Digat, B.

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WHY AND HOW TO DISTINGUISH THE PSEUDOJJJONASSOLACEARUM STRAINS, CAUSAL AGENT OF THE BACTERIAL WILT OF SOLANACEOUS AND MUSACEOUS CROPS IN THE CARIBBEAN ZONE

AgEcon Search: Research in Agricultural and Applied Economics

  Give to AgEcon Search    The World’s Largest Open Access Agricultural & Applied Economics Digital Library    This document is discoverable and free to researchers across the globe due to the work of AgEcon Search.    Help ensure our sustainability.         AgEcon Search http://ageconsearch.umn.edu aesearch@umn.edu          Papers downloaded from AgEcon Search may be used for non-commercial purposes and personal study only. No other use, including posting to another Internet site, is permitted without permission from the copyright owner (not AgEcon Search), or as allowed under the provisions of Fair Use, U.S. Copyright Act, Title 17 U.S.C. PROCEED~GS OF THECARIBBEAN FOOD CROPSSOCIETYSIXTH ANNUAL MEETINGST. AUGUSTINE, TRINIDADJULY 7.13, 1968VOLUl\:IE VIWHY AND HOW TO DISTINGUISH THE PSEUDOJJJONAS SOLACEARUMSTRAINS, CAUSAL AGENT OF THE BACTERIAL WILT OF SOLANA-CEOUS AND MUSACEOUS CROPS IN THE CARIBBEAN ZONEBy B. Digat*Bacterial wilt affects solanaceous and musaceoua crops in all the countriesof the Caribbean region. It is however difficult to estimate exactly the extent oflosses caused by this disease. In some territories, the pathogen is so virulent through-out the year that susceptible crops are difficult to grow or have entirely disappeared.In other countries the bacterium is virulent only during certain months of the year(wet season) and susceptible crops are grown during the period unfavourable tothe expression of bacterial virulence (dry season).The extent of losses caused by bacterial wilt is always a direct function ofbacterial virulence. If the virulence of the bacterium is subjected to the influeneeof the ecological factors (climate, soil, host-plant) it is at first conditioned by thenature of the bacterial population, i.e. by the nature of strains in the soil. Everystrain has a definite virulence power. It is, therefore, useful to distinguish betweenone strain and another.The purpose of this work is to describe certain specific characters which allowyou to recognize the Pseudomonas eolanacearum strains in the Caribbean zone.Of the specific characters only pathotype, biotype and serotype will be studied.PATHOTYPEThe Pathotype of a bacterial strain is the pathogenicity possessed by thatstrain towards one or several host plants.The pathotype gives only the relative value of the pathogenic potential of thestrain.When the pathotype of one isolate is studied, several difficulties arise sincethe quantity and the quality of the inoculum is dependent On the inoculationmetbods used for the host-plants.1. INOCULUMThe isouue must possess its highest virulence potent'ialTo obtain the highest level of virulence of an isolate it is necessary to makethe isolation on a selective medium (as KEL~AN'S medium) (ll) especially sincethe ratio of virulent to avirulent cells in inoculum influences the severity of bacterialwilt (2).The highest level of oirulence must be maintained before and d'uring the inoculationof the host plantThe best means of maintaining virulence of tho strains is to keep the isolateunder sterile distilled water (3, 4, 5).• Plant Pathologist-Institut Nutional de Ill. Recherche Agronornique (Franco).86CURRENTSTATIJSOFTHEDISTRIBUTIONOFTHEPSEUDOMONASSOLANACEARUMSTRAINS.'" 110.."....Ir;.,-. :?0......,..•...JO·I..~t.·CIdII"l,.iL.....~I..-:AI=m&I:I.tP::i_'\.......MlI'A;,I_."\)--bff-._-_..-~"ia:51~\AtICARIBBEAN- tr::.1IIV.~1'_"'lr.