j .•

The

rid

il ti vati n T enic illi m: pi:tio- _ ili UT ir. :rut ient c ""'l t

'""oluti ns .

J.irjrior T',esis in J.l"ycology

~iur' i tt ec to

T _e Vir inia Pol ,.tcchn.'..c In::;titute

By

J:" es F. '"" ea.rt .

1i' • tle Jr.aster f C"cicncc Ds::-ree in Plant :?~t,

and HorticuJ.t u·e

June 10 , 1 21

ppr -e •

..

. .

Snzy e ctivity of Xyl r~a n~~it ...... ta.

'11his · a_ e.t~ is a re:i;.ort of t -e results of an exporirrental .

study o:L the enc-y:rratic _:_.ctivities of t 11e furgu.s "''"yl ria :iir:itata

which ca' ses the 0lack root rot of the a:pple tree .

A pure cu1 t re o_ the fur•"l.J..S .wac secure, b,. trco-nsferin"· a l. o -

tio~ of a culturP, - r~ -iJusly isoJ~ter, to a pet~i {is~ ~J1~ in- t~F

funn:1rn to cleYelope c.nd t .e:ri tra.ncferinq: a :portion of the outer

edO'e of the .o-rowth to a tu1Je oi' a""2,,r . The growth was examin .... f at

tirce nd he:n l"O cont.:..rinatior r'leYe~0pec" it w.;.s then re ,y for

cul tur in rr in sa' dust .

Sawdust of ail ...... pple tree w·as r l~,ce" ir1 se1-en 'Erhlenmyer flasks

of 250 cc . capacity . The sawdust 1:vas then roistPne~ 7itr nistille

water and S!.11 the flasJ-s a-11C:. contents were stC'··~ilizea at tvrenty- fb:e

pounds p- ess11re '!·o·c a· out 45 ""inutes . 'l' e -PJas 1 '"C ...... nr- contents were

then c clcd <:vnd inoculater with tne f11n"L1S :Zro_ t e ~-,1 ·er-- en tnros

I!'en+ioned aboYe . "'t -Pirst it was -e .... 1arr to risti· ....,1is an;-

:irowth in t18 flus}rs but ::.fte:." a fP. ;vr-eJrs a v;11iti~l1 rr.ow.._h aipeared.

o:n tl1e SU..L"face cf the sa 1dust anc1 :f'in-"J ly the riyceliur c0 ll ·)e

seen sc tte;:-ed throuO'l1 out the sa.v1 nst .

wee· s t,,..e i'l_.s :s ''lerr-- er~ tiea .J.nc t A sa ., rnt mycr-liu,..,, ve.1.: '·:ief

out ave a ratiator tE,' ing care not to :. n the tpv· c=r_t1 e o -pr

about 30° C. Ji'te.c cryiri7 , trn r.:iatw"'L .. l vms rr:conrr" ir1 a "'"Ortar

with ...... pestle, care.bein- t~ken not to c use too uch ·eat to • e

P-e orated 'l)y f'riction . This rrouno , cr:y ate_·ial ·will be terPe ·, in

thL... d e _ , "r: celial r.ceal" . Fror1 t .. is "myci= 1 i~l re 1 n ·;ere rrac" e

ext.c...;.ctirrs 1-no·:m ...... s "r~7 celi-11 c"is:._e1·vior::::" . It consistC'f o.L afti:r..g

- 2 -

50 cc iar~ ( 35° - 40° C) cis+ill~r w~ter to ten

allo·,1in.,. it to ci~0st, usually ove1~-ni t dt -oJuene a~·ea to prn-

vent bacteri~l action . rI' he susperic1 cc ana the filter .rashea rith 50 cc warm (35° - 40° C) risti11e" '~ter .

The filtrate w~s t e "yc8lial eis:·prsie n" .

First 10 cc of the rcycelial e ispersio n w...,, ~ l .. cc· in 50 cc of a

1% starch (l'erks solu,ble ) and 1% Sucrose solution , the st_,, ch _,,r

SucroSP solution beinrr used as a s 0strate . L c1eeJ:: 1-'"~S also pre-

parea. sin"" 10 cc o::' ycelial c is~ e:.~:::i c, n in 50 cc c"L:,tiJle ·Is.tee.

