A PRELIMINARY SURVEY OF THE FUNGUS FLORA OF . " I. AQUATIC PHYCOMYCETES.

BY

MARGARET MARS'.1ALL HOLLAND /.I

THESIS SUBMITTED TO THE GRADUATE FACULTY OF THE

VIRGINIA POLYTECHNIC INSTITUTE

tN CANDIOACY FOR THE DEGREE OF

MASTER OF SCIENCE

IN

BIOLOGY, BOTANY OPTION

APPROVED: APPROVED:

DEAN OF AGRICULTURE ·~AJOR PROFESSOR

MAY 15, 1958

BLACKSBURG, VtRGINIA I I

PREFACE

A SYSTEMATIC INVESTIQATION OF THE AQUATIC FUNQI

OCCURRING IN THE SOILS OF HAS NEVER BEEN MADE.

TH~RE AR! A FEW RECORDS OF FUNGI COLLECTED HERE SY

VISITING MYCOLOGISTS, BUT NO INFORMATION IS AVAILABLE

CONCERNING TH£ HABITS, PREVALENCE, ANO DISTRIBUTION OF

THESE ORGANISMS IN THE STATE.

THE PRESENT CONTRIBUTION CONSIDERS BUT ONE AMONG

MANY HABITATS OCCUPIED BY THE AQUATIC PHYCOMYCETO·us

FUNGl 1 AND IS A PART OF A MORE COMPREHENSIVE INVESTIOA-

TION, UNDER THE DIRECTION OF PROF• WtLLIAM W. SCOTT,

OEALINQ WITH THE TAXONOMY, MORPHOLOGY, ANO DISTRIBUTION

OF THESE FUNGI. THE WRITER HAS BEEN PARTtCULARLY

fNTERESTEO IN THE CHYTRIDtACEOUS 1 BLASTOCLADIACEOUS 1

SAPROLEQNfACEOU8 1 AND PYTHIACEOUS SPECIES OCCURRING IN

THE SOIL ANO WATER OF CAVES IN SOUTHWESTERN VIRGINIA•

ALTHOUGH IT IS NOT POSSIBLE TO ACKNOWLEDGE THE

NUMEROUS INDIVIDUALS WHO HAVE CONTRIBUTED TO THIS

fNVESTtGATION, THE AUTHOR WISHES TO EXTEND HER SINCERE

GRATITUDE TO ALL THOSE WHO HAVE ASSISTED IN ANY WAY

TOWARD THE COMPLETION OF THIS STUDY. SINCERE APPRECIATION

IS EXPRESSED TO THE MEMBERS OF THE COMMITTEE UNDER WHOSE

DIRECTION THIS INVESTIGATION WAS CARRIED OUT. THESE

INCLUDED PROF. W. W. SCOTT, CHAIRMAN; DR. t. 0. WILSON, I J I

HEAD, DEPARTMENT OF BIOLOGY (RETIRED); OR. F. S. ORCUTT, HEAO, DEPARTMENT OF BIOLOGY; OR. A. B. MASSEY; OR. K. W. KING; AND DR. 0. E. NORBY. THE WRITER IS ALSO VERY GRATE- FUL TO MRr LARRY GRIFFIN FOR HIS ASSISTANCE IN THE

COLLECTION OF SAMPLES, ANO TO THE MANAGEMENTS OF THE

FOLLOWJNQ CAVERNS FOR PERMISSION AND AID IN COLLECTING

MATEP.IALSt , , LURAY

CAVERNS, SKY LINE CAVERNS, ANO .

APPRECIATtON IS ALSO EXTENDED TO THE VtRGINIA ACADEMY

OF SCIENCE FOR FINANCIAL ASSISTANCE, WHICH ENABLED THE

WRITER TO !XTEND THE SCOPE OF THIS tNVESTIGATION. THE

WRITER WISHES TO ACKNOWLEDGE THE ASSISTANCE OF OR• PAUL

Me PATTERSON, HOLLINS COLL£G£ 1 FOR THE tDENTIFICATtON OF

THE BRYOPHVTES AND DA. A. B. MASSEY FOR THE tOENTIFICATION OF THE FERNS• IV

TABLE OF CONTENTS

INTRODUCTION. • • • ••••• • •• • • • • • • • • • LITERATURE REVIEW • •• • •••• • • • • • • • • • • 5 MATERIALS ANO METHODS • • • • • • • • • • •• • • • • 10 FUNGI COLLECTED • • • •• • •• • • • •• • • • • • • t3 CHYTR llH ALES • • • • • • • • • • • • • • • • • • 13 I. NOWA~OWSKIELLA RAHOSA E. J. BUTLER. • • • • • f3 2. ? RHtZOPHLVCTIS SPP. • • • • • • • • • • • • 14 BLASTOCLADtALES• • • • • •••• • • • • • • • • 15 3, ALLOMVCES CYSTOQ!NUS EMERSON. • • • • • • • • 15

4. BLASTOCLAOIOPSIS PARVA (WHtFFEN) SPARROW. • • f6

SAPROLEGNtALES • • • • • • • • • • • • • • • • • JS 5. ACHLVA OUStA COKER. • • • • • • • • • • • • • 18 6. APHANOMYCES ~AEVIS DEBARY • • • • • • • • • • 19 7. SAPROL£QNIA FER~X (GRUtTH) THURET • • • • • • 2f

8. SAPROLEGNIA MIXT~ DEBARY. • • • • • Ill • • • • 22 LEPTOMITALES • • • • • • • • • • • • • • • • • • 24 9. APODACHLYA MtNtMA COK£ft AND lCtTNER • • • • • 24 lAGENIDIALES • 111 111 • • • • • • • • • • • • • • Ill 25 10. O~PtDtOPStS APHANOMYCIS CORNU. • • • • • • • 25 ti. 0LPIDtOPSIS SAPROLEQNIAE (BRAUN) CORNU • • • 26

PERONOSPORALES • • • • Ill • • • • • • • • • • •• 28 12. PYTHtUM AFERTILE KANOUSE AN~ HuMPHREVe • • • 28 • 13. PYTHIUM CAROLINIANUM MATTHEWS. • • • • • • • 29 14. PYTHf UM IRREOULARE 8Ul8MAN • • • • • • • • • 30 v

15. PYTHIUM MONOSPERMUM PRtNeSHEIM. • • • • • • • 32 16. PYTHIUM NA861t ITO AND TOKUNAQO • • • • • • • 34 17. PYTH!UM PAPILLATUM MATTHEWS • • • • • • • • • 35

f 8. PYTHIUM PULCNRUM MINDEN • • • • • • • • • • • 37 19. P'ITHIUM ROSTR6TUM BUTLER• • • • • • • • • • • 39 20. PYTHIUM VEXAN! OEBARY • • • • • • • • • • • • 40 SUMMARY ••• • •• • •••• • • • • • • • ••• • • • 42 BfBLIOGRAPHY • • • • • • • • • • • • • • • • • • • • • 43 VITA • • • • • • • • • • • • • • • • • • • • • • • •• 50 INTRODUCTION

ALTHOUGH THE FAUNA OF CAVES HAS BEEN STUDIED tNTEN-

81VELY1 RELATIVELY LITTLE ATTENTION HAS BEIN QtVEN TO THE PLANT LIFE PRESENT IN SUCH HABITATS• THll DISCREPANCY 18

DUE, FOR THE MOST PART, TO TH£ DEPENDENCE OF CHLOROPHYLLOU8

PLANTS UPON LIQHT. StNCE LIQHT ta ESSENTIAL FOR THE

QROWTH ANO DEVELOPMENT OF QREEN PLANT8 1 THE ABSENCE OY THESE FROM THE CAYE ENVIRONMENT MIQHT BE ANTICIPATED.

A FEW MEADER REPORTS ARE FOUND IN THE LITERATURE,

HOVEY£R 1 CONCERNING THE PRESENCE OF CERTAIN ALGAE, MOSSES,

AND LlVERWORT8 F~UNO IN CAVE8e THESE GENERALLY GROW NEAR

THE 'S ENTRANCE OR IN THE VICINITY OF ARTIFICIAL ILLUMINATION, AND MAY NOT BE REGARDED AS TRUE CAYE

INHA8fTANT8• MJ88 HARRING (1930), FOR EXAMPLE, HA8 REPORTED THE FOLLOWING BRYOPHYTES FOUND GROWING NEAR ELECTRIC LIGHTS

IN H'WE CAVERNe: MARCHANTl6 POLYMORPHA, AMPHIDIUM MONGEOT!I,

AMILY!TEJIUM JURALZKANUM, 8RACTYTHECIUM RUIABULUM, BRYUM

C6E8PITICU~, 8RYU~ RAPILLARE, FUNARI! HYGROMETR!CA1 AND LIPTOBRYUM PYRIFORME• IN ADDITION, SCHl8T08TJGIA 08MUNDAC§A HA8 BECOME TH! CLASSICAL EXAMPLE OF A GREEN PLANT STRUCTURALLY ADAPTED FOR GROWTH ANO DEVELOPMENT IN THE

8UIDUED LIGHT OF CAVEi (VEROOORN 1 1932). THE HIGHER CON- CENTRATION OF CARBON DIOXIDE IN CAVES 18 APPARENTLY ONE OF THE FACTORS MAKING tT POSSIBLE FOR MOSSES TO 8ROW UHDIR

THE8E CONDITION&. COINCIDENTAL WITH THE COLLECTION OF -2-

MATERIAL FOR THIS STUDY, THE FOLLOWtNQ BRYOPHVTES WER!

COLLECTED ANO IDENTtFIEDS FtSSIDEN! MINUTULUS SULL.,

LEPTODICIYUM RIPARIUM (HEDW.) WARNST., THUIDIUM VIRQINIANUM (BRIO.) LINOB., EURHVNCHIUM 8ERRUL!TUM (HEow.) KINDe •• ANO A&TOMUM MUHLENBERQIANUM (sw.) GROUT. ALSO FOUND. NIAR AN ELECTRIC LIGHT IN , WAI THE COMMON WOODLAND

FERN 0RYOPTERl8 tNTERMEOIAe As MIGHT BE EXPECTED SUCH 8PECtMENS ARE GREATLY ETIOLATED AND POORLY DEVELOPED OFTEN

TO THE EXTENT OF MAKINQ IOENTtFICATION DIFFICULT. SEED-

~tNG8 OF CERTAIN HIGHER PLANTS MAY SOMETIMES BE FOUND IN

TH£ REMOTE AREA8 OF CAVES• THESE ARE REMARKABLE FOR THE

EXTREME DEGREE OF ELONGATION EXHfBtTEO. SUCH SEEDLINGS

DIVELOP APPARENTLY FROM QERMINATINQ 8EEDS CARRIED •NTO

THE CAVES BY AIR CURRENTS OR BY ANIMALS, GROW AT THE

EXPENSES OF THEIR STORED FOOD, THEN WITHER AND DIEe

SINCE MANY OF THE FUNGI CAN COMPLETE THEIR DEVELOP-

MENTAL CYCLES tN THE ABSENCE OF LIGHT, IT MIGHT BE

EXPECTED THAT SUCH SPECIES WOULD THRIVE IN CAVE8• IN

THE CASE OF CERTAIN OTHER FUNGI, HOWEVER, LtQHT EXERTS

A DECIDED MORPHOQElltC OR A PHOTOSTATIC EFFECT OR, IN

SOME CASE8 1 MAY BE REQUIRED FOR THE FORMATION OF REPRO-

DUCTIVE STRUCTURE&. IT WOULD APPEAR LIKELY THAT SUCH

SPECIES WOULD BE ENTIRELY ABSENT FROM THESE HABITATS OR 1

IF PRESENT, WOULD BE ABORTIVE OR STERILE. PREVIOUS tNVESTIQATORS HAVE NOTED SUCH ABORTIVE FRUITING STRUCTURES -3-

AND OTHER REPORTS HAYE INDICATED THE FREQUENT PRESENCE IN

CA¥E8 OF STERILE MYCELIA• FOR MANY SAPROPHYTIC FUNQt,

HOWEVER, THE ENVIRONMENTAL CONDITIONS FOUND IN CAVES WOULD

S!EM TO BE IDEAL• THE UNIFORMLY HIGH HUMIDITY, TEMPERATURE OF s2-sso F., AND THE NEUTRAL TO 8Ll8HT ALKALINITY OF THE SOIL AND WATER ARE ALL FAVORABLE CONDITIONS FOR THE DEVELOP-

MENT OF MANY FUNQt. THE ABSENCE OF LIGHT AND THE RESTRICTED

SUPPLY OF AVAILABLE NUTRIENTS MfQHT BE EXPECTED TO LIMIT

THE VAAICTY OF FUNGI IN CAVES.

