Some pH Changes in Nutrient Solutions

Author(s): Emil F. Vacin and F. W. Went

Source: Botanical Gazette, Vol. 110, No. 4 (Jun., 1949), pp. 605-613
Published by: The University of Chicago Press
Stable URL: http://www.jstor.org/stable/2472666 .
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 SOMEPH CHANGESIN NUTRIENT SOLUTIONS
EMIL F. VACIN AND F. W. WENT

Introduction der othersit is necessaryto maintaina

In the courseof an investigationcon- narrower pH range.SincesolutionC is
cerningthe growthfactorsinvolvedin not renewed duringthe entireperiodof
germinationof orchidseed and seedling growth of orchid seedlings,it is funda-
growth,severalfactorswereencountered mentally important that the criticalpH
which caused fluctuationsin the pH range should not be disturbed significant-
value of the culturemedium.Sincethe ly by the absorption of chemicals from
changesin pH value weregreaterthan the solution, by the products of growth,
permissiblefor optimumgrowth,their andby externalmo]dor bacterialcondi-
sourceand correctionwerestudied. tions.
Generallythe nutrientsolutionC of lKNuDsoN (2) as well as commercial
Knudson (2) iS used in germinating growers expressed the opinionthat the
orchidseedsasymbiotically. critical pH range of the solutionwas be-
tween pH 4.5 and 5.5 for satisfactory
SOLUTION C
germination. Thisrangeforgoodgrowth
is shownin figureI.
Ca(N03)24H20 ........ I . OCG
(NH4)2S04 .............
gm. Immediately thequestionarises:What
o . 5oo gm.
MgS04 7H20 .......... o . 250 gm. is the buffering range of solutionC?Since
FeS04 7H20........... 0.02S gm. no published data were available,a buffer
MnS04 4H20 .......... ° °°7S gm. curvewas preparedand is includedin
KH2PO4 ............... o. 250 gm. figureI. This curvedefinitelyshowsa
tSucrose................ 20.0
Agar.................. I 8 .o
gm. weakbuffering actionin theoptionalger-
gm.
DistilledH20 .......... I COO . O ml.
mination and growth rangeof pH 4.5-
5.5.Thereforen it is unlikelythat the nu-
Germinationand growth of seedlings trientsolutionwouldremainwithinthe
continues,understerileconditions,from optimumpH rangeduringthe germina-
6 to I8 monthsbeforethe seedlingscan tion and growthperiodof I8 months.
be transplanted safelyto the morerigor- Basedon these initial observationsthe
ousatmosphere of thegreenhouse. Foran followingexperiments werepreformed to
optimumdevelopmentof seedlingsthe determine some of the pH changes that
nutrientsolutionshouldhavea sufficient mightsignificantly affectasymbioticger-
quantityof the necessaryelementsacces- mination and growth of orchidseeds.
sible for each successivegrowthperiod. Proceduresandresults
The availabilityto a plant of any par-
ticularchemicalin a nutrientsolutionis CHANGES INPH OFNUTRIENT SOLUTION
determinedby a criticalpH range,as C WITH GERMINATION AND GROWTH
shown by ARNONand JOHNSON (I). OFSEEDLINGS
WENT(3), workingwith tomatoplants, SolutionC waspreparedwithoutagar
foundthat undercertainconditionsthe by dissolvingeachchemicalseparatelyin
criticalrangemay be widebut that un- distilledwater.Immediately aftermixing
605
606 BOTANICALGAZETTE
[JUNE
the pH valuewas 4.92 at 25°C. At the Afterautoclavingthe solutionhada pH
end of I hour and 23 minutesit had valueof 5.46at 40°C.
changedto 4.74, and at the end of 3 The twenty flaskswere dividedinto
hoursand I3 minutesto 4.52. Afterthe two groups.No plantingsweremadein
agarwasaddedanddissolvedcompletely one group.The flasksin the othergroup
by heating,thepH valuewasadjustedto wereplantedwith seedsof Epidendrum
5.62at 40°C. A BeckmanElectrometric o'brienianum,whichhad been sterilized
pH meterwasusedin makingthe meas- in a solution of calcium hypochlorite
urementsto an accuracyof +0.02 pH, madeup of 70ml. of distilledwaterand5
and sufficienttime was alwaysallowed gm.of commercialchlorinated lime.This

