Effects of resistance in "Phaseolus vulgaris" on development of "Meloidogyne" species

126 Pages·1995·4.1 MB·English

by  Sydenham, Georgina Maria-Rosa Laura

EFFECTSOFRESISTANCEJNPHASEOLUSVULGARIS
ONDEVELOPMENTOVMELOIDOGYNESPECIES
By
GEORGINAMARIA-ROSALAURASYDENHAM
ADISSERTATIONPRESENTEDTOTHEGRADUATESCHOOL
OFTHEUNIVERSITYOFFLORIDAINPARTIALFULFILLMENT
OFTHEREQUIREMENTSFORTHEDEGREEOF
DOCTOROFPHILOSOPHY
UNIVERSITYOFFLORIDA
1995
UNIVERSITYOFFLORIDALBRARtES
ACKNOWLEDGMENTS
Iwouldliketoexpressmythankstothemembersofmyadvisorycommittee,
Dr.R.A.Dunn,Dr.R.McSorley,Dr.D.J.Mitchell,andDr.G.W.Simone,fortheir
supportandguidancethroughoutmydoctorateprogram IamgratefijltoDr.Dunnfor
hisfinancialsupportandtheopportunitiestotraveltoHondurasandBraziltopresent
myresearch. MythanksgotoDr.Simoneforhisencouragementtoenjoythese"best
daysofmylife." MyspecialthanksgotoDr.McSorleyandDr. Mitchellfortheir
continuousencouragement, theirexperienceandknowledge, andtheirtimespentin
discussionsdirectlyassociatedwithmyprojectsandphilosophicalviewsontherolesof
scientistsandeducatorsinaworldsociety.
Iwouldliketothankmyfamilyfortheiracceptanceandsupportofmydecision
tostayintheUSA,andfortheirwarmwelcomesonmyvisitshome. Thanksalsogoto
allmyfriendswhohavelistenedandencouragedmethroughoutthisdegree.Finally,I
wouldliketothankStephenRobinson,especially,forhistirelessreadingoffirstdrafts,
andforhisencouragement,support,laughter,andlove.
ii
1
TABLEOFCONTENTS
page
ACKNOWLEDGMENTS
ii
LISTOFTABLES v
LISTOFFIGURES vii
ABSTRACT ix
CHAPTER1:REVIEWOFLITERATURE 1
CHAPTER2:EFFECTSOFRESISTANCEINGERMPLASMOF
PHASEOLUSVULGARISONPENETRATIONANDDEVELOPMENT
OFMELOIDOGYNEINCOGNITARACE2ANDMELOIDOGYNE
ARENARIARACE1 9
Introduction 9
MaterialsandMethods 1
Results 14
GeneSystem1 14
GeneSystem2 17
Discussion 20
CHAPTER3:EFFECTSOFTEMPERATUREONRESISTANCEIN
NEMASNAPANDG1805BEANANDONDEVELOPMENTOF
MELOIDOGYNEINCOGNITARACE2 MELOIDOGYNE
ARENARIARACE1 44
Introduction 44
MaterialsandMethods 46
GeneSystem1andGeneSystem2 46
HighTemperatureExperiments1and2 48
Results 49
GeneSystem1 49
GeneSystem2 53
HighTemperatureExperiments1 57
HighTemperatureExperiments2 57
Discussion 5g
iii
page
CHAPTER4:RESEARCHSUMMARYANDCONCLUSIONS 85
APPENDIXA:RESISTANCEINGERMPLASMOFPHASEOLUS
VULGARISTOFLORIDAPOPULATIONSOFMELOIDOGYNE
SPECIES 87
Introduction 87
MaterialsandMethods 88
GeneSystem1 88
GeneSystem2 90
Results 91
GeneSystem1 91
GeneSystem2 92
Discussion 93
APPENDIXB:HISTOPATHOLOGYOVMELOIDOGYNEARENARIA
RACE1INFECTIONINRESISTANTANDSUSCEPTIBLE
PHASEOLUSVULGARISGERMPLkSU 104
Introduction 104
MaterialsandMethods 105
Results 106
LISTOFREFERENCES 109
BIOGRAPHICALSKETCH 115
iv
LISTOFTABLES
Table page
2-1pl.aGntasllinioncduelxatraetdinmgtohfMroeoltosiyosdtoegmysneofinBcloagcnkitVaalreanctei2neandNemasnapbean 24
2-2. NumbersofeggsofMeloidogyneincognitarace2extractedfromthe
rootsystemsofBlackValentineandNemasnapbeanplants,juveniles
hatchingfromtheeggsafter10days,andeggsperfemale 25
2-3. LeafareasofBlackValentineandNemasnapbeanplantsuninoculated
(NN)orinoculatedwithMeloidogyneincognitarace2(Mil) 26
2-4. EnergycontentofplanttissuesofBlackValentineandNemasnapbean
plantsuninoculated(NN)orinoculatedwithMeloidogyneincognitarace2
(Mil) 27
2-5. GallindexratingofrootsystemsofBlackValentineandG1805bean
plantsinoculatedwithMeloidogynearenariarace1 28
2-6. Numbersofeggs Meloidogynearenariarace1extractedfromthe
rootsystemsofBlackValentineandG1805beanplants,juvenileshatching
fromtheeggsafter10days,andeggsperfemale 29
2-7. LeafareasofBlackValentineandG1805beanplantsuninoculated(NN)
orinoculatedwithMeloidogynearenariarace1(Mai) 30
2-8. EnergycontentofplanttissuesofBlackValentineandG1805bean