NUMEROUS ACCOUNTS HAVE 8£~R PUBLISHED CONCERNING THE

FUNGI FOUND IN MfNE8 1 TUNNELS, CELLARS, AND SIMILAR MAN-

MADE HA8fTAT8• THE ORGANISMS ENCOUNTERED IN THESE HYPO•EAL ENVIRONMENTS DO NOT NEOE88ARILY REPRESENT THE KINDS OF FUN4t WHICH ONE WOULD EXPECT TO FIND IN NATURALLY

OCCURRING CAVES. CAVERNICOLOU8 FUNGI MAY BE DIVIOED 1 FOR

CONVENIENCE, INTO GROUPS. THOSE FUNGI, FOR EXAMPLE, WHICH

OCCUR NORMALLY ta CAVES AND THRtVE THEREIN BECAUSE OF

FAV0RA8LE ENVIRONMENTAL CONDITION8 MAY BE TERMED "TROQLO-

PHfLE8• (TROQLO • CAVE8 1 PHILEO •LOVE). THE SPECJES WHICH

ARE INTRODUCED INTO CAVES AND EXIST THERE ONLY TEMPORARILY

MAY BE TERMED HTROQLOXEM!S" (TROGLO • CAVE, XENO! • GUl8T).

THE LATTER ARE FOUND MOST FREQV!NTLY ON TtMBER, DISCARDED

FOOD, AND OTHER REJECTAMENTA INTRODUCED BY EXPLORERS OR

~EFT BEHIND BY CASUAL VISITOR&. -4-

THE PRESENT INVESTiCATtON IS CONCERNED PRIMARILY

WITH THE TROQLOPHflttS 1 THOSE FUNGI OCCURRING HATURALLV tN

THE SOIL AND WATER OF' CAV!!S. Tut WRITER IS PARTICULARLY fNTERESTED tN THE AQUATIC PHYCOMYC!TES 1 HENCE CONSfOERABLE

ATTENTION HAS BEEN GIVl::N TO THESE FUNQl ounrnn THE PRESENT

INVESTtGATION• -&-

LITERATURE REVIEW

OUR KNOWLEDGE OF CAYERNICOLOUS FUNGI IS LARGELY THE

RESULT OF INCIDENTAL 098ERVATtON8 AND COLLECTIONS MAD£ BY

QEOLOQleT8 AND ZOOLOGl8T8• THE POSITIVE tOENTIFtCATfON8

OF FUNGI FROM THE INTERIOR OF CAVES ARE FEW• Mo8T OF THE

FUNGI WHICH HAVE BEEN REPORTED ARE 09VtOU8LV TROQLOXENE&.

THESE INCLUDE WOOO-ROTTINQ FUNGt 1 COPROPHILOUS SPICIES 1

ANO OTHER SAPROPHYTIC 8PECtES QROWtNQ ON DISCARDED

MATERIALS AND ORIQtNATtNG FROM AIR-BORNE SPORES• IT 18

PROBABLE, HOWEYER 1 THAT THE PARASITIC FUNGI FOUND tNF!CT-

tNQ CAYE ANIMAL& ANO THOSE SAPROPHYTES OCCURRINQ IN THE

90tL AND WATER ARE TRULY TROILOPHILE8•

AMONG THE EARLY RECORDS IN THE LITERATURE ARI SEVERAL

WHICH APPEARED BEFORE THE TURN OF THE CENTURY• ACCORDING

TO KYRLE (1923), FRtE8 DESCRl8~D IN 1874 FOUR ~UNQt FROM

·FALKEN8TEINERH8HLE. THESE INCLUDED "YELLOW-BROWN MYCELIA

. OF RH!ZOMORPH!, A FELT-LIKE TUFT OF lg~TOMITUB LACT§U8, MMCOR MUCEOO QROWtNQ ON BAT DUlll, ANO THE FUNQU8

MXXOTRICHELLA SPELAEA•" HOVEY (1879) COLLECTED FUNQI FROM LURAY CAVERNS AND OESCRaBED ONE OF THE BLACK MOLDS A8 A

NEW 8PECIE81 MUCOR STALACTITts. THta SPECIES •• NOT

INCLUDED ON SACCARDo•s "SYLLOQE FUNGORUM" AND HAS NOT 8!£N

RECOGNtZEO AS A VALID SPECIES•

PACKARD (1879) COLLECTED SEVERAL FUNGI FROM MAMMOTH

CAVE WHICH WERE IDENTIFIED BY THE LATE PROFESSOR ROLAND -1-

THAXTER OF HARVARD UNIVERSITY. THESE INCLUDED AN ISOLATE

REFERRED TO AS 0ZONIUH AUBANTIACiUM ltNK, AND ISOLATES DESCRIBED A8 "A REDDISH BUTTON•SHAPED FUNQU81 A GREEN MOLD,

A LONG WHITE MOLD, AND A FUNGUS QROWtNQ ON THE HIND BODY OF

A CAVE CRICKETTe" IN THE SAME PAPER, PACKARD MENTtONEO AN

UNIDENTIFIED 8PECt!a OF PEZtZA AND A COLORLESS AGARIC FROM

WEYER'8 CAVE.

IN A SERIES OF TWO PAPERS CALL (t879 A, 1879 B) PU8Ll8H£D A LIST OF FUNGI COLLECTED IN MAMMOTH CAVE,

KENTUCKY. THESE WERE ALSO IDENTIFIED BY PROFESSOR THAXTER

OF HARVARD UNIVERSITY AND INCLUDED THE FOLLOWING SPECIES: COPRtNUS MICACEUS BULL., FOMES (POLYPORUS) APPLANATUS PERS., RH!ZOMORPHA MOLINARI& (?), MtCROASCUi b0NOIR08TRl8 ZUKAL,

LASMIDIUM CELLAR! FR., GYMNOASCUS SETOSUS EtDAM 1 !• UNCtNATU8

EIDAM 1 TWO ADDITIONAL SPECIES OF GYMNOA8CUS, A SMALL, REDOISH-

BROWN SPECIES OF P£ZIZA, ISAR!A (SPOROTRICHUM) OENSUM LINK,

THE LATTER GROWING ON THE DEAD BODIES OF CRICKETS (HAOENOA§CU8 ayBTERRANEU8), !• FL!VISSIMUM LINK, lABOULBENtA 8U@TERRANE6

ON ANOPHTHALMUS TELLKAMPFU, A NEW SPECIES OF PAPULOfPORA 1 A NEW SPECIES OF BOUDl£RA1 CARYCHIUM 8TVGJUM, AND MUCOR MUCEDO LINN.

FOLLOWING A VtStT TO THE UNITED STATE8 1 MAHEU (1926) PUBLISHED A LIST OF Hl8 EXTENSIVE COLLECTIONS FROM THE

KENTUCKY CAVERNS INCLUDtNG THE FOLLOWING CAVERNICOLOU8

FUNGI: COPRINU8 MICACEUI BULL., FLAMMULA 8P. 1 MVCENA sP., -1-

PAX!LLU8 PANUOIDES FR., CORlOLUS ELONQATUS BERK., RAOULUM

.!f!•• TRAMETES ODORATA (WULFe) FR., PEZIZA §!.•, ISARIA OENSA

ltNK GROWING ON THE CADAVERS OF H~DEHAECUS SUBTERRANEUS, AND

MYCELIAL STAGES BELONGING TO THE GENERA Svaaua, RHt%OMORPHA.

AND 0ZONtUM• MORE RECl?NTLV, BAILEY (1933) NOTED THE

PRESENCE OF M,aRASfOUfl flGRIPES SoHw., COPRINUS ATR!MENTARIUS

(BULL.) FR., A SPECIE& OF STEREUM• AND ONE OF THE BLACK

MOLD8 1 ALSO FROM THE CAVES OF KENTUCKYe

FROM HtS STUD1ES OF CAVE LlFE IN HUNQARV 1 GABHARDT

(1932) MADE REFERENCE TO ONLY TWO SPEC1ES OF FUNGI, THE

DISCOMYCETE, HYALINA FASCICULARE AND THE AGARIC 1 COPRINUS ATRAMENTARIUS (BULL.) FR.

WOLF (1938) S80LATED FROM SOIL SAMPLES, BAT EXCREMENT,

AND FRA88 FROM LEAF-CUTTING ANTS, COLLECTED FROM CAVES IN

YUCATAN, MEXICO, THE FOLLOWING FUNGI: AsPERGtLLUS CLAVATUa

DESM. AND PENICILLIUM 8Pe FROM BAT DUNQ 1 AND PyTNIUM SP.

FROM DAMP ROCK 8CRAPINQ8•

A LARGE CAVE SYSTEM tS USUALLV WELL VENTILATED ANO

AIR-BORNE SPORES OF FUNGI MAV BE CARRIED TO THE MOST REMOTE

AREAS OF THE CAVEe SUCH SPORES, IF DEPOSITED ON A SUITABLE

SUBSTRATUM, WILL GROW. THIS ACCOUNTS, PERHAPS, FOR THI ORfQIN OF THE FUNGUS OE&CRIBEO BY BtRO (1949) QROWINQ ON CANDLE IREA8E IN AN ENGLISH CAVE ANO tOENTIFIED AS

PENICILLIUM fTALICUM W£HM£R. AN EARLIER COLLECTION FROM ~-

THE 8AME CAVE AND FROM SIMILAR 8UB8TRATUM WAS MADE BY

WALTON (1944) WHO PROVED EXPERIMENTALLY THAT THIS FUNQU8

SERVED AS THE SOL! SOURCE OF FOOD FOR THE SPRINOTAIL

COLLEMBOLA• BtRD ALSO RECORDED FROM OTHER ENGLISH CAVES

A8PERGILLUS VER8ICOLOR (VUtLL.) TIRABOSCHI AND AN

UNIQENTIFIED SPECIES OF HELOTIUM. " -

HAZELTON AND GLENNIE (1953) AL80 MENTIONED THE

PRESENCE OF FUNGI IN BRITISH CAVES, ALTHOUGH THESE WERE

NOT IDENTIFIED. ONE TYPE WAS DESCRIBED AS "VERTICAL

QROWTH8 A FEW MILLIMETERS HIGH, ABOUT A CENTIMETER APART,

A FOREST OF MINUTE FUNGI QROWINQ ALL OYER A WIDE BENCH OF

AN OLD PA88AQE ABOUT FOUR FEET ABOVE THE PRESENT FLOOR,•

AND A SECOND ORGANISM, POSSIBLY A FUNGUS, WAS FOUND QROW-

IN8 ON BARE ROCK IN "SMALL CIRCULAR PATCHES, OFTEN WtTH

A YELLOWISH CENTER, SELDOM MORE THAN A CENTIMETER IN DIAMETER•" ANOTHER UNIDENTIFIED QROWTH, APPARENTLY THAT OF A WATER MOLD, WAS FREQUENTLY OBSERVED AT THE EDQE OF

RIM8TON£ POOLS.

Two 8PECfE8 OF CAVERNICOLOUS FUNGI REQUIRE SPECIAL MENTION HERE SINCE THEY HAVE SEEN R£PORTED ONLY FROM CAVES•

BOTH FUNGI WERE DESCRIBED BY PETCH (1934) AS PARASITES OF tN8£CT8• THEii SPECIES ARE HtRSUTELLA DIPTERICA PETCH,

PARASITIC ON TWE DtPTEROUI FLY, LERIA SERRATA, FROM CAVES

IN ENGLAND; AND STtBELLA KERVILLEI (QUEL.) ltNGELSH. 1 -9~

PARASITIC ON THE FIRST NAM£0 FUNGUS. BOTH FUNGI ARE

APPARENTLY THRESHOLD SPECIES, BUT AT TIMES MAY BE FOUND

IN THE tNTERtOR OF CAVES StNCE THE HOST FLV 1 NORMALLY A

THRESHOLD SPECIES, WILL FREQUENTLV MIGRATE DEEPER INTO

THE CAVE.