The three above so utions were testec af'ter about an hou-,.. aYJ"' t 1en

afte~ ~tincing over ni ht . C'~nsi ste" •

of a.aino- 3 cc Fehlin.o- 1 s A , 3 cc Fehlino- ' s B , 6 cc water sn alrnut

6 cc of.the Stbst_.nce to be testec1 u.:nC:.. e~~-in~:: fo1~ a o·t :five n:in1tes .

· ree ppt . for 0 t rch colution an· a e"'dish-yel ow r t. for sncrose

solution indicated the pl~esence of inve tase am: iast- e . The

check p.:o.ve re recucti n .

Cy3tase

Test for cellulose splittin• enz~o wa 0 ~ae as follows :- ~5 cc

cycelial rispereian were J ce~ in ~ra J fJ_c~ r a f 6 ! cc o-P' ,_, e • ·y

cellulose·~ STensian w~s ad e a luo sore t o luene . Thjs Nas

tested for re~uci~~ sugars by Fehli1: 1 ~ test at 24 r., 48 r . and

weekly intervals fo·r about three weP1'::S . rrhe reaction o: t 1 e ..:oJution

was also rlotenr:ined by titratin"' ac-ainst N/10 \i\ 2So4 an N/10 ra0fr .

t.!'or,-,. reduction w s shown j_n u.11 e:x:a inatiors ' 1 it e s 111t-i n s e

1 t . .._ · a·)o11t neutral . rr 1is s no crian - in :~e._.,cti n , the so n ion uein _,_

abecnc of any or _nic acids in the cli....::_e.t.:>io n W1C' tho rre ence of rc-,uc··

inf surars .

-3-

I11al t o. e .-

J.. teet for tho presence of the enzyn-e ·altoee was next r'ar,e .

To 20 cc of a 1'1; Daltose solution 10 cc "r:-~ celial c is:persici.T!n was ""' - ed

ancl 8 tu)eS ue-Y:e :.re a1'eC: . Te 1% altose solution w2.s a C'ec to eac1

of :four tubes an~ these ue1~e :i,,l ced in b ilin..,. water fo1~ ~i -e :nute ..-,

5 cc rryceJ ial a is_,_-ersi o n o.C.de:e ane eatin.o- c .nt inue, f'or ..t:'i -e ore

r.1inutes . The tubes ·were corled and t o luene affec as an ..... ntiSP!'tic.

T ese tubes were rarled "checks" . lialtose arc r·.·celial ris:;_rrsi r ,

•repared t be P'inninP" of parap-ra}!h 1 was ad ed to each of four tul) es,

t oluene u.c1 ded. anc cbeclrs and re r:uJ ars were incub ..;,tee t ~5-40° C.

Osazone test usini;r phenylhyclr Q\z ine -hycrochloride at first s1cwe •

or:ly !:'altos cr~rsta1s present . Tests wer.'e :rrarie at five aay inter,-als .

A ve"'y sli....,..1t forMation of dextrose crystals ( lucosazoPe) at t1- en

of the fifth day was evident in the resrulaJ.~ )Ut not in t:re control.

The 3d ai1d 4th (10th and 15th clay) test showed t e ""lucosaz ne to he

n10re :pronounced in the reO"ular but none were ::resent in t e cont :ol.

This showed t w. t t le enzy e !"'!alt ose , i.rhich h....,s the power of c owre::ctinn-

altose sugar to dextrose su ar, was rresent in the cis.i.:ersio n.

Enmlsin

1l1he test for e ulsin was next carrie ~ out . Four tubes oac o:f

o-L'!c....crclyliri , arbuti ar.d salicin were pre:pare ~ 9.C foll ws : Ten cc o:f

a 1'% solution of each o:f the abo·e s1 stances were placed in each

of 4 test tubes akin a total of 12 tubes for the 3 s1"'.:istanoes . T ··~

set was aaaea 4 cc :f "~e. ei.s~ e~~sion" c;una three tubes of eac :n e

tube was boiled forabout five ·i • es . To the ot .er tube of each

set 5 cc of c' istillea water wa::., ac1ced ei.s a control . r_ro 11 t 11 e tnhes

t o. .. 1Uene ·Nas ace ed . The .....,oove tu, es were incnb "t e' at ab ut 30° C . and

were tested at a out weekly intervals \!it l 111ehlin ' s solution . Jfter

-4-

the fi:1 st week tip two ~e ""Ul .... rs of .;;.rna 0 dylin c...rc arl;utin ·.reciuceri