plantsuninoculated(NN)orinoculatedwithMeloidogynearenariarace1
(Mai) 31
3-1. NumbersofmalesofMeloidogyneincognitarace2inrootsystemsof
susceptibleBlackValentineandresistantNemasnapcommonbean(gene
system1)incubatedatthepre-inoculationconditioningtemperaturesand
post-inoculationincubationtemperaturesof24°C,and28°C 62
3-2. NumbersofMeloidogyneincognitarace2infourdevelopmentalstages
inrootsystemsofsusceptibleBlackValentineandresistantNemasnap
commonbean(genesystem1)incubatedattemperaturesof30''Cor32°C 63
V
Table page
3-3. NumbersofMeloidogymarenariarace1infourdevelopmentalstages
inrootsystemsofsusceptibleBlackValentineandresistantG1805common
bean(genesystem2)incubatedattemperaturesof30°Cor32°C 64
A-1. ResistanceinPhaseolusvulgarisgermplasmtoMeloidogynespeciesand
races 95
A-2. OriginsandsourcesofPhaseolusvulgarisgermplasm 96
A-3. Sources Meloidogynespecies 97
A-4. ResistanceinPhaseolusvulgarisgermplasmcontaininggenesystem1to
aFloridapopulation Meloidogyneincognitarace3 98
A-5.ResistanceinPhaseolusvulgarisgermplasmcontaininggenesystem1toa
FloridaandaCaliforniapopulationofMeloidogyneincognitarace3 99
A-6.ResistanceinPhaseolusvulgarisgermplasmcontaininggenesystem1toa
FloridapopulationofMeloidogyneincognitarace2 100
A-7.ResistanceinPhaseolusvulgarisgermplasmcontaininggenesystem2toa
FloridapopulationofMeloidogynejavanica 101
A-8.ResistanceinPhaseolusvulgarisgermplasmcontaininggenesystem2toa
FloridapopulationofMeloidogynearenariarace1 102
A-9.ResistanceinPhaseolusvulgarisgermplasmcontaininggenesystem2toa
FloridapopulationofMeloidogynearenariarace2 103
B-1.Ethanol(EtOH)andtertiarybutylalcohol(TBA)dehydrationseries 107
B-2.Mineraloil,waxandparaffininfiltrationseries 108
vi
LISTOFFIGURES
Figure page
2-1. PenetrationanddevelopmentofMeloidogyneincognitarace2insusceptible
BlackValentineandresistantNemasnapcommonbeangermplasm(genesystem1,
trials1and2 33
2-2. PercentageofthepopulationofMeloidogyneincognitarace2infour
developmentalstagesinrootsystemsofsusceptibleBlackValentineandresistant
Nemasnapcommonbeangermplasm(genesystem1,trials1and2) 35
2-3. TotalplantdryweightsofBlackValentine(BV)andNemasnap(NS)
uninoculated(NN)orinoculatedwithMeloidogyneincognitarace2{Mil)(gene
system1,trials1and2) 37
2-4. PenetrationanddevelopmentofMeloidogynearenariarace1insusceptible
BlackValentineandresistantG1805commonbeangermplasm(genesystem2,
trials1and2) 39
2-5. PercentageofthepopulationofMeloidogynearenariarace1infour
developmentalstagesinrootsystemsofsusceptibleBlackValentineandresistant
G1805commonbeangermplasm(genesystem2,trials1and2) 41
2-6. TotalplantdryweightsofBlackValentine(BV)andG1805(G)uninoculated
(NN)orinoculatedwithMeloidogynearenariarace1(Ma\)(genesystem2,trials
1and2) 43
3-1. TotalpopulationsofMeloidogyneincognitarace2insusceptibleBlack
ValentineandresistantNemasnapcommonbeangermplasmatfourtemperature
regimesofpre-inoculation/post-inoculationtemperaturesof24/24°C,24/28°C,
28/24''C,and28/28''C(genesystem1,trials1and2) 66
3-2Effectsofpre-inoculationtemperatures(24^and28°C)onpenetrationand
developmentofMeloidogyneincognitarace2insusceptibleBlackValentineand
resistantNemasnapcommonbeangermplasm(genesystem1,trial1) 68
3-3. Effectsofpre-inoculationtemperatures(24°Cand28T)onpenetrationand
developmentofMeloidogyneincognitarace2insusceptibleBlackValentineand
resistantNemasnapcommonbeangermplasm(genesystem1,trial2) 70
vu
Figure page
3-4Effectsofpost-inoculationtemperatures(24°Cand28°C)onpenetrationand
developmentofMeloidogyneincognitarace2insusceptibleBlackValentineand
resistantNemasnapcommonbeangermplasm(genesystem1,trial1) 72
3-5. Effectsofpost-inoculationtemperatures(24°Cand28''C)onpenetrationand