DR. Te Ce BARR, JR. HAS REPORTED (PERSONAL COMMUNI- CATION) SEVERAL COMMON SPECtES OF FUNGI ON WOOD AND DUNG

FROM CAVES IN TENNESSEE• THESE INCLUDE MUCOR MUC!DOa

PHVCOMVCES NITEN!, XVLARIA POLYMORPH~, A SPECIES OF ISARtA

FOUND ON THE DUNG OF DAVE RATS, AND ~TOMOPHTHORA ON THE

CAVE CRICKET HADENOAECUS $UBTERRANEUSe BARR ALSO NOTED

THAT THE REO BEETLE, PSEUDOAN~PHTHALMUS, MAV BECOME SEVERELY PARAllTIZED av ...... LABOULSENtA --SP. -10 ...

M.l~ TER L'\LS AND METHODS

WtTHttt GILES, CRAIG, ROANOKE, ANO MONTGOMERY COUNTIES

OVER 255 CAVE' HAVE BEEN FOUND. ALL OF THESE ARE IN THE

LIMESTONE BELT OF THE APPALACHIAN VALLEY. SUCH CAVES

OCCAStONALLY CONTAIN SMALL BODltS OF STANDING WATER OR

UNDERGROUND STREAMS AND, I~ NOT, THE S01L FOUND THEREIN

13 QUITt WET. THIS 13 'TRUE EVEN DURING THE SUMMER MONTHS

WHEN THESE CAVES WERE SAMPLES. ALTHOUGH MOST OF THESE

CAVES HAYE BEEN KNOWN FOR YEARS, THEY ARE VISITED ONLY

BV SPELEOLOGISTS INTERESTEO IN THE EXPLORATION AND

SCIENTIFIC STUDY OF SUBTERRANIAN HABITATS. SUPERSTITION,

FEAR, AND GENERAL LACK OF CURtOSITY HAV! KEPT THE LOCAL

HUMAN INHABITANTS FROM VENTURING VERY FAR INTO THE INTERIOR.

THESE CAVES ARE GENERALLY UNDISTURBED BY THE C~SUAL VISITOR

AND THE ORGANISMS FOUND THEREIN MAV BE REGARDED AS

NATURALLY OCCURRING FUNGI.

A NUMBER OF WELL ... KNOVN COMMERCIAL CAVERNS WERE At.SO

VISITED DURING THIS INVESTIGATION. THESE WERE LOCATED IN

THE AREAS BORDERING THE • SINCE THESE

ARE FREQUENTED ev THE GENERAL PUBLIC, SUCH CAVERNS CONTAIN

ABUNDANT DISCARDED DEBRIS• THE ORGANISMS INHABITING SUCH

SUBSTRATA ARE NOT GENERALLY REGARDED AS NATURALLY OCCURRING

INHABITANTS OF THE CAVERNIOO~OUS ENVIRONMENT. IN COM-

MERCIAL CAVERNS CARE WAS TAKEN TO COLLECT MATERtAL FROM

THE LESS FREQUENTED AREAS IN ORDER TO LESSEN THE CHANCE OF

COLLECTING TROOLOXENIC CONTAMINANTS. -1 t-

SAMPLES OF SOIL AND WATER WERE COLLECTED FROM BOTH

COMMERCIAL CAVERNS AND NON-COMMERCIAL CAVES• SEVENTEEN

SOIL SAMPLES AND THIRTY-ONE WATER SAMPLES WERE OBTAINED

FROM FIVE COMMERCIAL CAVERNS LOCATED IN WIDELY SEPARATED

AREAS OF THE SHENANDOAH VALL!Ye NINETY-TWO SOIL SAMPLES

ANO THIRTEEN WATER SAMPLES WERE COLLECTED FROM THE CAVES

IN THE BLACKS&URQ AREAe

THE PROCEDURE USED IN THE COLLECTION AND ISOLATION

O~ OAVERNICOLOUS FUNQJ WAS ESSENTIALLY THE SAME AS THE

TECHNIQUES INITIATED BY BUTLER (1907) AND HARVEY (1925) 1 AND SINCE ADOPTED BY MOST STUDENTS OF THE SOIL AND WATER INHASITINQ PHVCOMYOETOUS FUNGle SOIL SAMPLES WERE COL-

LECTED IN STERILIZED 60 MLe GLASS BOTTLESe SUCH SAMPLES WERE OBTAINED BY SCRAPtNO THE SURFACE SOIL OR BY OIQQtN8

FRESH EXCAVATION8e WATER SAMPLES W!R! OBTAINED SY

PARTIALLY SUBMERGING STERILIZED 60 ML. QLA88 BOTTLES BELOW

THE SURFACE TO ALLOW THE WATER TO FLOW FREELY INTO THE CONTAINER. IN ALL CASES THE SAMPLES WIRE CAREFULLY LABELED AND RETURNED TO THE LABORATORY. THE TIME LAPSE BETWEEN

COLLECTION AND BAITING VARIED FROM SEVERAL HOURS TO

8£V~RAL DAYSe DURING THE LONGER DELAYS SAMPLES WERE

STORED IN A REFRIGERATED CONTAINER.

IN THE LABORATORY A ~ORTION OF EACH SOIL SAMPLE

SUFFICIENT TO COVER THE BOTTOM WAS TRANSFERRED TO A -12-

STERILE, DEEP PETRI-DISH. To EACH WAS ADDED APPROXtMAT£LY 75 cc. OF STERILE SOIL WATER ANO EACH DISH 80 PREPARED WAS

BAITED WITH HALVES OF SOILED HEMP SEED 1 SMALL PIECES OF

BOILED SNAKE 8KIN 1 SQUARES OF BOILED CELLOPHANE, A~D SMALL

FRAGMENT& OF STERILE, CHLOROPHYLL-EXTRACTED GRASS LEAVES. WATER SAMPLES WERE DISPENSED INTO STERtLE P!TRt-Dl8HES AND BAITED IN THE SAME MANNER• STERILE aGIL WATER WA8 OFTEN USED TO SUPPLEMENT THESE SAMPLES IF THE ORIGINAL MATERIAL WAS NOT SUFFICIENT TO FILL A 0£1P PETRI-DISH.

SAMPLES WERE MAtNTAINEO AT ROOM TEMPERATURE AND BAITS

EXAMINED MICROSCOPICALLY AT REGULAR INTERVALS• PR08A8L£

ANO POSITIVE l80LATE8 WERE TRANSFERRED TO FRE8H DISHES

AND REBAtTEO tN THE SAMI MANNER• SUCCEIStVE TRANSFERS

RESULTED IN UNtFUNQAL ISOLATES R£LATIYELY FREI OF

BACTERIAL CONTAMINATION. NO EFFORT WAS MADE TO OBTAIN

BACTERIAL-FREE CULTURES OF THE FUNGI STUDIED. THE

IDENTIFICATION AND MORPHOLOBtCAL OBSERVATtONS REPORTED IN

THE FOLLOWING lttltTION OF THIS PAPER WERE MADE FROM UNIFUNQAL MATERIAL MAINTAINED AS STOCK CULTURES• -13-

FUM G f COll ECT::::D

CH YTf-

t N I) t A 8 0 T • S l::R • , l: 14 f • f 9 07 •

MYCEL!UM COPIOUS, EXTRA-MATRICAL; HVPHAE MUCH BRANCHED,

IRREGULAR IN SfZE, MEASURING 3.3-5.0 p IN DIAMETER;

ZOOSPORANQIA CLAVATE TO SPHERICAL, TERMINAL OR INTER-

CALARY 1 M~ABURING 23.3-3f .6 µ tN WIDTH BY 63.3-19.9 µ

IN LCNQTH WITH A LONG APICAL PAPILLA OPENING BY AN

OPERCULUM MEASURING a.a µ IN OIAM~TER, APOPHYSATE, THE APOPHYSIS SPHERICAL TO CAMPANULATE ANO MEASURINQ to.o-

13.3 µ IN DIAMETER; ZOOOPORES ESCAPING FULLY OEVELOP£D

tN MASS, SPHERICAL, UNlFLAG£LLATE, CONTAINING A SINGLE

OIL DROP, MEASURING 6.7 µ tN DIAMETER; RESTING SPORES

NOT OBSERVED.

FOUNO ONCE ON A ROTTING STICK tN WATER, SHENANDOAH

CAVERNS, SHENANDOAH COUNTY, VtRGINIA•

RAMOS.A, WAS FIRST DESCRIBED ON ROTTING STEMS OF TRtTtCUM

VO!,OA,RE JN IHOIA (8UTLtrn 1 1907), AND WAS LATER REPORTED

OH VEGETABLE DEBRIS FROM HUNGARY (OOMJAN, 1936)1 ANO

FROM BRAZIL (KARLfNQ 1 1944). THE LATTER INVESTIGATOR HAS

FOUND THIS SPECIES ON SIMfLAR SUBSTRATA THROUGHOUT TH£

SOUTHERN UNtTED STATES (KARLING 1 1941 1 1942 A). SPARROW -14-

AND BARR (1955) HAVE RECENTLY FOUND THE SAME FUNGUS ON MORIBUND _...._..,.._CHARA BP. FROM NORTHERN MICHIGAN.

THE VtRQINIA ISOLATE AGREES IN MOST RESPECTS WITH

THE DESCRIPTIONS OF PREVIOUSLY DESCRIBED FORMS• RESTING

SPORES WERE NOT OBSERVED, NOT WERE THE PROLIFERATING

OELLS OF THE RHtZOMYCELIUM, AS NOTED BY BUTLER (1907).

NOWAKOWS_,l(f ELLA RAMOS A MAY BE 0 lSTINGU I SHED FROM OTHER

APOPHVSATE MEMBERS OF THE GENUS, BY THE POSSESS&OH OF

NON-PROLIFERATING ZOOSPORANQIA MEASURING 25-45 µ IN

DIAMETER, AND BY THE ABSENCE OF SETAE ON THE ZOOSPORANGIAL

WALL. IT ts OF INTEREST TO NOTE THAT, ALTHOUGH TH•s

ISOLATE MAY SE READILY CULTtVATEO, IT WILL GROW ONLY ON

WOOD CHIPS AUD FAILS TO DEVELOP ON OTHER BAITS USED IN

THIS STUDY.

2. ? RHIZOPHLYCTfS !!f.•

SOIL ANO WATER COLLECTIONS FROM A NUMBER OF CAVES

YIELDED A VARIETY OF MONOCENTRtC CHYTRIDS APPARENTLY

ALLIED TO RHIZOPHLYCTIS. SEVERAL CHARACTERISTIC TYPES

OF ZOOSPORANGIA WERE OBSERVED BUT TIME DID UOT ALLOW

FOR THE tNDUCTtON OF ZOOSPORE DISCHARGE. HENCE, IDENTI-

FICATION WAS NOT FEASIBLE. THESE ISOLATES APPEARED

PRIMARtLY ON SNAKE SKIN BAITS AND HAVE BEEN MAINTAINED

FOR FURTHER INVESTIGATION. OWING TO THE CONFUSED STATUS

OF TH£ SPECIES ANO TO OUR LACK OF UNOERSTANDINQ OF THE -as-

RHtZOPHLYTt8-COMPLEX 1 IDENTIFICATION ANO FURTHER STUDY OF THESE ISOLATES WOULD BE WELL WORTHWHILE.

