Antibiotics as Crop Protectants - ACS Publications - American

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A n t i b i o t i c s as

Crop

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Downloaded by NANYANG TECHNOLOGICAL UNIV on June 7, 2016 | http://pubs.acs.org Publication Date: September 18, 1986 | doi: 10.1021/bk-1986-0320.ch005

Arun K. Misra

Department of Biological Sciences, Morris Brown College, Atlanta University, Atlanta, GA 30314 Antibiotics, the miracle drugs, have a long history of being useful in agriculture. There has been increased interest in recent years in the use of antibiotics for the control of plant diseases. Some antibiotics that are toxic for use in the treatment of human or animal diseases may be used on plants. Antibiotics have been found useful for the control of bacterial, fungal, viral, and mycoplasmal diseases of a variety of crops and ornamental plants. Drug laws of various countries differ regarding use of antibiotics as plant protectants. Concern regarding uncontrolled use of antibiotics is appropriate, but more information is needed about the effectiveness and safety of the use of antibiotics for control of plant diseases. A n t i b i o t i c s are chemicals a n t a g o n i s t i c to l i f e . These are g e n e r a l l y produced by microorganisms and may be very e f f e c t i v e against m i c r o b i a l pathogens(1). Using a n t i b i o t i c s other than i n c o n t r o l l i n g diseases of humans has been c a l l e d non-medical or "non-pharmaceutical". The use of a n t i b i o t i c s i n food and a g r i c u l t u r e i s multifaceted(2-_5) aspect of t h e i r use with plants and animals, covered i n several i n t e r n a t i o n a l conferences, proceedings of which have been published(()-9 ) . The subject of t h i s review w i l l be the use of a n t i b i o t i c s as plant p r o t e c t a n t s . ff

M

1On leave of absence from Botany Department, L. N. Mithila University, C. M. Science College, Darbahnga, Bihar, 846004, India. 0097-6156/ 86/ 0320-0049506.00/ 0 © 1986 American Chemical Society

Moats; Agricultural Uses of Antibiotics ACS Symposium Series; American Chemical Society: Washington, DC, 1986.

A G R I C U L T U R A L USES O F ANTIBIOTICS

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Crop P r o t e c t i o n The use of a n t i b i o t i c s i n plant pathology, e s p e c i a l l y f o r the c o n t r o l of plant diseases i s a subiect of i n c r e a s i n g i n t e r e s t ( 1 0 ) . Nearly four decades ago, i n the f o r t i e s , use of a n t i b i o t i c s f o r plant p r o t e c t i o n was l i t t l e recognized. A n t i b i o t i c s were used against plant diseases only when they were found u n s u i t a b l e i n human medicine(11). Later, i t was found that many plant diseases p a r t i c u l a r l y those caused by fungi and b a c t e r i a were e f f e c t i v e l y c o n t r o l l e d using a n t i b i o t i c s (12). However, r e c e n t l y doubts have been expressed concerning the growing use of a n t i b i o t i c s ( 1 3 ) , e s p e c i a l l y because of p o s s i b l e residues i n vegetable products. There i s , however, l i t t l e evidence as to the d e l e t e r i o u s e f f e c t s of spraying a n t i b i o t i c s on crop p l a n t s . G l i o t o x i n , i s o l a t e d by Winding i n 1932 from Gliocladium fimbriaturn was found a n a t a g o n i s t i c to Rhizoctonia s o l a n i , thus h e l p f u l against the r o o t - r o t of potato and tomato. Brown and Boyle i n 1954(14-15) noted that p e n i c i l l i n was a c t i v e against the crown-gaTT bacterium. Zaumeyer (16)found that spraying with streptomycin was e f f e c t i v e against h a l o b l i g h t of beans caused by Pseudomonas p h a s e o l i c o l e . Zalaback(17) and Ark(18) used streptomycin to c o n t r o l bean b l i g h t , caused bv Erwinia amylovora, under f i e l d c o n d i t i o n s . Aureofungin was developed as a plant protectant against the fungal diseases of r i c e i n India(,19). S i m i l a r l y , b l a s t i c i d i n has been used i n r i c e c u l t i v a t i o n i n Japan f o r very long time(20). THe a n t i b i o t i c s used i n plant p r o t e c t i o n have been more s u c c e s s f u l i n c o n t r o l l i n g fungi than other types of plant pathogens. Aureofungin, cycloheximide, g r i s e o f u l v i n , ohyamycin and a host of others(see Table 1) have been used. Extensive reviews i n t h i s field(21.-26) are a v a i l a b l e . Aureofungin i s a heptaene a n t i b i o t i c anïï~~is extracted from Streptomyces cinnamoseous var. t e r r i c o l a . I t belongs to a new a n t i b i o t i c group among the heptaenes(27). I t i s a broad spectrum f u n g i c i d e , e f f e c t i v e against a wicfe v a r i e t y of f u n g i , and i s systemic i n a c t i v i t y . A golden yellow powder, i t i s unstable i n the presence of moisture and l i g h t , and needs to be stored dry and i n darkness. A host of fungal diseases have been c o n t r o l l e d by aureofungin(28-29). C i t r u s gummosis caused by Phytophthora c i t r o p h t h o r a may be cured by 20 mg/ml spray. Control of the D i p l o d i a r o t of mangoes and the A l t e r n a r i a r o t of tomatoes, by t h i s a n t i b i o t i c , e s p e c i a l l y during t r a n s i t and storage of f r u i t s i s noteworthy. Dipping mango f r u i t s i n 100-500 ppm of aurefungin prevents r o t t i n g f o r 11-20 days. Untreated f r u i t s r o t i n 2-3 days. In developing countries where r e f r i g e r a t i o n i s not common, t h i s method i s u s e f u l i n control l i n g post-harvest l o s s . The b l a s t disease of r i c e caused by P i r i c u l a r i a oryzae has been shown to be c o n t r o l l e d by a 7.5 g/hectare spray of aureofungin, four times at 12 days interval(30).Powdery mildew of apples caused by Podosphaera leucotricïïa can be e f f e c t i v e l y handled with aureofungin. Seed-borne i n f e c t i o n


