STUDIES IS THE SEXUALITY OF THE HETEROBASIBIAE % Hor&co L* B a rn e tt A THESIS P re se n te d t o th e G raduate School o f M ichigan S ta te C ollege o f A g ric u ltu re and A pplied Science in p a r t i a l f u lf i l l m e n t o f requirem ents f o r th e Degree o f D octor o f P hilosophy y Botany Department E a st L ansing, M ichigan 1937 ProQuest Number: 10008256 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. uest ProQuest 10008256 Published by ProQuest LLC (2016). Copyright of the Dissertation is held by the Author. All rights reserved. This work is protected against unauthorized copying under Title 17, United States Code Microform Edition © ProQuest LLC. ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 4 8 1 0 6 - 1346 ACKNOWLEDGMENTS The a u th o r l a in d eb ted t o Dr* E rn st A* Beosey f o r h e lp f u l s u g g e stio n s and c r i ti c i s m s th ro u g h o u t i. .o course o f th e i n v e s t i ­ g a tio n s and i n th e p r e p a ra tio n o f th e m an u sc rip t, t o Dr* G* W* M a rtin f o r i d e n t i f i c a t i o n o f specimens o f E x id ia , and to Mr* John B* B outien f o r c r i t i c a l re a d in g o f th e m anuscript* 109108 TABLE OF COIITEKTS Page I IHTnODUCTIOl MATERIALS ASD liLTaOIX' 1 C o llectio n s o f Spooioa iStudlod Ucrthodfi o f S ecu rin g 1 Culture© 3 C u ltu re !3odia %w3 6 r e s u lt :; o f b ^ h e w s ? G on.iinatlon o f B&sidiosparoo 7 Faerosooplc i^posarunoe of Cultures 22 Mlerofioepie Appsaraooo o f Ityoello, 14 P ro d u c tio n o f Oldie. 15 Oorj'.umxtiori o f Oidia 17 Production o f Fruit Bodloe In Culture IS Growth Ratos o f Cultures £1 Union o f Ifypbeo end F o r m tio n o f Clamp C onnections 22 S o x r o llty 23 A n r ic u la ria a u r ic u la - judae P a irin g s o f ISoaoeroro C u ltu re s from SameF r u it Body 20 P a irin g s o f Monoeporo C u ltu re s from D iffe re n t 34 Fruit Bodies B glftla g la a d u lo m P a irin g s o f ttonoepero C u ltu re s f t m Stsao F r u it Body 42 P airings o f Iforoeporo Cultures from D ifferen t F r u it 3odios46 E addla r e p ls a P a irin g s o f Baaoeporo Cuituareo f r o n Sano F r u it Body 54 P a irin g s o f Monospor© C u ltu re s from D iffe re n t F r u it Bodioe 54 B addla sa c o h a rln a Page 58 B x ld ia im c le a ta 58 A t t e s t s to Hybz*idiz© S pecies o f E x id ia 60 DISCUSSION 60 S W I 68 1 S tu d ie s ill th© S e x u a lity o f th e H e te ro h a sid ia e E3TR0DUCT101 I n th e p r e s e a t work a s tu d y was made o f v a rio u s sp e c ie s o f th e o rd e rs A u r lc u la r ia le s , T r e n e lla le s and D acryoraycotales. However, owing to th e absence o f clamp co n n ectio n s in th e sp e c ie s o f Dacryomyces, G uepinia and C alo c e ra t h a t w ere c o lle c te d , and owing t o th e f a i l u r e t o develop a s u it a b l e te c h n iq u e f o r s ta in in g t h e i r n u c l e i , th e s e x u a lity o f th e s p e c ie s o f t h i s l a s t o rd e r was n o t in clu d ed i n t h i s study# Spore germi­ n a tio n was s tu d ie d i n T reraella lu te s c e n s , b u t due t o th e f a i l u r e o f th e c u ltu r e s t o produce e x te n s iv e mycelium on th e v a rio u s media used no work co u ld be done concerning se x u a lity * The s p e c ie s from which mono­ spore c u ltu r e s w ere o b ta in e d and whose s e x u a lity was s tu d ie d w ere A a r ic u la r ia a u ric u la -ju d a e (P r» ) S c h rd t,, B xldia g lan d u lo sa ( B u ll.) F r . , E* r i o l s a (D ltm .) F r . , E. s a c c h a rin e F r . and B. n u c le a ta (S c h r.) B u rt. The l i t e r a t u r e co n cern in g th e H e te ro h a sid ia e , o th e r th a n from th e taxonom ic s ta n d p o in t, i s s c a rc e . S everal w r i t e r s , in c lu d in g B re fe ld ( 4 ) , Bangeard ( 1 1 ), M Sller (2 1 ), Shear and Dodge (SO), le u h o ff (2 4 , 2 5 ), G ilb e r t ( 1 4 ), end Eniep (1 7 ), have re p o rte d g erm in atio n o f sp o re s i n some s p e c ie s o f t h i s group, b u t few o f them (B re fe ld , Shear and Bodge, le u h o ff , and E aiep) have re p o rte d th e p ro d u c tio n o f ex­ te n s iv e mycelium. C y to lo g ic a l work has been p u b lish ed by le u h o ff (2 4 ), G ilb e r t ( 1 4 ), M h n er ( 1 9 ), Sogers (2 8 , 29) and Whelden (4 0 , 4 1 , 42, 4 5 ). MATERIALS AS© METHODS C o lle c tio n s o f S pecies S tu d ied The c o lle c tio n s o f th e s p e c ie s o f A u ric u la ria and B x ld ia used i n c u l t u r a l work and from w hich monospore c u ltu re s were o b ta in e d a re giv en in T able 1* 2 fa b le 1 C o lle c tio n number S p ecies A* a u r ic u la Judae P it ce o f C o lle c tio n Date A dirondaoks, 8/2 3 /3 5 R. Y*1 I n III 8 Host Spore s iz e F ir? 1 0 .5 -(1 4 .6 )-1 7 .5 x 4.8-< 5 * 4 )-7 L in co ln Kebr. 2 Received ll/7 /S 5 Decidu­ ous 11.8~(13.5)-15*6x 4 * 8 -(5 .2 )-5 .7 I? Chapel H il l M* C. 3 7 /6 /3 6 Oak 8 * 8 -(ll* 9 )-1 4 * 8 x 5 * 3 -(5 .6 )-6 .6 « Y Lincoln* H ebr.^ Received 10/28/35 Decidu­ ous 8 * 8 -(12*4)-14.4x 4 .4 -(5 * 2 )-6 t! TI Iowa C ity Iowa. “ 8 /2 /3 6 tt » V II « 9 /3 /3 6 Ash lU 3 -(1 3 * 3 )-1 4 x 4*8—(5*5)—6*2 n V III F t.G o11in s C olo.5 Received 1/5/37 F ir 13*2-(15)-17*5x 4*4-(5*4)-6*6 E* Lansing Mich* 11/4/34 Hickory E* gl&ndulosa I «4 10»5-(13*1)-15* 2x 4 ,8 -{ 5 .2 )-5 * 7 ft II M It 4/23/36 •» II V Lyons, Mich. 5/30/36 Oak I E. Lansing 3/28/36 Mich* U II Lyons, Mich 5/S0/36 Oak tt III E. Lansing Mich. 6 /4/26 H ickory E. sacch& rina I C ra y lin g , S ic h . 9/IO /3 6 White P ine E. n u c le a te I E. L ansing, Mich. 7/9/35 Maple E. re .c isa 1. 2* S* 4* 5. Hickory C o lle c te d by Dr* E. A* Bessey By c o u rte sy o f Dr* Leva B. Walker By c o u rte sy o f Dr. W, C. Coker By c o u rte sy o f Dr* G. W» M artin By c o u rte sy o f Prof* J . L. Forsberg A ll o th e r specimens were c o lle c te d by th e au th o r l a th e re c o rd s o f sp o re s i z e s , given in Table 1 , th e f i r s t and l a s t numbers i n each case r e p r e s e n t th© extrem es and th e numbers in p a re n th e s e s a r e th e means* One hundred sp o res from, each o f c o lle c tio n s I , IV, V, and T i l l were m easured, w h ile 20 sp o res each o f c o lle c tio n s I I I * Y I and T O w ere measured* I t i s in te re s tin g n o te th e d iff e r e n c e s i n th e s iz e s o f sp o res o f t h e c o lle c tio n s o f A* a u ric u la -ju d a e * The w id th v a r ie d o n ly slightly, b u t i n le n g th t h e sp o res v a r ie d c o n s id e ra b ly . C o lle c tio n s I and T i l l had th e lo n g e st sp o re s w ith r e s p e c tiv e mean le n g th s o f 14*6/^- and 15*0yu* The mean le n g th s o f sp o res from o th e r c o lle c tio n s ranged from 11 * 9/a t o 13*^1** A b e t t e r com parison between th e spores o f c e r t a i n c o lle c tio n s i s se en when th e d i s t r i b u t i o n o f spore le n g th s i s giv en in th e form o f a graph (T ex t F ig . 1 ) . Two f r u i t b o d ies w ere s e le c te d from c o lle c tio n V o f A* a u r ic u la ju d ae end d e sig n a te d as Ya and Vb. These had a lre a d y been removed from th e h o s t wood and th e d is ta n c e a p a r t could n o t be determ ined* From c o l l e c t i o n I I o f B. g len d n lo sa two f r u i t b o d ies ( H a and H b ) war© chosen from th e same lo g o n ly two inches a p a rt* The o n ly c o l l ­ e c tio n o f B* s a c c h a rin s was found growing on a s t i c k about 12 In d ie s long* Two f r u i t b o d ies were s e le c te d and d esig n a te d as l a and l b . Only on© f r u i t body was s e le c te d from each o f th e o th e r c o lle c tio n s . Method o f S ecuring C u ltu res !h e n f r e s h m a te r ia l was n o t a v a ila b le th e d rie d f r u i t b o d ies were m oistened w ith d i s t i l l e d w a te r and p laced on m oist p ap er to w e lin g in a p e i r i dish* A fte r a fear hours th e f r u i t bodies were examined w ith low power o f th e compound m icroscope to be su re t h a t th e y were producing a n abundance o f spores* At f i r s t a Cham berlain ra ic ro m n ip u la to r m s 4 45 40 30 Nuinter of Spores 35 15 10 8 9 10 j. 11 L 12 i 13 14 15 ± 16 17 18 Length o f sp o res in m icrons Text f i g . 1 . Graph showing d istr ib u tio n o f len g th s o f 100 spores each o f four c o lle c tio n s o f A. aur i cm1a - .1udae . 5 used to p ick th e spores from th e b a sid ia . However, t h is method soon proved to he u n sa tisfa c to r y from th e standpoint o f tim e and because o f o th er d isch argin g sp ores f a llin g on th e needle o f th e m anipulator before a s in g le spore could be picked o ff* A fter t h is th e spores were picked ou t from spore d ep o sits on agar p la tes* The m ost s a t i s f a c t o r y method o f o b ta in in g spore d e p o s its was s im ila r t o th e method u sed by Mounce ( 2 2 ). A p o r tio n o f a f r u i t body m s fa s te n e d t o th e cover o f a p e t r i d is h by means o f a drop o f th i c k Canada balsam* T h is p e rm itte d th e l a t e r a d d itio n o f a sm all drop o f w a te r to th e p ie c e o f f r u i t body w ith o u t d e s tro y in g th e adhesive prop­ e r t y o f th e balsam* The p e t r i d is h cover w ith th e p ie c e o f f r u i t body was th e n p la c e d o ver an a g a r p l a t e and "the spore® were allow ed t o f a l l upon th e agar* By slow r o t a t i o n o f th e cover a more o r l e s s even d i s t r i b u t i o n o f sp o res was o b ta in e d i n a c i r c u l a r area* Spores from f r u i t b o d ies produced i n c u ltu r e were suspended in s t e r i l e d i s t i l l e d w a te r which was th e n poured o v er ag ar p la te s * M ultispore cu ltu re s were obtained by c u ttin g out blocks o f agar co n ta in in g many spores and tr a n sfe r r in g them to t e s t tubes o f agar* F or mono spore c u ltu r e s v ia b le spores were in su red by allo w in g them to germ inate b e fo re th e y were p ick ed out* A t o o l f o r p ic k in g o u t s in g le sp o re s was fa sh io n e d from a sm all sewing needle w ith a v e ry sm all rounded eye* The m etal around th e eye was ground down somewhat and sharpened making a c a t t i n g edge* T his end o f th e n eed le was th e n b e n t a t an an g le o f about 45° and th e p o in te d end in s e r te d in to a m etal ho ld er* T h is made a v ery convenient to o l which could be s t e r i l i z e d i n a flam e and, when n o t allow ed to become red h o t, one such to o l la s te d a lo n g tim e* 6 fh© a g a r p l a t e w ith th e sp o re d e p o s it was p laced upon th e s ta g e o f a b in o c u la r d is s e c tin g m icroscope rood th e g e m in a te d spores lo c a te d by use o f th e h ig h power o b je c tiv e ( a t a m a g n ific a tio n o f about 96X). The eye o f th e s p e c ia lly p re p ared n eed le was p re sse d down around & w e ll- is o la te d germ inated s p o re , c u ttin g o u t a sm all a re a o f a g a r t o th e bottom o f th e p la te * At t h i s s ta g e th e spore could be observed i n th e a re a c u t o u t by t h e needle* T his p ie c e o f a g a r b e a rin g th e spore was l i f t e d o u t and w ith th e p o in t o f a second s t e r i l e n eedle th e to p p o r tio n o f th e ag ar b lo ck was pick ed o f f and tr a n s f e r r e d to a tu b e o f agar* The u se o f th e second n e e d le i n removing o n ly th e upper p o r tio n of th e a g a r blo ck r e ­ duced th e p o s s i b i l i t y o f tr a n s f e r r i n g o th e r sp o res w hich m ight s ti c k t o o th e r p a r t s o f th e f i r s t needle* The c u ltu r e s wore th e n p la c e d under b e l l J a r s in a m o ist atm osphere and in cu b ated a t room tem perature* C u ltu re Media Used During th e f i r s t p a r t o f th e in v e s tig a tio n s p o ta io -d e x tro se a g a r was used e x c lu siv e ly * h a te r , v a rio u s m o d ific a tio n s o f t h i s and s e v e ra l o th e r media were t r i e d i n o r d e r t o determ ine th e media b e s t s u ite d f o r v e g e ta tiv e growth end f o r th e p ro d u c tio n o f f r u i t bodies* in g media w ere te s te d * The fo llo w ­ p o tato -d ex tro s© ag ar w ith and w ith o u t p ep to n e, m alt a g a r w ith and w ith o u t p ep to n e, d e x tro -n a lto s e a g a r, prune e x t r a e t c o m meal a g a r, Csapek*s medium, powdered wood-ins.lt extract-pepton© medium a s d e sc rib e d by Arnold ( l ) , m oistened powdered wood o f m aple, h ick o ry and o ak , and au to cla v ed s tic k s o f basswood, m aple, h ick o ry and o ak . F o r th e l a s t medium, liv in g s tic k s o f wood about -}• to 1 inch in d ia m ete r w ere chosen* The b ark was removed from one s id e o f th e stic lc s which were p la c e d in la r g e t e s t tu b e s c o n ta in in g about two indues o f d i s t i l l e d w ater and s t e r i l i z e d a t 15 pounds p r e s s u re . T his fu rn ish e d 7 a c o n s ta n tly m o ist atm osphere I n th e tu b es and a range i n th e amount o f m o istu re in th e -rood. The b e s t media f o r v e g e ta tiv e growth o f a l l sp e c ie s were p o ta to d e x tro se a g a r ( b ro th o f 200 gnu p o ta to e s , 20 gau d e x tro s e , 15 guu a g a r, enough d i s t i l l e d w a te r to make 