BLASTOCLAD I ALES

3. A~LO~VCES CY8TOGENUS EMERSON, LLOYDIA, !,: 136. 1941.

HYPHA ABUNDANT, PSEUOOSEPTATE 1 MEASURING 23.3 p AT

TH! 8A8E TO 10.0 µ AT THE TIP; THIN-WALLEO Z008PORANGIA

TERMtNAL OR SUBTERMINAL IN A ROW, 08LONQ 1 MEASURING 26.6- 31.6 µ IN WIDTH AND 56.6-80.1 µ IN LENITH (AVERAGING 29.3 X 72.3 µ); RESTING SPORANQIA THICK-WALLED, PITTED, BROWN, DICIDUOU8, MEASURING 31.6-35.9 µ IN WIDTH AND 45.o-st.6 µ IN LENGTH (AVERAGING 33.3 X 49.3 p); GERMINATION OF RESTING SPORANOIA NOT OBSERVED; QAMETANGIAL PHASE LACKING.

l&OLATEO 5 TIMES FROM WET SOIL ON HEMP SEED FROM SMOKE HOLE ANO HOQ HOLE, GILES COUNTYJ AND FROM CLOVER HOLLOW, CRAIG COUNTY, VtRQtNtA. ALLOMYoga CV8TOGENU8

HA8 BEEN PREVIOUSLY OE8CRIB£D AS A SAPROPHYTE IN SOIL

FROM ARIZONA, VENEZULA, 8URMA 1 AND CHINA (EMERSON, 1941), AND 18 APPARENTLY WIDELY Qt8TRl8UTED IN SOILS•

THE CAVERNICOLOUS 180LATE8 AGREE WITH TM! DE9CRIPTtON OtVEN BY EMERSON (1941) FOR ALLOMYCE8 CY8TOGENU8• THE

MEASUREMENTS OF THE HYPHA£ 1 Z008PORANQIA 1 AND RESTINQ

SPORANQIA FALL WITHIN THE RANGE REPORTED FOR THAT SPECIES• FURTH£RMOR!1 !.• CVITOQENUS MAY BE DISTINQUfSHEO FROM -16-

OTHER SPECIES OF THE GENUS BY THE POS9ESStON OF D!CtOUOU8 RE$TINQ SPORANGIA,, BY TH£ COLORLESS CYTOPLASM,, AND BY A l.IF'E-CVCl.E WHICH CON8tST8 SOLELY OF AN ASEXUAL PHASE•

4. BLASIOC~ADtOPata PARVA (WHtFFEN) SAPRROW,, J. OF WA8H1NQTON ACAD. Set., iQ.1 sa. 1950~

THALLU8 UP TO 500 µ IN LENQTHJ HVPNAE DICHOTOMOUSLY 8RANCM£D1 OFTEN WITH ONE BRANCH 8UPPREl8EO, MEASURING

8.9-14.0 ~ IN DtAMETERJ ZOOSPORANGIA CLUB 8HAPED,

MEASURING 36.6-44.9 µ IN DIAMETER, WITH ONE APICAL AND

ONE AXILLARY PAP1LLAJ R£8TfNQ SPORES 8MOOTH-WALLED 1 NON-

PUNCTATE, SPHERICAL TO ovo10, MEASURING 25.0-33.3 ~ IN DIAMETER (AVERAQtNO 28.0 µ), LYING LOOIELY IN A SMOOTM-

WALLED CONTAINER, THI LATTER SPHERtC~L TO ELLIPSOIDAL M!A8URINQ 27.8-36.7 µ fN DIAMETER; GERMINATION OF THE RE8TINQ 8,0RE NOT 088£RV£D•

faOLATED ONCE ON HEMP 8££0 FROM WET SOIL, HOG HOLE•

GILES COUNT'Y1 VtRGINlA• BLA8TOOLADtOP818 PARYA HAS BEIN

PREYIOU8LV OS8CRt8ED AS A SAPROPHYTE lN SOIL FROM TEXAS (WHtFFEN1 1943) AND FROM CUBA (SPARROW, 1950). IT HAS

8EE.N REGARDED GENERALLY AS A FUNGUS RAt! IN tT8 OCCURRENCE.

THta l80LAT£ VAS OBTAINED ON HEMP 8EED, APPARENTLY AN UNUSUAL BAIT FOR tT8 t80LATION• PREVIOUS RECORDS OF THIS

8,101£8 INDICATE ITS ABtLITV TO GROW ON SNAKE SKIN ANO -17-

GRASS, BUT THErtE f S NO REPORT OF ITS OCCURR~NCE ON HEMP

SEED• THE FACT T~AT IT GROWi IN CULTURE OH HEMP SEED AND

THIS tSOLATE, AT LEAST, ts ALWAYS FOUND AMONG THE HEAVY

MYCELIAL QROWTH OF AN UNtDENTIFIEO IMPERFECT FUNGUS, MAY

WELL ACCOUNT FOR THE R£PORTtD RARlTV OF THIS FUNGUS.

THE MEASUREMENTS OF THtS ISOLATE ARE SLIGHTLY SMALLER

THAN THOSE CU Vim FOR 8.f.;,AS,TOCl.AD,IOP,S,l~ PARVA BY SAPRROW

( 1950). THIS SPEOIE8 es,, AT PRESE~lT, THE ONLY DESCRIBED

MEMBER OF THE GENUS. HOWEVER, SCOTT (PERSONAL COMMUNICA-

TION) OBTAINED A LARGE NUMBER OF ISOLATES FROM TROPICAL AND

SUBTROPICAL SOILS. THESE VARIED QREATLV WtTH RESPECT TO

SIZE AND NUMBER OF ZOOSPORANGIA AND NUMBER OF DISCHARGE

PAPtlLAE, AND IN TH~ StZE A~D COLORATION OF RESTtNQ

SPOr.Es. IT IS QUITE POSSIBLE, THEN, THAT THIO GENUS MAY

BE REPRESENTED ev SEVERAL SPECIES, AND THAT THE CAVERNl-

COLOUS ISOLATE MAY, INDEED, REPRESENT A NEW AND UNOESCRIBEO

MEMBER OF !LASTOCLADtOPStS.

THE GENUS BLASTOCLADIOPSIS MAY BE DtSTINQUt8H£0 FROM

THE CLOSELY RELATED GENUS BLASTOCLADfA ev POSSESSING NON-

PONCTATE RESTING SPORES WITH COMPLETELY SMOOTH WALLS AND

THESE NOT FILLING THE RESTING SPORANGIUM. FURTHERMORE,

THE MULTl-PAPILLATE NATURE OF THE ZOOSPORANGIA ANO THE

SUPRESSED DICHOTOMY OF THE THAt.t..US SERVES TO OtSTINGUISH

TH£Sl GENERA• -18-

SAPROLEGNtALES

5. ACHLYA DUSI! COKER, SAPROLEGNIACEAE1 P. 135. 1923.

HYPHAI STOUT, SPARINGLY BRANCHED, MEASURING 41.8- 78.5 µ tN DIAM!TERj QEMMAE ABUNDANT IN OLD CULTURES, IRREGULAR, TERMINAL OR fNTERCALARY; ZOOSPORANQIA ABUNDANT,

FUSIFORM, MEASURING 42.8-49.9 µ IN WIDTH BY 337.6-514.0 ~ IN LENGTH (AVERAGING 47.1 X 426.9 µ), RENcWED 8YMPODIALLV OR IN BASIPETAL SUCCESatoN; THRAU8TOTHECOID zooaPORE 018-

CHARGE FROM PRIMARY Z008PORANQIA 1 ACHLYOIO DtSCHARQE FROM SECONDARY ZOOSPORANGIAj ENCY.TED Z009PORE8 MEASURING 10.0 µ IN DIAMETER; OOGONIA ABUNDANT ON LATERAL BRANCHES, SPHERICAL

TO PYRtFORM 1 SMOOTH WALLED, PITTED ONLY UNDER ANTHERIDfAL ATTACHMENT, MEASURING 39.g_53.3 µ tN DIAMETER (AVERAGING 46.6 µ IN DIAMETER); 008PORE8 ECCENTRIC, SPHERICAL, 4-7 PER OOQONIUM, USUALLY 4, MEASURING JG.7-22.2 µ IN DIAMETER

{AVERAGING 19.3 µ IN DIAMETER), NOT ,ILLINQ THE OOGONIUM;

ANTHERtDIA MONOCLINOU8 1 ONE PER OOQONIUM 1 ANTHERIDIAL CELL

APPRE88£D; OOSPORE GERMINATION NOT OBSERVED.

ISOLATED ONCE ON HEMP SEED FROM WATER, SKY LINE

CAVERNS, WARREN COUNTY, VIRGINIA• ACHLYA DUBIA HAI BE£N 018CRl8ED PREVIOUSLY A8 A SAPROPHYTE IN SOIL FROM NORTH

CAROLINA, SOUTH CAROLINA, GERMANY, AND PUERTO RICO (JOHNSON, 1956 e). -19-

THE CAV!RNICOLOU8 l80LATE DIFFERS IN ANTHERlDlAL

ORIGIN FROM THOSE ISOLATES DESCRIBED BY JOHNSON, AND THE

Z008PORAN8lAL AND OOGONIAL MEASUREMENTS OF THIS ISOLATE

8UCIGEST A C.LOfl»R R£LATIO"f 8H tP TO ACHLYA CAROL IN tANA THAN

TO THAT OF ACHLYA DU8IA 1 ALTHOUGH THE THRAUITOTHECOIO

TYPE OF ZOOSPORAN81AL DISCHARGE 19 CHARACTERISTIC OF THE

LATTER SP£CJ!8• IT MAY WELL 8£1 THER£FORE1 THAT TfffS

ISOLATE 18 A FORM INTERMEDIATE BETWE£N THE TWO AFORE•

MENTIONED SPECIES. THE MOST DISTINCTIVE CHARACTERISTIC

OF A.• DU81A 18 ITS THRAUSTOTHECOID TYPE OF ZOOSPORE 018- 0HARGE FROM THE PRlMARY ZOOSPORANQIA, AND IT fS THIS

FEATURE WHICH DISTINGUISHES THIS SPECll8 FROM ALL OTHER

MEMBERS OF TH! GENUS•

6. APHANOMYCE8 LA£Vl8 DEBARY, JAHRB. wtss. BOT., £1 179. 1860.

HVPHAE SPARINGLY 8RANCH£0 1 UNDULATED, MEA8URING 5.0- 6.6 p IN DIAMETER; ZOOSPORANllA FtLAEMNTOUa, UNDIFFERENTIATED FROM YEOITATIVE HYPHAE, MEASURING 5.8 p IN WIDTH AND VARY•

INQ tN LENGTH; PRIMARY %008PORE8 ROD-SHAPED, MEASURING

16.7 ~ tN LENGTH; ENCYSTED ZOOSPORE8 MEASURING 7e5 p tN

OIAMETERJ OOQONIA TERMINAL ON SHORT LATERAL 8RANCH£9 1

SPHERICAL TO 8U88PHERtOAL, THIN WALLED SOMETIMES 8£COMING

IRREGULAR DUE TO THE DISINTEGRATION OF THE ATTENDANT

ANTHERIDIA, MEASURING 17.1-33.3 p IN DIAMETER (AVERAGINQ

22.6 ~); 008PORE8 8INQLE 1 HYALINE 1 CONTENTS GRANULAR, -20-

WtTH A LARQE CENTRAL OIL GLOBULE, MEASURING 15.7 µ IN

DIAMETER; ANTHERIDIA ONE TO SEVERAL, VERMIFORM, ANTHERIDIAL

STALK SIMPLE 1 OfCLINOUS OR MONOCLINOU8 1 FERTILIZATION TU8E8

ABSENT; OOSPORE GERMINATION NOT OB8ERVE0e

ISOLATED 5 TIMES ON SNAKE SKIN FROM SOIL AND WATER, STRALEY'• CAVE, GILES COUNTY; GRAND CAVERNS, AUGUSTACOUNTYj LURAY CAV!RUI• PAGE COUNTY; ANO SHENANDOAH CAVERNS, SHENANDOAH COUNTY; VtRQINIAe APHANOMYCES LAEVIS HAS BEIN PREVIOUSLY OE8CRIBEO AS BOTH A PARASITE ANO A SAPROPHYTE ON MANY DIVERS! SUBSTRATA FROM WIDELY SEPARATED ARCAS OP THE WORLD (SCOTT, t958). IT 18 APPARENTLY THE MOST

COMMON AND MOST WIDELY DISTRf8UTED MEMBER OF THE GENUS.

THE CHARACTERISTICS OF THtS CAVERNICOLOUS ISOLATE

AGREE WITH THE MODIFIED DE8CRtPTtON QIYEN IV SCOTT (1958).

APHANOMYCEa LAEYl8 MAY BE DISTINGUISHED FROM OTHER SPECIES

OF THE GENUS HAVING OOQONIA WITH SMOOTH WALLS OR WALLS.