~ehlir ' ~ •ut salici s~ wed n .e~uction at t is ti e . Te oile

and c cnt rol tubes showed no -f ec"ucti n . t the enc of t rec -re A ks incu-

bati-::n the s CLl ici~ tubes C"fft..e reC1c 1 ion r.it:b Wehli11""" ' s . rrr.ese tests

shov, that· the enzyr;e emulsin is present in the Xylaria Dig:itata FUY!o-1rn .

o ic a..se .

te.;:;t for the enzyr::e oxic1 Qse vms next mace . To 100 cc of . 01%

vanillin solution 20 cc of n;y-celial cisrersian were ·1aoeo -11d t e luene

was added to .... .revent bacteriul act ion . This was test e irTedia tely

ari-d at 3 da .. • inter -G.lS 1i'or a,)Qut t _ree e]·s , ""'or the -rre .. e ce of oxi: c:l,se.

The test was 'ade as folJows : .A :fev• cc of the above soluti n was pour-

ed in a test tube s :re acid nitrate of 0 0-,..cury rear-ent vms ad er anri the

tube ana controls heated for a few minutes . If oxieose is net pre -

sent thQ soluti n turned a re,, co r s owin.o- that the Tenill in ha -not

been oxidized to vanillic acid ; on the ot_er ~re , if t, e solutior

di not show ti1e red color the pre...1e:rice of oxif C\se w ich. oxiC'izes · 1e

vQnillin to · nillic aci6 is s~o\m . I n all t .e tests the re ence

of vanillin ias shown by t e rea color developin~ . T1is shows the

oxic o.se waf., not prerent in tl.,e ftui.r-us .

The ~ction of the enzy es upon Proteins and t1eir cleava~e pro-

ducts was next testec •

Protease

The ... resence of this enzyr:e can 1 e shown by it,.., acti n on fi1J ... in

' - :-e ·iously stainEC Vi ith car ·o rec and fixed in boiljno- water . If t 11('

enzy e is presert a rer coloe is liber:..utec" into t A sol11 t i ,., . K'o 1r

tubes ( 50 cc Cc;,f•~ci ty) were .. repare ..,S foll· VS :

1- control, 25 cc H20 ~ rpa fi1rin . 2- 10 cc yceli~l is· rEsio n , 15 cc H2o t re fibrin.

- 5-

3 - 10 cc . ., CP,lLil rispP. ,.,si on , 15 cc H 0 , cc IJ/10 H2

... o4 t- ,.,,,..,t fi ~riri

·2 4- 10 cc n " 15 cc H20, 1 c c 1r/10 F'... OH + Tf Tf

_t five day interTals an attempt Wdb .:.i.de to ecti· ate the ~ 0 nt of

l"'¢te lysis occur.ci:q ir t e ii.:ferent tu1rns , by car~" rir r t1-Je color

density . T):ie tubes , 1owever , d i e n t s11ow an~r c'i!' .... P'!'f'rCPS eue pe·"~1 aps

to tl"e exce s st ... in riot i-1a ·in ...... been t orou~ Lly v:a_, ea nt f t 1e f'ibrin

before p·"e:_arin~ the tubes )so the presi:o.nc e of t 1is enz:v1~e c uJ' not

be ete:..r ire Cl. .

ErE:']?Sin

The ethoa f Ir!a illP' t~rn test for Ere .. sin is to C' r :ri nstra te the

presence of try:ptopl1 0.ne in the dis1 e::sian . 10 cc .ycelial -i spensi on ..

w~s cded to 50 cc of a 1% solutioL ; ~eptoDe . To tl is soluti n

t &l uene was acJ.ded and the solut i "n VJaf, pL.cefl in inc1ib9. ti r . Tc sts , fo r

t r pto11ane o.t two day interY::.l s , :for ---1•011t t 1· ee VlAeY.s vere !!!a e.

