

161 S effect in Papillionacae — plants. CIPC inhibited the growth of

carrot leaves and main root but had no effect on the growth of the

lateral roots (Fig. 11 a, b). The application of the crude 161 preparation

led to suppression of the toxic action of the herbicide and even to a

certain stimulation of the growth and formation of main roots, with

concomitant inhibition of lateral ones Fig. 11 (1-4). The increase in the

yield of carrot roots was proportional to reduction in the concentration

of the crude preparation Fig. 10 (1-4), which corresponded with the

lenght and thickness of the roots (Fig. 13).

M icroorganism s as a fa ctor changin g the action.., 83 240 160 С D 20 Ш72-81 73 13X72-18 773

£

Fig. 4. The effect of crude substances from liquid culture of Pseudomonas sp. 161 on the growth of lucerne cultivated with CIPC (0,05 ppm)

с _ w ater with CIPC, b — w ater w ithout CIPC; dilution of crude preparation: 1 — 1:100, 2 — 1:1000, 3 — 1:10 000, 4 — 1:100 000

Fig. 5. The effect of crude substance from liquid culture of Pseudomonas sp. 161

on the dry mass of lucerne cultivated with CIPC (0,05 ppm) explanations as in Fig. 4 240 160

£

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Fig. 6. The effect of crude substance from liquid culture of Pseudomonas sp. 161 on the growth of seradella cultivated with CIPC (0,05 ppm)

я — w ater with CIPC, b — water without CIPC; dilution of crude preparation: 1 — 1:100, 2 — 1:1000, 3 — 1:10 000, 4 — 1:100 000

Fig. 7. The effect of crude substance from liquid culture of Pseudomonas sp. 161 on the dry mass of seradella cultivated with CIPC (0,05 ppm)

84 J. Sobieszczański

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с

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Fig. 8. The effe ct o f cru de substance fro m liq u id culture o f Pseudomonas sp. 161 ел the g ro w th c f seradella cu ltiv ated w ith C IPC (0,05 ppm )

a — wnt ; r wi t h С Г PC. о — w a t e r w i t h o u t C IP C ; dilution of crude preparation: l — 1:100, — i:;(),)0. :>’ — 1:10 000, 4 — 1:100 000, 5 — 1:1 000 000

Fig. 9. 'I he ef fect of cru;-e substance fro m liq u id culture o f Pseudomonas sp. 161 on the d r y mass o f seradella cu ltivated w ith C IPC ; explan ation s as in Fig. 8

Fig. 10. Effect of crude substances from liquid culture of Pseudomonas sp. 161 on the growth and develop­ ment of seradella cultivated with CIPC (0,05 ppm); November 20.72 —

Microorganisms as a factor cha nging the action. ₈₅ 240 160 80 % 80 160 240 19 Л 71-29 Ш 72 240 ICI 19I I 71-29 И 72 — Lateral roots -Carrot

I 2

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Fig. 11. T h e e ffe ct o f cru de substance fr o m liq u id culture o f P s e u d o m o n a s sp. 1G1 on the g row th o f ca rrot cu ltiv a ted with C IPC (0.05 ppm )

a — water with CIPC, b — w ater w ithout CIPC; dilution of crude preparation: 1 — 1:100,

2 — 1:1000, 3 — 1:10 000, 4 — 1:100 030, 5 — 1:1 003 000

Fig. 12. T h e e ffe ct o f cru de su bstance fr o m liq u id culture o f P seuclon io.ias sp. 161 on the dry mass o f ca rrot cu ltiv a ted w ith C IPC (0,05 ppm )

a — water with CIPC, t> — water w ithout C IP C ; dilution of crude p r . p u a lion : 1 — 1 :100, 2 — 1:1000, 3 — 1:10 000, 4 — ! 000

Fig. 13. Effect of crude substances from liquid culture of Pseudomonas sp. 161 on the growth and develop­ ment of carrot cultivated with CIPC (0,05 ppm); November 19.71 — June