1 l i t e r ) , end m alt e x tr a c t a g a r (15 gnu d e s ic c a te d m a lt e x t r a c t , 15 gcu a g a r , 1 l i t e r o f d i s t i l l e d w a te r ) . L i t t l e d iff e r e n c e was se e n betw een th e s e two media* The second medium was used alm ost e x c lu s iv e ly th ru o u t th e l a t t e r p a r t o f th e work and f o r a l l o f t h e p a ir in g s o f monospore c u ltu re s * The p resen ce o f 1 ga* o f peptone p e r l i t e r I n each o f th e media had no ap p aren t e f f e c t on th e v e g e ta tiv e growth* The p o o re s t v e g e ta tiv e growth was produced on d ex tro -m alto se a g a r , Czapek'g medium and on wood media* F or th e fo rm atio n o f f r u i t b o d ies th e b e s t media were p o tato -d ex tro s© a g a r w ith 1 ga* o f peptone p e r l i t e r , m alt e x tr a c t a g a r and p ru m -c o rn meal a g a r in th e o rd e r named* The most t y p i c a l l y shaped f r u i t bodies o f A* a u ric u la -ju d a e were produced on m o ist s tic k s o f basswood and hickory* RESULTS OF EXPERIMENTS G erm ination o f B asidiospores A* a n r ic u la -ju d a e , Kfoea d r ie d f r u i t bodies w ere re v iv e d , spore d isc h a rg e began a s e a r ly a s f iv e hours a f t e r th e y had resumed t h e i r normal shape and size * The f i r s t s p o re s , d isch arg ed from the f r u i t b o d ie s , germ inated v e ry p o o rly , on ly about 1-4;% a f t e r f o u r days on a g a r. Spores d isch arg ed l a t e r showed alm ost 100% g e m in a tio n a f t e r f o u r days on a g a r a t room tem perature* Most o f th e spores g e m in a te d w ith in 12 t o 48 hours a f t e r b ein g discharged* Spores from a f r u i t body which had been i n a d ry c o n d itio n f o r alm ost nine months germ inated poorly* At th e tim e o f d isch arg e th e spores were s in g le c e lle d , but b e fo re 8 g e rm in a tio n th e y o f te n , b u t n o t alw ays, te c me two o r t h r e e - c e l l e d , each c e l l c o n ta in in g a s in g le n u cleus ( p i . %t l ) . On a g a r th e sp o re s germ inated by means o f one o r r a r o ly by two lo n g germ tu b es* The germ tu b e u s u a lly a ro se from th e a p ic u la r end o f th e spore ( P i . I , 4 ) , b u t some a ro se from th e o th e r end o r from th e convex s id e o f th e sp o re (PI* I , 5)* The germ tu b e s o f te n rem ained unbranched f o r a d is ta n c e o f 75 t o lOGyu, a f t e r which th e y showed fre q u e n t branching* Some o f th e branches soon p e n e tra te d downward in to th e a g a r w h ile o th e rs grew on th e s u rfa c e o r tu rn e d upward t o produce a e r i a l hyphae. A fte r th e germ tu b e had re a c te d some le n g th th e c o n te n ts l a th e sp o re passed o u t in to th e tu b e , re v e a lin g c l e a r l y th e s e p ta i n th e spore and o ld e r p o r tio n o f th e tu b e (p i* I , 4 ) , Growth and branching co n tin u ed u n t i l abundant whit© mycelium was produced* B asidiospores w hich had f a l l e n on m o ist p ap er to w e lin g were examined a f t e r two days* Only about 10j£ had g e m in a te d and th e s e had produced c o a rse germ tu b e s s im ila r to th e germ in atio n o f sp o re s on agar* Some o f th e sp o res d isc h a rg ed from a f r u i t body f o i l back and c o lle c te d i n a m s s on th e f r u i t body i t s e l f * These were examined and s e v e ra l showed germ in atio n by th e p ro d u c tio n o f a s in g l e , s l i g h t l y cu rv ed , secondary sp o re on a s le n d e r germ tu b e ( p i . 17, 1)* sp o re s in d i s t i l l e d w a ter were examined a f t e r two days* B asidio­ The p ercen tag e o f g erm in a tio n was h ig h and most o f th e spores were d i s t i n c t l y tw oc e lle d * A few sp o re s produced s to u t germ tu b e s , b u t in most cases g erm in a tio n was by means o f a f in e germ tub© about lyuwide and 5-15 yu lo n g , from each c e l l o f th e spore* Each germ tu b e bore one o r more sic k le -sh a p e d o id ia a t i t s t i p (PI* I , 2)« S im ila r germ in atio n of sp o re s and p ro d u c tio n o f o id ia in w ater and in n u tr ie n t s o lu tio n were d e s c rib e d by B refeld (4 ) f o r A* aambucina. Measured alo n g th e chord co n n ec tin g th e t i p s th e o id ia ( p i . 17, 2) were 2 -3 jx lo n g , alth o u g h th e y were a c t u a l l y 4 -5 ^ u in le n g th . The th ic k n e s s a t th e m iddle was 1 . 5 ^ w h ile th e c u rv a tu re was such t h a t th e d is ta n c e from th e chord to th e o u te r convex s id e was 2-3/j. B. g la n d u lo sa . v ery re a d ily * Spores d isch a rg ed from f r e s h f r u i t bodies g e m in a te d Germ tu b e s a s lo n g as SO^u-were produced w ith in 12 hours a f t e r f a l l i n g on agar* The sp o res were s in g le c e lle d a t th e tim e o f d isc h a rg e b u t b e fo re g erm in ation some became two o r r a r e l y th r e e - c e lle d (P I. I , 10)* Most o f th e sp o res produced two c o arse g em tu b e s , one a t e i t h e r end o f th e spore ( P i. I , 1 2 ), o r one o f te n a ro se from th e convex s id e o f th e sp o re (PI* I , 13 ). a le n g th o f Some o f th e germ tu b es grew to b e fo re b ran ch in g and th e n branched fre q u e n tly * One c o l l e c t i o n was re v iv e d a f t e r b ein g i n a d rie d c o n d itio n in th e la b o ra to ry f o r e le v en months. t e n hours* These f r u i t b o d ies began to d isch a rg e spores a f t e r G erm ination o f sp o res was poor and much slow er th a n th o se from f r e s h ly c o lle c te d f r u i t bodies* The m ycolia produced, however, w ere j u s t as v ig o ro u s in growth as th o se o b tain ed from f r e s h f r u i t b o d ie s. Mien spores from a f r e s h f r u i t body were p laced in d i s t i l l e d w ater th r e e methods o f g erm in atio n were observed a f t e r two days. long g e m tu b e s v ery s im ila r to th o se on a g a r. Some produced A few produced a sh o rt sle n d e r germ tu b e b e a rin g a s in g le secondary spore (P I. I , 1 1 ). Many o f th e spores germ inated by means o f s e v e ra l (a s many as e ig h t) s h o rt s le n d e r g e m tu b e s (P I. I , 9AS B). At th e t i p s of th e s e germ tu b es one o r more s ic k lo - s h ped o id ia ( p i . I , 9C) were borne. These measured 3 . 5/x along th e chord co n n ectin g th e t i p s , w ith th e a c tu a l le n g th about 5/i, and in th ic k n e ss about l u . Only a few o id ia were seen a tta c h e d to th e germ 10 tu b es* S im ila r p ro d u c tio n o f o id ia by g e m in a tin g sp o res o f B« g lan d u lo sa was d e s c rib e d and i l l u s t r a t e d by B re fe ld (4)* on m o ist mood g e m in a te d poorly* Spores f a l l e n in a mass Some did so by means o f secondary sp o res and o th e r s by long c o a rse g e m tubes* B. re o isa * Spores from f r e s h f r u i t bo d ies germ inated w ith in 12 h o u rs a f t e r f a l l i n g on agar* G erm ination mas alm ost 1QQ/& a f t e r 36 hours* The s p o re s w ere s in g le - c e lle d when d isch arg ed and some remained s o , w hile o th e r s became tw o -c e lle d b e fo re g e m in a tio n ( P I . I I , 1 ). G e m in a tio n was by means o f one o r two c o a rse germ tu b e s , one a t e i t h e r end o f th e spore (PI* I I , 2 , 3 ) . About two o r th re e days a f t e r g e m in a tio n many o f th e young hyphae produced sm all s tr a i g h t o r s l i g h t l y curved o id ia , which were 5*5-4/* by 1.6yu in size* These were borne in sm all c lu s te r s a t th e t i p s o f th e hyphae o r on s h o rt l a t e r a l branches ( P I . I I , 3 ), Some s h o r t p e g - lik e branches b e a rin g o id ia were produced from th e main body o f th e spore* The p ro d u c tio n o f o id i a d id n o t p re v e n t th e growth o f th e hyphae ior th e subsequent fo rm atio n o f e x te n siv e mycelium* Son® hyphae bore no o id ia (P I. I I , 2) b u t continued growth t o form m y celia. A fte r 36 hours i n d i s t i l l e d w ater very few spores showed any s ig n s o f germ ination* These few produced c o a rse germ tubes* B* sacch arin s* Spores from f r e s h f r u i t bodies germ inated r e a d ily on a g a r and a f t e r 12 hours had formed s h o rt c o arse germ tu b e s . th r e e g e m tu b e s grew from each spore* th e tim e o f germ ination* One to Souse spores were tw o -c e lle d a t About 30 hours a f t e r th e sp o res had f a l l e n on a g a r, abundant sm all rod-shaped o id ia were observed on th e young hyphae (PI* V, 1 ) . They were borne i n th e same manner as th o se o f B* re c isa * The o id ia measured 3 .5-^ux 1 .5 -2 yu • B refeld (4) has d escrib ed and i l l u s t r a t ­ ed s im ila r p ro d u c tio n o f o id ia in sa c c h a rin a var* fo iia c e a * 11 B, p u c lea ta * Spores c o lle c te d from f r o s h f r u i t bo d ies showed le s s th e n X% g erm in atio n on ag a r in a l l c a s e s , and th o se from some f re s h f r u i t b o d ie s f a i l e d to g e m in a te a t a ll* However, spores from f r u i t bodfees grown i n c u ltu r e germ inated r e a d ily on a g a r T/ith th e p ro d u ctio n o f lo n g g e ra tubes* In d i s t i l l e d w ater some sp o res germ inated by means o f a s in g le secondary spore on a s le n d e r germ tu b e , (P I. V I, 2 ) , and o th e rs by means o f one (Pi* V I, 3) o r two g e m tu b es s im ila r to th o se formed by sp o res on agar* Spores which had f a l l e n on m oist wood around a f r e s h ly c o lle c te d f r u i t body germ inated poorly* Some produced secondary spores and o th e rs b ore sm all d u s t e r s o f sic k le -sh a p e d o id ia a t th e t i p s o f s h o rt s le n d e r germ tu b e s (PI* V I, 1 ) . Only one such germ tu b e was formed by each spore* These o i d i a measured 2*5-3*5y .alo n g th e chord between th e t i p s , about 4-4*5yuin a c t u s l le n g th and about 1 * 5 ^ In th ic k n e s s . B asid io sp o res in a 2% d e x tro se s o lu tio n produced secondary spores* O ther Species* On a g a r th e germ ination o f b a sid io sp o re s was v ery s im ila r i n Dacryorayces m inor P k ., C alooera cornea (B atsch) F r. and G uepinia s p a th u la r ia (Sohw*) Fr* Spores o f a l l th r e e ^ e e i e s were o n e -c o lle d when d isch arg ed from th e f r u i t body b u t th e y a l l became s e p ta te b e fo re germ ination* Those o f C. cornea ( p i. I l l , 6) end G* s p a th u la r ia became tw o -c e lle d and th o se of D. minor formed fo u r o r more c e l l s b e fo re g erm in atio n ( P I . I l l , 7 ). G erm ination began a f t e r about 12 hours on agar* One o r more coarse germ tu b e s were formed from each c e l l o f th e spore* These a re i l l u s t r a t e d f o r C* cornea ( P i. I l l , 1 , 2) 8111x1 f o r Sh m inor (FI* I I I , 9)* The g era tu b es soon branched abundantly and mycelium was formed d ire c tly * S im ila r form ation o f mycelium in D* d eliq u eso e n s was d e sc rib e d by G ilb e r t (1 4 ). In d i s t i l l e d w ater a 12 few sp o res o f Do m inor produced co arse germ tu b e s w h ile o th e rs bore sm all s p h e ric a l o i d i a on s h o rt ste rig z n a -lik e s tr u c tu r e s ( P I . I l l , 8 ) . Spores w hich had f a l l e n in a mass on m oist wood produced o id ia i n th e same manner. Such p ro d u c tio n o f o id i a was d escrib ed by B refeld (4) f o r sp e c ie s o f Dacryomyces. Spores o f T rem ella lu te s c e n s p e r s . germ inated v ery r e a d i ly on ag a r by re p e a te d budding ( p i . I l l , 12) which continued u n t i l th e r e was produced a sh in y opaque mass o f buds, ap p earin g much lik e a b a c t e r i a l co lo n y . Some o f th e s e buds germ inated and produced s h o rt hyphae, a p e r t o f which bore ovoid buds o r o id ia , b u t fo r some reason y e t unknown no d e f i n i t e m y c e lia l c u ltu re s were formed. B asidiospores which had f a l l e n back onto th e f r u i t body germ inated by means o f second­ a ry sp o res ( P I . I l l , 11). M acroscopic Appearance of C u ltu re s lo m acroscopic d iffe re n c e was noted between th e mono c a r yon and d icary o n m ycelia o f any s p e c ie s . The m ycelia o f A. a u r ic u la - ju d a e , E. g lan d u lo sa and E. r e c is a were so n e a rly a lik e t h a t in most cases i t was im possible t o d is tin g u is h between th e th r e e s p e c ie s by t h e i r appear­ an ce. The m ycelia o f a l l th ro e sp e c ie s were pure w h ile , o r brownish in some o ld e r c u ltu re s o f A. a u ric u la -ju d a e and E. r e c i s a , w ith abundant a e r i a l growth. The mycelium o f E. r e c is a was more v a r ia b le in appear­ ance th an t h a t o f any o th e r s p e c ie s . In s e v e ra l o f th e c u ltu r e s , bo th d icary o n and monocaryon, th e r e appeared s m a ll, r a is e d dark brown g e la tin ­ ous b o d ies about 2-5 mm. in diam eter ( p i. Y, 5 ). These were co n sid ered p o s s ib ly to be s t e r i l e o r a b o rtiv e f r u i t bodies and w ill be d iscu ssed la te r. An i n t e r e s t i n g s i t u a t i o n was found in th e m acroscopic appearance 13 o f c u ltu r e s o f B. g la n d u lo sa . A fte r th e mono sp