VL. OF1N. KOHADDEDTO lOOML.OF SOLUTION

C
FIG. I. Buffercurvesfor solutionC with 0.025Wo and o.6W0KH2PO4and maximumand minimumpH
limits for goodgerminationof orchidseeds and growthof seedlings.

for the electrodesto reachthe tempera- solutionwas shakenfor a few minutes

ture at which the measurementswere andthenfiltered.The resultantclearso-
beingmade.Solutionswhilebeingtested lutionwas usedfor sterilizingthe seeds.
wereunderconstantagitation. At the endof 5 minutesthe sterilizingso-
SolutionC wasdividedamongtwenty lutionwas removed,and the seedswere
soo-ml.Erlenmeyer Pyrexflasks,ISO ml. washedin sterilewaterfor I5 minutes.
per flask.The flaskswereclosedtightly Seedswereplantedwith a sterileloop.
with cottonplugs.They had beencare- All twentyflaskswereplacedin a special
fully washedin acid, rinsedthoroughly incubatorwith automatictemperature
withwarmwater,andfinallyrinsedwith controls;nightat 22° C. anddayat 27° C.
distilledwater,thus removingall traces A smallblowerslowlymovedthe air to
of acidandalkali.AllIdasks in anysingle maintainuniformtemperature through-
experiment wereautoclavedat one time out the incubator.Light intensity on
at a pressureof I 5 poundsforI 5 minutes. cloudydayswasapproximately I50 foot-
I 949] VACIN & WENT-NUTRIENT SOLUTIONS 607

candlesand on sunny days 500 foot- not contaminated withmoldor bacteria.

candles. Resultsareshownin figure3.
Periodicallyone flaskeach fromeach In sterilizationby autoclavingI liter
groupwas opened,and the germinating of solutionC withoutagarwas divided
seedsand seedlingswereremovedcare- among ten 2 so-ml. Erlenmeyerflasks.
fully fromthe agar.The agarwas then The solutionsin the flaskswereadjusted
heateduntil thoroughlydissolved,and
pH measurements weremade at 4o°C.
At the end of I90 days the pH value of
the agar on which the seedlingswere
grownhadchangedfrom5.46to 3.78.In
the next 87 days it droppedto 3.I2, a
total changeof 2.34 pH units. The pH
valueof the agaron whichno plantings
weremade changedonly slightly,from
the originalpH value of 5.46 to 5.23at
the endof 277days(fig.2).
This exploratoryexperimentshowed 100 200
several significantchangesin the pH NUMBEROf DAYSAFrER PLANTlMG
valueof solutionC whichresultedfrom FIG. 2. pH changes resulting from germination
(a) standingat room temperature,(b) and growth of seedlings of Epidendrum o'brienianums
autoclaving,and(c)germination of seeds on soluLionC.
and growthof seedlings.
l l l g b

CHANGES INPHRESULTING FROM 7

AUTOCLAVING
SOLUTION C WITHOUT AGAR.It was 6

apparentfromthe previousexperiment
thatit wouldbe advisableto compensate 5
for the changesin pH value resulting
from growthof seedlings.Only sterile 4
>

:n
, INITIALp H

nutrientsolutionscan be injectedinto
the sterilegerminatingflasks.Steriliza-
tion couldbe accomplished by heatorby ao G FlLTERED
o 40ML.OF SOLUTION
- -

