li."r Sxcl.I;~iw:\.t=-o;·~ Q! "t,L8 :cz'.~:.:·t :-0J:'cl."":! tl or; of ;·:.:tl-"!.e ':'ree .:>:tI'
an,::
1.. ts ::0;::'.i):":·3·~
Ot:-.0£
::,c:.
'\~eal'by
w: th
th;,"t of
;T:;::i t.ats
--
· i·
4'+
I) :'urPOSE
This was
hacit3.ts
3,
study in
~·uitable
identify as
ye~l..st
thei~ pro:ifera~-ion.
for
rr:[l:-~y yeast~s c.~s
yeast pop1J1atloY'ls
Growth in ar. er.vironrr.e:-!t that COtl,tai.ned many
th~t
~f;€
attempted to if:olate ar:d
posslbl1; frOp.l three serarate habitats.
Her.:! corr;raTcd were tho:::e foun·l in
and honey, all frolT; the 3a'ne
prop~'rty,
~hus,
~aplo:)
The
S8P. daffodils,
thi..s was a study ir. yeast
ecology.
:::1) Descriptior.
A ye.".l.Y seforce' the sta"!"'t of the ex?erir!lerlt t I'1:l;le sap was 6btaj ned frorr.
somf~ "':,ree~3
Tndia::a.
ex~
-:"l;
'1
p:"nce rJf rropt.:rty
is part 0: a "classIfied forest" in
The saD cOLtain.ed both bacteria. eJnd yeasts.
st i:l the
~;3.p.
Tr.~
ot·jec;tlvD of the
cOIr.pared with those :In
!3~u,dy ~Tas
''1 c£1diti~r:
tify yeasts of ttis hahitat.
wer~
t~at
o-tlv~r
to tbeir
;.earby ta"b::tats.
to
Five
d~
stiEct types
charact€r:~,;e
and
iden:if~catinr:.
ide~-
"the yeasts
These habitats were or.
the sarre property as the maple trees,
wer~"
C-:1C pla.ce yeasts
may te
yespor~si
tar from the
£'ou:::.d
l·r8.$
1!", the honey of a Lear::;:;
"hI
_' e as a vector ~1. or these yeas +. . 5. 1
(~aff'odils
£J..r from tf:e :3ap.
to make hOD€:Y.
Thus,
~.he ~€8S
Insects
The bees :nay obtair. nt)c-
!'::iybe the 'tJees would also obt20.ir: su-
could transport. the yeasts from tr:€' sc_p -tc
the r;or.ey a."1d even to the daffod:ls.
The 8ees couJ.d tra!1;""3port :tes.sts
h~.ve.
f~C:I1
r:'t:1S l)!."ocess could
~lso
''{o.!''J" i:l reverse.
the hive to the sap.
Tte ,;?ro??rty that thr: ye.:lstc ",ere taker, f-"'om is a classified forest of
:::::;.d~ar.u..
celle!·~t
Th~_.3
lar.d
j
s ke)t
i:~
It.s :Dtural sta-l:.e and so : t offe:.-:ed aI', ex-
l,lace where n:ttural r,r!.c:::-ob:ial ecology could be studiE·d.
;:'::1£', was
:::l
:~C'w pxper~e:lce
for me as yeast work -waG :l8H to
':'i1~;;.
:'he
2
T'erhalls at 3.r.other opp0Tturll ty. thc:;.(? ",1 11 ~e a cha.!";ce to co:niJle~e the work,
(;spp-c:ally
I:I)
tlF:~ ide:Ltlf:cat~on
of
al~
of :he species l.so1atr::d.
Fr'~C2c. ure
A) Isola-:'~o[: of yeasts f!."oTIl sar
;.;"3.S the.n usee.. tr..' k:".11 the cacterl:.L'l cuJ.o::~ os so
or,
P2.g8S
1-3.
3) Isolatio:: of ye8.sts fr,')m lla!,fodlls
were
::-::.d~~
of va.rious
conc8~:trat:.o~13.