are m ycelia had been in c u ltu r e f o r alm ost a y e a r , d u rin g which tim e fre q u e n t tr a n s f e r s had been made, i t was observed t h a t c u ltu r e s 23, 24 and 50 had ceased t o produce an abundance o f w h ite a e r i a l growth* Host o f th e hyphae were confined to th e s u rfa c e o r u n d er th e s u rfa c e o f th e a g a r. Only c u ltu r e s 23 and 24 a re i l l u s t r a t e d ( p i , V II, lAf B ), b u t c u ltu r e 50 was v e ry s im ila r i n app earan ce. P a irin g s were made between c u ltu r e s o f d i f f e r e n t ty p e s , u sin g 21 ( P i. V II, 1C) and 51 a s examples o f c u ltu re s w ith much f l u f f y a e r i a l mycelium. The d ic a ry o n mycelium produced from th e m ating 24 x 50 grew c lo s e to th e ag ar ( p i . V I I , 4 ), much i n th e same manner as d id each o f th e two mono c a r yon m y c elia. The dicaryon mycelium which r e s u lte d from th e p a ir in g 21 x 51 was f l u f f y w ith co n sid e ra b le a e r i a l growth ( P I . V I I , 2)* Hhen 21 was mated w ith 50 th e r e s u lt in g d icary o n mycelium was somewhat in te rm e d ia te in i t s manner of growth ( p i , V I I I , 3)* Although no d e f i n it e c o n clu sio n s can be drawn from so few r e s u l t s , th e y do seem t o in d ic a te t h a t th e changes in th e ty p e o f growth of th e monocaryon m ycelia in flu e n c e th e ty p e o f growth o f th e d icary o n mycelium produced when th e s e c u ltu r e s w ere mated w ith o th e r s . The mycelium o f B. n u c le a ta m s w hite and grew very slow ly. The growth f o r th e most p a r t was low and c lo se to th e a g a r o r was bunched up i n a mass w ith v ery few a e r i a l hyphae. The mono spore c u ltu r e s a l l produced th e same t . p e o f growth a t f i r s t . About s ix or seven months a f t e r th e y were s t a r t e d , i t was n o tic e d th a t a few o f th e s e c u ltu re s had produced s e c to rs which were more f l u f f y and w ith more a e r i a l growth. When such a s e c to r was tr a n s f e r r e d th e f lu f f y c o n d itio n o f th e mycelium p e r s i s te d . Such changes i n m y celial growth occured in f iv e nonospore c u l t u r e s , numbers 3, 17, 22, 25 and 26. This change was n ot c o r r e la te d 14 w ith any o th e r p e r c e p tib le c h a ra c te r o f th e mycelium n o r w ith e i t h e r sex u a l phase* U n fo rtu n a te ly , th e c u ltu re s were l o s t due to lo s s o f v ig o r and i t was n o t determ ined w hether th e f l u f f y c h a ra c te r was in ­ flu e n c e d by a g e n e tic a l f a c t o r o r w hether th e change was due m erely to p rolonged growth o r a r t i f i c i a l media* The d icary o n mycelium o f E» sac c h a rin e was v e ry s im ila r to t h a t o f E* r e c i s a b u t produced a more spreading growth and was c lo s e r to th e agar* The monospore c u ltu r e s v a r ie d c o n sid e ra b ly i n ap p earan ce, some b e in g more f l u f f y th a n o th e r s (Pi* Y, 3)* The masses o f o id ia produced i n c u ltu r e appeared as r a i s e d , sh in y , opaque a re a s which were w hite a t f i r s t b u t l a t e r tu rn e d dark brown ( P I, V, 2)* The m y celia o f Pacryomyces m inor, C alooera cornea and G uepinia s p a th u la r ia grew v e ry slo w ly , being q u ite c lo se to th e ag ar a t the edge and more bunched up and compact in th e c e n te r o f th e c u ltu re * The m ycelia o f a l l th r e e sp e c ie s v a r ie d from b r ig h t orange to yellow in c o lo r , and appeared more faded when grown in th e dark* T hick, rope­ l i k e , b ran ch in g u p r ig h t s tra n d s were produced in d icaryon c u ltu re s o f C. cornea* When f i r s t formed th e y looked v ery much lik e young f r u i t b o d ies b u t no spores were produced by them* M icroscopic Appearance o f M ycelia Abundant clamp co nnections were produced by dicaryon m ycelia o f A. a u r ic u la - ju dae ( p i , X, 7 ) , B» g lan d u lo sa (PI* I , 1 4 ), E« r e c is a (PI* I I , 5 ) , B. sa o c h a rin a (PI* I I , 6 ) , and E* n u c le a ta (PI* I I , 1 3 ). No clamp co nnections were p re s e n t in m u ltisp o re c u ltu re s o f P. m inor, C. cornea nd G* s p a th u la ria * The m ycelia o f a l l sp e c ie s branched f r e q u e n tly , e i t h e r a c u te ly o r a t r ig h t an g les to th e main hyphae. hyphae o f a l l s p e c ie s were q u ite narrow , ranging in most cases from The 15 1-4 p- i n w id th , w ith a few a s wide as 5 ^ • In g en eral th e d icary o n hyphae were somewhat w ider th a n th e mono c a r yon but th e d iffe re n c e was n o t d i s tin c t* The hyphae o f A. a u ric u la -ju d a e were s li g h t l y w id er th a n th o s e o f o th e r species* I n a l l sp e c ie s th e y were composed o f long c e lls * P ro d u ctio n o f O idia A* a u r ic u la -ju d a e , The fo rm atio n o f sic k le -s h a p e d o id ia on germ­ in a tin g b a sid io sp o c s i n w a te r h as been d escrib ed above* O idia s im ila r in shape b u t s l i g h t l y la r g e r were borne i n c lu s te r s on monocaryon m ycelia (PI* I , 6)* They measured 5 * 5 - 5 # along th e chord from t i p to t i p , abo u t 4 -6 * 5 y i n a c tu a l le n g th and about 1*7^ in th ic k n e s s . They were r a r e l y found i n o ld c u ltu r e s b u t were most abundant in young c u ltu r e s about one o r two weeks a f t e r th e spores had germ inated. Bo o id ia were observed on d icary o n m y celia. E. glandulosa* S ick le -sh ap ed o id ia in very sm all numbers were observed a few tim es in th e exam ination o f aonospore c u ltu r e s , e s p e c ia lly in number 50. They were f i r s t observed i n t h i s c u ltu re about s ix months a f t e r i t was s t a r t e d and fre q u e n t tr a n s f e r s had been ms.de in th e mean­ tim e . The o id ia were s im ila r i n s iz e and shape to th o se d escrib ed above in th e g e m in a tio n o f b a sid io sp o re s in w a te r. Bo o id ia were produced on d icary o n mycelia* S . re c isa * The p ro d u c tio n o f o id ia on v ery young monocaryon m y celia, s h o r tly a f t e r th e g erm in atio n o f b a s id io s p o re s , has been d escrib ed above. O idia were seen b u t r a r e ly in o ld e r monocaryon m ycelia and n ev er on d icary o n m ycelia* E. n u c le a ta . S ick le-sh ap ed o id ia m easuring 5* 3 -ej± along th e chord from t i p to t i p , about i* 5 -7 ,5 jx in a c tu a l len g th and 1 .4 |a i n th e th ic k - 16 n ess were found i n b oth young and o ld monospore c u ltu re s * The -were produced i n sm all c l u s t e r s a t th e t i p s o f branches ( p i , V I, 4 ) , c u ltu r e s produced a g r e a te r number th an o th e r s . Some The monospore c u ltu r e s ■which changed from a low growing type o f mycelium to a flu ffy - type a p p a re n tly a lso l o s t th e a b i l i t y to produee o id ia* O id ia were also abundant i n d icary o n c u ltu re s , b u t when examined m ic ro s c o p ic a lly none were found on hyphae which bore clamp connections* I t i s tho u g h t t h a t th e o i d i a were borne o n ly on monocaryon hyphae. E» s a c c h a rin a . The fo rm atio n o f s tr a i g h t rod-shaped o id ia on germ tu b e s o f g erm inating b a sid io sp o re s has been d esc rib e d above. When th e mono spore c u ltu re s were about th r e e weeks o ld most o f them showed th e p resen ce o f w h itis h sh in y opaque masses ( P i. V, 2 ) . M icroscopic exam ination rev ea led th e p resence o f very g re a t numbers o f s tr a ig h t rod-sh ap ed o id ia m easuring 7 -l^ u x 2 - 3 St a i n i ng w ith g en tian v io le t showed t h a t th e y were u n in u c le a te ( p i . I I , 8 ) , In th e s e c u ltu re s some o f th e o id ia were borne i n lo o se c lu s te r s a t th e t i p s o f hyphae ( p i . I I , 9) w h ile o th e rs were a p p a re n tly c u t o f f s in g ly . D icaryon c u ltu re s a ls o produced abundant o id ia which were o f th e same shape b u t more uniform in s iz e , m easuring 1Q-I4p.(mostly 10-1: ji) % 2-3 jx * These o id ia were observed to be borne d i r e c t l y on hyphae w ith clamp co nnections (P I. I I , 6) and a p p a re n tly were c u t o f f sin g ly * S ta in in g w ith g e n tia n v i o l e t re v e a le d t h a t th e s e o i d i a were b in u c le a te (pi* I I , 7 ), The p ro d u c tio n o f b in u c le a te o id ia on d icary o n m ycelia has been d e sc rib e d by Brodie ( 5 ) f o r C o lly b ia v e lu tip e s , by Vandendries (3 6 , 37, 38) f o r P h o lio ta a u r i v e l l a , T ra a e te s c ln n a b a rin a and P le u ro tu s p i n s i t i s , and by Dangeard (11) f o r Dacryomyces d e liq u e sc e n s. In th e l a s t sp ecie s th e b in u c le a te o id ia were borne on young f r u i t bodies b u t th e y each 17 d ivid ed t o form two u n in u c le a te o id ia b e fo re g e m in a tin g . In C alooera cornea and D&cryomyces m inor o id ia were produced in both m u ltisp o re and monospore c u l t u r e s . They were m ostly ovoid o r e l l i p t i c a l and measured 3-5 x 1 -3 p * They were borne s in g ly o r in sm all c lu s te r s ( P I . I l l , 4 ,1 0 ). on v e ry sm all p e g - lik e , sid e branches o f th e hyphae^ The n u c le a r number o f th e o id ia-w as n o t determ ined. M artens and V andendries (20) give a review o f th e p resen ce o f o id ia i n s e v e ra l B asidiom ycotes. G erm ination o f O idia A few o i d i a o f A. au ricu la-ju d & e showed g erm ination in th e c u ltu re in w hich th e y were produced and a lso i n a 2% d ex tro se s o lu tio n . Germ­ in a tio n was by means o f a f in e germ tu b e about 4-6 p long and 1p th ic k ( p i . I , 5 ). They f a i le d t o g e m in a te in w a te r and on a g a r. G erm ination o f o id ia o f E. g lan d u lo sa ( P I, I , 15) was v ery s im ila r to t h a t o f A. a u r ic u la -ju d a e . In E. nucle& ta s l i g h t germ ination o f o id ia was seen in th e c u ltu r e whore th e y vie re produced, in w ater and on m oist a g r r . G erm ination was by means o f a s in g le f in e germ tube 10p long and I p th ic k ( p i , I I , 1 2 ). In th e se th r e e s p e c ie s only th e f i r s t sta g e s in germ in atio n were observed and no mycelium was produced. The o id ia o f E. r e c i s a germ inated r e a d ily on ag ar (P I. V, 4) and produced e i t h e r secondary o id ia o r branching normal monocaryon mycelium. The u n in u c le a te o id ia o f B. sac char in a which were borno on monocaryon m y celia germ inated r e a d ily on ag ar to produce normal monocaryon m ycelia. The b in u c le a te o i d i a , borne on d icaryon m ycelia, a ls o g e m in a te d r e a d ily on ag ar and produced normal dicaryon m ycelia xvith clamp co n n ectio n s. The clumps were o fte n formed on th e g em tube n ear th e oidiura ( P i, I I , 11). B in u cle ate o id ia were p ick ed o u t end 37 s in g le - o id ia l c u ltu re s were o b ta in e d . When examined a l l o f them showed th e presen ce o f clamp cormcc- 18 tio n s . T his in d ic a te s t h a t th e two n u c le i p re s e n t in each oidium re p re s e n te d two com patible sex u al p h a se s. ( P I . I l l , 3) O id ia o f C alo cera cornea ^and Dacryonyccs m inor a ls o germ inated r e a d i l y on a g a r to produce normal mycelia* P ro d u c tio n o f F r u it Bodies in C u ltu re A. a u r ic u la -ju d a e » D icaryon c u ltu re s were grown on v a rio u s m edia and u n d er v a rio u s c o n d itio n s o f tem p eratu re and l i g h t , b u t mature f e r t i l e f r u i t b o d ies were produced i n only one c u ltu r e o f c o l le c tio n I I I and t r a n s f e r s from i t . This c u ltu re was o r ig i n a l l y s ta r te d f o r a monospore c u l t u r e , b u t soon a fte rw a rd m icroscopic exam ination o f mycelium re v e a le d th e p re sen c e o f clomp c o n n ectio n s. I t i s assumed t h a t two sp o re s b elo n g in g t o o p p o site sex u al p h s e s te d been picked o u t by m istak e . About s ix months a f t e r th e c u ltu re was s t a r t e d , a few s m a ll, rounded, l i g h t brown, g e la tin o u s f r u i t bodies appeared in a onem onth-old t r a n s f e r . Idiom examined, many young b a s id ia we re observed, some o f which bore f o u r e p ib a s id ia (F I. I , 8 ) . The b a s id ia were t y p i c a l i n appearance but some were s li g h tly curved* They were p ro ­ duced in a d e f i n it e l..y e r b u t d id not form so compact a hymenium as i n a normal f r u i t body* A com paratively sm all amount o f g e la tin o u s m a te r ia l was p r e s e n t, and th e f r u i t bodies were le s s tough th a n th o se growing under n a tu r a l c o n d itio n s . T ra n sfe rs from one o f th e s e f r u i t bodies continued to produce b a s id ia w h ile t r a n s f e r s from th e mycelium began to f r u i t a f t e r t h r e e to fo u r weeks on a g a r. In a f l a s k o f p o ta to -d e x tro se -p o p to n e ag ar the f r u i t bodies f i r s t appeared as sm a ll, s e p a ra te , r&isod bodies which I s t e r co alesced end formed a convoluted g e la tin o u s body about 1 cm. in d iam eter ( p i . IV, 4 ) . The f r u i t i n g stru ctu re; then continued