Environmental Factors that Affect Seed Germination
Alyzza Noreen O. Orogo BS Biology II AH
Jly !" #$%& !'%(
A scientific paper paper submitted submitted in partial partial fulfillment fulfillment of the requiremen requirements ts in General General Biology II Laboratory under Mrs. Winnie N. Camigla ! st sem. "#!$%"#!&
IN)*O+,-)ION
Germination is the resumption of gro'th and de(elopment after a period of dormancy )*oefnagels "#!+,. It may be defined as a series of e(ents 'hich ta-e place pla ce 'h 'hen en dry qu quies iescen centt se seeds eds imb imbibe ibe 'a 'ate terr re resu sulti lting ng in an inc increa rease se in metabolic acti(ity and the initiation of a seedling from the embryo. In order for germination to be initiated the follo'ing criteria must be meet )!, the seed must first be (iable )the embryo is ali(e and capable of germination,. )", Appropriate en(ironmental conditions such as a(ailable 'ater proper temperature o/ygen and in some cases light must be supplied. )+, 0rimary dormancy in the seed must be o(ercome )Arteca !112,. Germination 'hich is the beginning of gro'th of a seed depends on the interplay of a number of internal and e/ternal factors. In order to germinate a seed must first be (iable )ali(e, )3tern "#!!,. "#!!,. When conditio conditions ns are fa(orable for the gro'th of a particular seed germination )sprouting, begins. 4he ability of seeds to germinate is called (iability. 4he conditions fa(orable for germination include )!, a suitable temperature temperature )bet'een !&
℃
and "2
℃
, )", plenty of
moisture and )+, sufficient o/ygen dissol(ed in 'ater )Capco and 5ang "#!#,. Germination depends upon imbibition the upta-e of 'ater due to the lo' 'ater potential of the dry seed. Imbibing 'ater causes the seed to e/pand and rupture its coat and also triggers metabolic changes in the embryo that enable it to resume gro'th. )Campbell "#!!,. Basic necessities in order to commence
IN)*O+,-)ION
Germination is the resumption of gro'th and de(elopment after a period of dormancy )*oefnagels "#!+,. It may be defined as a series of e(ents 'hich ta-e place pla ce 'h 'hen en dry qu quies iescen centt se seeds eds imb imbibe ibe 'a 'ate terr re resu sulti lting ng in an inc increa rease se in metabolic acti(ity and the initiation of a seedling from the embryo. In order for germination to be initiated the follo'ing criteria must be meet )!, the seed must first be (iable )the embryo is ali(e and capable of germination,. )", Appropriate en(ironmental conditions such as a(ailable 'ater proper temperature o/ygen and in some cases light must be supplied. )+, 0rimary dormancy in the seed must be o(ercome )Arteca !112,. Germination 'hich is the beginning of gro'th of a seed depends on the interplay of a number of internal and e/ternal factors. In order to germinate a seed must first be (iable )ali(e, )3tern "#!!,. "#!!,. When conditio conditions ns are fa(orable for the gro'th of a particular seed germination )sprouting, begins. 4he ability of seeds to germinate is called (iability. 4he conditions fa(orable for germination include )!, a suitable temperature temperature )bet'een !&
℃
and "2
℃
, )", plenty of
moisture and )+, sufficient o/ygen dissol(ed in 'ater )Capco and 5ang "#!#,. Germination depends upon imbibition the upta-e of 'ater due to the lo' 'ater potential of the dry seed. Imbibing 'ater causes the seed to e/pand and rupture its coat and also triggers metabolic changes in the embryo that enable it to resume gro'th. )Campbell "#!!,. Basic necessities in order to commence
germination may be affected by some factors 'hich inhibit the gro'th of the seed. 4he researcher hypothesi6e hypothesi6e that if the optimum le(el for each of the basic requirements )e.g. 'ater temperature osmotic concentration and o/ygen, for germination is met then a high percentage of germination 'ith longer roots and shoot sho ots s cou could ld be yi yield elded. ed. 4h 4his is co could uld be der deri(e i(ed d fr from om th the e act acti( i(ity ity co cond nduct ucted ed 'herein best results of germination transpired from set ups 'hich pro(ided the soa-ed seeds 'ith enough medium 'hich enable it to sprout 'ith longer shoots and roots. 4he study aims to meet the follo'ing ob7ecti(es )a, -no' some physical requir req uireme ements nts for ge germi rmina natio tion n )b )b,, -n -no' o' the op optim timum um le( le(els els of th the e phy physic sical al parameters under 'hich normal germination can ta-e place and )c, -no' some chemicals that affect seed germination. 4he e/p e/perim eriment ent 'as conducted conducted at the Micr Microbio obiology logy Laboratory Laboratory of the 3outhern Lu6on 3tate 8ni(ersity in Lucban 9ue6on from ": th of ;uly up to +! st of ;uly "#!$.