MADE ROUQH BY THE DISINTEGRATION OF ATTENDANT ANTHERIDIAL

CELLS, BY POSSESStNQ NON-COILtNQ ANTHERIDIAL 8TALK8 1

SIMPLE Z008PORANllA1 AND BY THE SIZE OF OOGONIA AND OOIPORES• IT 18 OF INTEREST TO NOTE THAT, ALTHOUGH APHANOMYCES LAEVIS HAS NOT BEIN REPORTED PREVIOUSLY AS A CAVERNICOLOU8

SPECl£81 ANOTHER MEMBER OF THE QENUS 1 !.• 8TELLA!U8, HA8 BEEN ISOLATED FROM ORY SOIL COLLECTED IN AN ALEUTIAN BURIAL CAY! tN ALASKA (SCOTT, 1958). -2t-

7. SAPROLEQNIA FERAX (GRUtTH) THURET 1 ANN. Set. NAT. Ill, -r 4 : 2 r 4. r a50. HYPHAE ABUNDANT, IRREGULAR, SPARINGLY BRANCHED,

M£ASURING 18.3-42.8 p IN DIAMETER; ZOOSPORANOIA TAPERtNQ

TOWARD THE APEX, EXHIBITING INTERNAL PROLIFERATION, MEASURING 28.6-42.8 p IN WIDTH BY 371.3-470.8 p IN LENGTH

(AVERAGING 42.8 X 379.8 p); ZOOSPORES MEASURING 11.7-

13.3 p IN DIAMETER UPON ENCYSTMENT; OOGONIA SPHERtCAL 1 MEASURING 53.3-98.2 p IN DtAHETER (AVERAGING 73a9 p) SOME WITH HVPHAL-LIKE EXTENSIONS CONTAINING OOSPORES IN A

SINGLE ROW, OOGONIAL WALL PITTED; OOSPORES SPHERICAL TO

ELLIPSOIDAL, CENTRIC, NUMBERING 4-16 PER OOQONIUM

(AVERAGING 8), MEASURING 23.3-33.3 p IN DIAMETER (AVERAG-

ING 28.8 p); ANTHERIOIA PRESENT ON LESS THAN 10~ OF THE

OOQONIA 1 MONOCLINOUS, ONE PER OOGONIUM; OOSPORE

GERMINATION NOT OBSERVED.

ISOLATED ON HEMP SEED ANO SNAKE SKIN 3 TIMES FROM

WATER, GRAND CAV~RNSt AUGUSTA COUNTY 1 AND fNOLESS CAVERNS,

ROOKINGHAP.1 COUNTY, VtRCUNIA. SAPROLEGNIA F"~RAX HAS SEEN

C!SCRIBED PREVIOUSLY AS A SAPROPHYTE FROM WATER AND SOtL

f'ROM THE UtUTED STATES AND Eur~OPE (COKER ANO MATTHEWS,

1937} ANO IS APPARENTLY ONE OF THE MORE COMMON WATER MOLDS

TO BE FOUNO IN THESE HABITATS. -22-

As JOHNSON (1956 A) STATES, SAPROLEGNIA FERAX

(GRUITH) THURET, ANO THE FOLLOWJNG SPECIES, ~· MIXTA

OE8ARY 1 REPRESENT A COMPLEX OF TWO SPECIES WITH MANY

VAR1ANTS. THE PRINCf PAL OIFFERENCE BETWEEN THESE SPECIES fS APPARENTLY THE PRESENCE OF FUNCTIONAL OOOONIA WlTHIN THE

EMPTY ZOOSPORANGIA OF ~· ~~AX 1 AND THE LACK OF THESE IN !· MIXTA. JOHNSON (Loe. CIT.) REFERS TO ISOLATES, OTHER-

WISE SIMILAR TO !• MIXTA 1 WHICH PRODUCE OOGONfA WITHIN

DISCHARGED Z00SPORANGIA 1 AND ISOLATES CLEARLY CHARACTERtS- TIC OF -s. FERAX, WHICH LACKED SVCH OOGOHtA. OosPORE NUMBER IS MUCH TOO VARlABLE IN THESE SPECIES TO BE USED

AS A DISTINGUISHING FEATURE. SAPROLEGNIA FERAX, HOWEVER,

CAN BE Ot9TINCUISHED FROM OTHER RELATED SPECIES WHICH

LACK ANTHERIOIA ON MORE THAN HALF OF THE OOGONtA AND WHICH

HAVE OOGONIA THAT ARE NOT PRODUCED IN CHAINS, BY THE

P0$SESSION OF LARGE OOSPORES AND OOGONIA, THE LATTER WITH

EXTENSIONS CONTAINING OOSPORES ANO BY THE FORMATION OF

OOQONIA WITHIN OLD ZOOSPORANGIA• THE JSOLATES ASSIGNED TO -S. FERAX DURJNG THIS STUDY AGREE IN ALL RESPECTS WITH THE OESCRIPTION OF THE SPECIES AS MODIFIED BY COKER AND

MATTHEWS ( t 937) •

8. SAPROLEQNIA MIXTA DEBARY 1 BoT. ZEIT. 1 -41: 38, 54. t883.

HYPHAE ABUNDANT, IRR£0ULAR 1 SPARINGLY BRANCHED, MEASUR-

INQ 18.0-47.9 µ IN OtAMETER; ZOOSPORANGIA TAPERING TOWARD -23-

THI APEX, iXHIBITINQ INTERNAL PROLIFERATION, MEASURING

28.6-42.8 µ WIOE, 314.6-421.3 µ IN LENGTH (AVERAGING 42.5 X 352.7 p); Z008PORE8 MEASURING 11.7-13.3 p tN

DIAMETER UPON ENCYSTMENT; OOOONIA SPHERICAL, TERMINAL

OR INTERCALARV, NEVER FORMED WITHIN OLD ZOOSPORANGIA,

MEASURING 63e2 µ IN DIAMETER, OOQONIAL WALL PITT!OJ

009PORES SPHERICAL, CENTRIC, NUMBERINQ 6 PER OOQONIUM 1

NOT FILLING THE OOQONIUM, MEASURING 26.6-29.9 µ IN DIAMETER (AVERAGING 28.3 µ); ANTHERIDIA PRE8ENT ON ABOUT

35% OF THE OOQONIA, MONOOLINOU8 1 f PER OOQONIUMJ OOSPORE

QIRMINATION NOT 088ERYED•

f SOLATID ON HEMP SEED TWICE FROM WATER, NEV RtYIR

CAVE, GILES COUNTY, VIRGINIA• SAPROLIGNIA MIXTA HAS BEEN

DESCRIBED AS A SAPROPHYTE FROM WATER AND SOIL FROM THE

UNITED STATE81 GERMANY, ANO EUROPE (COKER ANO MATTHEV81 1937). Aa PREVfOU8LY 8TATEO (Pe 22) THE VALIDITY OF

THIS TAXON 18 QUESTIONABLE•

TH! CAVERNICOLOU8 ISOLATES AGREE WtTH THE MODIFIED

D£8CRIPTION QIV£N BY COKER ANO MATTHEWS (1937) FOR

SAPROL!QN!6 MIXTA IH ALL RESPECTS EXCEPT OOSPORE SIZE•

THE 008POR£8 OF THESE t80LATES AVERAGE SOMEWHAT LARGER

THAN THOSE OF PREVIOUSLY DESCRIBED FORMS•

IT IHOULO BE NOTED THAT THESE llOLATE8 WERE

PARA8tTIZ£D BV 0LPIDIOP818 8!PROL£QNIAEe THl8 MAY WELL

ACCOUNT FOR THE LACK OF OOQONIA WITHIN OLD ZOOSPORANGIAe THE StGNIFICANC! OF THE LACK OF OOGONIAL EXTEN810N8 AND TH! ABSENCE OF OOGONIA WITHIN THE OLD ZOOSPORANGIA MUST -24-

6£ CONSt0£'RE:D1 AT LEAST IN THESE ISOLATES,, SINCE tN EACH

CASE HAO THESE ACCESSORY STRUCTURES OEVELOPE0 1 THE FUNGUS

MIGHT WELL BE ASSfGN£D TO SAPROLEGNtA FERAX•

LEPTOMITALES

9. APODACHLYA MtNIMA COKER ANO lEtTN£R 1 J. ELISHA MITCHELL Sc r. Soc., M_: 313. 1938.

MVCELIUM FLACCID ON HEMP SEED; MAIN HYPHAE SLENDER,

SEGMENTED, BECOMING NARROWER TOWARD THE APEX, SEGMENTS

MEASURtNQ 6.1-10.0 µ tN DIAMETER av 84.3-166.5 µ IN

LENOTH 1 SEGMENTS OF OOQONIAL BRAMOH MEASURING 6.7 µ IN

DIAMETER SY 6.7-15.0 µ IN LENGTH; PROTOPLASM THSN EXCEPT

IN OOGONIAL BRANCHES; ZOOSPORANOIA NOT OBSERVED, ZOOSPOR-

ANQIA ARl$1NG FROM SINGLE LATERAL BRANCHES tN GLOMERULJ,

OVOID OR PVRIFORM 1 MEASURING AFTER DISCHARGE 12.8•17.6 µ IN

DIAMETER BY 17.6-26.2 µ tN LENQTH 1 (JOHNSON, 1955) ; OOGONIA

SPHERICAL WITH SMOOTH, THICK WALLS, MEASURING 12.9-15.7 µ fN DIAMETER (AVERAGJNG 14.9 µ); OOSPORE SINGLE, COMPLETELY

FILLINQ THE OOOONIUM 1 AT FfRST ECCENTRIC, BUT LATER THE

CONTENTS ROUND UP INTO A HYALINE BALL CLOSELY APPRESS£D TO

A SMALLER HYALtNE OIL DROPLET; ANTHERIDIUM ORIQINATtNQ

FORM A SUBOOQONIAL CELL AS AN OUTGROWTH, BECOMING EMPTY

BEFORE MATURATION OF THE OOSPORE•

ISOLATEO TWICE ON HEMP SEED, ONCE FROM WATER FROM

ENDLESS CAVERNS, ROCKltHlHAM COUNTY,, VIRQINIA1 AND ONCE -25-

FROM SOIL AROUND ROOTS OF DRYOPTERIS INTERMEDIA FROM LURAY CAVERNS, PAGE COUNTY, VIRGINIA• APODACHLYA MINIMA HA8 BEEN 0£8CRIBED PREVtOUSLY FROM A SLtBffTLY CONTAMINATED

STREAM FROM NORTH CAROLINA (COKER AND LEITNER, 1938) AND

FROM SOIL FROM PUERTO RICO (JOHN80N 1 1955).

THIS ISOLATE AGREES IN ALL RESP£0T9 WITH THE ORIQINAL

DESCRIPTION OF APODACHLYA MINIMA A8 GIVEN BY COKER AND . lEtTNER (1938). SPORANGIA, HOWEVER, WERE NOT OBSERVED IN

THIS ISOLATE• THEY HAYE BEEN OBSERVED AND DESCRIBED IN

DETAIL BY JOHNSON (1955) 1 WHO COMMENTS ON THE DIFFICULTY

OF OBTAINING Z008PORANGtA IN CULTURE AND ON THE OBSCURE NATURE OF THESE STRUCTURES. A• MINIMA MAY BE DtSTINGUtSHEO FROM RELATED SPECIES OF THE GENUS HAVING 8MOOTH•WALLED

008PORES AND OOQONIA BORN£ ON SHORT MONILIFORM SEGMENT& BY

THE P088E88ION OF ANTHERIDIA ORfQfNATING A8 LATERAL OUT-

GROWTHS OF THE 8UBOOGONIAL CELL AND BV THE RARITY OF

ZOOSPORANGtAL FORMATION.

LAGENIOIALES

10. 0LPIDIOP8!9 !PH6NOMYCl8 CORNU, ANNe Set. NAT., !§.l 148. 1872.

ZooaPORANGtA SMOOTH, SPHERICAL, OVAL TO ELONGATE, 1-4 PER SWOLLEN HYPHAL TIP, MEASURING 23.3-39.9 µ IN WIDTH AND 34.9•39.9 µ tN L£NQTH; WITH ONE OR TWO EJ1T PAPILLAE -26-

MEASURING UP TO 10.0 µ IN LENGTH AND EXTENDING THROUGH

THE HOST CELL WALL; RESTING SPORES NOT OBSERVED.