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Table I : Some Common A n t i b i o t i c s used f o r Plant P r o t e c t i o n Antibacterial :


A n t i b i o t i c C6, C e l l o c i d i n , Chloramphenicol, C i t r i n i n , Erythromycin, Gramicidin, Kanamycin, Novobiocin, P e n i c i l l i n , P h t o b a c t e r i omycin,Polymycin, Polymyxin, Rhizopin, Streptomycin, Agrimycin, Phytostrep, T e t r a c y c l i n e , and Vancomycin. Antifungal : A n t i b i o t i c P3, A n t i b i o t i c P9, Antimycin, Antimvcin, Aureofungin, B l a s t i c i d i n , Bulboformin, C a n d i c i d i n , C R R I - a n t i b i o t i c , Cycloheximide, Foliomvcin, N y s t a t i n , Oligomvcin, G r i s e o f u l v i n , Phytoactin, Polyoxin, T e t r i n , T r i c h o t h e c i n , B e n t u r i c i d i n , and Venturomycin. Antimycoplasmal

:

T e t r a c y c l i n e , Erythromycin, and Methacycline. Antiviral : Actinomycin D, A n t i b i o t i c 205-2B, B l a s t i c i d i n , Cycloheximide(actidione), Daunomycin DPB, Mithramycin, Mitomycin C, Pentaene G8, and T u b e r c i d i n .

by Helminthosporium oryzae(30) may be s i g n i f i c a n t l y reduce -d by overnight soaking of paddy seeds i n aureofungin s o l ution. B l a s t i c i d i n s are produced by Streptomyces grieseochro -mogens and i n h i b i t s e v e r a l species of b a c t e r i a and fungi (31). Pseudomonas i s p a r t i c u l a r l y vulnerable to b l a s t i c i d i n S. P i r i c u l a r i a oryzae causing the b l a s t disease of r i c e i s widely c o n t r o l l e d with b l a s t i c i d i n S i n Japan. I t i s appl i e d to the r i c e p l a n t s a f t e r i n f e c t i o n by the fungus has already ocurred(32), since the a n t i b i o t i c a f f e c t s the myce - l i a i phase more than the spore phase. I t would be d e s i r a ble to search f o r spore k i l l i n g a n t i b i o t i c s to c o n t r o l s o i l - i n h a b i t i n g microbes and to destroy the inoculum before i t i n f e c t s the crop. Numerous cases of the use of a n t i b i o t i c s ( e s p e c i a l l y : cycloheximide, ohyamycin, streptomycin, t e t r a c y c l i n e s , p e n i c i l l i n , g r i s e o f u l v i n , and polymyxin) against s e v e r a l b a c t e r i a l and fungal diseases are now known(33-35). In the United States of America, Merck s e l l s preparations of streptomycin and Upjohn s e l l s that of cycloheximide f o r the c o n t r o l of the diseases of ornamental plants(R.Burg,