86 J. S obieszczańsk i 240 г 160

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Fig. 14. The effect of crude substance from liquid culture of Pseudomonas sp. 161 on the growth of carrot cultivated with CIPC (0,05 ppm)

a — water with CIPC, b — w ater w ithout C IP C ; dilution o f crude preparation: l — 1:100, 2 — 1:1000, 3 — 1:10 000, 4 — 1:100 000, 5 — 1:1 000 000

Fig. 15. The effect of crude substance from liquid culture of Pseudomonas sp. 161 on the dry mass of carrot cultivated with CIPC (0,05 ppm); explanations as in Fig. 14

The results of the experiment В and С given in Fig. 14 and Fig. 15

substantiate the above described protective effect of the crude prepa­

ration against CIPC phytotoxicity.

DISCUSSION

The above data have revealed high complex interactions between

herbicides, bacteria and plants.

It is known that microflora, especially that of plant rhizoshpere

plays a m ajor role in their life; bacteria participate in the nutrition

of plants, regulation of their growth and development [8, 10, 12, 15];

they are influenced by root exudates which determine the predominance

of certain bacterial and fungal species in the environment of plants

[3, 4, 11].

The hërbicides disturb the natural bacteria-plant system. They may,

on the one hand, affect the activity and metabolism of bacteria and,

on the other hand, be subject to transformation by microorganisms

[1, 5, 6, 7, 14, 16]. As a result of the complex interactions a reduction

in the phytotoxic activity of the herbicide, may follow. The

possibility

o f detoxication of herbicide and other chemical compounds hc.s already

been demonstrated [3, 9].

The above observations confirm the assumption that the action of

herbicides can be differently modified by microorganisms and this

M icroorga n ism s as a fa cto r changin g the action...

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problem calls for further and deeper research and a patient tracing

o f biochemical transformations which take place in the herbicide-micro-

organism-plant system.

REFERENCES

[1] A l e x a n d e r M. , L u s t i g m a n B. K .: Effect of chemical structure on microbial degradation of substituted benzenes. J. Agric. Food Chem. 14, 1967, 410-413.

[2] B a l i c k a N., K o s i n k i e w i c z B., M u s i a ł M. , S t a n k i e w i c z M.: Deactivation of Rg seed dressing by Arhrobcater sp. 2 b. Acta microb. pol. 5, 1973, 1, 3-8.

[3] E g e r a a t P. W. S. M.: Pea-root exudates and their efefct upon rootnodule bacteria. Medelingen Landbouwhogeschool Wager.ingen, Nederland, 72-27, 1-90, H. Veenman and Zonen N. W., Wageningen 1972.

14] I s w a r a n K. V.: Rhizosphere root exudates and their effect on microbial activity. J. of the Post Graduate School 5, No. 2, December 1967, 212-218. [5] E l i a s a n A. A .: Effect of the carbon source on the amino acid content

in Candida yeast. Appl. Biochem. and Microbiology, Academy of Sei. of the USSR 1, 1974, 29-34 (in rus.).

16] K a u f m a n D. D.: Pesticide metabolism, Pesticide in the soil — Ecology, Degradation and Movement. Int. Symposium — Pesticide in the soil. Michigan University, East Leasing 1970, 73-85.

[7] K e a r n e y P. C., PI i m e r J. R.: Relation of structure to pesticide de­ composition, Pesticide in the soil, Ecology, Degradation and movement. Int. Symp. Pesticide in the soil, Michigan State University, East Leasing 1970, 65-72.

[8] K r a s i l n i k ó w N. A .: Wlijanije poczwiennych mikroorganizmow na rost i urożajnost rastienji. Wiestnik Moskowskogo Uniwiersitieta 3, 1962, 3-25. (Effect of soil microorganisms upon the growth and yelds of plants).

[9] N a i к M. N., J о с к s о n R. B., S t o k e s J., S w a b y R. J.: Microbial de­ gradation and phytotoxicity of Picloram and their substituted pyridines. Soil Biology and Biochemistry 4, 1972, 3, 313-333.

[10] P a n o s y a n A. K., A r t u n i a n R. S., M a r s h a w i n a Z. V .: The effect of metabolites of soil microorganisms on the growth and development of plants, Ztschr. Allg. Microbiol. 3, 1963, 49.