<>;·V-·~C~'Tt]"C·dI~]----".,----,---~ooc400II.........:lOO'-.......II...'".......NI.t•If•••,"e••,••L••'••T').O'IA,q0..,TM,,"rllle,,-if!C;;Ai?Ji~-~A~n:1_,4~\j.i_:.~~...~...._....~I.......ACJiIIEi:I..-:'...111~...,.••ltlll.ojIf','I'C'"~'A""""A'1111""'-'iJII;l.....•0••,.....,'_1'U'II•••lOGAWEST'INDIESAND~.~..._...9__.::....~....'t":'........-+-~•_110.C"'~I\,11....~..&...J.",,-'tJ~'D'il"'Illl'''Iv'''_-1lull'u,,,f'"'_1. -......",~"'''''''II_J;,.......~..~0..•I•0,~"-3,L,T-r\-if1....1oI.t.0",ICe.1I,!c§:J,"",........"'......"1\' 'II•••".•~~"_At'....lCGI."~"'."'.DC:.~1I :---~_._---\\ '\c_~~....V,O€..,.-"'':'~I(D.Jtr.."..)Ci.l'n)c;~I(~,~~~.~H~~~...<)"-~'-~--,'7'---.\.\'Ji\.L.I·"[-_.~o,J[Ic,(1tJ.l,",001---'"I II.i->">]I.~-I(:(..~c.nc(l..""",):..J,.E11,eo"-.JI I I,. I ICiJJli(iF,..4tNiftl I I I--Jl----, I ISCHEMATIC REPRESENTATION OF PRECIPITATION LINES PATTERNSobtained by immunoeclectrophoresis.+- -+-.+_.. --- -_. --'--. _. _._-. -- -~fROT~PE IS'EROT~Pt: II~-------~. --'-'--. -. --. -'-.- ..sfROT~Pf: mAg: Hole, cut in the agar, filled with somatic "0"antigen soluti on extracted from.a strainof PseuCbmonas solanacearum E. F. Sm.2. INOCULATION METHODSThe root- inoculation and stem-inoculation methods have been described byWINSTEAD AND KELMAN (6).3. HOST-PLANT RANGEFour groups of host-plants A, B, C, D are proposed, as shown in Table I. Accord-ing to the observed symptoms in these host plants, the pathotype of the isolatemay be restricted to one group or may belong to several groups.If the symptoms are severe ("fast wilt"), the designated letters for the pathotypeare A, B, C, D. But if the symptoms are mild (slow wilt) the designated letters arerespectively a, b, e, d.The isolate is classified as to pathotype only when it can produce in the group,symptoms of wilt. For instance, isolates from Guadeloupe bclong to the three patho-types A, a and b (see Table 2).II. BIOTYPEAccording to HAYWABD (7), the biotype or biochemical type is the power of aPseudomonas solanacearum strain to utilize or not:-the three disaocharides (maltose, lactose, cellobiose).-the three hexose-alcohols (mannitol, sorbitol, dulcitol).The method (7) allows ns to distinguish four biochemical types according asthe Carbohydrate oxidation is positive or negative,TABLE 1BIOTyrE1 2 3 4,CARBOHYDRATEMALTOSE ... ... - + + -LACTOSE ... ... - + + -CELLOBIOSE ... - + + -MANNITOL ... - - + +SORBITOL ... ... - - + +DULCITOL ... ... - - + +~The 23 strains of the Caribbean zone belong to biotype 1 or b,i,otype 3. (see Table 3).There is no direct relation.sh'i,p between the paf,hotype am.d the b'i,otype.Isolates from PUERTO RICO and MAB,TINIQUE have the pathotype A but COSTARICAN and HONDURAN isolates have the pathotype C. However, all these isolatesbelong to the same biotype 1. The homogeneity inside of some territories is remark.able. In PUERTO 111CO, all the isolates from tomato belong to the same biotype 1.In MABTINIQUE, all the isolates from both tomato and eggplant belong to biotype 1.The possibility exists of finding in the same territory two biotypcs as in GUADELOUP]lwhere biotype 1 and biotype 3 are present.87ITL SEROTYPEFor Pseudomonas solanacearum, ident.ities or non-identities among the strainsof the bacterium could not be clearly definited (8,9, 10, 11, 12, 13) and the validityof the notion of serotype Qt. serological type remained uncertain. This was dueespecially to the lack of sharpness of techniques used in the serological analysisof the Pseudomonas solanacearum. antigens.In a previous work (14) it was suggested that the Immunoelectrophoreticanalysis of Somatic "0" Antigens could permit the identification of one strainfrom another.The 23 Somatic "0" antigens of the strains from the Caribbean zone wereextracted and studied.One type of precipitating antiserum made from the "QUA To 4" strain permitsthe identification