T'1is wa8 doe y edcin...,. a :fe' ~rO)S o:!: ~cetic ecid to 'a ew c c

of the so 1 u ti n , b o i 1 i r -· it r ' t 1 en e. a f i r -· f P "- d r o

w ter . The soluti~r o-e, ~e no :est f'or tryptopbane . Tests were a l so

,aa e upon c3.Sein. Two n-rc-I"'S c::.sein aA ea tc 15 cc N/10 l'Ja OH, the

susrensiar.:. t oro11""hly sti'i.~red ciluted to 60 cc anf fi1te1~ea . 1 0 cc r r'P

enz~rr e dispe-csia.n was a ded to t e filtrg,te , t :nene ac1ce" - c incn-1 I

b tad s ~ ove for ~e tone . Tests were d e at two ·ay intor~als

as in .... e:Ptone solutirn e.n at t P en( of three 1veeks a rirJ: co .,., ·• '

s r;win"" t1 (' pre ;nee of tryptophane , was e ... ~icent. T is srows

the pre .::ience of the enzyr e e_ 01 sin.

rridase

iOr testir- t ::: ..tresence of this enz~ e t1e ction o~ t e ~ a..isper...;i .o..,,, u.n ""c""x:-: ..; : _.nn ..... s_,ara. .,.in VIcvS tester . ~ n e1t.... c ·irent wa ~et

up usin- fur 1 • ·e tul)es _Jt'A:f'are" '.:- follmrrn :

-6-

1- .. 2 P'"C . a la.'li"·-11 + 30 cc H20.

2- . 2 HH 30 H20 10 ".,.. u.lu.~ in i: cc ; cc r:ycrlial cis . Si n

-~ . 3 - . 2 O"r . c..1.sp~ra.O'i:n -t 30 cc H20

4- . 2 ,.,.r . " "\ .30 c c H20 t 10 cc yceJ ia.l ?ispecsian.

The.Je tnbP..., were testec with !\essler ' s rca"'ent _t four ~a~'" i ter-~·g,ls f r

the: for ation f rr: onia :'ron alarnin ana as ara~in . 1 one of "he tests

gave eviclence f tlle p::-e sence of arr.ic _se .

Sunrr-a.£1::·

Tests ca.I·~ied u,t to deterrr · ne t 10 r>nzyr a·'-ic acti ·i ty of t rn

fun 110 Xylariu. Di ·itata sho1.1ed the :preserce of

iastase ' InV· .ct_SP ,~ 0yt8.E.e ~ I. ltase ,'-Br ulsin anc "Rrepsin . '

Oxidase; protea.Se ann Lr id ...... ae -re re not f :n:U1C1 to l:i e nre ~Ent .

trian.o:u.l_r b'rster for tes,.:ir.'" ut 4 l p

fertilizer expe ril!.'er.1t s 1 s been v ell ccsc·ibee bJ-- '"'chreiner ~nc kinner

in vol. 10 , ro . 6 of thE Journal of t 1e .rl.ITeric n Qociety o:f .:.' O'ronomy .

It is the J::.iUri:;o:....e of t is paper t 1"i -e g, re: ort o; w i:k r r.ne 'l np:

this line, 1:..,in.,., inste .... c of fertLizerc ,_ nutrient s""lt soluti ns ana

~rowin,.,. t ·1e con-.ccn r1oulc Penic illim instead 0f la ts aL was done y

Schi."'eine· anr ckinner . In other words work siwi lar to t 1at carrier ut

in t e fi elC: With plants w ,...., carried out in the lahorc..tor~ uit t 1 e

co non rould Penicillium.