A)E*IA/S AN+ E)HO+S A. )he Need for 0ater
4hree )+, germination trays lined 'ith paper to'els at the bottom 'ere secured. It 'as then labeled as containers A B and C.
esults 'ere recorded in 4able !%A. B. )he Need for O1ygen
4hree )+, "$# mL ?rlenmeyer flas-s 'ith cor- stoppers and attached hoo-s 'ere secured. 4he flas-s 'ere mar-ed as A B and C. 4he follo'ing solutions 'ere placed in each of the follo'ing flas-s A "$ mL of "$@ * "$ mL of "$@ pyrogalllic acid B $# mL of "$@ * C $# mL distilled 'ater
0yrogallic acid and potassium hydro/ide is a caustic mi/ture so eye shields 'ere used and the liquids 'ere handled 'ith care. After'ards a piece of cotton net 'as obtained and 'as lined 'ith a moistened pad of cotton. 4'enty )"#, mongo seeds 'ere placed and tied to ma-e a bundle. 4hree )+, bundles of mongo seeds 'ere prepared. 4he bundles 'ere hanged to the hoo- fastened beneath each cor-. It 'as made sure that the bundles did not touch the solutions. 4he flas-s 'ere co(ered airtight. 4he flas-s 'ere left for a 'ee- at a room temperature. 4he number of mongo seeds that germinated in each set%up 'ere counted. 4he percentage of germination 'as computed using the formula
germination
=
number of seeds germinated × 100 total number of seeds
4he length of the roots and shoots 'ere measured in millimeters )mm,. 4he a(erage and other data 'ere recorded in 4able !%B. -. )em2eratre and Germination
A t'enty )"#, one%day old soa-ed 'ith mongo seeds 'ere obtained. It 'as rolled up in strip of moist paper to'el. 4he roll 'as put in a plastic bag and 'as sealed. 4hree bags 'ere prepared 'hich 'ere labeled A B and C respecti(ely. 3et%up A 'as placed in the refrigerator )about :
laboratory room )about "$ ℃
℃
℃¿
B 'as left upright in the
, and C 'as placed in an incubator set at +2
. 4he set%up 'as e/amined on the se(enth )2 th, day. 4he number of seeds
germinated in each treatment 'as counted. 4he percentage of germination 'as
computed. 4he length of the roots and shoots 'ere measured in millimeters )mm, and its a(erage 'as ta-en. 4he results 'ere recorded in 4able !%C. 4he gro'th
of
the
seedlings
'ere
compared.
4he
temperature)s,
that
fa(or)s,Dinhibit)s, germination 'as e/plained. +. Osmotic -oncentration and Germination
Nine )1, 0etri dishes lined 'ith paper to'el and mar-ed from A to I 'as obtained. 4en )!#, mL NaCl solution of the follo'ing concentration 'as placed to each 0etri dish A #.## )distilled 'ater, B #."$@ C #.$#@ E #.2$@ ? !.##@ < ".$#@ G $.##@ * 2.$#@ I !#.##@ 4'enty )"#, mongo seeds 'hich 'ere uniform in si6e and health 'ere placed in each plate. 4he set%ups 'ere left under room condition. n the second day the set%ups 'ere chec-ed by slightly opening the lid of plates 'ith
germinating seeds. 3olutions 'ere to be added if necessary. n the fourth day it 'as obser(ed. 4he number of seeds that germinated 'as counted. 4he percentage of germination for each treatment 'as computed. 4he lengths of roots and shoots 'ere measured in millimeters )mm,. 4he results 'ere recorded in 4able!%E )Camigla "#!!,. *ES,/)S AN+ +IS-,SSIONS A. )HE NEE+ FO* 0A)E*
Figre A.%.a.Soa3ed Seeds& -overed 4ith 0ater *eslt of Gro2 %
Figre A.%.5.Soa3ed Seeds& oist /ining *eslt of Gro2 %
Figre A.%.c.+ry Seeds& +ry /ining *eslt of Gr o2 %
Figre A.!.a.Soa3ed Seeds& -overed 4ith 0ater *eslt of Gro2 !