160
| A A

2'0
SUCCESSIVELY
a Seitzfilter.Thelattermethodwasdis- THRUSAMESEITZFILTERDISS
cardedowingto erraticpH changesupon FIG.3.-pH changes resulting from filtering solu-
filtering.In filtering,solutionC without tion C with Seitz filter in 40-ml. batches.
agarwaspreparedand dividedinto five
flasks,40ml.perflask.In thefirst40 ml. with eitherhydrochloricacid or potas-
filteredthe pH value was changedfrom sium hydroxideto coverthe pH range
4.92 to 6.5I. Withoutchangingthe filter 3-9. The flaskswerethen closedtightly
disk, the remainingfour flaskswerefil- with cottonplugsandautoclavedsimul-
tered,andthe pH valuesweremeasured taneouslyat a pressureof I5 poundsfor
beforeand afterfiltering.The solutions I 5 minutes.The changesin pH valuere-
usedhadbeenfreshlypreparedandwere sulting from autoclavingare shownin
 BOTANICALGAZETTE
608
figure4, curve2. Twosignificantmaxima
of changeoccur,onebetweenpH 3 and6,
the otherbetweenpH 6 and9. Thesere-
sults werereproducedmany times, and
every time the two maximaldeviations
at pH 5.5 and 8.o werefound.
Tests weremade to determineif un-
sterilizedsolutionC left standingwould
undergopH changessimilar to those
causedby autoclaving.Oneliter of un-
sterilizedsolutionC without agar was
5 6 7
STARTINGp H
9
FIG. 4. pH changesin solutionC without agar
resultingfrom (I) standingunsterilizedat 5° C. for
96 hours; (2) autoclavingsolution C with °.°25So
KH2PO4; and (3) autoclaving ion C with o.o6%
solu4
KH2PO4.
STARTlNGp H
FIG. 5.-pH changesin (I) solutionC, 0.02sSo
KHzPO4and without agar, when autoclaved;(2)
solutionC when agarwas added and pH measure-
ments madeat 40°C.; and (3) solutionC with agar
when autoclaved.
[JUNE
dividedamongten 2so-ml.Erlenmeyer
flasks,closedtightly with cottonplugs.
The solutionsin the flaskswereadjusted
to cover the pH range3-9. The flasks
wereleft in a cold roomat 5° C. for 96
hours,with no moldappearing.The re-
sults of pH measurements are shownin
figure4 (curve I ), showinga gradual
changetowardthe acid side. Moldfirst
beganto appearafterapproximately I20
hours.Comparisonof the data for the
g6-hourstandingtest (curveI) and of
the effects resultingfrom autoclaving
(curve2) indicatestheirsimilarity,auto-
claving producingthe changes more
rapidly.
SOLUTION C WITH AGAR.SolutionC
withoutagarwas prepared,testingpH
4.9Iimmediately aftermixing.Tenflasks
wereadjustedto coverthepH range3-9.
Aftertheseadjustmentsagarwas added
anddissolvedcompletelyby heating,and
pH measurements made at 40°C. Re-
sultsareshownin figure5, curve2. The
flaskswerethenreadjusted to theirorigi-
nal pH value beforethe agarhad been
added,afterwhichthey wereautoclaved
andpH measurements made.Resultsare
shownin figure5, curve3. CurveI was
addedto showthe pH changesresulting
from autoclavingsolution C without
agar.
SOLUTION C BUFFERED WITH0.6S0
KH2P04. The bufferingactionof solu-
tion C without agar was increasedby
usingo.6%KH2P04insteadof 0.025%o.
This concentration of phosphateis near
the maximumlevel toleratedby the
seedlings.TheeSectof theincreasedbuf-
feringactionis shownin figureI. The
bufferingeffectwithinthe good growth
limits of pH 4.5-5.5 is approximately
fourandone-halftimesas greatwith the
addedbufferas before.The pH changes
resultingfromautoclavingare shownin
figure4, curve3. In thepH range3-6 the
 25 ..

I 94 9] VACIN & WENT-NUTRIENT SOLUTIONS

609
changesare approximatelythe same,ir- distilledwaterand eachchemicalin the
respectiveof bufTerconcentration,but samepercentageas used in solutionC,
for the rangepH 6-9 autoclavingpro- and each solution was bufferedwith
duced smallerchangesi.n the solution TABLE 1
witho.6%0 KH2POthanin the onemore
weakly bufTered.Upon autoclavinga Temperature
whiteprecipitatewasformed,whichwas pH pH pH
(o C)

moreabundantabovepH 6. In the range 40............ 3.89 6.66

4 38
pH 6-9 theprecipitatewasgreaterin the 35 ........ 3.9o 4 39 6.72
morestronglybufferedsolution. 30.......... 3 .9o
3 .9 I
4 39 6.78
6.83
4-43
VARIATIONS INPH VALUE RESULTING 20............ 3 .9 I 4 45 6.90
3.92 6.98
FROM TEMPERATURE CHANGES. Solution I3I0 .....
............
4 47
7 .02
3 93 4-49
C withoutagar was heatedslowly,and
TABLE 2
EFFECT OF AUTOCLAVING ON PH VALUES OF SOLUTIONS