growth 19 and a f t e r some tim e i t had extended t o th e s id e o f a 5 cm* f l a s k (FI* W , 5 ). The l a t e r growth was somewhat r a is e d b u t was n o t so convoluted as th e e a r l i e r growth* The b a sid io sp o re s were normal in s iz e and shape. Both monospore and insult isp o re c u ltu r e s were o b t ined from th o se s p o re s, b u t no f r u i t bo d ies were formed in any o f th e muI t ispo re c u ltu re s nor i n th e d ic ary o n c u ltu r e s produced by p a ir in g two com patible monospore c u ltu r e s . T ra n sfe rs from a f r u i t i n g c u ltu re were made to s t e r i l i z e d s t i c k s o f basswood, h ic k o ry , oak, ash and maple* About fo u r months l a t e r a c u ltu r e on basswood produced sm all b u t ty p i c a l l y shaped f r u i t bo d ies (FI* IV, 6)* About a month l a t e r s im ila r f r a i t bodies were formed on two d i f f e r e n t s tic k s o f hickory* ifo sp o res o r b a s id ia were found on any o f th e f r u i t bodies on wood* E. gXandulosa* Bo sig n s o f f r u i t body p ro d u c tio n were seen in any o f th e d icary o n c u ltu re s* A few dark gray g e la tin o u s b o d ies 1-2 mm* in d iam eter appeared i n monospore c u ltu re number 5* examined and many young b a s id ia were seen* They were Some o f th e s e were s e p ta te , m ostly by a s in g le o b liq u e septum b u t a very few showed two o b liq u e s e p ta form ing a f o u r - c e lle d b&sidium* i n shape and measured 14-20 x 7 -9 ^ * The b a s id ia wore m ostly a ty p ic a l A few o f th e tw o -cel le d b a s id ia showed tvao s h o rt e p ib a s ld ia (Pi* 1 , 16) but no b a s id ia produced fo u r e p ib a sid ia * A few t y p i c a l b a sid io sp o re s m easuring 9-12 x 4 -5 * 5 ^ were seen ( p i . I , 10)* Ho c u ltu r e s wore o b tain ed from th e s e spores* The mycelium o f c u ltu r e 5 was examined c r e f u ll y f o r th e presence o f clamp co n n ectio n s b u t none were found* observed were u n in u c le a te . S u clear s ta in s were used b u t a l l c e l l s Sorae t r a n s f e r s from t h i s c u ltu re produced & few s m a ll, g ra y , g e la tin o u s bodies but no b a s id ia o f them* e re found in any Monospore c u ltu r e 85 a ls o formed a few sm all g e la tin o u s bodiea 20 i n which a g ro a t number o f young b a s id ia were p r e s e n t;but no spores were soon* These appear t o be ca ses where f e r t i l e b u t q u ite a ty p ic a l and weak f r u i t bo d ies were produced by monocaryon m ycelia. E. r e c i s a . A few t y p i c a l f o u r - c e lle d b a s id ia w ith long e p ib a s id ia (1 1 . IX, 4) were observed i n one m u ltisp o re c u ltu r e on p o ta to -d e x tro s e peptone agar* Bo b a s id io s p o re s were seen* The f r u i t i n g body in t h i s case was m erely a brown soraowh t g e la tin o u s la y e r on th e ag ar and was p a r t i a l l y overgrown by d u ll brown hyphae* I n many o f th e d icary o n c u ltu r e s and a ls o i n s e v e ra l o f th e xaonoc&ryon c u l t u r e s , th e r e appeared rounded, dark brown, g e la tin o u s b o d ies ( P I . V, 5) c lo s e ly resem bling normal young f r u i t b o d ie s . They ranged from 2-9 mm* i n d ia m eter, and were u s u a lly la r g e r in th e d icary o n c u ltu r e s . ?«hen examined th e y were found to b© r a th e r tough in t e x t u r e , much l i k e t h a t o f a normal f r u i t body. Among th e s le n d e r hyph. e p re s e n t, th e r e were a number o f round o r oblong c e l l s which r e e nbled young b a s id ia , b u t none were ev er seen to be s e p ta te . I t was th o u g h t th a t th e s e g e la tin o u s bodies re p re se n te d a b o rtiv e f r u i t bodies which remained s t e r i l e , p robably b ee ,u se o f u n fav o rab le c o n d itio n s . Such c u ltu re s were su b jec te d to v a rio u s environm ental co n d itio n s b u t no reco g n izab le b a s id ia were observed. E. n u c le a ta . A n ru ltisp o re c u ltu re showed s e v e ra l sm all f r u i t bodies a f t e r about fo u r months on p ru n e-co rn meal a g a r. c u s h io n -lik e bo d ies about 1-3 m * in d iam eter. showed a l r g e number o f t y p ic a l b a s id io s p o re s . These were c l e a r , Exam ination o f th e se Many young club-shaped b a s id ia and a sm a lle r number o f m ature f o u r - c e lle d b a s id ia w ith fo u r lo n g e p ib a s id ia were p r e s o r t (P I. I I , 14, 15). In most cases th e e p ib a s id ia were lo n g e r th a n th o se p re s e n t in w ild f r u i t bodies and freq u en tly th e fou r on a baaidiura w r o unoTued In length* The lon ger ep ib o o id ia were o fte n ones© o r tw ice soptat©* T ransfer* o f mycelium from t h e f r u i t i n g c u ltu re s produced f r u i t b o d ies w ith m ature b&eidiu w ith in 14 days on p o ta to -d o x t ro oo-£© ptone agar* F ru it b o d ie s war© a ls o produced on p o ta to -d e x tro s c ou t peptone) and on n a l t agar* g- r (w ith ­ Ifioaosporo c u ltu r e s were s t a r t e r frora th e sp o re s h om e in c u l t u r e , and o u t o f th o IS com patible p a ir in g s between tho ao m sp o ro c u ltu r e s seven produced f r u i t bodies* Bo s ig n o f any c e n t r a l c r y s t a l s tru c tu re * c h a r a c t e r is ti c o f th e w ild f r u i t b o d ie s, was soon in any o f th o f r u i t bodice in c u ltu re * Bo f r u i t to d ie s wore produced in any c u ltu r e s o f E« sa coharlna* In each o f two m x ltisp o ro c u ltu re s o f C alocora oornoa a sm all y e llo w , rounded g e la tin o u s body m s formed* Theso were v ery a ty p ic a l f o r f r u i t b o d ies b a t b oth produced forked b a c id ia (pi* I I I , 5) and spores* Growth R ates o f C u ltu res D u p licate m alt a g a r plate© o f noiltieporo ctilturo© o f th e d if f e r e n t sp e c ie s were sad© and incu b ated a t room tostforator© ("ranging from 21£4° C}* A second s o t o f d u p lic a te c u ltu re s wcro p laced i n th o ic e box where th e tem p e ra tu re ranged fro n 10° t o 15° C* The r e s u l t s o f t h i s cap& rlaeab a re shown in Tablo 2* T able 2 R adiol Growth Ratos o f Dicaryon g y o o lla R adiol growth In 15 days R adial gro^’t h r a t e j o r day 0 o 51°~24°C 10 - I S C no* Of c o ll e c t i o n Lrooios A* a u r ic u la I judue 20 ran.* 5 rasa# 1*5 m * 0*5 rata* l» gliiiidulosa I 44 ram* 5 m* 2*0 rr» 0*5 ran* E* r e c i s a I 22 ran* s li g h t 1*5 ::rs* E* m o lo a ta I s lig h t 2l°~24° C 10°-1S°C Bo g ro a t d iff e r e n c e was found i n g en e ra l between growth r a te s o f monocaryon and d ic a r yon m ycelia o f any s p e c ie s , b u t in most cases th e growth o f die: ryon m y celia was somewhat f a s t e r th a n t h a t o f monoc a r yon n y ce lia * The growth r a t e s o f some o f th e monocaryon c u ltu re s a re g iv en in Table 3* D u p licate p la te s wore grown a t room tem p eratu re on m alt e x tr a c t agar* Thor© w ere, however, co n sid e ra b le d iffe re n c e betw een growth r a t e s o f d i f f e r e n t monocaryon c u ltu re s* This i s n o tic e ­ a b le when a com parison i s mad© between c u ltu re s 7 and 9 o f A. a u r ie u la ju d a e , c u ltu r e s 50 and 23 o f B* g lan d u lo sa and c u ltu r e s 256 and 263 (PI* V, 3D, B) o f E* aacoharina* Union o f Hyphae and P ro d u ctio n o f Clamp C onnections I n B* saccharins, one f u s io n between young hyphae from germ inating D oSidiospores (PI* I I , 10) was seen SO hours a f t e r th e sp o res had f a l l e n on a g a r, o r about 20 hours a f t e r germ ination* In B* r o c is a one such u n io n between young hyphae was seen 48 hours a f t e r th e sporos f e l l on a g a r, and i n B. g landulosa ( F I . I , 17) one was observed fo u r days a f t e r th e d isc h a rg e o f th e spores* In th e s e th re e sp e c ie s clamp co n n ectio n s were formed w ith in a few hours a f t e r two com patible mono­ caryon m ycelia net* These o b se rv a tio n s were made on young m ycelia from g erm in atin g spores* The p ro d u c tio n o f clamp connections in A* a u ric u la -ju d a e seems to be delayed somewhat, f o r in c e r ta i n m u ltisp o re c u ltu re s clamps were f i r s t observed 11 to 18 days a f t e r th e spores had germinated* I t was a ls o observed t h a t i n a p a ir in g between two com patible monocaryon m ycelia clamp co n n ectio n s were n o t found u n t i l th e two m ycelia had in te rm in g le d q u ite w ell* Table 3 R ad ial Growth R ates o f Monocaryon C u ltu res no. o f c u ltu re S p ecies R d i a l Growth r a t e a f t e r 7 days 9 mm. R ad ial Growth Rate p e r day 1 .3 ran, A. au ricu la-ju d ae 4 w 7 11 n 1 .6 n n 9 6 « 0 .9 n tt 101 10 tt 1 .4 it 23 16 tt 2 .3 tt « 28 12 tt 1 .7 n tt 30 13 R 1 .9 tt « 50 8.,5 » 1 .2 tt n 53 13 « 1 .9 n tt 55 14 n 2 .0 « tt 52 14 « 2 .0 tt II 57 13 n 1 .9 tt E. r e c isa 4 7 » 1 .0 tt 6 7 tt 1 .0 w 256 9 n 1 .3 « n 257 14 « 2 .0 tt « 263 15 « 2.1 tt R 265 10 n 1 .4 II E* glandulosa « E. sae ch u rin a S e x u a lity I t i s s t i l l a q u e s tio n as to what c h a r a c te r is tic s a re b e s t to u se as a sta n d a rd in d eterm ining th e tr u e dicaryon mycelium i n th e B aa id io ay c eto s• Bose (s) quotes Oort (27) as follow s* "As a c r i t e r i o n f o r d ip lo id m ycelia th e o ccurrence o f clamp connections can on ly be of a re s tric te d use. Usod in connection w ith th e c h a r a c te r is tic ?A d ip lo id h a b it and th e d ip lo id f r u i t bo d ies i t rem ains, o f c o u rs e , a v a lu a b le c r ite r io n " * Because th e sp e c ie s s tu d ie d in th e p re s e n t -work showed no o th e r d is tin g u is h a b le d iffe re n c e between d icary o n and monocaryon m y c elia, and because th e p ro d u c tio n o f f r u i t bodies was r a r e , th e p rese n c e o f clomp co n n ectio n s alo n e was ta k e n as an in d ic a tio n of t r u e d ica ry o n mycelium. By s ta in in g m y celia, b o th th o se w ith and w ithout clamp c o n n e c tio n s, e a r ly i n th e course o f t h i s in v e s tig a tio n th e c o r r e c t­ ness o f t h i s assum ption was determ ined f o r th e sp e c ie s in q u e s tio n . D ried f r u i t b o d ies were re v iv e d , or f r e s h ones wore used when a v a ila b le . Spores were c o lle c te d on ag ar p la te s and s in g le germ inated sp o res pick ed o u t i n th e manner d escrib ed above. A fte r th e c u ltu r e s had grown s u f f i c i e n t l y in t o s t tu b e s th e y were examined m ic ro s c o p ic a lly f o r th e p resen ce o f clamp connections* Tho monospore c u ltu r e s were numbered c o n s e c u tiv e ly , g iv in g th o se from each f r u i t body a d i f f e r e n t s e t o f /T ab ic numbers* The number o f each f r u i t body corresponds to th e number o f th e c o lle c tio n o f th a t sp e c ie s and i s in d ic a te d by a Homan num eral. R arely a c u ltu r e became contam inated and was discarded* T h is f a c t accounts f o r some o f th e m issing c u ltu re s in a s e t . The monospore c u ltu r e s from each f r u i t body were th e n p a ire d in a l l p o s s ib le com binations. T his was accom plished by p la c in g b i t s of m ycelia o f two c u ltu re s a t th e s id e s o f an ? g ar s la n t in a 15 mm. t e s t tube* The p a ir in g s were th e n p laced under b e l l - j a r s i n a m oist a t ­ mosphere and l e f t a t to o n tem perature* A fte r about two weeks th e two m ycelia had in te rm in g le d th o ro u g h ly and a t t h i s tim e a sample was removed from th e lin e o f c o n ta c t o f th e two m y celia. A l i t t l e d ilu te aqueous s a f r a n in was u s u a lly added to make th e hyphae show more c le a r ly and th e samples were th e n examined f o r th e presen ce o f clamp o o n m ctio n s 25 by us© o f th e o i l immersion lens# The p resen ce o f clamp connections in such, a p a ir in g v/as ta k e n to in d ic a te t h a t th e two monocaryon m ycelia -wore co m p atib le, o r t h a t th e y belonged to d i f f e r e n t sex u al phases# Since o n ly two se x u a l phases were found i n each f r u i t body, th e absence o f clamp connections was ta k e n , i n g e n e ra l, to mean t h a t th e two m ycelia belonged to th e same s e x u a l phase* However, o th e r f a c to r s may be p r e s e n t which in flu e n c e th e c o m p a tib ility o f two monocaryon m ycelia# In s e v e ra l cases s h o rt b ranches wer© seen a t s e p ta and th e s e may have been a b o rtiv e clamp c o n n e c tio n s, b u t u n le ss com plete and d e f in ite clamps wore seen th e r e s u l t s w ere co n sid e re d as n e g a tiv e . Then i r r e g u la r r e s u l t s wore ob­ ta in e d th e p a ir in g s were re p e a te d , but when the r e s u l t s wore r e g u la r o n ly one m ating was made between any two monospore c u ltu re s# In a l l ta b le s th e + s ig n in d ic a te s th e presence o f clamp c o n n e c tio n s, o r t h a t th e two c u ltu r e s were co m p atib le, rind th o - s ig n in d ic a te s th e absence o f clamp connections* So s p e c ia l work was done w ith th e phenomenon o f a v e rsio n b a t m ention i s m de where i t was d e f i n it e l y