Figre A.!.5.Soa3ed Seeds& oist /ining *eslt of Gro2 !
Figre A.!.c.+ry Seeds& +ry /ining *eslt of Gr o2 !
Figre A.$.a.Soa3ed Seeds& -overed 4ith 0ater *eslt of Gro2 $
Figre A.$.5.Soa3ed Seeds& oist /ining *eslt of Gro2 $
Figre A.$.c.+ry Seeds& +ry /ining *eslt of Gr o2 $ )a5le %#A6 Seed Germination and the Effect of 0ater
)reatment s
Gro2 Nm5er Soa3ed seeds& oist lining Soa3ed seeds& -overed 4ith 4ater +ry seeds& +ry lining
Nm5er of Seeds Germinate d N7%( % ! $
8ercentage 9:; of Germination
Average /ength 9mm; of *oots
Average /ength 9mm; of Shoots
%
!
$
%
!
$
%
!
$
!+
!!
!$
F&.& 2
2 +
!# #
$&.$ :
"+
22. &
++.+ F
"1.$ F
:F.& 2
#
!:
+
#
1 +
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#
+$.1 +
:!
#
:+
2+.+
!$
!:
!$
!##
1 +
!# #
$
".21
F.& 2
$
2.21
:.&
B. )HE NEE+ FO* O<=GEN
"$ mL of "$@ * "$ mL of "$@ pyrogallic acid
$# mL "$@ *
$# mL distilled 'ater
Figre B.%.Seed
Germination and the Effect
of O1ygen *eslts of
Gro2 %
Figre B.!.Seed Germination and the O1ygen
Effect of
*eslts of Gro2 !
"$ mL of "$@ * $#$# mLmL distilled "$ mL of "$@ 'ater pyrogallic acid *
Figre B.$.Seed Germination
and the Effect of
O1ygen *eslts of Gro2 $ )a5le %#B6 Seed Germination Effect of O1ygen
and the
mL distilled 'ater "$$# mL of "$@ "$ $# mL "$@* * mL of "$@ pyrogallic acid
)reatments
Gro2 Nm5er !( m/ of !(: >OH ? !( m/ of !(: 2yrogallic acid (' m/ of !(: >OH (' m/ of distilled 4ater
Nm5er of Seeds Germinated N7!' % ! $
8ercentage 9:; of Germinatio n % ! $
Average /ength 9mm; of *oots
Average /ength 9mm; of Shoots
%
!
$
%
!
$
!
#
!!
$
#
$$
&
#
#
2
#
:."2
!1
!1
#
1$
1$
#
&."$
#
#
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:.:"
#
!1
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!1
1$
!## 1$
"2.1
+#.F
+&.#+
"2.$
"+.&$ :$.!#
#
$
-. )E8E*A),*E AN+ GE*INA)ION
Figre -.%.a.Seeds in " ℃ *eslt of Gro2 %
+
Figre -.%.5.Seeds in !( ℃ *eslt of Gro2 %
Figre -.%.c.Seeds in $@ ℃ *eslt of Gro2 %
Figre -.!.a.Seeds in " ℃ *eslt of Gro2 !
Figre -.!.5.Seeds in !( ℃ *eslt of Gro2 !
Figre -.!.c.Seeds in $@ ℃ *eslt of Gro2 !
Figre -.$.a.Seeds in " ℃ *eslt of Gro2 $
Figre -.$.5.Seeds in !( ℃ *eslt of Gro2 $
Figre -.$.c.Seeds in $@ ℃ *eslt of Gro2 $
)reatments
Gro2 Nm5er
Nm5er of Seeds Germinate d N7!' % ! $
8ercentage 9:; of Germination
Average /ength 9mm; of *oots
%
!
$
%
!
$
%
!