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AGRICULTURAL USES OF ANTIBIOTICS

personal communication). Interest i n the use of aureofungin i s s t i l l continuing i n some l a b o r a t o r i e s (36^) . A c t i d i o n e has r e c e n t l y been reported to be e f f e c t i v e against s e v e r a l s o i l f u n g i ( 3 7 ) . I t has a l s o been noted that some a n t i b i o t i -cs may supress V e r t i c i l l i u m d a h l i a e and protect peppers against i n f e c t i o n and a l s o stimulate seed germination and growth of the plants(38-39). Bacillomycin(40) has been found e f f e c t i v e against Helminthosporium turcium(41), which i n f e c t s s e v e r a l c e r e a l crops. Antimycin A i s e f f e c t i v e against A l t e r n a r i a s o l a n i spores(42), karumin i s e f f e c t i v e against Rhizoctonia s o l a n i ( 4 3 ) . Nikkomycin i s being used to cure trees of Dutch elm disease i n Hamden, Connecticut(44). I t i n h i b i t s the formation of c h i t i n and stops the myceTia of Ceratocys t i s ulmi from growing normally. There i s a long l i s t of a n t i b i o t i c s that have been t r i e d against the b a c t e r i a l pathogens of plants(45-47). Pseudomonas, Xanthomonas, Agrobacterium, Aplanobacterium and other genera have been found to be i n a c t i v a t e d by streptomycin, t e t r a c y c l i n e , o x y t e t r a c y c l i n e , p e n i c i l l i n , agromvcin, and vancomycin; e s p e c i a l l y i n the crops of cherry, maize, bean, poplar, cotton, r i c e , c i t r u s , apple, plums and s e v e r a l plants of h o r t i c u l t u r a l importance such as geraniums(48-51) Tomato canker caused by Xanthomonas can be c o n t r o l l e d by the a p p l i c a t i o n of t e t r a c y c l i n e ( 5 2 - 5 3 ) . Streptomycin r e s i s t a n t s t r a i n s of b a c t e r i a have Been found on peach, tomato and peppers(54), and the mixture of two a n t i b i o t i c s has helped to stop tïïe build-up of r e s i s t a n c e i n the patho -gens i n some cases(55-56). The s i l v e r y disease of sugarbeet caused by CorynëTSacterium i s i n s u f f i c i e n t l y c o n t r o l l e d by m e r c u r i a l compounds, but i s completely e l i m i n a t e d when seeds are dipped f o r s e v e r a l hours i n a s o l u t i o n of s t r e p tomycin. Most of the work with a n t i b a c t e r i a l a n t i b i o t i c s seem confined t o Europe, India, Japan and New Zealand. Treatment of tomato seeds with a n t i b i o t i c s c o n t r o l s b a c t e r i a l pathogens(57). Xanthobacidin has been s a i d to be a c t i v e against Xanthomonas and other s p e c i e s . Seed-borne b a c t e r i a l tumors i n tobacco may a l s o be t r e a t e d with chloramphenicol and t e t r a c y c l i n e ( 5 8 ) . Bacterium f o r crown-gall(with T i plasmids) can be i n h i b i t e d by spraying t e t r a c y c l i n e s on plants that are i n f e c t e d with Agrobacterium. Screening of the e f f i c a c y of a n t i b i o t i c s against b a c t e r i a l plant pathogens i s continuing(59-60). The involvement of plasmids i n c o n t r o l l i n g r e s i s t a n c e of plant pathogens to a n t i b i o t i c s has now been w e l l studied(58). Caution i n using a n t i b i o t i c s against b a c t e r i a l plant patïïogens i s very important, to avoid r e s i s t a n c e build-up i n the environment. The use of a n t i b i o t i c s f o r the c o n t r o l of plant v i r u s diseases (6^1 ) i s of i n t e r e s t . Several a n t i b i o t i c s have been t e s t e d f o r i n h i b i t i o n of r e p l i c a t i o n of v i r a l n u c l e i c a c i d and/or p r o t e i n synthesis w i t h i n the host c e l l . Chloramphen i c o l , cycloheximide, actinomycin D and others are the most used a n t i b i o t i c s ; and the disease caused by tobacco mosaic