observed# The p rese n c e o f a v e rsio n i s in d ic a te d i n th e table© by a d o t (•)# A« auricula-*judae P a irin g s o f Monospore C u ltu res from th e Same F r u it Body Twenty-two mono spore c u ltu re s ‘.ere o b tain ed from f r u i t body I , These were p a ire d i n a l l p o s s ib le com binations and th e r e s u l t s a re g iven in T able 4# I t i s observer t h a t th e monospore c u ltu re s f e l l in to two groups, based upon th e p ro d u c tio n of clomp connections in c e r ta in com binations* phases"* a# Throughout t h i s pap er th e se groups a re c a lle d "sex u a l These two sex u al phases were designate- g e n e tic a lly as A. and VJhen members o f th e same sex u al phase were p a ire d none produced 26 Table 4 A» au ricu la-ju d as* R esu lts o f p a irin g 22 jjioiooispore eultm res o f f r a it body I . r— / .4 5 6 8 9 26 18 20 23 21 3 7 l — — — — — — — 4 4 -h + + 1 4 — +■ ■+ 4 5 4 4 i- 4- — 7 r 10 12 13 15 — 4-. — 4 4 4 4 4 4 4 4 4 4 — i ■16 17 19 22 24 -4 4 4 — 4 4 4 4 4 4 4 4 4 4 4 18 — — — — 4 4 t 4 4 4 4 20 4 H- + 4 -f 4 + — 4- 4 4 4 21 — — — 4 — — 4 4 4 4 4- 4 4 4 23 — 4 4 4 4 4 25 4 4 4 4 4 4 + 4 ■+ -h -1- 4 -h ■4" +- 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 5 6 —— 8 9 10 12 13 15 — — - - - 16 — — — — 17 — — — 19 22 — — I 24 — 27 clamp c o n n e c tio n s. When m m bors o f A -were p a ire d w ith a , clamp c o n n e ctio n s were found i n a l l b u t s i x o f th e 112 p a ir in g s . n in e te e n monospore c u ltu r e s were o b ta in e d from f r u i t body I I I and w ere p a ir e d i n a l l com binations* 5* The r e s u l t s a re given in Table The f r u i t b o d ies produced by d icary o n mycelium grown from sp o res o f f r u i t body I I I were d e sig n a te d a s S e v e ra l mono spore c u ltu re s were o b ta in e d from th e a and te n were p a ire d in a l l com binations* r e s u l t s a r e shown in T able 6* The A version was p r e s e n t in some p a ir in g s b u t l i t t l e c o r r e l a t i o n i s seen between i t s p resen ce end th e absence o f clamp connections* E leven mono sp o re c u ltu r e s o f f r u i t body 17 wore p a ire d i n a l l com binations and th e r e s u l t s a re shown in T able 7* I t was observed t h a t s e v e ra l o f th e p a ir in g s showed r a t h e r d i s t i n c t aversion* In th e s e p a ir in g s thro® r a t h e r d i s t i n c t ty p e s o f p a ir in g s were observed* (1 ) 22 p a ir in g s showed a space o f a v e rs io n between th e two m ycelia (PI* 17, 3A, B ). (2 ) two showed a v e rsio n i n w hich th e gap m s f i l l e d w ith a growth o f d icary o n mycelium (p i* 17, 30)* (3) 31 p a ir in g s showed even in te rm in g lin g o f th e two m ycelia (PI* 37, 8C)* Clamp c o n n ectio n s wore found i n o n ly one c u ltu r e o f ( 1 ) , in b o th c u ltu r e s o f (2 ) and i n a l l b u t two c u ltu r e s o f (3)* The r e s u l t s in d ic a te t h a t i n th e s e p a ir in g s th e re m e a r a th e r c lo se c o r r e la tio n between th e p resen ce o f a v e rs io n and In c o m p a tib ility o f tho c u ltu re s* T able 8 shows th e r e s u l t s o b ta in e d when 11 Bonosporo c u ltu r e s o f f r u i t body Va w ere p a ire d in a l l com binations* was seen o n ly i n some o f ih© incom patible p a irin g s * S lig h t a v e rs io n The r e s u l t s o f p a i r in g seven monospore c u ltu re s o f Vb a re given in T able 9* T able 10 Table 5 A* aairic u la-lu d aa . B a sa lts o f p a irin g 19 monospora cu ltu r e s o f f r u i t bodfer III* h _ _____________________ 5i f' ---- \ 101 103 105 107 109 110 111 L13 L14 L17 118 104 112 115 116 120 121 122125 L01 4 4 4 4 4 4 4 4 4 -h 4 4 4 4 4 4 L03 — — 4 4 4 4 4 4 4 4 107 4 4 4 4 4 4 4 4 10 S 4 4 4 4 4 4 4 4 J no — — — — — 4 4 4 4 4 4 4' 4 111 — — — — 4 4 4 4 4 4 113 — — — 4 4 4 4 — — 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 L05 114 117 118 > 104 4 4 4 4 4 112 115 116 — - — - 120 — — — 121 — — 122 — 125 29 i o^ tO Table 6 At au rieu la-.iu d ae. R esu lts o f p a irln g ten monospore eoTCures o f f r u i t body I I I F j• s aoro or lo o s i W i i o i present* / S. / /---301 302 303 306 307 309 304 508 310 4 4. 4 4 SOI • 302 . — — 303 306 — . ■+■ 4 4 4 4 — 4 4 4 4- 4 4 4 • — 307 4. 4. — 309 4 4 4 4. 4 4. 304 — — 305 — — 308 — 310 At au riclu a-4u d ae. R esu lts o f p a irin g 11 monospo^e o u itu res o f f r u it bodjr IF* •* wore or lo o s aversion p resen t. / ✓— - _ ,S' S\ ia 156 159 160 4 4 4 4 • — t 4 4. 4 4 4 ■ • • -4 4 4 4 4 % 4 4. 4 4 4 4 4 4. 4 4 + 4 4 4 4 • • • • • • La 152 155 157 158 161 153 4 . . • L51 L52 L55 L57 158 ( L61 153 154 <156 159 160 - * t • • 9 so ta b le 6 A* a a ricu la -iu d a g . R esu lts o f p a irin g 11 aosaoapore cu ltu r e s o f f r u it bo.____________ 1 201 7 16 3 A. 4 6 8 9 — 205 — 4 — + — — — — — — — 4 204 — — — 4 206 — — 4 207 §08 203 23.1 4 — — — + — — 4 4 — — 4 — — — — — — — — — — fable 16 Table 17 A* aurIcula-judae * Results o f pairings between fru it bodies 1 and 7b, A -^ _ _3 7 . 4 a A* aur i eu1a~iudae« Resiftt s of pairings m h r i j p a fru it bodies I and 71* T A , 3 6 a I 7 4 6 401 — — — — 251 — — — — — — — 253 — — — — 403 — — — — 252 — — — 405 — — — — 256 — — 402 4 — — — Table 18 Table 19 km a u rleu la -jo d a e. B a sa lts o f p a ir in g s between f r u it bodies I sod H I . A * au ricu la-.ju d ae* B e su lts o f p a irin g s between f r u i t bodies I end T ill- I A a a i 3 7 I A a l 4 6 7 4 — — — — 501 — 4 4 6 4 552 — — — — 502 + 4 4 4 554 — — — — 503 -5 4 4 4 356 — — — — 504 — 4 4 4 505 — 4 4 4 551 3 Table 21 Table 20 A* e a rio u le-lu d a e• B a sa lts o f p a irin g s aetseen f r u it bodies I I I and IT* A* aurloul&-.1u -A 15L 352 353 354: X. 151 152 153 154 201 4 4 4 4 251 4 4 4 205 4 4 4 4 253 4 4 4 4 4 203 4 4 4 4 252 4 4 4 4 204 4 4 4 4 256 4 4 + 4 Table 28 Table 29 km atarim la* Judas* R esu lts o f p a irin g s between f r u it bodies IT and T il* A* auricula».1udae« R esu lts o f p a irin g s between f r u it bodies IT and T i l l . A2 , _.> —j 151 152 153 154 . IT a ---------- a2 — L 151 152 L53 154 551 4 4 4 4 501 — 552 4 4 4 4 502 — 554 4 4 4 4 503 556 4 4 4 4 — — — — — — — 4 — 504 — — — — 60S — — — — — Table SO Table 31 A» au r i c u l a - idae. R esu lts o f p a irin g s f r u it bodies 7 e sand 7b* A* a u r ic u la -ju d a e * R e su lts o f p a ir in g s between f r u i t bodies 7a and 71. Va A3 Va a3 201 305 203 401 + + ■f *4 7b 10 2 + + + 403 - f 405 + + + 204 + + + ■+ + A,- A 5 VI -f- 5 ‘ 201 >05 203 204 + + 251 + + -f- + 252 + + + -t + + + 252 256 + + Table 38 Table 32 A* aaarlo u la-ju d ae. R e s u lts o f p a ir in g s oefawen f r u i t b o d ies Va and 711 • A* a u ric u la -J u d a o « R e s u l t ^ p a i r i n g s between f r u i t bodies 7a and T i l l . Va a3 201 205 203 204 ^3 a3 i. L 201 305 303 204 351 -h + + + 501 352 -h + + "h 502 354 + -5 + + 503 — ” t~ -1- — 356 + + -f + 504 — — — + 505 — — — + — — + — — — , Table 35 Table 34 A» eur leular-.ludae « R esu lts o f p a irin g s between f r u it b od ies f l l and V I. b od ies V II and V III. A 6 V II a 6 > 3a 354 352 35 6 251 253 252 256 Aq V II 351 4 4 4 4 501 354 4 502 352 4 4 4 4 4 4 4 4 4 a6^ 356 4 V III 4 H" 4 4 4 4 4 4 4 4 4 4 4 505 + 4 4 4 t 503 4 504 4 4 Table 36 A* «arierimenfcB showed t h a t th e low numbers o f clamp connections were l i m i t ­ ed t o p a ir in g s betw een c e r t a i n d e f i n it e sex u al p h a se s, namely x Ag and ap x To determ ine th e q u a n tita tiv e d iffe re n c e s i n th e numbers o f clump c o n n e c tio n s, p a ir in g s were made between a l l com binations o f se x u a l p h a se s, u sin g th r e e mono spore c u ltu re s from each* were th e fo llo w in g s These c u ltu r e s 2 1 , 23, 24, 27, 28 and 30 from Ila * and 50, 53, 55, 51, 52 end 57 from l i b . B its o f th e two monoearyon m ycelia were p la c e d on m alt a g a r p la te s a t & d is ta n c e o f about 1 cm* a p a r t and in cu b ated a t room te a p e ra tu re * In o rd e r to male© th e c o n d itio n s u n i­ form , i t was n ecessary to know th e ex act tim e a t which th e two m yoelia cante i n c o a t ct* T his was accom plished by p la c in g s t e r i l i z e d h alv es o f cover g la s s e s in to tho ag ar in an u p rig h t p o s itio n between th e two rayceliu o f a p a irin g * The m ycelia were allow ed to grow up a g a in s t th e cover g la s s which was th e n removed* Care was ta k e n n o t to d is tu r b th e xnycolia any more th a n necessary* At th e end o f 48 hours a f t e r th e removal o f th e in te rv e n in g cover g la s s e s , th e p a ir in g s wore examined f o r ©lamp connections* The ag ar p l a t e c o n ta in in g a p a ir in g was p laced on th e s ta g e o f th e m icroscope 49 and examined w ith th e o i l in n er8io n le n s a t th e l i n e o f c o n ta c t between th e two raonooi’. ryon m y celia . A drop o f aqueous s a f r a n in v &s u s u a lly added t o nako th e hyphae more d i s t i n c t* Areas in which th e hyphae co u ld be se en c l e a r l y and in which th e number o f hyphae was about th o average were chosen and counts were made o f th e olaaop co n n ectio n s seer; i n th e e n t i r e f ie ld * b o th on th e su rfa c e o f th e ag ar and be lor/ th e s u rfa c e as f a r as th e fo c u s o f th e Ions would reach* Pour co u n ts were ra de from one s e t o f p a ir in g s and s ix were made from a l a t e r se t* The r e s u l t s a re shown in Table 4U* The numbers in th e ta b le r e p re s e n t th e average o f th e t e n c o u n ts. In T able 47. which i s a summary o f Table 46, th e a v e ra p s f o r each com bination o f sexual phases a re given* The numbers i n th e ta b l e re p re s e n t th e average number o f clamp connections p e r m icro sco p ic f i e l d f o r n in e p a ir in g s . C e rta in d i s t i n c t d iff e r e n c e s between th e se x u a l phases a re ev id en t* The com binations Aq x a q , A*> x ag a3S^ Ag x &q showed approxim atgly th e seme number o f clomp c o n n e c tio n s, av erag in g IS*6 , 15*0 art! 15.0 r e ­ s p e c tiv e ly * The com bination/, Aq x Ag and a 3 x ag showed a d ecid ed ly s m a lle r number o f d a m p s, av erag in g 6*4 and 5*5 re sp e c tiv e ly * The number found i n p a ir in g s o f Aq x &g was in te rm e d ia te , averaging 1 0 .3 , S t a t i s t i c a l d a ta comparing th e d i f f e r e n t com binations o f sex u al phases a re g iv en in T able 48* S t a t i s t i c a l l y / s ig n if ic a n t d iffe re n c e s were found between th e fo llo w in g p a ir s o f com binations j Aq x Ag and Aq x a g , Aq x Ag and Aq x a q , Ai x Ag and aq x Ag, Aq X Ag and Ag x ag , aq x Ag and aq x ag , Aq x ag and aq x a g , and Aq x a s and Aq x aq* Ifo s ig n i f i c a n t d iff e r e n c e s were found between Aq x Ag and aq x e g , A2 x aq and Ag x ag , and Aq x aq and Ag x eg* 50 Table 46 S . glan d u los*. Average number o f clamp connections (fo r te n count*) per m icroscopic f ie ld in p airin g* between f r u it bodies I la and l i b . A1 al a2 21 23 24 27 28 30 51 52 57 50 5.5 5.1 7.5 Ilw8 L3.1 13.1 13.9 14.3 12.4 a 2 53 7.3 6.1 7.1 14.6 16.4 11.4 14.7 13.5 14.6 55 4.8 7.9 6.7 11,3 12.6 13.1 12.7 12.0 8.8 51 11.6 12JB 9.2 5.2 7.5 5.0 a 2 52 10.010.5 8.8 6.5 5.0 5.0 57 9.8 10,5 9.6 5.3 4.9 53 27 15.3 17.110.9 al 28 11.913.212.2 30 123 16J912.5 Table 4? E. glandulosa* SuHffltar: o f fab le 46* A1 al a2 A2 6 .4 1 3 .0 1 3 .0 a2 1 0 .3 al 1 3 .6 5 .5 51 T able 48 S t a t i s t i c a l S ig n ific a n c e o f th e D ifferen ces between th e Means o f Numbers o f Clump C onnections i n "various Combinations o f Sercual Phases o f F r u i t Bodies I l a and lib * C o m b in a tio n s o f S e x u a l Phases A c tu a l A ctual d i f ; 6 D ifferen ce 6 A y x A2 and Ap x ag 0 .5 2 5 S. 9 7 .4 3 --1 x * 2 an>^ A1 x a l 0 *7 9 5 7 .2 9 .0 6 A1 x '-2 an£* a l x Ag 0 .6 1 4 6 .6 10*75 A i x Ag and Ag X &g 0 .6 9 8 6 *6 9 .4 6 x Ag a n d x ag 0 .4 4 9 0 .9 2 .0 0 & i x Ag a n d x ag 0 .5 7 6 7 .5 1 3 .0 2 Ap x ag a n d x ag .0.480 4* 3 10* v0 x 0.879 3.3 3.75 x ag and ap x Ag and a 2 x a<> 0.785 Ap x &p a n d Ag x ag 0.927 6* 0 0 .6 0.06 S tan d ard E rro r o f D ifference* Attem pts were made to determ ine e x a c tly where th e d iff e r e n c e in th e numbers o f clamp co n n ectio n s lay* The number o f clamps in r e l a t io n t o th e number o f s e p ta was counted on d icary o n m y eelia. I t m s found t h a t th e number o f septa, w ith o u t clamp connections r; ,s very sm all and no d i s t i n c t d iff e r e n c e was noted between any o f th e d icary o n m yeelia observed* I t was a ls o th o u $ t p o s s ib le t h a t a d iffe re n c e m ight e x i s t in th e r a te o f d ip lo id iz a tio n o f one monospore c u ltu re by .n o th er* The nxetliod employed to determ ine t h i s was s im ila r to t h a t used by D u ller ( 3 ) . Sloaospor© c u ltu r e s o f 55, 55, 52 and 5? were grown 52 Table 49 g la n d u lo sa . Presence o f clamp co n n ectio n s a f t e r seven days a t given d is ta n c e s from th e f i r s t p o in t o f d i p l o i d i s a t i o n and growth r a t e s o f d i c a r yon myoelie* C u ltu r e s P re se n c e o f