$
!F
1#
F#
!#
#
#
#
1.2"
+
2.#$
+$.#$ $&.+$ :!.+1
!&
"#
" ℃
"#
"#
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!## !##
# &#
!$
!2
!2
2$
F$
"1
Average /ength 9mm; of Shoots
!!.!$ 1.+: +:.1"
!( ℃
F$
+$.F
:$."1 +#."F ":.& "!.$+
$@ ℃ )a5le %#-6 Seed Germination and the Effect of )em2eratre
+. OSO)I- -ON-EN)*A)ION AN+ GE*INA)ION Figre
+.%.A. '.'': and B. '.!(: of %' m/ Na-l *eslts of Gro2 % Figre +.%.-. '.(': and +. '.@(: of %' m/ Na-l *eslts of Gro2 %
Figre +.%.E.
%.'': and F. !.(': of %' m/ Na-l *eslts of Gro2 %
Figre +.%.G. (.'': and H. @.(': of %' m/ Na-l *eslts of Gro2 %
Figre +.%.I. %'.'': %' m/
Na-l *eslts of Gro2 %
Figre +.!.A. '.'': and B. '.!(: of %' m/ Na-l *eslts of Gro2 !
Figre +.!.-. '.(': of
and +. '.@(: of %'
m/ Na-l *eslts
Gro2 !
Figre +.!.E.
%.'':
and
F. !.(': of %' m/ Na-l *eslts of Gro2 !
Figre
+.!.G. (.'': and H. @.(': of %' m/ Na-l *eslts of Gro2 !
Figre +.!.I. %'.'': %' m/ Na-l *eslts of Gro2 !
Figre +.$.A. '.'': and B. '.!(: of %' m/ Na-l *eslts of Gro2 $
Figre
+.$.-. '.(': and +.
'.@(: of %' m/ Na-l
*eslts of
Gro2 $
Figre
+.$.E. %.'': and F.
!.(':
of %'
m/
Na-l
Gro2
*eslts of
$
Figre
+.$.G. (.'': and H. @.(': of %' m/
Figre %'.'': %' m/
Na-l *eslts of Gro2 $
+.$.I. Na-l
*eslts of Gro2 $
)a5le %#+6 Germination and Effect of Osmotic
Seed the -oncentration
)reatments
Gro2 Nm5er +istilled 0ater '.'':
Nm5er of Seeds Germinate d N7!' % ! $
8ercentage 9:; of Germination
Average /ength 9mm; of *oots
Average /ength 9mm; of Shoots
%
!
$
%
!
$
%
!
$
"#
!#
1$
1$
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$2.+
22.:
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!.(': (.'': @.(': %'.'':
+IS-,SSION
A. )he Need for 0ater
three groups. It could be infer to the fact that it ga(e the optimal amount of 'ater for the mongo seeds. 4ruth be -no'n that 'ater is one of the initial necessities before a seed could germinate. According to Miller McEonald H'ater is a basic requirement for germination. It is essential for en6yme acti(ation brea-do'n translocation and use of reser(e storage material. In their resting state seeds are characteristically lo' in moisture and relati(ely inacti(e metabolically. 4hat is they are in a state of quiescence. 4hus quiescent seeds are able to maintain a minimum le(el of metabolic acti(ity that assures their long%term sur(i(al in the soil and during storage. A lac- of 'ater during the germination process can reduce the germination percentage due to 'ater stress )Eoneen and MacGilli(ray !1:+= *an-s and 4horp !1$&,. ?/posure to e/cess 'ater results in the production of a substance 'hich reduces o/ygen supply to the embryo and ele(ates inhibitory substances in the seed 'hich reduce germination )At'ater !1F#= *eyde-er !122,.
B. )he Need for O1ygen
4he second e/periment 'as to test the effect of o/ygen in the germination of the mongo seeds. It made use of * pyrogallic acid and distilled 'ater. 4he * and pyrogallic acids role 'as to absorb o/ygen carbon dio/ide and 'ater 'hich 'ill inhibit the germination of the mongo seeds. 4he 'ater on the contrary pro(ided enough o/ygen for the suspended mongo seeds.