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virus(TMV) the most t r e a t e d . In most of the cases(62)the work i s s t i l l at the t h e o r e t i c a l or experimental l e v e l and the p r a c t i c a l use of a n t i b i o t i c s f o r c o n t r o l of v i r a l d i s eases has not been e s t a b l i s h e d . P r i n c i p l e s i n s e l e c t i n g a p a r t i c u l a r a n t i b i o t i c have not been d e f i n e d . Mostly a broad spectrum, r e l a t i v e l y inexpensive, non-phytotoxic one should be s e l e c t e d . Those with growth s t i m u l a t i n g a c t i v i t y f o r the host plant are p r e f e r r e d ( ^ 3 ) . Much work i s needed i n t h i s area, but mention may be made that a n t i b i o t i c s are known that cure the l e a f - c u r l disease of tomato, and a l s o r e s u l t i n l a r g e r s i z e and number of tomato f r u i t s ( 6 0 ) . C e r t a i n v i r a l diseases of tobacco, potato, cucumber, tomato and other crops have been t r e a t e d with a n t i b i o t i c s , but s t i l l many of the plant pathogens(especially v i r u s e s ) causing widespread and severe damage have not been c o n t r o l l e d successfully. Viruses such as bromegrass mosaic, broadbean mottle, c h i l i mosaic, cowpea yellow mosaic, cucumber mosaic, pea streak, potato v i r u s X, soybean pod mottle, tobacco mosaic, tobacco n e c r o s i s , tobacco tumor, tomato l e a f - c u r l and tomato spotted w i l t have been t r e a t e d with a n t i b i o t i c s such as actinomycin D, b l a s t i c i d i n S, a c t i d i o n e ( c y c l o h e x i m i d e ) , miharamvcin A, ohvamycin, polyoxin A, pentaene G8, chloramphenicol, c i t r i n i n , daunomycin, dextromycin, formvcin, kan^ amycin, mitomycin C, mithramvcin, r i b a v i r i n and t u b e r c i d i n (ICI, 64). An a n t i b i o t i c that i s r e l a t e d to r i b a v i r i n (known i n animal v i r o l o g y ) , c a l l e d t a i z o f u r i n has r e c e n t l y been t r i e d i n Sao Paulo, B r a z i l , against tomato spotted w i l t virus(TSWV) and i s s a i d to be an e f f i c i e n t a n t i - v i r a l drug (65). S i m i l a r l y i n India, DPB(code name f o r an a n t i b i o t i c ) i s u s e f u l i n c o n t r o l l i n g the tomato l e a f - c u r l v i r u s , and a l s o increases the s i z e of the tomato f r u i t s ( 6 6 ) . C y t o v i r i n i s a wide spectrum a n t i b i o t i c against plant v i r u s e s , and has proved e f f e c t i v e against the v i r u s diseases of the crops l i k e r i c e , c i t r u s and sugarcane(Table 2). Mycoplasma-like organisms(MLOs) and r i c k e t t s i a - l i k e organisms(RLOs) are i n a c t i v a t e d more e a s i l y than v i r u s e s by a n t i b i o t i c s , since they have membranes l i k e b a c t e r i a and are a f f e c t e d by a n t i b i o t i c s more d i r e c t l y during membrane biogenesis(62). T e t r a c y c l i n e treatment has been very e f f e c t i v e against s e v e r a l MLO-diseases, e s p e c i a l l y i n egg p l a n t , sandal, mulberry dwarf, sugarcane stunt, and grassy shoot. Grapevine n e c r o s i s , hop c r i n k l e and beet yellows are s a i d to be caused by RLOs. C i t r u s greening and other s i m i l a r diseases may be c o n t r o l l e d with t e t r a c y c l i n e , p e n i c i l l i n and aureofungin(68). It i s o f t e n s a i d that when a suspected v i r u s disease may be c o n t r o l l e d by a n t i b i o t i c s ( T a b l e 3), the cause of the diseases must be mycoplasma and never a v i r u s . This a r b i t r ary statement segregating v i r u s e s from mycoplasma has many times been h e l d v a l i d , but there remain s e v e r a l other i n s tances where a p p l i c a t i o n of a n t i b i o t i c s to the host plant has reduced the pathogenesis of v i r u s e s to a considerable degree(69-70).