of only 3 different patterns among these 23 antigens.Serotype I (see schematic diagram)The "GUA To 4" autiserum precipitates its somatic corresponding antigen(extracted from the GUA To 4 isolate) according to two arcs a and B.This pattern is the most common. Twenty somatic antigens precipitate accordingto this pattern:PUERTO RICOGUADELOUPEPR 65PR 69FR 81PR 211 BFR 212GAU To 1GUA To 2GUA To 3GAU FT 1QUA TA 1GUA TA 2QUA To 4MARTINIQUE MAR To 1MAR Au 1MAR Au 2MAR Au 3COSTA RICA B 139-BBDCOLOMBIE H 249Serotype II (see schematic diagram, Figure 1)This pattern includes not only the two arcs A and B but a supplementary arc,originating from the A arc with the same shape and length as the Bare.Only one isolate belongs to this pattern:FRENCH OUIANA (OUY TO 1)Serotype III (see schematic diagram)In this pattern, three specific precipiting bands arc joined to form a continuouscurve, located above the A and Bares.Two somatic antigens give this pattern:HONDURAS (SFRPUERTO RICO (PR 80.88DISCUSSIONThe specific characters of the Pseudomonas solanaceann» strains are summarizedin Table 3.The geographical distribution of these strains in the Caribbean zone as Beenin the attached map shows that the pathogen is uniformly distributed, but thatthe pathotype C (virulent strain for the Musaceous crops) is rather continental:GUATEMALA, HONDURAS, COSTA RICA, PANAMA, COLOMBIA, VENEZUELA, BRITISHGUIANA (except for its existence in TRINIDAD which is close to the VENEZUELANMainland).It is very likely that the pathotype A (which is virulent for Tomato, Potato,Eggplant and Pepper) is present in all territories of the Caribbean. Its presencecould explain the weak development of the Solanaceous vegetables in this zone.The lack of biotype 4 and the infrequency of biotype 2 (only present inCOLOMBIA) are noticeable,A more comprehensive study of the serotypes is in progress and will leadto a better understanding of the origin, distribution and properties of some Pseudo-m.onas solanacear'um strains,TABLE 1Host Plants Groups for the determInation of Pathotypes of Pseudomonas SolanacearumIsolatesA (a) B (0)IC (e)lD(d)(Vegetable Solanaceous) (Tobacco Group) (Musaceous)TOMATO (Lyeopersioon TOBACCO (Nieoliana BANANA (Musa. spp.) PEANUT (Arachiseeculentum Mill) tao<tcum [_.) hypogae<t L.)POTATO (Solanum HELICONIA caribea SESAME (Sesamet·uberosurn L.) indieurn L.)EOOPLANT «Solan'um HELICO.NrA psittacorum GINGER (Zingibermv!onoena L.) officinale Rosc.)PEPPER (Capsicum HELICONIA spp.frutescens L.) STRELITZIA spp.TABLE 2Pathotypes of the isolates of Pecudomonas Solanoccarum E. F. Sm. in Guadeloupe~ TOMATO TOMATO TOBACCOHOST-PLANT from (fast wilt) (slow wilt) (slow wilt)GROUl.'A (a) * t tB (h) § § tC (e) § § §D (d) + § •PATIIOTYPE A a bDegrees of virulenr;e: * strong (fast wilt).t medium (slow wilt).t weak (no wilt).§ avirulonce.89TABtm aSpeelflcal Ch"r"eters of the Pseudomonaa Sol"nllee"ruID StraJnsIn the C"dbbenn ZoneTERRl'l'ORY orI ARIIA OF IHOST-PLANTINUMBER OJ!' j AUTHORS, DATH OJ!' IPATHO· rBIOTYFE SIlBO-ORiGINE SAlO'LlNO THE ISOLATE ISOLATION TYPE TYPEPUlUlTO-RICO ? TOMATO PR 65 J. e. FllRllZ andA. O. MONLLOR 1962 (1958) A 1 I? TOMATO !'II 69 " " A 1 I? TDMATO PR 80 " .. A 1 III? TOJdATO PR 81 '. " 1963 (1959) A 1 I? TOMATO PR 211B .. " ? A 1 I? TOMATO PB 212" " ? A 1 I--- -GUADIILOUPE Duclos TOMATO GUA TO 1 B. DIGAT 1965 A 3 ILongueteau TOMATO GUA TO 2 A. ESCUIllE 1964 A 1 IVx Habitants TOMATO OUA TO 3 B. DIOAT 1960 A 1 IPte Noire POTATO OUA PT 1 .. " 1965 A 1 ICspeeterre TOBACCO OUA TA 1 " " 1067 b 1 IRoujol TOBACCO GUA TA 2 " " 1907 b 1 IBeauport TOMATO GUA TO 4 .. " 1965 B 3 IIURTHIIQUE Basse-Pointe TOMATO MAR TO 1 B.DIGAT 1965 A 1 IBasse-Painte EGGPLANT MAR AU 1 .. .. 1967 A 1 ISaint·Pierre BGOFLANT MAR AU 2 " .. 