Carbon dio/ide is a product of respiration and 'hen gas e/change is poor can accumulate in the soil resulting in an inhibition of germination )Arteca "##:,. 4he results sho'ed that the mongo seeds suspended in the flas- 'hich contained *" continued to germinate and gi(e rise to more roots 'hile the mongo seeds suspended in the flas- 'hich contained pyrogallic acid )C&*&+, and 0otassium hydro/ide )*, 'ere not able to germinate. C&*&+ together 'ith * absorbs the o/ygen carbon dio/ide and 'ater needed for respiration. Because the chemicals absorbed e(erything needed for respiration the mongo seeds 'ere not able to germinate. 8nli-e the other flas- filled 'ith distilled 'ater the seeds 'ere able to germinate because nothing hinders the upta-e of o/ygen )Lestran et al. "#!:,.
-. )em2eratre and Germination
4he third e/periment 'as to test seed germination under different temperature. 3et up A 'as placed in a refrigerator to ha(e an en(ironment of :℃. Group ! had !F )1#@, seeds that germinated 'ith an a(erage length of 1.2" mm for the shoots. 4here 'ere !& )F#@, seeds that sprouted in Group " 'ith an a(erage length of + mm for the shoots. All of the third groups seeds had germinated )!##@, in this set up 'ith an a(erage length of 2.#$ mm. It could be obser(ed that the all the roots from the mongo seeds did not sprout. 3et up B 'as placed inside the laboratory to ha(e a room temperature of "$℃. 4he first group got "# )!##@, germinated seeds 'ith an a(erage length of
+$.#$ mm for the roots and !!.!$ mm for the shoots. Group " yielded "# )!##@, germinated mongo seeds 'ith an a(erage length of $&.+$ mm and 1.+: mm for the roots and shoots respecti(ely. 4he third group only got !" )&#@, seeds that germinated 'ith an a(erage length of "1 mm for the roots and +:.1" mm for the shoots. 3et up C 'as put inside an incubator to ha(e a temperature of +2
℃.
4here
should ha(e been no gro'th of seeds on this set up but an error occurred for the incubator 'as turned off and thus the aim to -eep the set ups at the said temperature for se(en days 'as not achie(ed. Group ! had !$ )2$@, seeds that germinated 'ith an a(erage of :!.+1 mm for the roots and +#."F mm for the shoots. Group " obser(ed that there 'ere !2 )F$@, seeds that had germinated 'ith an a(erage length of +$.F mm for the roots and ":.& for the shoots. 4here 'ere also !2 )F$@, seeds that sprouted in the third group 'ith an a(erage of :$."1 mm for the roots and "!.$+ mm for the shoots. Among the three set ups 3et up B should ha(e the best result of the germination percentage and the length of the shoots and roots since it is the closest one to the optimum temperature that a mongo seed needs in order to germinate. 4emperature regulates the rate of germination germination percentage and subsequent seedling gro'th. In general the germination rate is lo' at reduced temperatures but increases as the temperature rises to an optimum le(el beyond 'hich there is a reduction due to seed in7ury. n the other hand the germination
percentage may remain constant o(er the middle part of this temperature range if enough time is allo'ed for germination to occur )Arteca "##:,. Mongo seed is a 'arm season plant and 'ill gro' 'ithin a mean temperature range of about "# to :# ℃. It is sensiti(e to lo' temperature and is -illed by frost. 0oelhman )!12F, suggested that mean temperatures of "# to "#
℃ may
be the
minimum for producti(e gro'th 'ith mean temperatures in the range of "F to +#℃ being optimumJ With temperatures abo(e "F ℃ increases in transpiration and respiration could offset benefits from increases in photosynthesis and retard plant gro'thJ Germination is inhibited by lo' temperature. In a germination study the rate of germination declined slo'ly belo' "$ ℃ dropped off sharply belo' !: ℃ and (irtually ceased belo' !!.$ ℃ )3imon et al. !12&,.
+. Osmotic -oncentration and Germination
4he last e/periment 'as conducted to determine the effect of osmotic concentration on the germination of the mongo seeds. 0etri dish A 'hich had #.##@ of NaCl had "# )!##@, germinated seeds for Group ! 'hich ha(e $:.1 mm and 22.:$ mm a(erage lengths for the roots and shoots respecti(ely. Group " got !1 )1$@, germinated seeds 'ith an a(erage length of $:.$F mm for the roots and &1."& mm for the shoots. 4here 'ere also
!1 )1$@, seeds that germinated in Group + 'ith an a(erage length of $2.+: mm for the roots and 1!.#$ for the shoots. 0etri dish B 'hich had #."$@ of NaCl had "# )!##@, seeds that germinated for all of the groups. nly that the a(erage length for the roots and shoots of the Group ! are !.&$ mm and $.:# mm respecti(ely.
respecti(ely.