SOLUTIONS ADJUSTED BEFORE AUTOCLAX'ING TO PH VALUE OF

SOLUTIONS
H20+0.025°o KH2PO4
ADDED TO
3-° 4.o 4-5 5 so 6.o 6-S 7.0 8.o 9.o
5-5

(NH4)2S04 -O . 02 -O . °4 -0. I0 -o. I8

-O. 2I -O. 2I -°. 2I -O. 27 -O. 63 .79
MgS04 7H20 ...... O O +o.o8 o.o8
+O. OI o o O -O. IO - o.67
MnS04@ 4H20 _ O . OI + O. I4 +0 . 02 +0. I9 +O. I4 o +O. OI -O. OI -O. 27 -o.88
Sucrose . . . C O o -0.09 -O. IO -o c8 -o o8 -o. I4 -°-49 -I .32
FeS04-7H20 ... .. +0. 02 -O. 2I -o . 50 - o.85
-o.o8 -o o8 o -O . OI -O . 30 -I . 09
Ca(N03)24H>0 .... -O.OI +o.o4 +o.o3 +0. IO o
-O. 92 -I .24 -I .69 -2.30 3 .o6
SolutionC ....... -O. OI -O. 22 - o 54 -0.92 -I . I5 - o 53 -I . 44 -I.7I I.99 -o.60

pHmeasurements weretakenat different 0.02s% KH2PO4.The followingis the

temperatures. Table I gives the results. compositionof one of the specialsolu-
Thedifferences arenot significant.
It can tions:
be assumedthat, whensolutionC with- Ca(N03)24H20.......... I . 000 gm.
out agaris testedforpH value,the tem- KH2P04 ................ 0.025 gm.
peraturefactor does not producesig- DistilledH20............ I 000 . O ml.
nificantvariations.No appreciablevaria-
tionsresultingfromtemperature change After each of these solutions was
wereobservedwhenthisexperimentwas adjusted to a range of pH values
repeatedusing solution C containing (table 2), each was autoclaved.For
agarin a temperature rangeof 30°-40°C. solutionsof (NH4)2S04, MgS04 * 7H20,
MnSo4* 4H20, and sucrose,autoclav-
INVESTIGATION OFEACHCHEMICAL ing producedvery small changes in
OFSOLUTION C pH value. However,for FeS04 * 7H20
Two differentfactorsmay be respon- solutionoriginallyadjustedto pH 3 to 6,
sible for the changein pH value after autoclavingproducedchangescompara-
autoclavingsolutionC in the pH ranges ble to those producedin the complete
of 3-6 and6-9 as shownin figure4, curve solutionC, andonlyvery smallchanges
2. Specialsolutionswereprepared,using whenthe originalvalueswerepH 6 to 8.
 BOTANICALGAZETTE
6IO
In Ca(N03)2-4H20 solution
at pH 3-6 very smallchangesoriginally nally acid, no precipitateforms after
[JUNE