!.r2.thin two days, the plates
~.J'e!'e
fulJ
-
s(;.p8.!:'ate c:olD::ies ~)y ~L:.·e.:t~{:'nc. a:,d re~~treClki.::.s 0nto 3A'Q plates,
gr.-.:.:::ts of yea:.:;t,~ f::::-o!f! daff'J;} lIs
Honr:-]
trick
~'Ta:
gt·:t·l:ir~g
obtai.led f:::'om
th.:.s v:'scnus
8.
:.LTC
r,ivc or;. the
s'J.bsta.:.c-~ O!~-t,~
Five differe::f; 8,:7lourt:-: of hor:ey
::'oi-tic-'.:ly, yea.3ts grew o:lly
cO.'":t~d CAd
~J:
~.:!:·oth
sa/'~e
plates
ur-
D~
-
a:.-:'8
~!l
It was 3.ctu.al1y
a measurable amo·,.l.">1t.
~)ur8
hOl:ey.,
I found that COl1r.1erci.s,lly. ho-r;ey is
8.T:.d put. in brntr: to shclke.
the c.est r18th..,d of c'.Jtain:!_g ye:lsts frorr:
from honey
p:!"operty.
the plat'3 w],th 10ml of tLe honey hrcth r-1att'd.
hgaterl te' f53°:~ for 3C mi..n. to) kill a!'lY yeasts.
';ro~:er.
on .pages 4-5 of the aprend~. x.
we:t'e rlated clor..,:; with
enoll(;h, the c,)ld h'.Jr.E-:Y j-ielded yP.e::.::.;.ts.
w:c;re
rhotom.:ic):'o-
coptai ~~l(:d. i.'1 pages
7-15
If
Sev'2!''"-1
ho~ey.
conc~1;;.tri:itio:.s 'Jf
rhot~micTograp~s
broth
of yeasts
tIl the ~ppe:1dix.
Tsolat:"o:1 of yea.sts fro1"! .1.pples
used previously.
:r~:
is yeast ex-t.r:1.ct- malt E.'xt:-.:''ict ag3.r whi8h
cor,t~2ns
4
(;>er Ii ter; 3gms. ma1 t extr:"{ct, Jgms. ye:].~t extract, 5gms. peptoJ'Tl€. 10c;ms.
gl ucose. and. 20gros. a.gar. 5 The same cOClcentr"tions were plCl.ted onto apple
juice agar
a IT.ore
Yi
1. th pc;-;.icll1in.
The
differe~~t rrl0Qiu.':1s
were usnd in hoper;. that.
]'(l.edi1l1T. :night heIr isolqte yeasts.
~,~pecific
Cn the apple jUiC8 1{;'1r, tlle only growt'J waS bach'rta1.
'J"'vtair.e5.
0:'1
the
yp~ ac:a.Y.' -:)!it
t.he~
I{,)ll'. d cot
Yeast" were
G!'o~;r wher~ trans:ferrf~d
This cot;ld have bee:1 dlle to use err old (2 m0~lth) :J'-1 sla!~ts.
to YN sl a~ tt.
','Ie were already
a 1 ittle behInd schedule so ;.:['2 left t!-'le arples and conL:.nued wl.t:1 thc other
yeasts,
s) Culture pJ:-eparation
All of -::.he isolate'!
tj
me there wer,,:
yea3t:-~ ~,.;ert~
3 tubes of each 0::
restrBaked to IDal::tBi!1 a
?~
the;. put Or.
s~ l.nts
yeast lsola.tes.
fres~-: col1ec-t~_0L.
0:
Y;,! agar.
At t!1ts
Ther;e Here :;erioiica"'Lly
The col1E'ctlon was kept .in tte
refrigerator.
F) Ide':"'_t:i"f3.catio~:
slide.
tinct t3"tr',,,s,
-~.,he
di"fference be-:ne twice the ,size.
n.y color alo:1e there i.ere
2) Sue;:tL' test~;
h
'!'he procpdure foll."")wed was that of ::"ootier,'-
were p:epareJ in
-
tU.883
(fil ter s-:'er:'lized).
of Yeast
"l"t;')'~eY'
-'
• t.> .' ~··'<::e
.-''-.....