clam ps a t g iv e n d is ta n c e from f i r s t p o in t o f d ip lo i d i z a t io n d ip lo id iz e d by 1 cm. 2 cm. 3 cm. 4 cm. 53 by 28 + H- — — 28 by 53 4- — — — 53 by 30 4 _____ - _____ 30 by 53 + — — — 55 by 28 — — — + — 4- — 55 by 30 -J-h 4- — — 30 by 55 4- — — — 52 by 28 + — — — 28 by 52 i- — — — 52 by 30 + — — — 30 by 52 4 — — — 57 by 28 + — — — 28 by 57 + 4- — 57 b y -30 4- — — — 30 by 57 i- — — — 28 by 55 R a d ia l growth o f d icaryon m ycelium in sev en days 12 mm. 12 mm. 15 mm. 14 mm. 13 mm. 14 mm. 12 mm. 16 mm. 53 on a g a r f o r 13 days and a t t h i s t i n e one s e t was d ip lo id iz e d by p la c in g b i t s o f c u ltu r e 28 a t t h e i r edges* iz o d by c u ltu r e SO* A second s e t o f c u ltu re s v/as d ip lo id - C o n v ersely, b i t s o f c u ltu r e s 53, 55, 52 and 57 v/ere u sed t o d ip lo id iz e 13-day c u ltu r e s o f 28 and So* Seven days l a t o r sn c .ll sam ples o f mycelium were ta k e n a t d is ta n c e s o f 1 , 2 , 3 and 4 cm* from th e p o in t o f f i r s t d ip lo id iz a tio n and pl...ced on fro s h a g a r p la te s* A fte r now growth had s t a r t e d th e y were examined f o r clamp connections and th e r e s u l t s a r e reco rd ed i n T able 49* I t I s scon a t once t h a t i n every ca se clump co n n ectio n s were p re s e n t a t a d is ta n c e o f 1 cm* T h is d ie t.n o © , however, i s s h o r t enough t h - t t h e i r p resen ce can bo accounted f o r i n e v e ry c a s e by th e r a d i a l growth o f th e d ic a ry o n rayed ia* grovrbh r a t e a a re a ls o g iven i n T able 49* These I t i s a ls o e\*ident t h a t th e r e w ere o n ly s l i g h t d iff e r e n c e s between th e growth r a t e s o f th e d icary o n m ycelia* In o n ly th r e o o ases -./ere clump connections found a t a d ista n c e . g r e a te r th a n 1 am* Those were 53 d ip lo id iz e d by 28, which showed clamp co n n ec tio n s a t a d is ta n c e o f 2 eta*, 28 d ip lo id iz e d by 55 and 28 by 57, of b o th rtw hich ©hawed clamps a t a d is ta n c e o f 3 cm* from th e f i r s t p o in t o f d ip lo id is a tio n * At p r e s e n t no e x p la n a tio n can be given f o r th e s e d iffe re n c e s * I t was a ls o th o u g h t t h a t th e q u a n tita tiv e d iffe re n c e found i n tho numbers o f clamp co n n ectio n s raight be due to a d iffe re n c e in th e exact tim e o f th e f i r s t u n io n o f th e two m ycelia f t e r th e in te rv e n in g cover g la s s was removed* I t seems t h a t th e r e may be & g r e a te r a t t r a c t i o n betw een c e r t a i n c u ltu r e s ( i f such an a t t r a t i o n e x is ts a t a l l ) and as a r e s u l t th e u n io n o f m ycelia may ta k e p la c e sooner th a n i f a le s s o r a t t r a t i o n i s p re sen t* C onsequently, a g r e a te r number o f clamp connec­ t i o n s would be form ed w ith in th o 40 hours t h a t e lap sed between th e 54 rem oval o f th e c o v e r g la s s and th e co u n tin g o f th e clamps* T h is , how ever, i s p u re ly s u p p o s itio n a l and i s o f fe r e d m erely as a p o s s ib le e x p la n a tio n f o r th e differen ce® found in th e numbers o f clamps d e scrib ed above* A s, y e t , no s u c c e s s fu l a tte m p ts have been made to dem onstrate th e p re se n c e o f such an a t t r a c t i o n between com patible inycelia# E* r e o is a P a ir in g s o f Mono spore C u ltu re s from th e Same F r u it Body Twenty monospore c u ltu r e s o f f r u i t body I were p a ire d i n a l l com binations and th e r e s u l t s a re g iven i n T able SO* Ten monospore c u l t u r e s from f r u i t body I I were p a ir e d , as were te n c u ltu r e s from f r u i t body H I* The r e s u l t s sir© shown i n T ables SI and 52 re sp e c tiv e ly * In a l l f r u i t b o d ies th o r e s u l t s o f p a ir in g s were v ery re g u la r* Each showed th e p rese n ce o f two d i s t i n c t sex u al p h a se s, showing t h a t E* r o c j s a i s t y p i c a l l y b ip o la r* R e s u lts o f P a ir in g Monospore C u ltu re s from D iffe re n t F r u it Bodies Four monospore c u ltu r e s o f f r u i t body I was p a ire d w ith fo u r c u ltu r e s from each o f f r u i t bo d ies I I and I I I , Th© r e s u l t s , which a r e shown i n Table® 53 and 54 r e s p e c tiv e ly , showed th e p resen ce o f clamp co n n ectio n s i n ev e ry p a irin g * I n th e p a ir in g s between c u ltu re s o f I I and I I I (T able 5 5 ), clamp connection® were ag ain found i n every p a irin g * Such couplet© c o m p a tib ility between th e monospor© c u ltu r e s o f d i f f e r e n t f r u i t b o d ies shows t h a t no two o f th e sex u al phases w ere id e n tic a l* As i n A, a u ric u la -ju d a e and in B* g lan d u lesa th e c o m p a tib ility f a c t o r s a re c o n sid e red t o e x i s t as m u ltip le allelom orphs* d e s ig n a tio n s o f th e se x u a l phase® a re given in th e t a b l e s . The g e n e tic s ! 55 Table 50 g» re o ise u R e s u lts o f p a ir in g 20 raonosp o re c u ltu r e s o f f r u i t bo^y !♦ ✓V. 1 2 5 8 — — 1 2 5 8 *---9 10 12 16 18 20 21, 22 24 4 11 15 17 19 23 4 4 4 4 4 4 + 4 4 4 4 4 + 4 4 4 4- 4 25 4 4 -4 4 + 4 4 + 4 9 4 4 4 4 4- 4 10 — — — — — — — 4 12 4 16 4 18 — — — — 4 — — — 4 4 4 4 4 4 4 4 4 4 4 22 — — 4 — 4 4 4 4 24 4 4 4 4 20 21 4 4 4 4 4 4 4 4- + 4 4 4 + -b 4 4 4 4 4 4- 4 4 4 4- 4 + 4 4 + 4 4 4 + + 4 11 15 17 19 23 25 — — — — — — — — — — 56 Table 51 5* re o ia a , R esults o f p a irin g ta n monospore c u ltu re s o f f r u i t b o ^ I I , A1 al /\- ■f 51 52 55 56 62 53 54 58 59 60 — — — — 4 4 4 4 4 51 — — — 4 4 4 4 4 52 55 — — 4 56 62 4 4 4 4 — 4 4 4 4 4 4 4 4- 4 4 53 — — — — 54 — — — 58 — — 59 — 60 Tabl© 52 E» racism* Results of pairing tea momepor© eulturos of fru it body III* A2 a2 / r ' 101 106 109 102 103 104 105 107 108 110 — — 4 4 4 4 4 4 4 101 106 109 — 4 4 4 4 4 4 4 4 4 4 4 4 4 4 102 103 104 — — — — 105 — — — 107 — — 108 — 110 57 Table 55 Table 54 E« re o lg a . R esults o f p a irin g s between f r u i t bodies I and Xl« A * E> r e c l s a * R e s u lts o f p a ir in g s b e tw e e ifT ru it b o d ies I and 22I« a > 1 5 4 11 51 4 4 4 4 1 a2 II 55 4 8-1 53 4 54 4 4 4 4 4- 4 4 -t 4 4 a A ---- v— — III S ir 4 4 4 106 4 4 -4 4- 102 4 4 4 4" 103 4 4 4 4 E. r e o ls a . R e s u lts o f pairing® between f r u i t bodies IX and 111* II al j , 51 55 53 54 L01 4 4 4 4 L06 + 4 4 4 102 + 4 4 “ f* 103 4 4 4 4 4 n 101J -h Table 55 A1 5 58 Em sa c c h a rin a Two f r u i t b o d ie s o f th e o n ly c o l l e c t io n wore chosen and d esig n a te d as l a and lb* S e w n iaonospore c u ltu r e s o f l a were p a ire d in a l l com­ b in a tio n s and th e r e s u l t s a r e g iv en i n T able 56* Tho r e s u l t s o f p a ir in g 16 monospore c u ltu r e s o f lb a r e g iv en i n Table 57* The mono spore c u ltu r e s o f each f r u i t body f e l l in to two sex u al phases and showed norm al b ip o la r ity * F our monospore c u ltu r e s (two from each sex u al phase) o f l a were p a ir e d w ith f o u r c u ltu r e s o f lb* e v e ry p a irin g * C lasp co n n ectio n s were found In The r e s u l t s a r e shown in T able 58* A gain, a s in B» g la n d u lo sa , com plete c o m p a tib ility was found t o e x is t between mono sp o re c u ltu r e s from two f r u i t bodies growing v ery c lo s e to each o th er* E* n u c le a te A few m u ltisp o re c u ltu r e s were o b ta in e d from one w ild f r u i t body, b u t owing t o th e v e ry p o o r g erm in atio n o f th o sp o re s no monosp o re c u ltu r e s w ere se cu red a t t h i s tim e* Monospore c u ltu r e s , however, w ere o b ta in e d from sp o re s borne on f r u i t bo d ies in cu ltu re * These c u ltu r e s grew v e ry slo w ly and s e v e ra l o f them e i t h e r became contam inated o r d ie d o u t because o f lo s s o f v ig o r . E leven c u ltu r e s were p a ire d in a l l com binations (w ith th e e x c e p tio n o f a few p a ir in g s ) and th e r e s u l t s a re g iv en i n Table 69* due to lo s s o f c u ltu re s * The b lan k s in th e t a b l e re p re s e n t no p a i r in g C u ltu re s 3 , 14, 15, 17, 26 and 27 f e l l in to one se x u al p h a s e , w h ile 1 2, 13 and 22 f e l l in to a second sex u al phase* A ll members o f th o f i r s t phase v/ere com patible w ith a l l members o f th e second* C u ltu re s 25 and 50, however, were e x cep tio n s to th e normal p o l a r i t y i n t h e i r re a c tio n s * They were incom patible w ith a l l o th e r 59 Table 56 E« s a o c h a rira u R e s u lts o f p a ir in g seven monosp o re c u ltu r e s o f f r u i t body la* A a — >» 201 202 205 203 204 207 208 — — 4 4 4 4 201 — 205 4 4 4 4 4 4 4 4 203 — — — 204 — — 207 — | 202 Table 58 E. sa c c h a rin s^ R e s u lts o f p a ir in g s between f r u i t bodies l a and Ib» A la a lb 201 202 203 >04 251 4 4 4 4 A1 252 4 4 4 4 256 4 4 4 4 26C 4 4 4 4 208 Table 5? E. s a c c h a rin e * R e s u lts o f p a ir in g 15 nonospore c u ltu r e s o f f r u i t b o # lb* A1 ^ ---------------------------— ---- al 'V- 251 258 253 254 255 257 258 259 264 265 256 260 262 263 266 251 4 4 4 252 4 4 4 4 4 252 4 4 4 4 4 4 4 254 — — — — - — 4 255 4 257 259 — — — — 4 — — — 4 — — 4 264 — 4 265 4 258 256 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 — — — — 262 — — — — — 263 — 260 266 60 Table 59 E* nucleate* R esults o f p a irin g 11 faonospor® c u ltu res* $ » mature f r u i t bodies and baaidiospores* 3 3 14 15 17 26 27 12 13 22 25 30 H- © + — 14 — — — — — — — - — — — © 15 — — -t- © © — — 17 — — + H- © — — — + e + — — © — 12 — — — 13 — — — - — 26 27 + 22 25 — 30 Table 61 T able 60 R e s u lts o f p a ir in g s between E* y.landulosa.an d E. r o o is a . R e s u lts o f p a irin g s between E» r e c ia a and E« s a c c h a rin e . E. r e c is a A c E. r e c i s a — 'l ----' 5 10 4 11 ~ A ~o ui 1 — — — — — — — 2 — 3 — — — — ui 5 — — — c £ , 5 4 11 — — — <_> flS 1/1 0 202 — — — — 203 — — — — LU 204 — — — — j C. 20] — o SC *00 K - 1 nS — 61 mono sp o re c u ltu r e s and a ls o w ith each o th e r . I t i s n o t p o s s ib le to t e l l from th e s e r e s u l t s w h eth er th e s e groups r e p re s e n t th r e e sex u al p h a se s o f a q u a d r ip o la r .s p e c ie s , o r w hether B* n u e le a ta i s t y p i c a l l y b ip o la r and t h a t c u ltu r e s 25 and. 30 were m erely ex c e p tio n s to th e norm al p o l a r i t y , t h e l a t t e r view seems th e more p ro b a b le . I n t a b le 50 , th o © s ig n r e p r e s e n ts th e p ro d u c tio n o f f r u i t b o d ie s w ith m ature h & sid ia .~»d b a s id io spores* I t i s i n t e r e s t i n g t o n o te t h a t none o f th e p a ir in g s i n w hich c u ltu r e 12 was involved produced f r u i t bodies* A ttem pts to H y b rid ize S p ecies o f E x ld ia Four monespore c u ltu r e s o f B. g lan d u lo sa I were p a ir e d w ith fo u r monospore c u ltu r e s o f E. r e c i s a I* i n sa y p a irin g -. Ho clamp co n n ectio n s were found The r e s u l t s a re given i n f a b le 60* P a irin g s were a ls o made betw een c u ltu r e s o f B. s a c c h a rin a I& and B. r e c is a I* r e s u l t s a r e g iv en i n fa b le 61. i n any o f th e p a i r in g s . The Again no clamp co n n ectio n s were found Ho In d ic a tio n s were observed o f th e presen ce o f f r u i t bodies o r any k in d o f g e la tin o u s bodies* DISCUSSION The torn® used by d i f f e r e n t a u th o rs in d e s c rib in g th e s e x u a lity o f th e Baa id io n y c e te s v ary to a c e r ta i n ex ten t* The term "soxes" h a s been u sed by a cumber o f in v e s tig a to r s to ap p ly to th e d i f f e r e n t groups in to w hich th e monospore c u ltu r e s f a l l , based upon th e p ro d u ctio n o f clang? co n n ectio n s when th e y a re p a ird l w ith o th e r monospore c u ltu re s o f th e Sum© f r u i t body, "box” , in i t s fundam ental s e n s e , seems t o in p ly th e f u s io n o f n u c le i a t some s ta g e in th e l i f e o f th e fungus. Thor© i s some evidence to in d ic a te t h a t th e u n io n of n u c le i and th e p p ro d u c tio n o f b a s id ia do n o t always fo llo w th e th e fo rm a tio n o f clamp c o n n e c tio n s. im 9B o f m ycelia ;-nd Brunswik (6 ) found t h a t c e r ta in 62 d ic a ry o n m y celia o f Coprirats f r i o s l l a r i s i n g from th e union o f two mono c a r 2/0n m y celia produced f r u i t bodies w ith m ature b a s id ia and spores w h ile u n d er th e same c o n d itio n s o th e r d icary o n m ycolia d id not* (1 2 ) found th e some c o n d itio n s i n C» s p h a ^ ro s p o ru s . Dickson A s im ila r s i t u a t i o n m s found i n E. n u c le a ta i n th e p r e s e n t work* C o n sid erin g th e fu n g i i n g e n e ra l, th e term "sex es" i s most common­ l y u se d t o d i s t i n g u i s h two in d iv id u a ls , on© o f which and t h e o t t e r a s female* c t s as m l© T h is co n cep tio n in c lu d e s th e p assag e o f n u c le i from o n ly on© in d iv id u a l (m ale) t o th e o th e r (fem ale)* f u n g i, however* t h i s i s n o t th e case* I n many B i l l e r (9 ) has shown t h a t , i n Coprim ts la g o p u