occurred,
but, whenthe originalvalueswerein the autoclaving,and (b), as shown later,
rangepH 6-9, the changeswere com- whencomplexironsalts are substituted
parableto thosein the completesolution forferroussulfate,thereis no appreciable
C.Allsolutionsoriginallyat pH g changein pH valueresulting
showed claving. The complexiron fromauto-
significantchangesafterautoclaving. phosphates
Thesedatasuggestan explanationfor whicharepresumablyformedwhenthe
the two-maximaldeviationcurve originalsolutionC is autoclavedseemto
inthe changeof thepH valueof found be muchless effectivefor the growth
C resulting from autoclaving, solution theplant thanothercomplexiron of
when salts.
plottedas a functionof the originalpH Thisis indicatedby the smallamountsof
ironcitrate or iron tartrateneededto
preventiron chlorosis,when compared
withferroussulfate.In KNUDSON'S solu-
tionC,theironsulfateshouldbereplaced
byone of the organicironsalts.
At the originalpH 7 and8 the dropin
valueafterautoclavingis connectedwith
theformationof a flocculose
precipitate,
apparently of calciumphosphate.Since
suchhighpH valuesare not withinthe
optimal growingrange of orchidseed-
lings,no correctivemeasuresare neces-
sary,althoughsomeattemptshavebeen
STARTI
NG p H made to discoverthem.
FIG. 6.-pH changes
resultingfromautoclaving
solutionC without agar; (2) solutionC
(I)
when ANUTRIENT SOLUTION NOTAFFECTED
FeSO4 is replacedby Fe2(C4H406)3; (3)
when Ca(NO3)2is replaced with Ca3(PO4)2
solutionC BYAUTOCLAVING
KNO3; and (4) solution5. and
It hasbeenshownabovethat the iron
value. andcalciumcompounds of solutionC are
Whereasat pH 3, 4, 4.5, 6, and9 altered
the mediumbecameonly slightlymore affect sufficiently
during autoclavingto
acidafterautoclaving, the pH valueof the solutionover
at pH 5.5,7,and8 the
entireoriginalrangeof pH 3-9. This
theacidityincreasedmorethanonepH
unit.
This two-maximal undesirableeffectwas overcomeby sub-
deviationcurve stitutingother iron and calciumcom-
wasfoundin everyexperiment,indicat-
ing
thatin thetwomaximalrangeseither pounds,as shownby the followingdata.
acomplex IRON COMPOUNDS. A numberof iron
cationformationoccurs,thus
reducing the ionizationandtherebylow- compounds were employedto replace
eringthe pH value, or a precipitateis FeS04
*7H20in solutionC. Specialiron
formed of a strongcationwith a rather solutions
wereprepared(table3). In each
weak instance
anion. The former explanation tion the same millimoleconcentra-
seemsto holdin the caseof the depres- of iron was maintainedas in the
sion
at pH 5.5, wherethe presenceof original
solutionC.
ferrous sulfate apparentlycauses this From the data of table 3 it may be
drop.
For (a), whensolutionC is origi- seen
that solutionsof ferrousphosphate,
ferric
ammoniumsulfate,ferriccitrate,
I 949] VACIN & WENT NUTRIENT SOLUTIONS
andferrictartrateshowedno significant substitutedin solutionC. Thevariations
pH changesresultingfromautoclaving. in pH value resultingfromautoclaving
When0.025 gm. of ferroussulfateis re- when the above changesin solutionC
placedby o.o28 gm. of ferrictartratein weremadeareshownin figure6, curve3.
solutionC, the millimoleconcentration The changesin pH valuein the original
of ironis approximately the same,thus rangeof pH 6-9 werereducedsubstan-
still maintainingthe desirableratio of tially. In the originalrangeof pH 3-6,
3: I of ironto manganese.
Whenthis sub- however,a significantchangeoccurredas
stitutionwasmadein solutionC, no sig- comparedwith solutionC, the pH value
nificantvariationsin pH valueresulting risingafterautoclavinginsteadof drop-
from autoclavingwere observedin the ping.Howthis changein calciumsource
originalpH range3-6 (fig. 6, curve2). will affectthe availabilityof ironto the
CALCIUM COMPOUNDS. Sincein solu- growingseedlingsis now beinginvesti-
tion C the effectof autoclavingon cal- gated.
TABLE 3
PH CHANGES RESULTING FROM AUTOCLAVING WHEN DIFFERENT IRON
COMPOUNDS ARE USED

SOLUTIONS ADJUSTED BEEORE AUTOCLAVING TO PH VALUE OF

SOLUTIONS
H02+0.025% KH2P04
ADDED T O
3 .o 4-° 4-5

FeS04 7H20 .... .. +0 .02 -O. 2I -O.