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Table II :

A n t i b i o t i c s Used against Plant Viruses


Viruses

Antibiotics

Bromegrass mosaic

Actinomycin, B l a s t i c i d i n

Broadbean mosaic

Actidione/Cycloheximide

C h i l i mosaic

DPB (chemical name unknown)

Cowpea yellow mosaic

Actinomycin

Cucumber mosaic

Blasticidin

Egg plant mosaic

Actidione

Pea streak

Actidione

Potato v i r u s X

Actinomycin, B l a s t i c i d i n , Miharamycin, Ohvamycin, Polyoxin A

Soybean pod mottle

Actinomycin

Sunhemp mosaic

Pentaene G

Tobacco mosaic

Actidione,Actinomycin D, B l a s t i c i d i n , Chloramphenicol, C i t r i n i n , Daunomycin, Dextromycin, F e r r i m y c i d i n , Formycin, Imanin, Kanamycin, L a u r i s i n , Miharamycin, Mitomycin C, Naramycin, Ohyamycin, Pentaene G8, Polyoxin A, Puromycin, Streptomycin.

Tobacco n e c r o s i s

A c t i d i o n e , Chloramphenicol

Tobacco tumor

Chiorampheni c o l , Daunomyc i n , Mithramycin, T u b e r c i d i n

Tomato l e a f - c u r l

DPB (chemical name unknown)

Tomato spotted w i l t

Taizofurin

A n t i b i o t i c s from higher forms of L i f e I s o l a t i o n and c h a r a c t e r i z a t i o n of a n t i b i o t i c s from microorganisms has been attempted f o r several decades. There has r e c e n t l y been i n c r e a s i n g i n t e r e s t i n e x t r a c t i n g a n t i m i c r o -


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Table I I I : Therapeutic drugs against Mycoplasmal Diseases of Plants Disease

Drug

Application

Host-Plant

Dwarf

Tetracycline, c h l o r - , oxy-, dimethyl-,and other d e r i v a tives

root immersion f o l i a r spray girdling

mulberry carrot tomato potato rice

Yellows

Methacycline, dipping, hydroChlorampheni- ponics, spray, c o l , Tetracyc- i n f i l t r a t i o n line

Stunt

Tetracycline

root immersion

corn

Phyllody

Doxycline

f o l i a g e dip

aster

Little-leaf

Tetracycline

foliar

legumes tomato

Greening

Tetracycline

sprays

citrus

Decline

Oxytetracyc-line

transfusion

pear trees

Spike

Tetracycline

girdling

sandal

Yellowing

Tetracycline

trunk i n j e c t i o n

coconut

spray

aster chrysanthemum celery tobacco

b i a l compounds from higher plants(71-73). Lichens, algae, angiosperms and various types of marine organisms are being used as sources of a n t i m i c r o b i a l compounds(74). The emphasi s i s obviously on o b t a i n i n g a n t i b i o t i c s usëTul i n human medicine but searches may a l s o be c a r r i e d out f o r chemical compounds e f f e c t i v e against plant diseases. There are r e p o r t s that plant v i r u s i n h i b i t o r s occur n a t u r a l l y i n p l a n t s , and thev could be p r o t e i n s , glycoprote - i n s , polysaccharides, phenols etc(7_5). E x t r a c t s of mosses, e s p e c i a l l y Sphagnum(76), algae(77_) and Cassia of the family Leguminosae(78) are e f f e c t i v e i n i n h i b i t i n g tobacco mosaic virus(TMV), But much more work i s needed to develop v i r i c i des that may be sprayed s a f e l y and economically on crop plants i n the f i e l d . Often medicinal plants known from f o l k - l o r e are picked up and t h e i r e x t r a c t s t e s t e d against known plant v i r u s e s by mixing them with the inoculum and doing h a l f - l e a f experiments. Each h a l f of the l e a f i s rubbed with v i r u s suspension,