In t e f llmvin iscussion t 1e t ria 1ple syste r i:::: fu.J ly ex-

pl ined ana outlined .

r_r ree nutrient stock solutions vere prere-rea bused upon the esseri-

tL.1 ele ents JIT , Y, e P. ,.,,or P. a stool: s lution f CaH4 (P04 )2. R20

was r · area b1 a i so..) .... 1 vin IJ' 1 . 776 o;rarrs 0£ the salt in lOOOcc ist ille ~ water . T e ... 1 tion carried os}h~te ( p 2e5 ) 1000 pp' • For

:r a stock solution f Nuro 3 wa.s :prepared by ( issoli.-inP 5 raITs of t ;.e

salt in 1000 cc , t is ives ~ ccncentr tiori of nitro~en ( in terns of HR~ )

of 1000 pp~ . In li~e erner as source of l tes (K20) 1 . 852 ra'DS

of r~so4 w~s eissol·ed in lOCO co of water ~ivin~ a c - centr~tinn cf

1000 pp ~hese etock soluti rs were filute? to te~ ti e~ i ~-in · a K O. 2

con centre.ti n of 100 pr.; of t"1e nut ient elcr: ent . 1J:ne s lut i rs uere

~raded in 10 percent st~aes ri inry ~ totQl if 66 tPcts int e e e~irent .

To bear in· ·nc: all t1 e...,e C'i.J....r.P..'ent ra+i s is ::1rost an i· sNi"l-)iJit;-

eo "" di a a sl' owin"" t11e r i ffe:r:ent points etc . , 1"'as 1ee:r:: p era re ,

This triangle represents c.;.ll t 1"'e os ..... i"le !'atios cf 10% st "'es .

Eacb extrer:e :poirit of t1"'e e n ·1ate·.a1 tris.nrrle ref""C~Prts 100 % of

-2-

P205

, IIB3 orK20. Teter "lOC :.·c~ce:ritn refer~ t t-ie si.xi m: rl.nt·ty

of solution used ~ I •• lOC cc of a lOC p . p.r. scJ~+i n . Ttc sires

of t e triano-le v.re i •if ed into 10 ec11al ,,~rt.:i arri lines corriectin

these point"' 11&Ye l)een drawn. For c m;enience t .e intersi::>ct ons of tlrnse

lines ve bePn rn be re • T'""'hle 1 ,..,.i es the ra. ti 0'.'!is for the t ree

constituents . By refei~ence to the c me.ti tue:nts f :::oints in ta')le 1 d

studyini: Figure 1 the ra.tirs fo1· CLTI;- rortion o~ the t".'iangle ca e ce-

te • inea .

Sup ose we consiaer tJ1 e 1.ine rep:ceser:1tinp: t'h.e lL.Se oft e trian"·Je,

then the. point vvhich represents lOO;'b K20 (Point 56) will -=--ye no NH3 nor P

2o

5, 1:· 1 e1i·ise the ~oint~ repr0':::leritinrr 100% NR3 (Point 66) an

10a% P2o5

(P int 1) will -aye reither of the otller consti ti ents present .

It is plainl · SePr t ~t tJ·e e:...teere l ojnts oft a trian!)'le re· _esent

10o% solutions and th "'.: s the lines rro toward the oppoei te base t ey

decrea~e in lo% sta~es .

u pose we ta~e points :rearer to eit .er -Pt e c:orrers , + ere ·i 1

be a _1iabcr perce:rit .... '""e of ne constit ier1t ana a lower rercE'nte.""e o.L t 1e

others . Take poi. t 58 for exaLple , re ra e , 80~ K20 ~na 20% liffi3 ;

No . 3 P-ives 9Cf/o p2o

5, 10% NH

3; on the otlf"r hana if rointP are ·a'eri near

the cer-ter of tl·e trian-""le a r ore e u'""l ratio is sl'own . Point 33 P.:i\es

30'% p2o5 , 4CP/o NH3 nd 30% K20 •

..!or the E ~e of con ei ierce it w ... s deciaecl in r'""':.inn- t1'i3 test

to use sr all 51 liweyer flasks ( 150cc). ± n ar para tus conrir. ctin°· flas :s

conte.i:r:in' .. t et ree pr ierly ril1Jter (100 PPM) salt solutions to 'nrnrcttes

was prerx:ired . The oiffe.:ent a' ou1ts of tlle vari~us sol··' i 1-is were then

added directl;:, to t c fJ a ks ~rd t'·es 'U"" e m1r:be".'ec, c ri...,ecuti"' oly . lOOcc

oft; e '8 solutions rer·esented io05b. ~lerefo~e , ii (as 'or- oint5 ) it

was desired t fl'et a ixtu- e of so% P205, 107& :NR3 ~nc1. lO'fa 20 it was

sir1ly a case of dra inp 80 cc of tr.c P20 5 solution , 10 cc oft e m3

- 3-

soluti"n ::..nc:. 10 cc f t 10 K20 ...,oluti r into t 1f -'"-'::_ ·~k . T i:::: s st"'!.' f

using 100 cc as :cep.cesenting 10Cf% faci itnted -atterc coroi 'r i·aJly.