Cultures for ir:.ocu13.tio!"l
(""l"fcQ'
in-~""I-. J " " cQn+-...
,,,"--"",,.. · · 0
~~
-I"
,--1u('O<::"'L>
-'
.... ,-,
L"-
Aftpr gro'(i"th, +J:/-2se cultures '"rere used to inocula-: o
7
~;terlilzed~' •
tuccs containec 2.?% 0:f thE:
;,/h
were 8.bou-~
Wj.S
IT,;;'
The
r...-.l ,.
-',----,;
wIde.
(- ~ •
~
;'(lslt:v~ rrhile ~+ was or.ly WE'~lk~y ~}oSiti·/E-.(
,!,:~.e key us';.:' if. id.c::~-:'1::;_c~~2.0-::
~':al tose
[Y'0wth tes~~
:e11:_):-/10::(:
=r~rtr.ri toJ_
[~ovtb
(+'
, ,
(+' R::cfi:1oSE-~
~
, I
:~rt
,I
~
?o.rt
"
hay€ ~UJ.'tLr:T -:c-;.,;t:s ;::0::12. u
J)
C::-owtt te f) t
(-)
(+) Cc~lOb:.OSt_~
,
;
Tart
.L.
test
~1!()t()7.icrogr~-..phs
I
,
\-
~
?art F
~:.l..rt
-:'hi~--, 'h"3.8 :1.5 f2T 3,::;
,,+',
T
~,Jt
'; __ tL,'
?1:'JCf-;d-':.lT~: "'::_eca'ls,~'
I
\
,
,
(
1 ",
ref8TS tc :;, C')-:";"~,_8 of' w:::.;r; -:'h:lt they ;'l?-;'{ be dl.sti-'lg·~.lisht.c:'."'"'-
'::':;.e ts t.h::>.t _~_n
••
~ 1,,;,,
was uses...
~he s~. :"des
'7
,
~esult.s
frO::-i S'J.LaY
:;l'.1cose
~A?;T
"l)ay rca.J.
.."i.- ,
12-29 1-12
~:C')-i.-t:~
"':ubes
:':0.1 :'-·)SB
:\:<:1r..r~l.,t.ol
12-29 1-12 12-29 1-12
1+
1+
2+
,~+
2+
3+
2+
,,+
.-
J+
3+
2+
}t
3+
3+
,'1·t
J+
3+
3+
2+
;+
1+
)'+
3"t
3+
,"+
3+
J1'
3+
3+
3+
3+
3+
J+
2+
]'T
2+
1+
3+
1+
J+
,"+
3~
1+
3+
2+
3+
3+
1+
3+
2+
-
1+
1+
1+
3+
2+
3'"
J+
3+
1+
2+
3+
2+
3+
2+
'h
-
3+
,'1+
!: -'j
3+
3+
}l
2+
3+
r:-1C
2+
1+
:':-12
2+
J+
3+
:-:-13
2+
---
3+
3+
3+
3+
3+
,,".-...
2+
2+
3-3
.::;-'}
-r:
c:
-'
""-?
3-7
3+
2+
:;+
J+
1+
3+
3+
~..:.
.-'
3+
3+
3+
2+
1-·
2+
3+
3+
2+
.I'
"
The ye3.st:3 ",ere inoculat.ed
:)r-!.
31-
J+
)'T
3+
2+
,"+
',~b.e!1 exa;:,.i~e.i aJ63.il:
;t
.,,
}~
A
-'
? ..
r.
;+
.,
J+
..,.~
"
'"
"
1+
1+
2+
,1+
2.,.
"
2+
J+
2-t
c
2+
c
2-+
c
..,-
A
3+
3+
3+
3+
1+
J+
2+
2+
J+
2+
J+
J+
3+
3+
"
3+
..,...
.-'
3+
3+
J-t
12-19 so th2....t read.:"'rlgcl ·,.,ere t:.tkeL a:'ter
gro',';, it was prssw;;.ed to be bacter::'a,
\
,,+
;+
to '!'a!:t
12-29 1_1"
2+
2+
J-2
12-29 1-:"2
1+
:·.-h
1
12-29 1-12
Ra!f~ f', :15,:;:
1+
2+
~.