1967 A 1 IMorne-Rollge EOOFLANT MAR AU 3 .. " 1967 A 1 I---~TRllIIDAD ? MUSA sp. a H6 A. C. HAYWARD 1957 0 1 ?(TRI BA 1)La REUNION MUSATrinidad "Mysore" TRI BA 2 B. :OIOAT 1068 0 3 ?GovernmentCocoa propa-illation unitLas Lomas lII.ITSA"Moko Fig" TRI DA 3 .. " 0 3 ?Matura MUSA"Glantestate Cavendish" TRI DA 4 .. .. 0 3 ?Matura MUSAestate "Lacatan" " " C3 ?Sangre MUSAGrande "Horse"EI Reposo" plantain" TRI BA /; ., " C 1 ?StationSangre MUSA TnI DA 6 " " 01 ?Grande "Mysore"Sainte-Marie? TOMATO a 610 W. J. DOWSON 1959 A 1 ?(TBI TO 1) J. A. SFENOE 1957Aranguez TOMATO TRI TO 2 D. D10AT 1968 A 1 ?MarketGardenEstateCaura Valley TOMATO TRI TO 3 .. " 1968 A 1 ?St. HelenaPlaroo TOMATO Till TO 4 " .. 1968 A 1 ?BGG·PLANTSangre1968 1 ?Grande TOMATO TR1 TO 5 " .. A"El Reposo"Stationl/RllNOH GUIANA Matoury TOMATQ aUY TO 1 H. J'LOCH and 1965 A 3 II(Slcama) B.DIGAT 1968Cayenne TOMATO GUY TO 2 B. mGAT 1968 A 1 ?SuziniKourou TOMATO GUY TO 3 B. maAT 1968 A ? ?(Beae)COBTA RICA Coto VaUay BANANA B. 139-B I. W. BUDDENHAGEN 1959 e 1 IBANANA B n n .. " c 1 IBllLICONIA D u tJ .. " 0 1 Iasp------HONDURAS U1ua Vallay lIAN.il.NA SFR I. W. BUDDENHAaEN 1961 C I III------COLOMBIA ? BllLIGONJA H 249 I. W. llUDDENHAGEN 1900 a 1 ICARlBIilA90REFEHENCES1. KELMAN, A. 1954 "The relationship of Pathogonicity of Pseudomonas solanacearu17l tocolony a.ppearance in a tetrazolium medium." PhJytopathology 44: 693-695.2. AVERll.E, C. W. AND KELMAN, A. 1964. "Severity of bacterial. wilt as influenced by thoratio of virulent to avirulent cells oiPseudosnona« solanaceaT'um in inoculum." Phytopatlwlogy54: 779-783.3. KELMAN, A. AND JENSEN, J. H. 1951. "Maintaining virulence in isolates of Pseudomonas80IarUJCearum." Phylopatlwlogy 41: 185-187.4. KELMAN, A. 1956, "Factors Influence viability and variation in cultures of Pseudomona.~8olanacearum. Abate." Phytopathology 4(,: 16-17.5. KELMAN, A. and PERSON, L. H. 1961. "Strains of Pseudomonas 80lanacearum differingin pathogenicity to tobacco and poanut." Phytopathology 51: ] 58-161.6. WINSTEAD, N. N. and KELMAN, A. 1952. "Inoculotion techniques for evaluating resistanceto Pseudomonae solanacearum" Phytopathology 42: 628-684.7. HAYWARD, A. C. 1964. "Characteristics of Pseudomonas solanaccarum" J. Appl. Bact. 27(2): 265-277.8. OXABE, N. and GOTO, N. 1954. "Studies on .Pseu,domonu8801unaeearu.m VI A comparativestudy of the colony type organisms F and Op. Shlzuo~:a Univ. Eac, Ag", Rcpt, 4: 41. 60.9. HARBISON, D. E. and FREEMAN, H. 1961. "Bacterial wilt of Potatoes, II. Serologicalrelationship of two strains of Pseudomonas 801anaeear'um and a culture of Corynebacteriumsepedonieum" AU8traiian J. oj Agr. Research 12: 872-877.10. PEREZ, J. E. 1962. "The use of Fullers's Formamide method in the serological identi-fication of Pseudomonas 80Ianacearum." The J. oj Aqr, oj Uni», Puerto-Rico 46: 144--·153.11. QUINON, V. L., ARAGAXI, M. and ISHII, M. 1964. "Pathogenicity and sorologiosl relation.ship of throe strains ofPseudomonas 8olanar.earum in Hawaii." Phyf,opathology 54: 1096-1099.12. MORTON, D. J .• JENKINS, S. F. JR. and Dl;XES, P. D. 1965. "Serological relationship amongraces I, 2 and 3 of Pseudomonas solanacearum;' (Abstr.) Phytopathology 55: 1069.13. MORTON, D. J., DUKES, P. D. and JENKINS, S. F. JR. 1966. "Serological relationship ofreces I, 2 and 3 of Pseudomonas 801anacearum.'' Plant D1:S. Reptr. 50: 275-277.14. DIGAT, B. ot Escrrnra, A. 1967. "Reconnaissance du Flotdsscmont bacterien aux AntillesFraueaiaea." Phytiatrie·Phytopharmac'ie 16: 187-197.91

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WHY AND HOW TO DISTINGUISH THE PSEUDOJJJONASSOLACEARUM STRAINS, CAUSAL AGENT OF THE BACTERIAL WILT OF SOLANACEOUS AND MUSACEOUS CROPS IN THE CARIBBEAN ZONE

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