s, two com patible m ycelia may u n ite and th e r e would f o llo w an exchange o f n u c le i* so t h a t each monocar yon m ycelia becomes d ip lo id iz e d by th e o th er* I t seems more reaso n ab le to co n sid e r th a t i n such fu n g i* s e x u a lity i s p r e s e n t b ut w ith no d i s t i n c t i o n between m aleness and fe a & le m ss . For th e s e re a s o n s , " se x e s” does n o t seem th e most a p p ro p ria te term to u se I n r e f e r r in g t o th e d i f f e r e n t groups o f monoo&ryon m y celia. O th er term s have been u sed f o r th e s e same groups. Brodi© u ses th e term 'se x u a l g ro u p s", Dickson (12) u se s " p a irin g groups", w hile Bessoy (2) u s e s th e term " se x u a l p h a se s" . T his l a t t e r term , "sex u al phases" i s u sed th ro u g h o u t th e p r e s e n t p ap er i n th e same sense t h a t "sexes" has been a p p lie d to many o f th e B asidiom ycetes. Many i n v e s tig a to r s have used th e terra " f e r t i l e " to d e s c rib e th e p re se n c e o f clamp co n n ectio n s in a p a ir in g o f two monoc&ryon m ycelia and th e t e r n " s t e r i l e " when no clomp connections were produced in such a p a irin g * The t e r n " f e r t i l e " , however, seems to apply more a p p ro p ri­ a t e l y t o t h e p ro d u c tio n o f s p o re s , o r a t least, to th e form ation o f f r u i t 63 b o d ie s . F ollow ing Smith and Brodie (S I) and Beesey ( 2 ) , in th e p re s e n t p a p e r th e term "com patible*’ ie used t o d e sc rib e two monocaryon m ycelia w hich u n i t e , w ith subsequent p ro d u c tio n o f d icary o n mycelium ( in d ic a te d ) by th e p re sen ce o f clamp c o n n e c tio n s ). The term '■incom patible" i s u sed to d e s c rib e inonocuryon m y celia, which when p a ir e d , do n o t produce d ic a ry o n mycelium. The r e s u l t s o b ta in e d from p a ir in g s o f mono spore c u ltu r e s o f th e same f r u i t body o f e .ch s p e c ie s in d ic a te t h a t A. a u ric u la -ju d c .e , E. glandulow a, E, r e c i s a and B. saccharins. a re t y p ic a l ly b ip o la r . The p a ir in g s o f monospore c u ltu r e s o f one f r u i t body o f E. n u c le a te d id n o t show normal b i p o l a r i t y and i t i s n o t known w hether th e s p e c ie s i s norm ally b ip o la r o r q u a d rip o la r. The f a l l i n g o f th e monospore c u ltu r e s o f a fungus in to a c e r ta in number o f sex u al p h ases based upon c o m p a tib ility i s co n sid ered to be * ie t o th e p re se n c e o f one o r two p a i r s o f f a c to r s which fo llo w th e sim p le M endelian r a tio s * f a c to rs " * Kniep (17) has c a lle d th e s e " c o p u la tio n B u tle r (10) q u o tes BrunswUe (7) as f o llo w s * "Brunswik has in te r p r e te d id e n t i c a l phenomena p r e f e r r in g to th e p resen ce o f Kniep*a c o p u la tio n f a c t o r s / i n h is experim ental m a te ria l by assuming th e o p e ra tio n o f s t e r i l i t y fa c to rs * He assumed t h a t autogamy i s th e fundam ental p ro c e ss i n b o th monoecious and d io ecio u s fu n g ij t h a t i s , b o th have th e same gen o ty p ic c o n s titu tio n t s f a r as sex i s concerned. H e te ro th a llis m i s determ ined by th e a d d itio n o f in h ib itin g o r s t e r i l i t y fa c to rs , nd xaut& bility and allelom orphism a rc lin k e d w ith th e s e r a t h e r th a n w ith th e r e a l sox f a c to r s " . ?,b o th e r th e c o n d itio n o f se x u a l phase® i s to bo co n sid ered as due to th e p resence o f " s t e r i l i t y f a c to r s " o r due to th e absence o f f a c to r s which may be c a lle d " c o p u la tio n 64 f a c to r s " o r " c o m p a tib ility fa c to rs " * seeme to be m ainly one o f term ­ inology* S c a tte re d i r r e g u l a r i t i e s were found in th e r e s u l t s o f th e p a ir in g s o f monospore c u ltu r e s from f r u i t bodies I and V III o f A. a u r ic u la ju d a e . These w ere re p re s e n te d by c o n s ta n t in c o m p a tib ility between c e r t a i n monospore c u ltu r e s b elo n g in g to d i f f e r e n t sex u al p h a se s. Such i r r e g u l a r i t i e s o r d e v ia tio n s from th e normal p o l a r it y a re s im ila r to c o n d itio n s re p o rte d by Y andendries (3 5 , 36) f o r Faneolus catrrpanulatus, F . s e p a ra tu s , Coprinus mieadeug and heptoporus in b e r b is ; by Kniep (1 7 , 18) f o r Schlzophyllum coM ins} by Brunswik (6 ) f o r Coprinus p ic a c e u s ; by Dickson (12) f o r C. sph. e ro a p o ru s; and by Z a ttl e r (44) f o r C o lly b ia v e lu tip e a . Concerning th e p a i r in g o f monocaryon m ycelia from th e same f r u i t body o f C, sp h aero sp o ru s, Dickson (12) s a y s, " I t is co n sid ered t h a t th e two p a ir in g groups a re due to a s in g le f a c t o r d iff e r e n c e , but th e evidence i s n o t s u f f i c i e n t to show w hether th e v a rio u s degrees o f s t e r i l i t y e x h ib ite d a re duo to th e presence o f in c o m p a tib ility f a c to r s and, i f s o , how many such f a c t o r s a re concerned"* Yandendries (36) s ta te s t h a t th e ten d en cy tow ard " s t e r i l i t y " between th e two "sexes" o f Leptoporus in b e rb is i s p ro b ably due to "une d e fic ie n c e des r ^ a lis a te u r s so x u e le s" , Kniep (17) a t t r i b u t e s such d e v ia tio n s from normal p o l a r it y to q u a n tita tiv e gene ohanges. I t h as boen dbserved in th e p re s e n t paper t h a t w ith in th e s.-me s p e c ie s com plete c o m p a tib ility was found between th e th re e c o lle c tio n s o f E. rd an d u lo sa, th e th r e e c o lle c tio n s o f E. r e c is a and between f iv e o f th e seven c o lle c tio n s o f A. a u ric u la -ju d a e ( I I I , IV, V, V I, V III)* Two o f th e c o lle c tio n s o f A. a u ric u la -ju d a e were from N ebraska, two 65 from lo m and one from U orth C a ro lin a . In a d d itio n t o th e c o m p a tib ility betw een c o l l e c t i o n s , com plete c o m p a tib ility w„.s found between two f r u i t b o d ie s o f th e sane c o l l e c ti o n o f A. a u ric u la -ju d a e (Va and Vb). This was a ls o t r u e f o r two f r u i t bo d ies o f £ . sac char in a ( l a and lb ) growing on th e same s t i c k w ith in 12 inches o f each o th e r , and a ls o f o r two f r u i t b o d ies o f E. g lan d u lo sa ( I l a and l i b ) growing b u t two inches a p a r t. Complete c o m p a tib ility between two o r more f r u i t bo d ies growing e i t h e r a t some d is ta n c e a p a r t o r n e a r one a n o th e r has been re p o rte d by Eniep (17) f o r Sohi zo phyllua comraune, A leurodlseus po ly g o n iu s, C o lly b ia v e lu tlp e s , C. conlgena, C. c i r r h a t a , A m e lla r ia mo I le a and C oprinus f im e ta r lu s ; by Brunswik (6) f o r C. f im o ta r iu s , C. coiaatug, C. n lv e u s , C. p ic a o e u s , 0 . lagopus and C. f r i e s i i ; by Hanna (15) f o r 0 . lag o p u s; by Newton (26) f o r C. ro s tr u p ia n u s ; by Yandendries (5 2 , 33, 54, 56, 59) f o r C» r a d ia n s , C« m icaceus, Faneolus com panulatcs, Traxaetes suaveolens and Hypholoma s u h la te r itiu m ; by Sfcmnce and Macrae (25) f o r h e n z ite s s a e p l a r ia , L. tr a o e a and Trem etes am ericana; by Mounce (zz) f o r Foraes p in io o la ; and by Arnold (1) f o r Marasmius c lo n g a tip e s . V andendrics (54) concluded from h is work w ith Coprinus m icaceus t h a t p a ir in g s between c o lle c tio n s o f th e same re g io n were com patible and t h a t , i n g e n e ra l, p a ir in g s between very d is ta n t c o lle c tio n s were in co m p atib le. S ev eral ex cep tio n s were found to t h i s g e n e ra l r u l e . Complete in c o m p a tib ility was found in A. a u ric u la -ju d a e when p a ir in g s wore made between f r u i t body I and each o f f r u i t bodies I I I , IV, VI and V II. P a r t i a l in c o m p a tib ility was found between f r u i t bodies I and Va, I and Vb, I and V I I I , I I I and V I II , IV and V I I I ; and Va and V III. F r u it bodies I and V III were c o lle c te d from c o n ifero u s h o s ts 66 and had lo n g e r sp o re s th a n th o o th e r f r u i t b o d ie s, which were c o lle c te d from deciduous h o sts* I t i s suggested t h a t th e v a r ia tio n in th e sp o re le n g th s o f th e d i f f e r e n t f r u i t b o d ie s , o r th e d iffe re n c e in th e h o s ts , o r b o th , may be a s s o c ia te d w ith g e n e tic d iff e r e n c e s in th e f r u i t b o d ie s g ro a t enough to in flu e n c e th e c o m p a tib ility o f th e xnonoccryon m ycelia* S im ila r ca se s o f com plete o r p a r t i a l in c o m p a tib ility between f r u i t b o d ie s o f th© same s p e c ie s have been re p o rte d by Kniep (17) f o r Schiaophyllum o o a m e and ColXybia c i r r h a t a ; by Brunswik (6) f o r C oprinus com atus; by T andendries (34) f o r C* m icaccus; and by Mourn© (22) f o r Pones p in ic o la , S e v e ra l d i f f e r e n t th e o r ie s have been advanced i n an a ttem p t to e x p la in th e se x u a l phenomena i n th e Basidioraycetes* Among them a re th© th e o ry o f sex u a l m u ta tio n s , th e th e o ry o f r e l a t i v e s e x u a lity advanced by Hartmann ( I S ) , th e th e o ry o f m u ltip le “s e x e s" , and th e th e o ry o f m u ltip le allelo m o rp h s advanced by Kniep. Banna (IS ) s ta te s t h a t " th e sex f a c t o r s f o r a given sp e c ie s may bo undergoing fre q u e n t m u ta tio n s w ith th e r e s u l t t h a t new sex u al s tr a i n s are c o n tin u a l ly a p p e a rin g ". I n speaking o f th e "com plete i n t e r f e r t i l i t y " between d i f f e r e n t geographic ra c e s o f Coprinus r o s t r u p i &nus, Hewton (26) says, w h ile each s t r a i n i s b is e x u a l, th." sp ecies as a whole must be reg ard ed as m u lti s e x u a l". Kniep (1?) considered t h a t th e com plete i n t e r c o n p a t i b i l i t y between two f r u i t bodies was due to q u a n tita tiv e gone changes and t h a t m u ltip le allelom orphs were Involved* Ho a ls o s ta te d t h a t in c o m p a tib ility between f r u i t bodies o f th e same s p e c ie s may be caused by too g re a t a q u a n tita tiv e d iffe re n c e i n th e g en es, o r may be due to secondary f a c to r s . Kniep1s th e o ry o f m u ltip le allelo m o rp h s stems t o be th e b e s t f o r in te r p r e tin g th e r e s u l t s o b tain ed 67 i n th© p r e s e n t work* C o n sid erin g t h a t th© c o m p a tib ility f a c to r s e x i s t as m u ltip le a lle lo ia o rp h s , th© q u e s tio n a r i s e s w hether c e r t a i n p a ir s o f f a c to r s produce a s tro n g e r degree o f c o m p a tib ility th a n o th e r s , an oppor­ t u n i t y 'to stu d y t h i s q u e s tio n aro se when a l l monospore c u ltu re s of B» g la n d u lo sa I l a were found to be com patible w ith a l l monospore c u l t u r e s o f l i b and a g r e a te r number o f clamp connections were found i n c e r t a i n com binations th a n in o th ers* The assum ption i s made in th e s e ex p erim en ts t h a t th© g r e a te r number o f clamp connections in ­ d ic a te s a s tro n g e r deg ree o f c o m p a tib ility between th e two monospore c u ltu r e s involved- Th© r e s u l t s given in T able 45 would th en in d ic a te a much s tr o n g e r d eg ree o f c o m p a tib ility between Aj and a ^ , Ag end &g. Ay and a g , and a j a id Ag, th e n e x is te d between Ap and Ag, and ag* and I f th e numbers o f e-lamp connections can be c o n sid e re d , i n t h i s c a s e , a s r e p re s e n tin g d iff e r e n c e s between members o f an allelo m o rp h ic s e r i e s , i t i s found t h a t and ©g a re th e most s im ila r , w ith Ag o n ly s l i g h t l y l e s s s im ila r . and Th© g r e a te s t d iffe re n c e would be between A^ and a ^ , w ith Ag e q u a lly d if f e r e n t from and ag- It m y , t h e r e f o r e , be concluded t h a t i n B. glandulosa th e c o m p a tib ility f a c t o r s may show v a ry in g degrees i n th e s tre n g th o f c o m p a tib ility when th e y a r e paired* D ip lo id iz a tio n experim ents w ith mono spore c u ltu re s of E» g landulosa showed t h a t in o n ly th ro e o u t o f 16 cases was th© d ip lo id iz a tio n rat© d i s t i n c t l y g r e a te r th a n th e growth r a te o f th e d icary o n mycelium in th e sum© p a irin g * The d iff e r e n c e s in th e numbers of clamp connections produced i n v a rio u s p a ir in g s , th e r e f o r e , cannot be due ( a t leafct in a l l 68 c:.soa) to a d i f f e r e n t i a l d ip lo id iz a tio n ra te # I t is suggested t h a t th e r e may be a g r e a t e r a t t r a c t i o n between c e r ta i n monocaryon m ycelia th a n betsr en o t h e r s , b u t as y e t , no means have been found to demon­ s tra te t h i s . SUMMARY I* A* a u r ic u la - ju d a e , E» g la n d u le sa , B* r e c is a and E. n u c le a ta ■were found to bo t'h e t a r o t h a l l i c n and b ip o lar# 2. P a irin g s between monospore c u ltu re s from seven f r u i t bodies o f A# a u ric u la -ju d a e showed t h a t f iv e o f them were com pletely com patible w ith each o th er* Two f r u i t b o d ies showed v ary in g degrees of in c o m p a tib ility w ith th e other® and between them selves* 3* Complete c o m p a tib ility was found between fo u r f r u i t bodies o f B» g la n d u lo sa , even between two w hich were growing only two Inches a p a rt* 4. A d iff e r e n c e was found in th e number o f clamp connections formed in p a ir in g s in v o lv in g c e r t a in com binations o f sex u al phases o f two com p letely com patible f r u i t bodies o f 5* glandulowa* 5. Complete c o m p a tib ility was found between th re e f r u i t bodies o f E« re c is a * 6. Complete c o m p a tib ility was found between two f r u i t bodies o f £ . sac chi;xina growing w ith in 12 in ch es o f one another* ?. The th e o ry o f xaultipl© allelom orphs seems to serv e as th o b e s t e x p la n a tio n o f th e r e s u l t s obtained* 69 LITERATURE CITED 1* °f A rnold, Jean D. A com parative stu d y o f c e r ta in s p e c ie s Marasmius and C o lly b la i n c u ltu r e . "Kycologla 27- 388-417. P i s . 