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one h a l f r e c e i v e s untreated v i r u s , while on the other h a l f v i r u s i s mixed with plant e x t r a c t . Numerous plant e x t r a c t s are being screened and u s e f u l compounds have been i s o l a t e d . However, v a l i d a p p l i c a t i o n i n plant p r o t e c t i o n has not been e s t a b l i s h e d , although several drugs f o r use i n human medicine have already been developed(78). Some angiosperms l i k e Acalypha i n d i c a has been shown toHSe a c t i v e against plant pathogenic fungi l i k e A l t e r n a r i a ( 7 9 ) . The biomedical potent i a l of the sea has a l s o r e c e n t l y a t t r a c t e d considerable attention(80-81) and searches f o r plant p r o t e c t i n g a n t i b i o t i c s are underway i n the marine environment. Legislation Almost every country has c e n t r a l i z e d drug r e g u l a t i o n s with regard to p e s t i c i d e s and drugs, i n c l u d i n g a n t i b i o t i c s , i n plants and animals. Discussions about the unwarranted use of a n t i b i o t i c s i n human medicine and plant p r o t e c t i o n stem many times from discrepancies i n the drug laws of d i f f e r e nt nations, a thorough account of which has been presented i n several works(TO). A very s i m p l i e f i e d account of the present s i t u a t i o n regarding the use of a n t i b i o t i c s i n plant p r o t e c t i o n reveals that there are extremes of 'no' ( l i k e i n USA and Western Germany) through l i b e r a l y e s ( a s i n Japan and I n d i a ) . Most other countries have adopted a middle path, where a n t i b i o t i c s are allowed i n animal feed, but not i n plant p r o t e c t i o n , or v i c e versa. Reasons f o r t h i s are probably n o n - s c i e n t i f i c . Much of the fear of the unwanted use of a n t i b i o t i c s may be removed a f t e r we have f u r t h e r information i n the f i e l d . The USSR and West Germany r e g u l a r l y p u b l i s h l i s t s of chemicals that are allowed to be used as p e s t i c i d e s and f o r spraying on plants(8283), but a n t i b i o t i c s do not appear i n them. The a g r i c u l t u r a l uses of t e t r a c y c l i n e s have r e c e n t l y been discussed. In the UK i t has now been r e a l i z e d that a f r e s h look i s needed at the problem of using a n t i b i o t i c s i n a g r i c u l t u r e (84). It i s thus necessary to t e s t more and v a r i e d a n t i b i o t i c s against plant pathogens, under c o n t r o l l e d experime n t a l c o n d i t i o n s , before reaching a f i n a l opinion i n the matter. f

f

Growth Promotion In a d d i t i o n to being used as cure against plant diseases, a n t i b i o t i c s may a l s o be used as agents to stop preharvest f r u i t drop, or as a b s c i s s i o n agents to c o l l e c t f r u i t s i n c i t r u s crops(85). Some a n t i b i o t i c s help enhance growth of the crop plants i n a d d i t i o n to c o n t r o l l i n g t h e i r diseases (4). Hence, an i d e a l a n t i b i o t i c that may act against pathogens, increase crop p r o d u c t i v i t y and o f f e r other d e s i r a ble properties without l e a v i n g longterm residues should e a s i l y f i n d approval with drug l e g i s l a t i n g agencies(86).


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Conclusion A n t i b i o t i c s are commonly used as drugs f o r humans and a n i mals p a r t i c u l a r l y against b a c t e r i a l i n f e c t i o n s . There has been i n c r e a s i n g i n t e r e s t i n the use of a n t i b i o t i c s f o r the c o n t r o l of plant diseases since these compounds may o f f e r more e f f e c t i v e and/or safer a l t e r n a t i v e s to chemicals p r e s e n t l y used to c o n t r o l plant diseases. The debate over the p o t e n t i a l biomedical consequences of a n t i b i o t i c s and the need to impose some kind of r e s t r a i n t on t h e i r usage i n a g r i c u l t u r e has become very intense l a t e l y . Caution i s appropriate but the present concerns may be unfounded and excessive. The use of a n t i b i o t i c s has been i n c r e a s i n g i n a g r i c u l t u r e , because of t h e i r obvious b e n e f i t s . Their use seems to pose no obvious harm to the environment. I t i s thus b e t t e r to r e f r a i n from making i l l - f o u n d e d arguments, and to put more e f f o r t i n determining what sort of a n t i b i o - t i c s can be s a f e l y and e f f e c t i v e l y used i n crop p r o t e c t i on. Acknowledgment s Dr. Luther S. Williams and Dr. Joe Johnson provided necess -ary encouragements and f a c i l i t i e s to acomplish the task. Dr. Nipen K. Bose helped with pertinent l i t e r a t u r e on the subject. Mr. S i e b r i n Simon helped with improving the l a n guage of the manuscript. Literature

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