FeP04 4H20 ....... . o +O.C4
FeNHa(so4)2- I 2H20 o -o . o3 -o . :
FeC6HsO7 3HaO. . . -o.o3 +0. OI
Fe2(cJH4o6)3 2H20 . +0 . 0 I +0.02 +0.< IoO

o7

ciumnitrateis apparentlyresponsible for PROPOSED NUTRIENT SOLUTION 5.-

thepH variationsin the originalrangeof Basedon the information obtainedfrom
pH 6-9 as indicatedin table 2 as com- the above experiments,a new solution
paredwith figure4, curve2 othercal- knownas solution5 wasprepared.
cium compoundswere considered.Ter-
SOLUTION 5
tiary orthophosphateof calcium was
usedinsteadof calciumnitrate.The solu- Ca3(P04)2 .............. 0. 2¢0 gm.
KN03................. 0.525 gm.
bility of this compoundat 25° C. iS 0.02 KH2Po4 .............. o . 250 gm.
gm. per I00 ml. of water.This should MgS04 7H20.......... o. 25e gm.
supplythe necessaryamountof calcium (NH4)2S04... . ... 0.50C gtn.
that shouldbe availablereadilyto the Fe2(C4H406) 3 2H20..... 0.028 gm.

growingseedlings.To maintainapproxi- MnS04 4H20.......... 0.0075 gm.

Sucrose............... 20.00 gm.
matelythe samemillimoleconcentration Distilled H20 .......... I OC0 . 00 ml.
of nitrateprovidedby calciumnitratein Agar(flake)............ I6 .00 gm.
solutionC, potassiumnitratewas used.
The ratio of potassiumto calciumwas The changesin pH values resultingfrom
adjustedto 5:I. Insteadof usingI gm.of autoclaving this solution are shonvnin
calciumnitrateper liter, 0.200 gm. of figure 6, curve 4. These differencesare
Ca3(PO4)2 ando.s2s gm. of KNO3were smaller than those for solution C. Fur-
 (+) cysteine

6I2 BOTANICALGAZETTE
[JUNE
ther studies of solution 5 are continuing, glutamic acid was added, the value
especiallyin relationto the germinationof droppedI.50 pH units; for cysteine,
orchidseeds and the growthof seedlings. 2.00; andforasparticacid, I.42. If these
solutionsare not then readjustedto the
CHANGES IN PH VALUE RESULTING FROM originalpH 6, the rateof growthfollow-
ADDITION OFAMINO ACIDS ing may be significantlyatTected.Such
TOSOLUTION C solutionswould drop in pH value as
When any one of several amino acids growthcontinuedand thus reachmore
in a concentrationof o.os%Owas added quicklythe limits wherethey couldno
to solution C, to which had been added longerefficientlysupportgrowth.Since

TABLE 4
PH CHANGES RESULTING FROM ADDITION OF o.o5% AMINO ACIDS TO SOLU-
TION C CONTAINING o.o5 P.P.M. OF FOLIC ACID AND o.5 P.P.M. THIAMIN

Initial Adjusted pEl after

Solutions
pH pH autoclavillg

C.............................
Solution 4 75 6.oo 5 5o
C+folicacid+thiamin..........
Solution 4 72 6.o2 5.5o
1(+) glutamic acid 3. 25 6.oo 5.60
+ dlnorleucine 4 7o 6.oo 5 53
.= 1(-) leucine 4. 8I 6.oo 5 5°
E3 glycine AP. 4.89 6.co 5 48
,, 1 (-) hydroxyprollne 4.72 6.oo 5.42
2 79 6.oo 4 37
+ dlserine 4.82 6. CI 5e5o
t 1(-) cystine 4 II 6.oo 5.65
ce k 1(-) histidine 4 34 6.oo 5.92
.e) 1(-) tyrosine 4 7o 6.co 5.42
O d (-) arginine 4. 7I 6 .o I 5-58
+ dlvaline 4.78 6.oo 5 58
_> dltryptophane 4 48 6.o2 5 65
= dlalanine 4. 2I 6.oo 5 65
.° d lysine 4.68 6.oc 5 49
:5 dl,B-phenylalanine 4.52 6.oo 5.52
O dlaspartic acid 3 33 6 .o I 5 54
v) <dlmethionine 4.82 6.oo 5 4I