35, 36. 1935 2. P. B essey, E rn st A. A te x t-b o o k o f mycology* 495 pp. 139 fig s * B la k is to n ’ s Son and C o ., P h ila d e lp h ia * 1935. 3* Rose, S. R. S e x u a lity o f Polyporus o s tr ie f o rr a is and P o ly g tic tu s k irs u tu s * 4* La C e llu le 42* 249-266* 1 pi* 1933. B re fe ld , O scar. Busidiorayceten. I I , P roto b u sid io rr, c e te n . B ntersuchungen aus dein Q e sa m tg e b ie te d e r Mykologie 7* 1-178. P i s . 1 -1 1 , 1888. 5* B rodie, H* J* Hymenoniycetes. The occurrence and f u n c tio n o f o ld ia i n the American Jo u rn a l o f Botany 23? 309-327, P ig s. 1-41. 1936. 6. Brunsw ik, H. Untersuchxmgen fiber d ie G eschlechts - und X o rn v erh E ltn isse d e r HyiT«enoinycetengattung Coprinus* Rot&niseh© Abhandlungen herausgegeben von Dr. K. Goebel 1 (5 ): 1-152, Figs* 1 -3 . 1924. 7 ..... .................. t{ liber d ie S e x u a lita ts v e rh A lin is s e b e i den R asidiom yceten. Verb. Zool. Bot* G e s e ll. Wien. 73: 153-104* 1924 C ited by B u tle r (10)* 8. B u lle r, A. H. Reginald* The b io lo g ic a l s ig n ific a n c e o f co n ju g ate n u c le i i n Coprinus lagopus and o th e r Hyinenonycetes. Nature 126: 686~u89. F ig s . 1 -7 . 1930. 9. , — . Researches on fu n g i Vol. 4. 329 pp. 4- p i s . 149 f i g s . Langroans, Green and C o., London and Nevr York. 1921 70 IQ* B u tle r , L# F* 66n* on apple** C o rticiu m c e n trlfu g am , a h e te r o t h a lli o p a th o - J o u rn a l o f A g r ic u ltu r a l R esearch 41* 269-294. P i. 1 Fig* 1 - 2 . 1930 11. D angeard, P . A* B asid io m y cetes. 12* Memoi r e s u r l a re p ro d u c tio n s e x u e lle des Le B otanist© 4? 119-181. F ig s . 1-24. 1894-95. D ickson, H. S tu d ie s on Coprinus sphaerosporus I . The p a i r i n g b e h a v io r and th© c h a r a c t e r i s t i c s o f v a rio u s h a p lo id and d ip lo id s tr a in s * 13* A nnals o f Botany 48* 527-547* p i . 11* Gr&taa&nn, E r n s t, 1934* Comparative morphology o f fu n g i. T ra n sla te d by C a r r o ll HI* Dodge* 701 p p . 406 f i g s . McGraw - H i ll Book Co*, Sew York. 1928* 14* G ilb e r t, E. I . B asidiom ycetes» C y to lo g ic a l s tu d io s o f th e low er T ra n sa c tio n s o f th© W isconsin Academy o f S c ie n c e ., A rts and L e tte r s 20* 387-397. 15. Hanna, M, F , F ig . 1* 1921. The problem o f sex i n C oprinus lag o p u s. Annals o f Botany 39* 431-467. 16* p . 29. 1925* Hartmann, M* R e la tiv e Sexual i t At und ih re Bedoufcung fS r c in e allg em ein e S e x u a litfita - und ©in© allgem eine B e fru e h ts th e o rie . f la tu n r is s . 19* 8—19, 31—37* 1931* 17. 221 f i g s , 18, Kniep# Hans. C ited by Bose ( 3 ) . P ie S exu& litSt d e r n ied ero n Pfl&naen. 544 pp. Gustav F is c h e r. Jena 1928* — - „ V ererbungserscheinungen b e i P ilz e n . B ib lio g ra p h ic s G en etica 6* 371-478* Figs* 1—15* 1929 19* Kuhner, R, C o n trib u tio n a 1* etude des Hyraenomycetes ©t sp ecialam en t des A garic ace s . 1926. Le Botanist© 17* 5-207. 57 f i g s , 4 p i s . 71 20* M artens, p . e t R. V undendries. ©t diploid© chez p h o llo ta a u r i v e l la . l& cy cle co nidion h ap lo id e La C e llu le 41 s 335-388. 3 p i s . 1933. 21# MSI l o r , A* P ro to b as idiom yceten. Botan. M itt. a . d . Tropen 8* 1 -1 7 9 . P i s . 1—6 . 1895. C ited by G&umann (1 3 ). 22* Mounoe, Iren© . The b io lo g y o f Pomes P i n i c o la (gw .) Cooke. Canada Departm ent o f A g ric u ltu re B u lle tin 111. Hew S e r ie s . 75 pp. P ie . 1-10* 1929 23. , and Ruth M acrae. The b eh av io r o f p a ire d roono- sporoue myc e l i a o f Lena it® s a a e p ia r la (7 ;u lf) F r . , L« tr a b e a ( p e r s .) F r . , L* th erm o p h ila F a lk , and Trametes am ericana Qverh. Canadian J o u rn a l o f R esearch , S e c tio n G* 14s 215-222* 1936. 24. H euhoff, W* Z y to lo g ie und sy s te m tis o h e S te llim g d e r A u ricu l& riaceo n und Troiaellaoooa* B o taaisch e A rchiv S* 250-287. P i s . 1 -4 . F ig s* 1 -7 . 1924. 25. --..i.—— ( T re m e llin e a e )• 26* Die P ila e M itto le u ro p a s. Band I I . Die G a lle r tp ilg e I l l u s t r a t e d . 1935-1956. Sewton, Dorothy S* B is e x u a lity o f in d iv id u a l s tr a i n s o f C opriaus r o s tr a p la n u s « Annals o f Botany 40 1 105-128* PI* 6 . Figs* 1-17* 1926. 27. O o rt, A. J* P . Die S e x u a lltftt von C oprim s f iia e ta r iu s . R ec ev ll des Travaux B otaniques Bleerland&is 27 * 86-148* p is* 4 -6 . Figs* 1 -5 . 1930* 28. Gazette 29 C ite d by Bos© ( 3 ) . R ogers, D. p . A e y to lo g ic a l stu d y o f T u la s n e lla . B o tan ical 94* 86-106. F ig s . 1 -79. 1932* a ■i B a s id ia l p r o l i f e r a t i o n through claiiip-form at ion i n a new S eb acin a. Myoologia 28 j 347—882*33figs. 1936. 72 SO* S h ear, C, L* and B® 0* Dodge* The l i f e h is to r y o f P ila c r e fa g ln e a (Fr®) B* Is Br® Jo u rn a l o f A g r ic u ltu r a l Research 30* 407-417. PIS* 1-2* 1926. 31. S m ith, A. H* and H. J . B rodie. C u ltu re ! c h a ra c te rs and p a i r in g r e a c tio n s o f monosporous m ycelia and development o f th e f r u i t body o f P h o lio ta (Flaxnmula) polychroa® B o tan ical G azette 96* 533- 646. SO f i g s . 1935. 32. V an d en d ries, Rene. Reoherches ex p eriiaen tales su r l a h i p o l a r i t e a e x u e lle d©6 B asidiom ycetee« B u lle tin de l a S o ciete Royal© Bot&nique d© B elgique 57 * 75-78. IS 24. S3. C o n trib u tio n n o u v elle a 1* etu d e de l a s e x u a lite des 3&sidiomycete s* La C e llu le 35* 129-157* pis® 1 -2 . F ig s . 1 -4 . 1925. 34® . . . . Le coi^portraent sexuel du Coprinus rolcaceus dans ees r a p p o rts avec l a d is p e r s io n do 1 'e s p e c e . B u lle tin de l a S o c ie te Royal© Botanique de Belgique 60* 62-65. 1927. 35l® ii .......... . d iss e m im tu s Pers® So*— — — . La b ip o la r it© s e x u e lle chez Coprinus I b id . 62* 133-136® 1930. H ouvelle in v e s tig a tio n s dans le domain® se x u e l des Hymenomycet es® B u lle tin T rim e s trio l de l a S ociete liycologique de Prance 48* 130—165® 57. ......... . l e genre Traraetes. 38. Pigs* 1—7* 19S3® C o n trib u tio n a 1* etude de 1& se x u e llo dans Ibid* 50* 98-110* Pis® 5 -6 . 1 t e x t f i g . .La chez P le u ro tu s p i n s i t i s * 1934. p o la r ity sexuell© o t l e regime co n id ien Ib id . 50* 203-212. F ig s . 1-10. 1934 73 39* Lee a f f i n i t e s s e x u e lle de Hypholoina g u b la te r ttiu m . 40* I h e ld e n , Soy M* T rem ella* 41* 8-11* I b id . 50* 213-217. F ig s . 1 -4 . 1934. C y to lo g ic a l s tu d ie s in th e Trem ellaceac I . Ily co lo g ia 26s 415-435. F I s . 47-49. 1 t e x t f i g . — — —. — — — I I . E x id la . 1934. M ycologia 27* 41-57. P is* 1935* 42* F ig s . 1-3* 43* --------- — * S ebacina. M yeologia 2?j 503-520. 1935. — — —• g lo b o sp o ra n . sp* 44. H I. O b servations on th e cy tology o f S eb acim Rhodora 37* 121-127. p i* 331. 1 t e x t f i g . Z a ttle r * F ritz * V ererbungsstudien an H u tp ilzen . Z e i t s c h r i f t f tl r B otanik 16s 433-499* 1924* 1935* E x p la n a tio n o f F ig u re s i n P la te 1 F igs* 1 -8 , A» a u ric u la -ju d a e * 1, b a s id io s p o re s ; 2 , A, b a s id io s p o re g erm in atin g in w a te r by means o f o id ia , B, o i d i a produced by sp o res i n w ater? S , 4 , b a s id io spores g e rm in a tin g on a g a r; 5 , g erm in atin g o i d ia ; 6 * monooaryon hyph&e b e a rin g o i d i a ; 7 , d io ary o n hypha showing clamp connec­ t i o n s ; 8 , basldium from f r u i t body i n c u ltu re # E. glandulosa* F ig s . 9-17, 9 , A, B, b a s id ie s p o re s germ in atin g in w ater* C, o i d i a produced by s p o re s in w a te r; 10* b a s id io spores* 11 , b a s id io sp o re p ro d u cin g secondary sp o re ; 12 , 13, b a s id io - ep o res g e m in a tin g on a g a r; 1 4 , d icary o n hypha showing clamp c o n n e c tio n s; 1 5 , g e m l m t ng o id i a ; 16, tw o -c e lle d b&sitlium w ith two e p ib a a ld ia , produced by monocaryon mycolium; 17, u n io n o f two young hyphao from germ inating b a s id io s p o re s , arrow s p o in t tow ard spores* plate x Explanation o f Figures in P late II Figs* 1 -5 , E« re e ls a # 1, b& sidio spores 5 2 , b a s id io - spor© g e m in a tin g on a g a r ; 3 , same a s 2 b u t showing produc­ t i o n o f o i d i a on young hypha©? 4 , f o u r - c e lle d basidium produced by d io ary o n mycelium; 5 , d ieary o n hypha showing clamp connection# Figs* 6 -1 1 , F.. sacch arin e# 6, dio ary o n b e a rin g o i d i a , a ls o showing clamp connections? 7 , b in u c le a te o i d i a from d ie a ry o n mycelium? 8 , u n in u c le a te o id ia from raonooaryon mycelium? 9 , roonocoryon hypha b e a rin g o id ia ? 10 , u n io n o f young hypha© from two b a s id io spores? 11 , b i - n u c le a te aidlum g e m in a tin g on ag a r showing clamp co n n ectio n on th e ger.n tube# Figs* 12-15, £ . n u cleata* 12, germ in atin g o id ia ? 1 3 , d ie a ry o n hypha showing clamp connection? 14, 15, f o u r - c e lle d b a s id la showing s e p ta te e p ib a s id ia from f r u i t body i n c u ltu re * PLATE IE C333 Explanation, o f Figures in P late III F igs* 1—6, C qloeera cornea* 1, 2, b a s id io s p o re s germ­ in a tin g on a g a r j 3 , o id ia g erm in atin g on a g a r,} 4 , hypha from mono sp o re c u ltu r e p ro d u cin g o id ia ; 5 , basidiuro from f r u i t body i n c u ltu re s 6 , b a s id io spores from f r u i t body in c u ltu re * 8, Figs* 7 -1 0 , Dacryomyces minor* 7 , b a sid io e p o re s b u s id io s p o re s pro d u cin g o id ia in -water; 9 , b a s id io spor< g e m in a tin g on a g a r; 1 0 , hypha from monospore c u ltu r e p ro ­ d u cin g o id ia * Figs* 1 1 -12, T rem ella lu tesce n s* 11, b a s i d i o tp o re g e m in a tin g by means o f secondsry sp o re ; 12 , b a s id io spore g e m in a tin g on a g a r by budding* plate 6 nr Explanation o f Figures in F la ts I? A ll f ig u r e s A« a u r ic u la -J u d a s , 1* b a s id io s p o re p ro d u c in g seco n d ary spore* 27502 2 , o id ia produced by g e r a im tln g . baaldteapore© i n ^ a t o r , X7SQg S, p a ir in g s o f uasoa&oro c u ltu r e s fro a f r u i t body fif, X 7/8j a* a v e rs io n , 155 x 1 5 7 , B, a v ersio n * 157 x 151, G, ©von in te rm in g lin g o f saycolia* 157 x 165, B, aversion, with gap f i l l e d with d io a ry o n r y e e l i m , 154 x 157; 4 , young f r u i t bodies i n 12day a g a r c u ltu re * X-i/Sj 5 , siaao c u ltu r e a s 4* th ro e tsceka l a t e r , 2 4 /5 ; 6 , f r u i t b o d ies growing os. bassaood s tic k * X4/&. PLATE 1SZ E x p la n a tio n o f F ig u res i n P la te V Figs* 1-5* K» saco h arin a* 1 , b a s id io s p o re g e m in a tin g on a g a r and p ro d u cin g o i d i a , X4GQf 2 , monocaryon c u ltu r e s showing s h in y m asses o f o i d i a , A, c u ltu r e f iv e weeks o ld , B, c u ltu r e th r e e weeks o ld , X ?/8| 5 , monocaryon n y c e lia , A, 265, B, 265, G, 2 5 ?, B, 256, F ig s . 4 , 5 , E» r e c l s a . 4 , o id ia g e m in a tin g on a g a r , some p ro d u cin g mycelium and o th e r s producing secondary o i d i a , X?5Qj 5 , s t e r i l e g e la tin o u s to d ie s produced by, A ,d ie ary o n mycelium, B, G, m noearyon m y celia, X?/8* ■B H P L A T E SI E xplanation o f Figures i n p i to YI A ll f ig u r e s E. n u cleate., 1, b a sid io s p o re g e m in a tin g on ja o is t wood and p ro d u cin g o i d i a , X700; 2 , b a s id io s p o re form ing secondary sp o re i n w a te r, X7S0j Z, b a s id io spore form ing lo n g g e m tu b e i n w a te r, 7,750; 4 , monocaryon hyphao p ro d u cin g c l u s t e r s o f o id ia , X750. PLATE m Explanation o f Figures in P late ¥11 A ll f ig u r e s K» g lu n d u lo sa , 1 , Etono spore c u ltu r e s , a, 2 3 , B, 24, C» 21, X |-f 2 , dioaryon agree lim a formed by p a i r ­ in g monospor© c u ltu r e s 21 and 51, X i/5 j 3 , d ieary o n mycelium farm ed by p a ir in g roonospore c u ltu r e s 21 and 50, X 4/o; 4 , d io ary o n mycelium formed by p a ir in g mono spore c u ltu re s 24 and 50, X4/5. PLATE m