[11] R i v i e r e J.: Rhizosphere et croissance du blé. Ann. Agronom. 14, 1963, 619-653.

[12] S o b i e s z c z a ń s k i J.: Role of microorganisms in life of cultivated plants. I. Quantitative and qualitative changes in the microflora of the rizosphere of rye and winter vetch during the vegetation period. Acta microb. pol. 14, 1962, 161-182.

[13] S o b i e s z c z a ń s k i J.: Role of microorganisms in life of cultivated plants. II. Effect of microorganisms from the rye and vetch rhizosphere and from root-free soil on the development of plants. Acta microb. pol. 14, 1965, 183-202.

[14] S o b i e s z c z a ń s k i J.: Studies on the Role of Microorganisms in the Life of Cultivated Plants. III. Origin of the bacterial substances stimulating the growth of plants. Acta microb. pol. 15, 1966, 67-84.

88

[15] 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.: Biologically active substances produced by bacteria of some cultivated plants rhizosphere. Proceeding of the Int. Symposium on Plant Stimulation, Sofia, 1S63, 335-843.

[16] S o b i e s z c z a ń s k i J.: The influence of herbicides on the production of gibberlin-like substances by Pseudomonas sp. and Arhrobacter sp. Cooloq. Int. Medelingen Faculteit Landbouw. Wetenschappen, Gent 35, 1970, 2, 681-688. 1970.

J. SO B IE S Z C Z A Ń S K I

DROBNOUSTROJE JAKO CZYN N IK ZM IEN IAJĄCY D ZIAŁANIE HERBICYDÓW N A ROŚLINY

Instytut Przechowalnictwa i Technologii Żywności, Zakład Mikrobiologii Technicznej i Bioinżynierii,

Akademia Rolnicza we Wrocławiu

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

W pracy przedstawiono wyniki wskazujące, że metabolity drobnoustrojów mogą ograniczać lub znosić fitotoksyczność herbicydu (CIPC). Doświadczenia szklarniowe wielotygodniowe i kilkumiesięczne prowadzono z uprawą lucerny Kleszczewska, seradeli Miejscowa i marchwi Nantcjska. Nasiona tych roślin moczono w różnych rozcieńczeniach surowego preparatu 161. Preparat ten otrzy­ mywano z płynnej hodowli Pseudomonas sp. 161 przez ekstrakcję jej octanem etylu. Następnie 10-krotne podgęszczenie ekstraktu. Nasiona po zabiegu opłuki- wano wodą wodociągową i wysiewano po 50 sztuk do kuwet z piaskiem wzboga­ conym solami mineralnymi i 0,05 ppm CIPC. Kontrolą były nasiona moczone w wodzie. Podłoże zaś wzbogacano 0,05 ppm CIPC lub nie stosowano herbicydu. Herbicyd hamował wzrost i rozwój roślin, a u motylkowych uniemożliwiał w y­ twarzanie brodawek. Te ujemne efekty były znoszone przez surowy preparat, a w wielu wypadkach uzyskiwano dzięki niemu jeszcze pobudzenie roślin do wzrostu, przyśpieszenia rozwoju, krzewienia, wytwarzania strączków. Ciężar suchej masy roślin z obiektów doświadczalnych był również większy.

E. С О Б Е Щ А Н Ь С К И МИКРООРГАНИЗМЫ К А К Ф АКТОР ИЗМЕНЯЮЩИЙ ВОЗДЕЙСТВИЕ ГЕРБИЦИДОВ Н А РАСТЕНИЯ Институт хранения и технологии пищевых продуктов, Отдел технической микробиологии и эиоинженерии, Сельскохозяйственной академии во Вроцлаве Р е з ю м е В труде представлены результаты исследований доказывающие, что ме­ таболиты микроорганизмов могут ограничивать или выносить фитотоксичность гербицида (CIPC). На протяжении многих недель и нескольких месяцев про­

M icroorga n ism s as a fa cto r changin g the action..,

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