ISOLATED ONCE AS A PARASITE OF APHANOMYCES LAEYl8

FROM WATER, SKY LtNE CAVERNS, WARREN COUNTY, VIRGINIA•

0LPIDIOP8f8 APHANOMYCIS HAS BEEN DESCRIBED PREVIOUSLY AS A PARASITE OF APHANOMVCES !f.• FROM FRANCE, DENMARK, GERMANY, INOIA, AND THE UNITED STATES (KARLING, 1942 B)

ANO IS APPARENTLY OF COMMON OCCURRENCE ON THIS HOST•

THE CAVERNtCOLOUS ISOLATE AGREES WITH THE MODIFIED

DESCRIPTION GIVEN FOR TH! ISOLATE BY KARLINQ (1942 8)

A~THOUGH HE FAILED TO INCLUDE THE Z008PORANGIAL MEASURE-

M!NT8 OF HIS ISOLATE•

0LPIOtOP818 APHANOMVCIS MAY OTHERWISE BE Dt8TINGUISHED

F.RQ~ OTHER SPECIES OF THE GENUS BY THE HOST ON WHICH tT GROWS. WHETHER OR ;\!OT HOST SPECfFtCtTY rs A VALID CRI-

TERIGN FOR THE fDENTIFIOATION OF SPECIES ts QU~8TtONABLEe

U~Tf~ f~RTHER STUDIES ON MEMBERS OF THtS GENUS ARE lNJTtATED ANO MORPHOLOGICAL OfSTfNCTtONS CLARIFIED, MOST

SP!CJE8 tNCLUOING THIS AND THE FOLLOWfNG ISOLATE MUST

REMAIN PROBLEMATaCAL.

''· 0LPIDIOP81& SAPROLEQNIAE (BRAUN) CORNU, ANN. See. NAT., -151 145. 1872. ZooaPORANQIA SMOOTH, HYALINE, SPHERICAL TO ELONQAT!,

4-12 PER SWOLLEN HYPHAL TIP OF THE HOST, MEASURING 14.9-33.3 ~ -27-

IN DIAMETER WITH 1-2 EXfT PAP,LLAE EXTENOtNQ TO SURFACE OF

HOST CELL; RESTING SPORE NOT OBSERVED.

ISOLATED TWICE AS A PARASITE ON SAPROLEGNIA .. PP. FROM .-.....-.....;...... --...... - WATER, NEWCASTLE ~lURDER HOLE, CRAIG COUNTV 1 ANO FROM WATER,,

New RIVER CAVE, GILES COUNTY, VtRGINIA. OLPfOfOPSIS

SAP~OLECNIAE HAS BEEN DESCRIBED PREVtOUSLV AS A PARASITE OF SAPROLEGNIA AND ISOACHLYA FROM GERMANY, FRANCE, Rusa•A,,

DENMAfH<, UNITED STATES, JAPAN, ENGLMJ01 ANO RUMANJA

(SPARROW, 1943).

THE ZOOSPORANGtAL MEASUREMENTS OF THIS ISOLATE AR£

WITHIN THE RANGE GIVEN BY KARLJNQ (1942 B) ANO SPARROW

(1943) FOR 0LPIDIOPSJS SAPROLEGNIAE. fT 19 INCLUDED HERE

BECAUSE OF ITS REGULAR SMOOTH WALLED ZOOSPORANCSA AND

BECAUSE OF THE HOST ON WHICH IT GROWS. WITHOUT RESTING

SPORES, THE fSOLATE CANNOT BE DEFtNJTELV DISTINGUISHED FROM ft• fNCRASSATA. As MENTIONED PREVIOUSLY THE IDENTIFI- CATION OF SPECIES tN THIS QENUS BASED ON HOST SPECIFICITY

8 ~UESTIOMA8LE. THIS t$0LATE, THEN, MAV ONLV BE INCLUDED

HER£ TENTATIVELY• -28-

PERONOSPORALES

12. ?!1HIUM !FERTILE KANOUSE AND HVMPHREV, PAPER8 MtcH. AcAo., !r 127. 1927.

HVPHA! BRANCHING, MEASURINQ 3.3-f0.0 µ tN DIAMETERJ

ZOOSPORANGIA UNBRANCHED, TERMINAL OR INT£RCALARY, MEASUR-

ING 3.3-S.0 µ IN WIDTH BY 83.2-527.0 ~ IN LENGTHI

Z008PORE8 FORMED IN A VESICLE MEASURINQ 23.3-35.7 ~ IN

DIAMET!R 1 ZOOSPORE8 MONOPLANETIC 1 MEA8URINQ 8.3-10.0 ~J

SEXUAL REPRODUCTION UNKNOWNI CHLAMYOOSPORES NOT OB8ERVE0e

ISOLATED ON HEMP SEED, GRASS, AND SNAKE SKIN 6 TIME8 FROM WET SANDY SOIL, CLAY, AND WATER; Hoo HOLE, GILES

COUNTYI EARHART'S CAVE 1 MoNTQOMERY COUNTY; AND LURAY CAVERNS, PAQE COUNTY, VIRGINIA• PYTHIUN AYERTIL§ HAS BEEN OE8CRIB£0 PREVIOUSLY AS A PARASITE OF ...... GossVPtUH ___SP • FROM ANGLO-fQVPTIAN SUDAN, OF VAUCHERIA s~. FROM THE

UNITED STATES, AND OF VIOLA TRICOLOR FROM THE NETHERLANDS

(MIDDLETON, 1943). THIS t8 APPARENTLY THE FtRST REPORT

OF THl8 SPECIES OCCURRING AS A SAPROPHYTE FROM 801L AND

ALL CAV£RNICOLOU8 ISOLATES OF PvTH!UN WHICH POS8E8S

FILAMENTOUS ZOOSPORANGIA AND NO SEXUAL REPRODUCTION HAYE 8E£N tNCLUDED UNDER THE BINOMIAL PVTH!UM AFERT!LE• MIDDLETON (1943) STATES THAT ttf• !FERTILE MAY EITHER BE -29-

MAtHTAtNEO AS A DISTINCT SPECIES, LACKING A SEXUAL STA8E,

OR AB A 81ffOMIAL WHICH INCLUDES A VARIETY OF RELATED

SPECl£8•·•~••••; TRANSFERENCE TO ANOTHER BINOMIAL WOULD

BE MANDATORY ONCE THE SEXUAL STAGE APPEARED tN THE 8PICIE8

OF THE LATTER CATEGORY•" StNCE MOST SPECIES OF PYTHtUM

CAN BE D19TINQUISHED ONE FROM ANOTHER ONLY OH THE BASea OF

TH!IR SEXUAL REPRODUCTIVE CHARACTERISTICS AND SINCE THE ZOOSPORANGIA OF l!• !FERTILE 18 NOT MARKEDLY DIFFERENT FROM THAT OF OTHER FILAMENTOUS ZOOSPORANGIAL FORMS OF THE GENUS,

TNE AUTHOR CONSfOER9 THIS TAXON TO BE AN AGGREGATION OF

UNRELAT£D IMPERFECT ISOLATES.

THE TWO ISOLATES INCLUDED HERE WHICH VERE OBTAINED

FROM PAGE COUNTY, VtRQtNIA WERE THE ONLY ISOLATES OF

PYTHIUM AFERTILE WITH A LtMIT!D HYPHAL GROWTH• THEIR HYPHAE NEYER REACHED A LENGTH OF MORE THAN 500 p, WHEREAS, ALL OTHER ISOLATES OF f• AFERTILE EXHIBITED

EXTEN81VE HYPHAL DEVELOPMENTe THt8 18 THE ONLY MARKED

DIFFERINCE IN THE ISOLATES INCLUDED UNDER THIS BfNOMIALe

13. PVTHIUM CAROLINIANUM MATTHEWS, STUDIES ON TH! GENUS

PYTHtUM, P. 71. 1931 •

. HYPHAE 8MOOTH, MEASURING 3.3-s.o ~ IN OfAMETERJ ZOOSPORANttA 8AMERICAL TO 8UBSPHERICAL, SHOWING INTERNAL

PROLIFERATION, MEASURING 16.7-30.0 ~ IN DIAMETER (AVERAGING 24.7 ~)J ZOOIPOR£8 NUMEROUS FORMED IN A STALKED VE81CAL ARISING FROM A SHORT APICAL PAPILLA; SEXUAL REPRODUCTION UNKNOWN. -30-

ISOLATED ON HEMP SEED TWtCE FROM CLAY ANO WET SANDY

SOIL, VICKER ROAD CAVE, MONTGOMERY COUNTY, VIRGINIA.

PYTHtUM CAROLINfANUM HAS BEEN PRE-tOUSLV DESCRIBED AS A

PARAS I TE OF SP I tlOOVRA !.!!.• FROM NORTH CAROL INA (MATTHEWS, 1931) ANO AS A SAPROPHYTE FROM VEGETABLE DEBRIS IN THE

UNITED STATES (MIDDLETON, 1943).

THESE ISOLATES AGREE WITH THE MODIFIED DESCRIPTION OF

PYTHJUH CAROLINIANVM GIVEN BY MIDDLETON (Loe. CIT.) EXCEPT

THAT THE RANGE IN SIZE OF THE ZOOSPORANQIA IS GREATER IN

THES£ ISOLATES THAN IN THOSE DESCRIBED BY MIDDLETON. LtKt PVTHIUM !FERTILE, f.• C!ROLtNIANUM HAS NO KNOWN SEXUAL REPRODUCTIVE STRUCTURES. HOWEVER, THE LATTER

SPECIES HA9 A ZOOSPORANQIUM WHICH 18 MARKEDLY DIFFERENT

FROM ALL OTHER PVTHIUM SPECIES. FOR THIS REASON, IT IS

REQARD~O AS A DISTINCT SPECIES AND NOT AS A BINOMIAL WHICH IS APPLIED TO A VARIETY OF RELATED FORMS• f• CAROLINIANUM MAY BE READILY RECOGNIZED ANO DISTINGUISHED FROM OTHER

SPECl£8 OF PVTHlUM HAVING SPHERICAL, PROLIFERATING

ZOOSPORANGtA Af'JO NO KNOWN SEXUAL REPRODUCTfON BY THE

POSSESSION OF A STALKED VESICLE ARISING FROM THE SHORT

APICAL PAPILLA OF THE ZOOSPORANQIUM. 14. PVTHIUM IRREQULARE BUtSMAN, MED. PHYTOPATH• LAB., -IJ: 1-1 s. 1927.

HVPHAE LOBULATE, FREELY BRANCHED, MEASURING 2.9 µ IN OtAMtTER: ZOOSPORANGIA SPHERICAL TO ovoto, TERMINAL OR -31-

INTERCALARY, NOT PROLIFEROUS 1 MEASURtNQ 22.9-30.0 µ IN DtAMETER (AVERAGtNQ 26.0 µ); OOQONIA SPHERICAL TO IRREGULAR, MEASURING 17.1-21.4 µ IN DIAMETER {AVERAQING 19.S µ); 008,0R!S SINGLE, APLEROTtc, MEASURING s.6-17.1 µ IN DIAMETER (AVERAGING 13~2 µ), CONTAINING A SINGLE OtL

GLOBULE; ANTHERIDIA I PER OOGONIUM 1 MONOCLtNOUS 1 SMALL,

APPLIED APICALLY TO THE OOQONIUM; OOSPORE GERMINATION NOT

OBSERVED.

ISOLATED ON HEMP SEED AND SNAKE SKIN TWICE FROM VET

ROCKY 80tL1 CLOVER HOLLOW, CRAIG COUNTY, VIRQINIAe PYTHIUM

!RR§GULARE HAS BEEN DESCRIBED AS A PARASITE ON 43 GPECIEB

OF FLOWERING PLANTS FROM THE UNITED STATES, NETHERLANDS,

HAWAII, ENGLAND, AND THE UNtON OF SOUTH AFRICA (MtDOLETON, 1943).