o.os p.p.m.of folicacidando.s p.p.m.of the solutionshadto be sterile,it wasnec-

thiamin,changesin pH value wereob-
servedas recordedin table4. Thesesolu-
tions wereused in studyingthe rate of
growthof Rhizoctoniafoundin the roots
of certainCymbidiumorchids.However,
only the observedvariationsin these
solutionsareconsidered in this paper.
The additionof only o.os%O of amino
acidto the solution,as shownin table4,
in someinstanceshad significanteffects
uponthe pH value.For example,when
essaryto autoclavethem.Afterautoclav-
ing the cysteinesolution,the pH value
droppedfrom6.o to 4.37. It was neces-
saryto readjustthe solutionto pH 5.50
by steriletechniques.Varioussourcesof
amino acids produced different pH
changes,so that it becamenecessaryto
test carefullyforpH valueat everystep
in the experiments.This preliminary
work indicates that amino acids are
definitelybetterbuffersthanphosphates
I 949] VACIN & WENT NUTRIENT SOLUTIONS 6I3

in the goodgrowthrangeof pH 4.5-5.5. whataminoacidswillbe satisfactorynot

This workis being continuedto deter-
minewhichof the aminoacidsmay act
as goodgrowthfactorsas well as satis-
factorybuffers.
Summary
I. The culturemedia used generally
for the asymbioticgerminationof orchid
seedsareso weaklybufTered that, as the
seedlingsstart to grow, the pH value
drops below the level for optimum
growth.Evenwhenthe phosphatebuffer
wasincreasedtwenty-fourfold,the buf-
feringactionwas still weakin the opti-
malgrowthrangeof pH 4.5-5.5,indicat-
ing that phosphatebuffersare unsatis-
factory.An investigationindicatedthat
aminoacids are excellentbuffers.This
work is being continuedto determine
onlyas buffersbutalsoas growthfactors.
2. Autoclavingthe culturemediawith-
out agarproducedsignificantchangesin
pH value.Similarchangesin pH value
were observed,only at a slowerrate,
whenthe solutionsstoodfor sometime.
These resultsare equallyapplicableto
ordinary nonsterilizedculture media.
The changeswhenthe originalpH value
was 3 to 6 weredue to the presenceof
ironsulfate,whichshouldbe replacedby
oneof the organicironsalts;whenit mras
betweenpH 6 and 9, the changeswere
due to the presenceof calciumnitrate.
Based on these studiesa new nutrient
solutionis proposed.
KERCKHOFFBIOLOGICALLABORATORIES
CALIFORNIA
INSTITUTEOFTECHNOLOGY
PASADENA4, CALIFORNIA

LITERATURE CITED
I. ARNON,D. I., and JOHNSON,
C. M. Influenceof germinationof orchid seeds. Amer. OrchidSoc.
hydrogen ion concentrationon.the growth of Bull. I5: 2I4-2I7. I946.
higher plants undercontrolledconditions.Plant
Physiol. I7: 525-539. I942. 3. WENT,F. W. Effectof the rootsystemon tomato
2. KNUDSON,L. A new nutrient solution for the stemgrowth.Plant Physiol. I8 :5 I-65. I943.

INHIBITIONOFPHOTOPERIODIC INDUCTION IN
XANTHIUM BY APPLIED AUXINI
JAMES BONNER AND JOHN THURLOW

Inkoduction longerthan the critical(morethan ap-

It has long been knownthat many proximatelyI 52 hours of light per 24
plantsgrowvegetativelyat a slowerrate hours)to a regimeof photoperiods short-
when subjectedto short photoperiods er than the critical but, in addition,
thanwhensubjectedto longphotoperi- apicaldominance is lessenedor eliminat-
ods. In cockleburtXanthium),a short- ed. Sinceboth reducedgrowthof stems
day plant with regardto inductionof andlessenedapicaldominance aresymp-
flowering,not only is vegetativegrowth toms which may be associatedwith
abruptly checked when the plant is lessenedauxinsupply,it is naturalto in-
moved from a regimeof photoperiods quire as to the auxin relationsof the
Report of worksupportedin part by the Her- plant during photoperiodicinduction.
I

manFraschFoundationfor AgriculturalChemistry. Assaysof auxinin a varietyof speciesby