~.fter the 66 flasl:s hc.u been :.. re_are , nur-berec a d rlli...,..,.er ;ith

cotton they were tlen sterilizec in the autocla.we st a lrj 1 onn s pressure

for ,ll ut 15 n-inutes . -~fter a lowing- the to c ol tJ- ey we1·e a.11

·noeuls,ted ·ith the Pe- icilliu pinop1ili~ . xar-ina ti ns ana note~ as in

the rapidity , c c.uracter , c lor etc . of rowt"' were ta1'"ATI at r'ail-r in -., -

terv""'ls but y the fir.al ei o-'1t of . O'Vt rlll he ni\ en in t is paper .

In the meantirie a 1votal of 66 · eir:J-iino· 1Jottles har" 1rneri prepare ,

rJroperly ·nu....,..,berea , 'li·ei(l'hed !1nd tr..e wei.o- ts ec r <=ic on a c1 i tahJ e or .•

flasl ........ , •r icl: jn s r-e cases consistE'd f lr. ct a..,, ~id ia..·0 f ,.,.rowth ,

t:1eir respective weir:hing bottles ana t11e ') ott es heatea. in a a_ryiri.c.r

oven . He t w:..s a1::--liee urtil no rrore Foisture was )resent in t e t ibes

and then they were J:laced in ressicators . 'ei~_.;_r.:""d 'e:- t_1 n rate ft e

bottle"' :::.nc c oritents end t ~e se wei ('Y'ht s Vi" re ·"'ec reed in t'1e f r "C!l-

tionecl a1)ove , in their ?esrective -~lacec . T,1e iffererce between t"e

\7eiP". ts of tho er.ipty tubes 11 the tubes contair-ina the tr~· myceliUT" ~a ·e

the mi hts of funPllS for the res~ ecti ·e flas'rs . Fi,ml!'c 2 ("i...-·es t};.e

\Veitnts f yce i !"1 aevclopinr- in the cl; fferent flasl-s, ir fO"!'.'

trian le s i -,, n in :Pi ure 1 .

T sun:"J.rize the results cf t e eY"'erir:erit it is nece::::117 t ~lre

a cal·ofnl stucy f 1i'i"'ure 2 . I n ora.rr to ceter ire · }•ic11 lers f the

outsice trian,.,.le Gave t e best {'rrowtl" ,a rera"·es o~ t "'e fun,..,,'l."' weiv1•ts of

e c _ o the t ee outer leo-s were r °' - e anc t _e wei hts were as follows :

0 ide 1 to 56 g-ave ave a e of . 0075 ('Y'r~ s f 1r0 us wt -ice 56 to 66

'.:'3.Ve a ·e al""e Of • 00 75 i:rra S fU.UJ:"LlS ,..,.rowt 1 , Si( e 66 t 0 } rra •9 a 'E'ro..1"''8 Of

. 0381 I" r:1s :fur us o-_cowth . T11i.s s1 wee. t,"~t t e 1e~ 1f t C' trianr-le c ,..,t-

-4-

T le weir- ts of t e E' ·s f t e rext

tri n"'· e 11ithin ·were then &YAra~::ed anr'' tne veiP-~11-~· ·1ere ar:: .(' llows :

QiC:e 5 to 47 "'3.Ve . 0708 rams funrrns ''T -rt., sice 47 to 54 o-a·e .lL'-:-5 ''ra s

funm1s l"l'rowth . iC: e 47 to 54 , which contair:v !:i-"'her percen ~"'" of K20

in this c_i,Se . This latter n:ve:-carre i-es 1. lit _e etter Wo"':'.'~inp· l:l-:.::iis

n ixt11 .. e o:f elJ three constituents an" t e: .. e-"' re

ives a better test .