Euring first de(elopmental stage salinity caused considerable delay and reduction in seed germination and seedling gro'th characteristics ho'e(er the underground part )roots, affected more ad(ersely as compared to the upper aerial part )stem, under high salinity )K ?C!#.# dsDm,. 4he roots absorb 'ater and nutrients from soil therefore radical length pro(ides significant clue to the response of plants to salinity stress )Mehmet Eemir "##+= Moose and Mumm "##F= Muhammad and Ma7id "#!+,. 4o/ic le(el of Na and Cl% ions produced an outside osmotic potential that a(oids 'ater upta-e or due to increased dormancy of seeds under salinity stress )Munns and ;ames "##+, )3ehrra'at et al. "#!+,. 3alts in the soil 'ater may inhibit plant gro'th for t'o reasons.
ability of the roots to e/clude salt thus increasing the upta-e rate of salt and its accumulation in shoots )Munns !1F#,.
S,A*= AN+ -ON-/,SION
3ome factors that affect seed germination 'ere tested through four e/periments. 4he first 'as to determine the need for 'ater of the seeds during germination. Among the three set ups 4ray B 'hich 'as for the soa-ed seeds 'ith moist lining had germinated !+ )F&.&2@, seeds 'ith an a(erage length of $&.$: mm for the roots and ++.+F mm for the shoots. Group " got !! )2+@, seeds that germinated 'ith an a(erage length of "+ mm and "1.$F mm for roots and shoots respecti(ely. 4he third groups seeds had germinated all )!##@, 'ith an a(erage length of 22.& mm for the roots and :F.&2 mm for the shoots. It ga(e the best result in terms of the percentage of germination and the a(erage lengths of the shoots and roots of the three groups. 4he moist en(ironment ser(ed as the optimum 'ater requirement for the already soa-ed seeds. 4he second one 'as conducted in order to determine the need for o/ygen during seed germination.
a(erage length of +&.#+ mm for the roots and :$.!# mm for the shoots. 4he said flas- had gi(en the best result among the three set ups. ?nough o/ygen is needed in order for the seed to germinate. 4his happened through respiration 'herein o/ygen 'as deri(ed from the distilled 'ater. 4he third acti(ity 'as set to -no' 'hat temperature best suit seed germination. 3et up B 'as placed inside the laboratory to ha(e a room temperature of "$ ℃. 4he first group got "# )!##@, germinated seeds 'ith an a(erage length of +$.#$ mm for the roots and !!.!$ mm for the shoots. Group " yielded "# )!##@, germinated mongo seeds 'ith an a(erage length of $&.+$ mm and 1.+: mm for the roots and shoots respecti(ely. 4he third group only got !" )&#@, seeds that germinated 'ith an a(erage length of "1 mm for the roots and +:.1" mm for the shoots. It had gi(en off the best result among the three set ups. >oom temperature ser(ed as the optimal temperature requirement for the seeds to germinate because lo' temperature temporarily inacti(ate the production of en6ymes and high temperature -ills the said en6ymes. 4he last e/periment 'as conducted to determine the effect of osmotic concentration on the germination of the mongo seeds. 0etri dish A 'hich had #.##@ of NaCl had "# )!##@, germinated seeds for Group ! 'hich ha(e $:.1 mm and 22.:$ mm a(erage lengths for the roots and shoots respecti(ely. Group " got !1 )1$@, germinated seeds 'ith an a(erage length of $:.$F mm for the roots and &1."& mm for the shoots. 4here 'ere also !1 )1$@, seeds that germinated in Group + 'ith an a(erage length of $2.+: mm for the roots and 1!.#$ for the shoots. It had gi(en the best result among the 1 set ups. 4he lo'er
the salinity the better for the germination of seeds because salt reduces the ability of the seeds to imbibe 'ater. 4herefore if the optimum le(el for each of the basic requirements )e.g. 'ater temperature osmotic concentration and o/ygen, for germination is met then a high percentage of germination 'ith longer roots and shoots could be yielded.
/I)E*A),*E -I)E+
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