'.!.- .... .(.
SucrOEB
J+
1+
~'~-~!
12-29
,
~ello:i'Jsc;
2+
.1
/
=:':rytl:.l.~':' t'~l
~.~ C1r!Q 2~-t
2+
.)'
C'
~
21-
1+
days,
2-+-
A
Si;lca 3-7 iid Eot
microscop::'cally, it 'lfas i:1deBd cacte:r-ia,
(
ra.ffi~os,,~.
':'~:1e
next
~;te~ ~':{)-11d.
be to cont:!.Due to parts A a!"!.d C
~t~'1d
d.o tb0:
V~' ::!oncluslo!':
asout yeasts.
There 1s :jot
3. :T.aSSiVE
compl:!.cated this 1 S th8-t \'arj ,")us
tification.
cultures
-
!~,ust
r:e~ded,
be userU
~Eld
had
},f
?~
ttP.,n
C);.
J"e3.zts, 'h,ut I :::::houJd
is:i.ve d ~_ffe!>er!'~ tech:::iques i!1 iden-
SOUTeS's
It would have been bent
all tbe :fl'3.ter:.a13 '
amo':.1Tt wr~
"!:
~1 an~ed
thj::; out
pr..;cis~:;lJr,
ktlm...-r.
definl te pyocAdure before: startel.l.
?er:n3.ner:t ';111 tcre.s sto-J.lf 1:·e kep:'
sepa!'!~e i!"~
c::.se ;:.
-
•
Yeasts from Maple Sap
!·!-1
:'age 1
8
•
Yeasts from Haple Sal'
Page 2
,,
o
o
()
Yeasts f:rom Haple Sap
~1-10
Page;
•
•
o
()
C
Yeasts fron Drtffodi]s
D-l
-
«)
•
Yeasts from Honey
~-5
Fage 7
-
U
Yeasts from Daffodils
D-2
Page
5
,:
-
Yeasts from Honey
3-4
Page 6
-
~8A.S"t~
from
'1-6
~0!18y
Yeasts frop'o Eoney
J-6
Page
9
,.
~",.
... f: ,
;1
~
•
'.
.." , . ,...
•
,,
•
,
,
,"
or
Yeasts fro" Honey
3-6
Page 10
•
•
,.
\",.... }~oi
"
•
'>-_.'
•
Yeasts from Eoney
,,-6
Page 11
'
....
;.
•
". ,
l ";' •
,
"
"'
•
!
Yeasts
fro~
1-6
?age
~2
Honey
•
Yeasts f!'ol'l Honey
3-6
Page
13
•
Yeasts from Honey
3-6
,.
-t,.
")'-T."';7)~'~"'?~
-
• ' ........
1
Pl. . ,.tff, H.J. et 301.
?ress, 1978. :? 211 •
Th~
_ ... L.....
.....J....;
Li:::'e of 'ff;astb
Garn:1rldge, Eass: Ea.l"V2.rd Unl v.
.,
. . Icdder J. edt
The Yeasts
T.o~donl X()rtb-~ol13:nd
?u:'lishir:.c Gom:::)ar-.y,
197::'. F. 38.
4
!';crse, ~oLer A. ed.
SOT;;,;.a~.l T,~~·~iv. ~res~, 1972.
<
- I..c-d.der,
6
7
reid.,
"
0
10
11
'"\'"
""oJ.
r • ..
:p.
Ieid.
p. 42
.;... ,-,-~.,
p • ltL.•
:::icl.,
p.
Tl-.~ f'~
rred.ators, ar:d Jis€3.-se L-:-
_:~o:~:
?-o.
J .A., ?nd R.J. "Pc:."
::::"s..: ';:.8r ~ubl':s~i":_!~':': c;).,
:'()dder~,
~ee
?ests,
-- ---
r. ;9.
::ar:~et-L,
.\~eric3.~-.
:-:cney
l~5.
to 1.::<: Yeasts
Ym::k: