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Evaluation of Seed Soaking Times on Germination Percentage, Germination Rate and Growth Characteristics of Pistachio Seedlings

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Ali Esmailpour at Iranian Pistachio Research Institute
Ali Esmailpour
Patrick Van Damme at Czech University of Life Sciences Prague
Patrick Van Damme
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ActaHortic.1109.ISHS2016.DOI10.17660/ActaHortic.2016.1109.17
XXIXIHC–Proc.Int.Symp.onNutCrops
Ed.:M.Wirthensohn
107
Evaluation of seed soaking times on germination
percentage, germination rate and growth
characteristics of pistachio seedlings
A.Esmaeilpour1,2andP.VanDamme2
1Iran’sPistachioResearchInstitute(IPRI),POBox:77175-435,Rafsanjan,Iran;2GhentUniversity,CoupureLinks
653,Ghent9000,Belgium.
Abstract
Intraditionalpistachiocultivation,priortosowingseedsthelatteraresoakedin
waterfordifferenttimesasfarmersclaimthatthiswillcontributetogermination.To
ourbestknowledge,thisisthefirstattempttoevaluatetheeffectofdifferentwater-
soakingtimeperiodsongerminationinpistachio.Forthispurpose,seedsofpistachio
(P. veraL.‘Badami’)weretreatedwithwaterfor1,6,12and24h.Wemonitored
germinationpercentage,germinationrate,meantimeforgerminationrate,and
seedlingheightatdifferentstages,numberofleaves,shootandrootdryweight,and
trunkdiameter.Ourresultsindicatedthatgerminationpercentageandrate
significantlyincreasedandseedlingheightincreasedinseeds,whichweretreatedfor
24h.Although,plantrootdryweightsignificantlydecreasedinseedsofthis
treatment,meantimesforgermination(MTG)valueswerenotfoundtobestatistically
significantbetweentreatments.Weobservedthatthe6hsoakingtreatmentledtoan
increaseinshootandrootdryweight,andtrunkdiameter.Alsoleafnumbersandroot
lengthvalueswerehighestinthe1hsoakedseeds,althoughdifferenceswerenot
significant.Positiverelationshipwasobservedbetweentheplantshootlengthand
plantleafnumber.
Keywords:meangerminationtime,Pistaciavera,lengthgrowth,plantdryweight
INTRODUCTION
Pistachio species belong to the Anacardiaceaefamily.Pistaciavera L., i.e., cultivated
pistachio,is the most importantspecies in the genusthat are all deciduousand dioecious.
Pistachioisanimportantfruittreegrowninthesub-tropicalareasintheworldwithhighest
production figures for Iran, Turkey, Syria and USA. Seed germination is the common
procedure for large scale propagation of pistachio seedlings. Seedlingsaregraftedto
desirable cultivars in nursery or field conditions to establishthepistachioproductive
orchards(Panahietal.,2002).
Seed quality is affected by several factors and seed germination percentage,
germination rate and seed vigor index have an important role in determination of seed
quality (Black and Bewley, 2000; Rajjou et al., 2012). To begin the germination process in
dried seeds, priming, pre-soaking, scarification, stratification and chemical treatments are
necessaryand the typeof treatment used depends on species, cultivarsandseedfeatures.
Pistachio species have different seed germination patterns; mostofthemhavedormancy
andneedspecialtreatmenttobreakdowntheirdormancy(Panahi et al., 2002). Acid
treatmentandgibberellicacid(GA3)areoftenusedtobreakdownespeciallythickseedcoats
(RahemiandBaninasab,2000)inwildpistachiorootstocks.Khanetal.(1999)reportedwild
pistachioseedstreatedwithKNO3at1%solutionsignificantlycompletedearlygermination
in135days,increasedthegerminationpercentageandproducedseedlings with higher
lengthanddiameterwhensoakedfor24h.Soakingseedsinpolyaminesisausefulmethod
to induce lateral root in pistachio seedlings (Sedaghat and Rahemi, 2011). Studies on
germination of citrus seeds indicated that osmotic stress causes decline of seed water
content,whichresultsininhibitionofseedgerminationandseedlinggrowth(Zekri,1993).
Thereisno dormancy in P. veraseedsand additionallythey haveahighgermination
108
percentage after being soaked in water (Panahi et al., 2002), thedurationofsoakingis
howeveraveryimportantparameterinachievingthemaximumgerminationpercentage.As,
too lengthy periods of soaking cause bacterial decay of the embryo, prior to the actual
germination, and inadequate duration causes inactivation of the enzyme facilitating the
germination. Thus, both of these conditions cause a decrease in seed germination
percentageandrate,andanoveralldeclineinthesubsequentseedlinggrowthparameters.
The objective of this study was to evaluate the effects of seed soaking with varying
durationsinwatertofind,optimizeandfinallystandardiseadurationtimeforoptimalseed
germinationfeaturesandsubsequentgrowthparameters of seedlingsofthemostcommon
pistachiocultivar,whichisusedaspistachiorootstockinIran.
MATERIALSANDMETHODS
TwoexperimentswereconductedinagreenhouseattheBioscience Engineering
Faculty, Ghent University (51°3’N, 3°42’E). Certified seeds of pistachio, PistaciaveraL.
‘Badami’ were obtained from the Iranian Pistachio Research Institute (30°39’N, 55°94’E),
Rafsanjan,Iran.Seedsizevariedfor17.9mm(length),10.6mm (width), 11.4 mm (thick)
and0.87g(weight).
Experiment1
Seedsof‘Badami’pistachiocultivarwerepre-treatedwithdistilledwaterfor1,6,12
and24hassoakingtimetreatments.Thenallpre-treatedseedsweredisinfectedwith0.01%
Captan,abroad-spectrumfungicidefor20min(Panahietal.,2002).Disinfectedseedswere
placed in petri dishes on Whatman filter paper and moistened with distilled water. Then
seedswerekeptinlabconditionsfor21daysandtheirrigationofthem wasperformedby
distilled water regularly. The germinated seeds were recorded every day and germination
percentagecalculatedonthe21stday.
Allgerminatedseedsweresownimmediatelyin4-Lpotscontainingsandandorganic
materialandkeptingreenhouseconditions. Plant cultural requirements were done
according to good agricultural practices during the experimental time. After 30, 40 and
50daysofsowingdate, shootheightandrootlengthweremanuallymeasuredwitharuler,
shootdiameterdeterminedwithadigitalcaliperandseedling(shoot and root) dry weight
wasdeterminedafterdryingat85°Cfor48hinanelectricoven. Germination percentage
andrate,mean of germinationtimeandvigorindexwere calculatedaccordingtofollowing
equations(EllisandRoberts,1981):
- Germination percentage (GP): GP (%) = SNG/SNO×100; where SNG is the number of
germinated seeds and SNOisthenumberofexperimentalseedswithviability,
respectively.
- Germinationrate(GR):GR(%)= SNM/SNG×100;whereGRisthegerminationrateand
SNMisthenumberofgerminatedseedsonagivenday.
- Mean of germination time (MGT): MGT is an index for germinationrate.MGT=
∑ (nd)/∑n (day-1);wherenisnumberofgerminatedseedsindayd,∑nistotal
germinatedseeds,Ddayofcounting.
- Vigorindex(VI):VI=GP×seedlingdryweight(g)(Abdul-BakiandAnderson,1973).
Experiment2
The similar seeds of ‘Badami’ pistachiowereputinpetridishes.Threepetridishes
were considered for each treatment and control. Polyethylene glycol (PEG) solutions were
preparedin0.2,0.8,and1.5MPaconcentrationsand5mLofeach solution was added to
petri dishes containing seeds. Also distilled water was applied to control petri dishes. All
petridisheswereplaced in lab conditions and every24hthenumberofgerminatedseeds
wascountedandincaseofmoisturedeficiencydistilledwaterwasused.
Statisticalanalysis
Treatmentswere arranged in a randomized completedesign with fivereplicatesper
treatmentsinexperiment1andthreereplicatespertreatmentsinexperiment2.Atwo-way
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analysisofvariancewasusedtotestfortreatmentdifferencesandeffects.Meanswere
comparedbyaDuncantest.AllanalyseswereperformedinSPSS20(IBMCorporation,USA).
RESULTSANDDISCUSSION
Experiment1
1.Germinationpercentage(GP).
Thegerminationpercentage ofseedsincreasedwithincreasing thesoakingtimesfor
pistachioseeds(Figure1).Seedssoakedfor24hwasthemosteffectivetreatmentonGPand
treatmentfor1hshowedthelowestpercentage.Statisticalanalysisshowedthattherewere
significant differences between these two treatments (Figure 1). These results are in
agreementwiththeresultsofSedaghatandRahemi(2011)thatreported application of
polyamines for 12 h compared to 6 h increased percentage of germination capacity in
‘Ghazvini’pistachioseeds.While,Khanetal.(1999)showedthatnormalwatersoakinghad
better germination percentage in 48 h duration compared to 12, 24 and 36 h duration in
wildpistachio.
Figure1. Changesingerminationpercentageindifferenttreatments(h)of‘Badami’
pistachiocultivar.
2.Meangerminationtime(MGT).
Meangerminationtimesignificantlydecreasedbyincreasingsoakingtimedurationof
seedsto12h.Althoughsoakingseedsfor24hsignificantlyincreasedcompared to 12 and
6htreatments(Figure2).Therewasnosignificantstatisticaldifferencebetween1and24h
treatmentsin ‘Badami’pistachiocultivar(Figure2).SedaghatandRahemi(2011)indicated
thatmeangerminationtimeofpistachiowasenhancedbysoakingtimeofseedsin
polyamines.
Pre-soaking times (Hour)
1 6 12 24
Germination %
0
20
40
60
80
100
ab
ab
b
a
110
Figure2. Changes in mean of germination time in different treatments(h)of‘Badami’
pistachiocultivar.
3.Germinationrate(GR).
Germinationrate(%)wasalsoaffectedbysoakingtimeduration in this experiment.
AsGPsignificantlyincreasedinseedswhichweresoakedinwaterfor24hcomparedamong
othertreatments,soakingofseedsfor6hhadthelowestpercentage of GP in ‘Badami’
pistachio cultivar (Table 1). There were significant differences between soaking times on
germinationpercentageinthiscultivar.
Table1. Effects of soaking of seeds on vigor index, germinationrate,leafnumber,root
length, shoot dry weight, plant dry weight and plant diameter for ‘Badami’
pistachiocultivar.
Treatments
(h) Vigor
index Germination
rate Leaf
number
Root length
(cm) Shoot dry weight
(g) Plant dry weight
(g) Diameter
(mm)
1 85.4 a 25.3 b 14.5 a 23.2 a 0.763 a 0.998 a 4.7 a
6 76.6 a 20.4 c 14.0 a 11.7 a 0.886 a 1.130 a 5.4 a
12 54.7 a 25.4 b 13.5 a 14.1 a 0.758 a 0.975 a 4.7 a
24 59.3 a 33.2 a 18.0 a 11.8 a 0.607 a 0.743 a 4.0 a
4.Vigourindex(VI).
The results of analysis showed that prolonging seed soaking times decreased the
vigourindexinpistachioseeds,althoughtherewerenosignificant differences among
treatments (Table 1). These results are in agreement with the finding of Oskouie and
Divsalar(2011)whichshowedthatvigourwasnotaffectedbydifferent level of nitrogen
fertilizeronmotherplantsofrapeseed.
5.Seedlingheight.
Observationof experimentshowedthattherewerenosignificantdifferencesinplant
height (cm) between different treatments in different periods after sowing the seeds
(Figure3),although,thereweremoreheightvaluesinthe24-htreatmentafter40daysin
‘Badami’ pistachio cultivar (Figure 3). These results are in agreement with the results of
SedaghatandRahemi(2011)thatreportedapplicationofpolyaminesfor12hcomparedto6
hincreased the shoot lengthin ‘Ghazvini’ pistachio seeds. While, Khan etal. (1999) found
Pre-soaking times (Hour)
1 6 12 24
MGT (Day)
0
2
4
6
8
10
12
14
16
b
c
a
a
111
that normal water soaking for 48 h duration produced long seedlingscomparedto12,24
and36hdurationinwildpistachio.
Figure3. Changesinshootlengthindifferenttimesafterseedsowingin different soaking
timesin‘Badami’pistachiocultivar.
6.Rootdryweight.
There were significant differences in root dry weight between 6and24soaking
treatments. Seeds which treated for 24 h had the lowest root dryweightamongother
treatments(Figure4).Applicationofpolyaminesanddistilledwater (control) for 12 h
comparedto6hdecreasedthe dry weight of roots in ‘Ghazvini’pistachioseeds(Sedaghat
andRahemi,2011).
There were no significant differences among different seed soaking times for leaf
number,rootlength,shootdryweight,plantdryweight,andtrunk diameter (Khan et al.,
1999)ofpistachioseedlings(Table1).
Figure4.Changesinrootdryweightindifferentsoakingtimesin‘Badami’pistachiocultivar.
Pre-soaking times (Hour)
161224
Shoot length (cm)
0
2
4
6
8
10
12
14
16
18
20
30 days
40 days
50 days
a
a
aa
a
a
a
a
a
a
a
a
Pre-soaking times (Hour)
1 6 12 24
Root dry weight (g)
0,00
0,05
0,10
0,15
0,20
0,25
0,30
b
a
ab ab
112
Experiment2
Evaluationsofthisexperimentshowedthattherewerenogerminatedseedsinanyof
thePEGsolutiontreatmentsexceptthecontrol.Aswecouldnotrecordanydataonthispart
ofourexperiment,wewerenotabletocomparethePEGtreatments with control.Inother
words,osmoprimingcausedbyallPEGtreatmentspreventedthegermination of pistachio
seeds in this experiment. These results are in agreement with thefindingofZekri(1993)
that indicated in germination of citrus seeds osmotic stress causes decline of seed water
content, results in inhibition of seed germination and seedlinggrowth.Althoughsome
references indicated that increasingPEG concent rationsdecreased germination percent in
canolacultivars(Shahverdikandietal.,2011)andinZiziphuslotus(Maraghnietal.,2010).
CONCLUSIONS
Theresultofthisgreenhouseexperiment revealedthat soakingseedsofpistachiofor
24hindistilledwaterimprovedgerminationcapacity,germination rate and plant length
growthcomparedtoothertreatments.Although,effectsofthistreatmentonrootdryweight
androotlengthweredecreasedcomparedto1and6htreatments, a positiverelationship
wasobservedbetweentheplantshootlengthandplantleafnumber. Thus, it is
recommendedtousedistilledwaterfor24hforPistaciavera(differentcultivars)seedsthat
lack dormancy to achievethe bes t germination parameters and plant length growth rates.
Theauthors suggestedevaluatingtheeffectsofPEGasprimingorosmoprimingtreatments
onotherpistachiocultivarsandspeciesseeds.
ACKNOWLEDGEMENTS
WewouldliketothanktotheIran’sPistachioResearchInstitute(IPRI)andAgriculture
Research, Education and Extension (AREEO) of Iran to provide the funding credit of first
authorasaPh.D.studentatGhentUniversity,Belgium.
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... Щодо передпосівної теплової обробки насіння, отримані нами результати співпадають із даними експериментів, де прогрівання насіння та його намочування забезпечило суттєве підвищення енергії проростання (Macdonald & Asotie,1999;Noorhosseini et al., 2018). Вважають, що при намочуванні насіння з наступним підсушуванням до вихідної вологості відбувається активація ферментів та можливі морфологічні зміни в структурі насіннєвих покривів, що покращує схожість насіння (Esmailpour & Van Damme, 2014;Lin & Sung, 2001). ...
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  • S Sedaghat
  • M Rahemi
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