THta ts APPARENTLY THE FIRST REPORT OF THE OCCURRENCE

OF PYTHIUM IRREQULARE AS A SAPROPHYTE FROM SOILe GILMAN

(1957) INCLUDES THIS SPECIES AS A SOIL INHABITING FUNGUS AND REFERS TO AN ISOLATE REPORTED EARLIER BY YOUNKIN (1938).

THE LATTER INVESTIGATOR REISOLATED FROM EXPERIMENTAL SOIL

PLOTS A SPECIES OF PVTHIUM WHICH HE TENTATIVELY A881QNED TO

THIS BINOMIAL• HAWEYER, THE IDENTITY OF THE ISOLATE IS

QUE8Tf0NA9LE SINCE YOUNKIN FAILED TO INCLUDE A DESCRIPTION

OF THE FUNQUS. FURTHERMORE THE FUNQU8 WAS ORIGINALLY

ISOLATED FROM PARA81TtZED ROOTS OF WATERMELLON 8EEDLINQ8 -32-

AND WAS NEVER 1 AS FAR AS CAN BE DETERMINED, ISOLATED IN

NATURE FROM SOIL.

THE CAVERNICOLOUS ISOLATES OF PvIHIUM IRREQULARI

DIFFER FROM THOSE DESCRIBED BY MIDDLETON (1943) IN THE

AVERAGE SIZE OF OOQONIA ANO ZOOSPORANGIA• MEASUREMENTS

OF THESE STRUCTURES ARE WITHIN THE RANGE GIVEN FOR fHE

SPECIES, BUT THE AVERAGE DIAMETERS ARE MARKEDLY LARGER•

THE AVERAGE DIAMETER OF 008PORE81 HOWEVER, ARE 80M~WHAT

SMALLER THAN THAT GIVEN FOR ~REVIOUSLY DESCRfBED ISOLATES.

PyTHtUM IRREGULARE 18 THE ONLY MEMBER OF THE GENUS

WITH APLEROTIC 008PORE8 ANO IRREGULAR OOQONIA 1 WHICH IN

THE LATTER CASE, ARE NOT MADE IRREIULAR BY THE FUSION

OF THE ANTHERIDIAL CELL WITH THE OOQONIAL WALL. THE

VARIATION IN SIZE BETWEEN THESE ISOLATES ANO OTHERS

ASSIGNED TO THIS SPECIES 18 NOT CONSIDERED SIQNIFICANT

ENOUGH TO WARRANT THE ERECTION OF A NEW TAXON.

15. PYTH!UM MON08PERMUM PRINQSHEIM 1 JAHRB. Vt88• BOT., -t : 284. 1858. HVPHAE MEASURING 2.a-s.o "' IN DIAMETER, WtTH SWOLLEN

BUD-LIKI LATERAL OUTQROWTH8 1 BRANCHING FREELY: Z008PORANGIA

BRANCHED OR UNBRANCHED, UNDIFFERENTIATED FROM THE HYPHA£ 1

DELIMITED BY SEPTA, TERMINAL OR INT£RCALARV 1 MEASURING

3.3-5.0 µ IN DIAMETER, 83.3-642.6 ~ IN LENGTH; ZOOSPOR£8 6-12 IN NUMBER, to.o-ta.o µ IN DJ4METER; OOQONIA SMOOTH, SPHERICAL, TERMINAL ON SHORT BRANCHES, 11.4-20.0 µIN -33-

DIAMETER (AVERAGING 16.1 µ); OOSPORES SINGLE, PLEROTtC, 8.6-15.7 µ tN DtAMETER (AVERAGING 14.5 µ), CYTOPLASM

GRANULAR GIVING THE APPEARANCE OF SMALL GLOBULES CLOSELY

APPRESSED; ANTHERIDIA ANOROGENOUS OR MONOCLtNOUS, I PER

OOQONlUM; OOSPORE GERMINATION NOT OBSERVED.

ISOLATED ON HEMP SEED 3 TIMES FROM WET ROCKY SOIL,

CLOVER HOLLOW, CRAIG COUNTY; AUNT NELLIE'S HOLE,

MONTGOMERY Co UN TV; AND NEWCASTLE MURDER HOLE, CRAIG

COUNTY, VtRQINIA, PYTHIUM MONOSPERMUM HAS BEEN REPORTED

PREVIOUSLY AS A SAPROPHYTE IN ANIMAL DEBRIS FROM GERMANY

AND FRANCE, ON PLANT DEBRIS FROM GERMANY, IN SOIL AND

WATER FROM fRELANO ANO THE UNITED STATES, ANO AS A

PARASITE ON HORDEUM ~ULGARE, L£P10$UM SATIVUM, ~tCOTIANA

TABACUM, 0RYZA !ATIVA, PERSEA AMERICANA, RtCHARDIA

AETH&OPICA• SACCHARUM OFFICINARUM, SPINAC•A OLERACEA,

ANO ZINGIBER OFFICINALE (MtODt..ETOt4 1 1943).

ALTHOUGH EACH OF THE THREE ISOLATES INCLUDED HERE VARY

MARKEDLY ONE FROM THE OTHER, ALL THREE MAV BE ASSIGNED TO

THIS TAXON WITHOUT QUESTION• THE ABOVE DESCRIPTION

JNCLUOES THE COMBINATIVE VARIATIONS OBSERVED AND RESULTS

tN A SOMEWHAT BROADER CONCEPT OF THE SPECIES. AMONG THE

ISOLATES OBSERVED, LITTLE VARIATION OCCURS IN THE HYPHAE 1

OOQONIA 1 AND OOSPORE CHARACTERISTICS AND THESE FEATURES

ARE QUITE StMILAR TO THOSE OF PREVIOUSLY DESCRtSED t80LATES. -34-

THE ANTHERCDI~ HOWEVER, DIFFER AS TO ORlGIN. Two ARE

ANDROGENOU8 1 THE THIRD IS MONOCLINOUS. PREVIOUS INYESTt-

GATORS FAtL TO DISTINGUISH BETWEEN ANDROGENOUS ANO MONO-

OLINOUS ANTHERIDIA WITH RESPECT TO PVTHtUM MONOSPERMUM•

THE ZOOSPORANQIA OF THESE ISOLATES DIFFER MARKEDLY FROM

ONE ANOTHER ANO FROM PREVfOUSLY DESCRIBED ISOLATES. THE

MEASUREMENTS QfVEN BY MtOOLETON (1943) FOR THE ZOOSPORANGIA

OF PYTHtUM MONOSPERMUM ARE 2-5 µ x 80-170 µ.. Two OF THE

ISOLATES MENTIONED HEREIN MEASURE 5.0 µ X 93.3 µ. ANO 3.3 µ. X

83.3 µ RESPECTIVELY. THESE MEASUREMENTS ARE WITHIN THE

RANGE DESCRIBED BY MIDDLETON ALTHOUGH O(FFERtNG FROM ONE

ANOTHERe THE THtRO CAVERNICOLOUS ISOLATE IS EVEN MORE

STRICKING IN ZOOSPORANGIAL CHARACTERISTICS, MEASURING

5.0 µ tN DIAMETER AND UP TO 642.6 µ. tN LENGTH. WITH

RESPECT TO THE MODIFIED SPECIES DESCRIPTtON GIVEN ABOVE•

THESE THREE MAY SE ASSIGNED WITH CONVICTION TO PVTHIUM

"ONOSPERMUM DESPITE THE MARKED VARIATIONS IN THE

ZOOSPORANGIAL AHO ANTHEltDIAL CHARACTERISTtcs.

16. PVTHIUM NACAll ITO AND TOKUNAG0 1 J. FAC. AGR• HOKKAIDO IMPEfh UNIV., -32: 201. 1933. HYPHAE MEASURING 3.3-4.2 µ IN DIAMETER; ZOOSPORANGIA

SPHERICAL, NUMEROUS, TERMINAL OR INTERCALARV 1 SHOWING INTERNAL PROLIFERATION, MEASURING 20.a-2s.o p IN DIAMETER (AVERAGING 23.0 µ); ZOOSPORES FORMED IN A SPHERICAL VESICLE -35-

BORNe ON A SHORT EVACUATION TUBE; OOQONIA SPHERICAL,

MEASURtNG 20.0 µ IN DIAMETERJ OOSPORE APLEROTIC, SINGLE, SPHERICAL, THIN WALLED (0.8 µ), MEASURING 15.7 µ IN

DIAMETER; ANTHERIOIA NOT OBSERVED; OOSPORE GERMINATION

NOT OBSERVED.

ISOLATED ON HEMP SEED ONCE FROM MUDDY SOIL, VICKER

ROAD CAVE, MoNTGOMERY COUNTY, VtRQINfA• PVTHIUM NAGAll

HAS BEEN REPORTED fREVIOUSLY AS A PARASITE OF 0RVZA SATIV!

FROM JAPAN ANO FRO~ THE UNITEO STATES (MIDDLETON, 1943).

THIS IS APPARENTLY THE F1RST REPORT OF THIS SPECIES

OCCURRING AS A SAPROPHYTE FROM SOIL.

THE CAVERNtCOLOUS ISOLATE DIFFERS BUT LITTLE FROM THE

DESCRIPTION OF PVTHIUM NAGAll GIVEN BY MIDDLETON (1943).

OOQCNtA WERE OF RARE OCCURRENCE IN CULTURES OF THIS FUNGUS•

WHEN THESE WERE OBSERVED ANTHERIDIA WERE FOUND TO BE LACK- ING. NEVERTHELESS, f• NAQAll CAN BE READILY DISTINGUISHED FROM ALL OTHER RELATED SPECIES HAVING SPHERtCAL PROLIFERAT-

INQ ZOOSPORANGIA AND APLEROTIC OOSPORES BY THE POSSESSION

OF CHARACTERISTICALLY THIN WALLED OOSPORESe

17. PVTHIUM PAPILLATUM MATTHEWS, J. ELISHA MITCHELL Set. Soc., ...... 43: 229. 1928 •

HYPHAE LOBULATE, MEASURING 2.7-6.7 µ IN DtAMETER;

ZOOSPORANQIA FILAMENTOUS UNDIFFERENTIATED FROM THE -36-

VEGETATIVE HYPHAE• TERMINAL OR INTERCALARY 1 MEASURING

6.7 µ WIDE X 183.2 µ LONG; ZOOSPORES 8-16 PIR VE8lCL£1

MEASURING 9.9 ~ IN DIAMETER; OOQONIA SPHER&CAL TO sua-

8PHERICAL, OCCASIONALLY CATENULATE, M£A8URtNQ 12.9-15.0 ~ IN DIAMETER (AVERAGING 14.3 µ); OOSPORE8 PLEROTIC EXC!PT

AT OOQONIAL NECK, WITH A StNQLE OIL GLOBULE, MEASURING

12.3-14.3 ~ IN DIAMETER (AVERAGING t3.6 µ); ANTHERIDIA

NOT FORMED; 008POR£ GERMINATION NOT OBSERVED.

ISOLATED ON HEMP SEED ONCE FROM VET ROCKY SOIL 1

NEWCASTLE MURDER HOLE, CRAIG COUNTY, VtRGfNIAe PYTH!UM

PAP!LLATUM HAS BEEN DESCRllED PREVIOUSLY AS A SAPROPHYTE

FROM THE SOIL ANO FROM THE ROOTS OF HbROEUM VUhGARE FROM

THE UNITED STATES (MIDDLETON, 1943).

THIS ISOLATE OF PYTHIUM PAPILLA!UM POSSESSES REPRO-

DUCTIVE STRUCTURES WHICH MIA8URE SOMEWHAT SMALLER THAN

THOSE RECORO~D FOR TH! 8PECll8 IN TH! MODIFIED O!SCRtP-

TION GIVEN BY MIDDLETON (Loe. CIT.). NEITHER DOES THIS

t80LATI FORM PAPILLATE OOQONtA. IT DOIS, HOWEVER, POasrsa

ALL OTH!R CHARACTERISTICS USED IN THE SEPARATION OF ...P• PAPILkATUM FROM CLOSELY RELATED SPECIES AND MAY BE 018-

TINQUISHID FROM TH08£ SPECIES HAVING UNDIFFERENTIATED

FILAMENTOUS ZOOSPORANGIA BY P088ESSINQ APANDROU8 1 PLEROTIC

OOIPORES• -37-

18• PYTHIUM PULCHRUM MINDEN, MYKOL• UNTERSUCHe BERe R. FALCK, J_: 146. 1916.

PYTHIUM EPIQYNUM J-t6HNK 1 MYCOLOQIA, ~I 505. 1932.

HYPHAE MEASURING s.o-6.7 µ IN DIAMETER; Z008PORAN~IA

SPHERICAL TO PYRIFORM, TERMINAL OR INTERCALARY 1 23.3-33e3 µ IN DIAMETER (AVERAGING 29e3 µ), OCCASIONALLY CANTENULATE

WITH 2-3 ZOOSPORANGIA tN A SERIES; zooa~RES ABOUT 24 tN NUMBER, 13.0 µ IN DIAMETER, FORMING IN A VESICLE WHtCH ARl8£8 FROM AN APICAL OR LATERAL EVACUATION TUBE; OOQONIA

SMOOTH, SPHERICAL, t8.6-24e3 µ IN DIAMETER (AVERAfUNQ

21.s ~); 008PORES SINGLE, APLEROTIC, ts.1-20.0 µ IN DIAMETER (AVERAGtNQ 18.4 µ), CONTAINING A SINGLE LARGE OIL ILOBULE AND A SMALL LAT!RAL REFRINQENT soov;

ANTHERIDIA HVPOGYNOU8 1 USUALLY I PER OOQONIUM; 008,0R!

QERMINATtON·NOT 088ERVEDe

ISOLATED ONCE ON HEMP SEED ~ROM WET CLAY SOIL,

VICKER ROAD CAVE, MoNTQOMERY COUNTY, VIRGINIA• PYTHIUM

PMLQHBUM HA& BEIN REPORTED PREVIOUSLY AS A SOIL SAPROPHYTE

FROM THE UNtTED STATES (MATTHEWS, 1931; ffiHNK 1 1932, 1933;

MEREDITH, 1938)1 ON ANIMAL DEBRIS FROM GERMANY (MINDEN,

1916)1 ON PLANT MATERIAL FROM THE UNITED STAT!8 (MtOOLETON,

1943)1 ANO AS A PARASITE (THROUQH INNOCULATION) ON PHA!EOLUS

VUL86Rlla P18UM SATIVUM, AND !§.!.t:1A!.l (H8HNK 1 1932), AND ON

MIPICAGO 8ATIYIA (BUCHHOLTZ ANO MEREOtTFt 1 1938) FROM THE UNITED STAT!a. -38-

THE ISOLATE REPORTED HERE DtFFERS FROM THE ORIQl~AL

OESCRtPTION IN THE P089E8810N OF SMALLER Z008PORANGIA1

OOIONIA, AND 008PORE8 1 AND IN THE ABSENCE OF BOTH MONO-

CLINOU8 AND DICLINOU8 ANTHERIDIA• IN THESE RESPECTS IT

AGR!EI WITH THE DESCRIPTION OF THE FUNGUS ISOLATED BV

HgHNK (1932) AND REFERRED TO BY HIM AS A NEW SPECIES,

PJTH!UM EPtQYNUM• HOWEVER, AS MIDDLETON (1943) HAS

8U8QESTED, THE DfFFERENCES IN SIZE OF THE REPRODUCTIVE

STRUCTURES 18 NOT SUFFICIENT TO ALLOW SPECtFtC 8EQREQA-

TION. FURTHERMORE, 80ME STRAINS OF f.• PULCHRUM FAIL TO PRODUCE ANTHERIDIA OTHER THAN THOSE OF THE HYPOQYNOU8

TYPE. StNCE IN ISOLATES OF BOTH 8PECIE8 1 THE Z008PORANGIA

AND OOQONIA ARE SIMILARLY IHAPED, THE Z008PORANOIA ARE

FREQUENTLY PRODUCED IN CHAINS, AND THE OOSPORES ARE

APLEROTtC, THERE 18 APPARENTLY LITTLE JUSTIFICATION FOR MAINTAINING -P. EPllYNUM AS A SPECIES DISTINCT FROM -P. PU~Ck"UM. THE PRESENT ISOLATE IS A88tQN£D THE LATTER

BINOMlAL AND MAY 8E 018TINQUl8HEO FROM ALL OTHER SPECIES

WITH NON-PROLIFERATING Z008PORANQIA AND WITH APL!ROTIO

008PORE8 IN P088E8SINQ HYPOQYNOUS ANTHERIDIA. IT MAY AL80 BE 8IPARATEO FROM f• R08TRATUM AND f• HYPOGYNUM, TWO CLOIELY RELATED SPECIE&, BY THE PRESENCE OF APLEROTIC

008POR!S• -39-

19. PVTHJUM ROSTRATUM 6UTLER 1 MtM. DEPT. ACRe fNOIA BOT., 1: t. t 907.

HYPHAE MEASU~l~G 3.3-5.0 µ t~ DtAMETER; ZOOSPORANQIA SPHERICAL, TERMINAL, NON-PROLtFEROUS, MEASURtNQ 13.3-f6.7 µ tN DIAMETER (AVEAAGINQ 15.7 p); ZOOSPORES 16 IN NUMBER, MEASURING 5.0 X 8.3 µ; OOGONtA SPHERICAL TO SUBSPH!RICAL, TERMINAL OR INTERCALARY, MEASURING 13.3-16.7 p IN DIAMETER

(AVERAGING 14.7 p)J OOSPORES SINGLE, PLEROTIC 1 WITH A

SINGLE OIL GLOBULE; ANTHERIDIA MONOCLINOUS 1 I PER OOQONIUM;

OOSPORE GERMINATION NOT OBSERVED.

ISOLATED ON HEMP SEED ONCE FROM CLAY SOIL, EARHART'S

CAVE, MONTGOMERY COUNTY, VIRGINIA• PYTHIUM ROS~R!TU~ HAS

BEEN PREVIOUSLY DESCRIBED AS A PARASITE ON 28 SPECIES OF

FLOWERINQ PLANTS FROM THE UNITED STATES, HAWAtt, ANO

GER~ANY ANO AS A SAPROPHYTE FROM BOIL FROM FRANCE ANO THE

UNITED STATES (MIDDLETON, 1943).

THE CAVERNICOLOUS ISOLATE VARIES FROM THE MODIFIED

DESCRIPTION GIVEN BY MIDDLETON (1943) FOR PVTHIUM RO$TRATUM

IN THE POSSESSION OF CONSISTANTLV SMALLER REPRODUCTIVE

STRUCTURES. FURTHERMORE, THE ANTHERIDIA WERE NEVER SESSILE

AS HAS BEEN FREQUENTLY REPORTED FOR THIS SPECIES. THE

ABOVE MENTIONED DIFFERENCES ARE NOT, HOWEVER, CONSIDERED

SIGNIFICANT ENOUGH FOR THE EXCLUSION OF THtS ISOLATE FROM

THE SPECIES. -40-

20. PYTHtUM V£XAN8 DEBARY, JOURe SOT., 142 tos. 1876.

HYPHAE MUCH BRANCHED, MEA8URINQ 2.9 p IN DIAMETER;

ZOOSPORAN&tA 8PHERICAL 1 TEPMINAL OR INTERCALARV,MEASUR- INQ 16.7-18.3 µ IN DIAMETER (AVERAGING 17.3 µ); ZOOSPORE8 MANY IN NUMBER, MEASURINQ 7.2 µ; OOGONIA SPHERICAL, TERMI-

NAL ON SHORT LATERAL 8RANCHE8 1 MEASURING 12e2-17t2 µ fN DIAMETER (AVERAGING 15e2 µ); OOSPORES APLEROTtc, WITH A llNQL! RESERVE GLOBULE, MEASURING 8.6-14.3 µ IN DtAM!TER (AVERAGING 12.3 µ); ANTHERl01A ANDROGENOus, I PER OOQONIUMJ OOSPORE GERMINATION NOT OBSERVEDe

ISOLATED ON HEMP SEED TWICE FROM WET SANDY SOIL,

TONY 1 8 CAVE, GILES COUNTY, AND CATAWABA MURDER HOLE,,

ROANOKE COUNTY, VIRGINIA• PYTHIUM V§XAN8 HAS BEEN FOUND GROWING AS A PARASITE ON A LARGE NUMBER OF PLANTS FROM

THE UNtTED STAT!&, HAWAII, THE UNtON OF SOUTH AFRICA,

MALAYA, INDO-CHINA,1 NETHERLANDS, JAVA, INDIA 1 ANO GERMANY•

IT HAS ALSO BEEN REPORT!D IN SOIL FROM ENGLAND, IRELAND 1 FRANCE, AND THE UNITED STATES (MIDDLETON, 1943).

TH! MEASUREMENTS OF REPRODUCTIVE STRUCTURES ~F THESE

CAVERNJCOLOUI 180LATE8 ARE CONSISTANTLV SMALLER THAN THOSE GIVEN IN MtDOLETON's MODIFIED DESCRIPTION OF PYTHIUM VEXAN8,

&UT IN AU. O'THCR RESPECTS THESE ISOLATES AGREE WITH THOSE

PREVIOUSLY DESCRIBED• -41-

PYTHf UM VEXANS MAV BE OISTfNSUISHEO FROM OTHER RELATED

SPECIES HAVING SPHERICAL NON-PROLtFIROUS IOOSPORANGIA ANO

APLEROTtC 008PORES BY THE POSSESSION OF ANDROQENOUS 1

STALKED ANTHERIDIA• -42-

SUMMOtRY

A PRELIMINARY SURVEY OF CAVES LOCATED IN THE

BELT OF THE APPALACHIAN VALLEY OF VIRGINIA, HAS REVEALED

THE PRESENCE OF NUMEROUS SOIL AND WATER INHABITING SPECIES OF AQUATIC PHVCOMYCETE8• A REVIEW OF THE LITERATURE INDI- CATES A PAUCITY OF INFORMATION DEALING WITH THESE ORGANl8M8•

THE CAVE& SAMPLED AND THE METHODS USED IN THIS INVESTIGATION

ARE BRIEFLY DESCRl8EOa

FROM NINETEEN CAVES, 162 SOIL AND WATER 8AMPL£8 WERE

COLLICTED FROM WHICH 33 ISOLATES HAVI BEEN OBTAINEOa IN

ADDITION A NUMBER OF MONOCENTRIC CHYTRI09 WERE 088ERVED 1

BUT HAVE NOT BEEN FURTHER IDENTIFIED. EACH SPECIES

SSOLAT!O HA8 BEEN STUDIED AND DESCRIBED UNDER UNIFUNGAL

CULTURAL CONOITtONSe THE OCCURRENCE ANO Ot8TRf8UTION OF

EACH 8P£CIE8 18 CITED AND ANY VARIATtON8 IN MORPHOLOQY OR

DEVELOPMENT ARE DISCUS8£De QUESTIONS DEALING WtTH THE

TAXONOMY AND NOMENCLATURE ARE CONSIDERED IN !ACH CAIE•

THE CAYERNICOLOU8 ENVIRONMENT MAY BE REGARDED AS A

NEW STATION FOR EACH SPECIES REPORTED HEREIN• THREE 8P£CIE8

OP' PyTHIUM, !• AFERTU.E, f• IRREQUL!RE, AND f• NftOAI 1,, ARE RIPORTED FOR THE FIRST TIME AS SAPROPHYTIC INHABITANT8 OF

THE SOIL• -43-

818L IOGRAPHY

BAILEY, VERNON. 1933. CAVE LIFE IN KENTUCKY. AMER•

Mf OlANO NATURALIST, J.i! 385-635.

BARY, A. DE. 1860. EtNIGE NEUE SAPROLEGNIEEN• JAHRBe WISS. BOT., g: 169-192.

1876. RESEARCHES INTO THE NATU~E OF THE POTATO-

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NEW MILTOth

YOUNKIN, S. G. 1938. PYTHfUM IRREQULARE AND DAMPING-OFF OF WATERMELLONa. PHYTOPATH., -2a: 596. The vita has been removed from the scanned document ABSTRACT

A PRELIMINARY SURVEY OF THE FUNGUS FLORA OF CAVES. I• AQUATIC PHYCOMVC!TES • ., MARIARET MlRIHALL HOLLAND

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