developmentofMeloidogyneincognitarace2insusceptibleBlackValentineand
resistantNemasnapcommonbeangermplasm(genesystem1,trial2) 74
3-6. TotalpopulationsofMeloidogynearenariarace1insusceptibleBlackValentine
andresistantG1805commonbeanatfourtemperatureregimesofpre-
inoculation/post-inoculationtemperaturesof24/24°C,24/28°C,28/24°C,and
28/28''C(genesystem2,trials1and2) 76
3-7. Effectsofpre-inoculationtemperatures(24°Cand28°C)onpenetration,and
developmentofMeloidogynearenariarace1insusceptibleBlackValentineand
resistantG1805commonbeangermplasm(genesystem2,trial1) 78
3-8. Effectsofpre-inoculationtemperatures(24°Cand28°C)onpenetration,and
developmentofMeloidogynearenariarace1insusceptibleBlackValentineand
resistantG1805commonbeangermplasm(genesystem2,trial2) 80
3-9. Effectsofpost-inoculationtemperatures(24°Cand28"C)onpenetrationand
developmentofMeloidogynearenariarace1insusceptibleBlackValentineand
resistantG1805commonbeangermplasm(genesystem2,trial1) 82
3-10. Effectsofpost-inoculationtemperatures(24^and28°C)onpenetrationand
developmentofMeloidogynearenariarace1insusceptibleBlackValentineand
resistantG1805commonbeangermplasm(genesystem2,trial2) 84
viii
AbstractofDissertationPresentedtotheGraduateSchool
oftheUniversityofFloridainPartialFulfillmentofthe
RequirementsfortheDegreeofDoctorofPhilosophy
EFFECTSOFRESISTANCEINPHASEOLUSVULGARIS
ONDEVELOPMENTOFMELOIDOGYNESPECIES
By
GeorginaMaria-RosaLauraSydenham
December,1995
Chairman: Dr.R.A,Dunn
MajorDepartment: EntomologyandNematology
Root-knot nematodes, Meloidogyne spp., are major limiting factors in the
production ofcommon bean, Phaseolus vulgaris. Use of resistant P. vulgaris
germplasm has potential for management ofroot-knot nematodes. Mechanisms of
resistanceoftwogeneticresistancesystemsandeffectsoftemperatureonresistance
expression were studied using a susceptible cultivar 'Black Valentine' and either
M
resistantcultivar'Nemasnap'(genesystem1)with incognitarace2,orgermplasm
M
G1805(genesystem2)with arenariarace1. Mechanismsofresistanceandeffects
ofpre-andpost-inoculationtemperatures(24°Cand28°C)wereevaluatedatspecific
harvestdatesbydeterminingnematodepopulationswithinrootsandtheproportionof
eachpopulationwithindistinctdevelopmentalstages.
Sincesimilarnumbersofjuvenilespenetratedrootsofresistantplantscompared
withsusceptibleBlackValentine, pre-penetrationmechanismsofresistancewerenot
expressedineitherNemasnaporG1805. Resistancewasexpressedbypost-penetration
ix
mechanisms, which resulted in smallerpopulations ofnematodes and their delayed
developmentinresistantplantscomparedwithsusceptibleplants.
Resistancewaseffectiveatalltemperatureregimesexamined,withsmaller
populationsofnematodesinrootsofNemasnapandG1805comparedtoBlack
Valentine.Pre-inoculationtemperature,perse,didnotmodifyresistanceexpressionin
M
NemasnaporG1805tolaterinfectionsby incognitaoxM.arenaria,respectively.
However,post-inoculationtemperatureshadpronouncedeffectsonthedevelopmentof
Meloidogynespp.inallgenotypes.Themorerapiddevelopmentofnematodestoadult
stagesatthehighertemperatureof28°Ccomparedwith24°C,inboththeresistantand
thesusceptiblegermplasms,indicatesthattheeffectofpost-inoculationtemperatureon
nematodedevelopmentisdirect,ratherthanviaresistanceexpressionofeithergene
system1orgenesystem2. Resultsofstudiesathighertemperaturesdemonstratedthat
resistanceinNemasnapandinG1805alsowasstableatSOTand32°C.
TestswithFloridapopulationsofroot-knotnematodesidentifiedpotentially
M M
virulentpathotypesof incognitarace3and javanicaintheirresistantP.vulgaris
hostscontaininggenesystems1and2,respectively. Thusitiscriticaltoassessthe
performanceofresistantgermplasmtoawiderangeofgeographicallylocalized
populationsofnematodespecies.