Points 40 , 48 and 4S' (r;ith OY1;e exception ) si'ows the 11.i hest wei.rr __ t of

t -€ fun'"'11,;J -;;.TI a· e enclose' iri cotte• lires in FiPUre 2. rrhis s 1nws tha t

the a~ea of the best r vth is n~arest t e E20 corner of t e trian7le an

t at the solutions for t1:esP points co:ritain "l;.i,crher ... e· cer..taf""€)S cf K20

ar..d rrn3 and less p 2o5 • Point 54 shows t 11e hiP' est i.,veip- t but sorr-e

unaccou tecl r".ista.l::e has been :r.ic..c e anc is not inc uCer in t11e corr,_ris ns .

Qchreine ..... ana ckirnor foun" t~at the ncrral '"'T'O\Vth of wreat in

nutrient soluti ns ( nn b~sis of r .... een .. ,ei O'}it ) was best in YJUJ"'lers 41, 50 and

-5-

j__~~U.-.~~L-~~~~'>l!L-~~~~..lif,l!!o'--___l~~--!!E-1--~~~~bb /'I H3

Fi ".lire 1- ~a1•ilc~teral tria m lar dial"."'ra s , ir.P' points :rmJ':'bi:->re ; re -

p esent i · t e 66 nut riE'r .... salt c ' ina. ti 1.., in 10 E' c C"'nt sta es .

... " . -6-

.Ratio R2.tio or :percentai:re --Point c Point Co Dosi t ion.

o. P205 No. P205- :NH K 0 3 2

1 100 0 0 34 17Q 50 20 2 90 0 10 35 r;;o 60 10 3 90 10 0 6 30 70 0 4 80 0 20 37 !"'O 0 80 5 80 10 10 38 20 10 70 6 80 20 0 '?9 20 20 60 7 70 0 "'0 40 20 !?0 50 8 70 10 ;: 0 41 20 40 40 9 70 20 10 42 20 50 ;,o

10 70 '?0 0 43 20 60 20 11 60 0 40 44 20 70 10 12 60 10 30 45 20 80 0 13 60 20 20 46 10 0 ~o

14 60 30 10 47 10 10 80 15 60 40 0 4-8 10 20 70 16 50 0 50 Ll.0 10 '7Q 60 ~.,

17 50 10 4-0 50 10 40 50 18 50 20 30 51 10 50 40 19 50 30 20 52 10 60 ,...o 20 50 40 10 53 10 70 ~o

21 50 50 0 54 10 80 10 22 40 0 60 55 10 90 0 23 . 40 10 50 56 0 0 100 24 40 20 40 5'7 0 10 90 25 40 30 30 58 0 20 80 26 40 40 20 59 0 '30 70 27 40 50 10 60 0 40 60 28 40 60 0 61 0 50 50 29 30 0 70 62 0 60 40 30 30 10 60 63 0 70 '30 31 30 20 50 64 0 80 20 32 30 30 40 65 0 90 10 33 30 40 30 66 0 100 0

Table 1- The 66 possible ratios of the three nutrient salt constituents,

P2o

5, NR

3 and K20 given in 10 percent sta es .

. ... I' • - 7-

. 0038 . 0~83 ( 3 )

. 0046 . 0596 . 0358 ( 6 )

• 0028 • 0582 • 0784 • 0706 ( 10 )

. 0069 . 0 752 . 1083 . 0482 . 0491 ( 15 )

T0105 00523 . 0905 .1030 . 1175 . 0411 ( 1:1 ) .

• 0098 • 0669 .125f:: .14.06 . 1351 . 1320 • 0385 ( 28 )

. 0154 . 0819 . 0974 . 1187 . 1301 . 0537 • 0414 ( '7 6 )

"' ,,,. I

. 0112 . OPOB . 110§, < 1 56'71 . 1465 . 1396 . l'",78 . 0512 ( 45 ) ,,,.. I

,,," I . • 0094 . 0913~.J..4_:0_ .!..160~_1. 1179 . 1481 . 1454 . 1423 . 1926 . 06'76 (55 )

0007

K 0 2

. r:z9 •.0u69 . 0016 . 0047 . 0012 . 0057 • 0066 • 0007 (66 )

FR'3