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VIRUS AND VIRUS- LIKE DISEASES OF ROSES IN NEW ZEALAND
A thes i s on s tudies conduc ted in the Dep artment of Hort icul t ure and Plant Hea lth in part ial fulfi lment of the req uiremen t for a Do c t or of Phi losophy Degree at Mas sey Univers i ty .
'
by
P . C . Gardner June 1 9 8 3
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p 1 Para . 4 , line 4 : p 1 Para . 6 , l ine 2 : p 2 Para . 2 , line 1 0 : p 15 Para . 1 , line 2 : p 1 5 Para . 8 , line 2 : p20 Para . 5 , line 3 : p31 Para . 2 , line 1 : p40 Para . 4 , line 2 : p5 4 Para . 7 :
p 5 7 Para . 1 , line 1 : p 7 0 Para . 1 , line 3 : p9 1 Para . 5 , line 1 : p92 Para . 1 , l ine 2 :
ERRATA
Rosa not R .
d iseases not disorders
' Iowa State Univ - 6 0 (ISU-6 0 ) though t not though
vi rul i fe rous not viuliferous for not in
diseases not disorders
amaranti color not amaranthicolor De lete this EaragraEh
electrophore s i s not electorphores is TobRSV not TSV
cond i t ion not condi t i ons indicator not test
i ABSTRACT
Roses in New Zealand we re surveyed for t he presence of symptoms which mi gh t be caused by vi rus or vi rus- like graft t ransmi s s ib le agent s . Repre s entative s ample s ( 2 2 1 ) o f all symptom type s and many apparen tly symptomle s s plants we re indexed by a number o f me thod s . Prunus necro t i c ringspo t virus was found to be widespread , oc curing i n plants b o t h with and wi thout symptoms . Apple mo saic virus was detected in one plant and arabis mosaic virus was detected in s ome plants o f one cult ivar c lone .
Prune dwarf virus , tob acco s t reak virus , tobacco ringspot virus , and s t rawberry latent rings po t virus we re no t de tected .
Two graf t t ransmi s s ib le virus-l ike diseases of rose flowe r s are described . One o f them , rose pe tal fleck , was widespread in both obvious ly affected and symptomless plants . The o ther , rose co lour b reak , was largely confined to s ome glasshouse cut flower cul t ivars and a few garden cult ivars .
i i SUMMARY
Two hundred and twenty one rose samples showing a wide range o f symp toms whi ch might be caused by virus o r virus-like gra ft t ransmi s s ib le agents we re indexed for t ransmi s s ion and pe rpetuat i on on Rosa mul ti flora 'Iowa State University 6 0 ' unde rstocks.
Al l samples we re indexed on herbaceous hos ts and by the enzyme-linked immunosorbent assay ( ELISA) serological technique for prunus necro tic ringspot vi rus ( PNRSV) , apple mosaic virus (ApMV) , prune dwarf virus
( PDV) , danish p lum line pat te rn vi rus ( DPLPV) , tobacco s t reak virus ( TSV) , t obacco ringspo t virus ( TobRSV) , s t rawbe rry latent ringspo t vi rus ( SLRSV) and arabi s mos aic virus (ArMV) .
A range o f selected s amples we re also indexed on ' Golden Queen ' peach seedl ings , apple unde rs tocks , 'Shi ro fugen' che rry and Rosa multiflora
' Burr' .
The ELISA te chni que was modified to detect heterologous s t rains o f PNRSV and a me thod was developed to detect the presence o f any one or more o f three vi ruses in any one or mo re o f ten plant s s imultaneously .
One plan t only, wi th heavy go ld leaf blotching, was found t o be infected wi th ApMV . All o the r p lan ts tes t ed wi th mosaic type leaf sympt oms or ro se wi lt type decline and dieback o f ma ture plan ts were found to be infec ted with PNRSV as we re a number o f apparently sympt ornless plants . Sympt oml ess ArMV was fo und on some pl ants of one cul t iva r clone only . No o the r viruses were detected .
The symptoms in mature plants a t t ributed to 'rose wi l t virus ' could
invariab ly be associated with the presence o f PNRSV b ut the proli ferat i on sympt om in maiden plan t s could no t .
In infected plan t s PNRSV reached a higher t i t re in mo re me tabol ically act ive and younge r tis sue than in older tissue but virus could no t be detected in emb ryos excis ed from seeds .
Four symp tornless plan t s o f supposedly 'hi gh health' s t atus o f each o f 274 rose cultivars were indexed . Fourteen cult ivars we re posit ive for PNRSV and one cultivar positive for ArMV.
iii
Approximately 200 cultivars o f s o-called "old and s pecies type " roses were indexed. Seventeen cultivars were positive for PNRSV and all
tests for other viruses were ne gative .
Six clones o f R. multiflora unders tock , in commercial use for some year s , were all at least in part infe cted wi th PNRSV b ut wer e ne gative for other viruses .
A po lyacrylamide ge l electrophores i s separation o f ribonuc leic acid ( RNA) extracted from PNRSV showed a multipartite genome wi th RNA with mo lecular we i ghts of ab out 1 . 4 , 1 . 0 , 0 . 7 and 0 . 3 + 106•
Two virus-like dis eases of flowers were des cribed . Rose petal f leck ( RPF) and rose co lour b reak ( RCB) were transmitted by grafting both separate ly and together to healthy rose plants . No causal agent was detected for these dis orders .
Rose petal fleck was widespread occuring in both obvious ly infected and apparently symptomless cultivars . A number of s o-called "old and spe c ies type" roses cons istently had flecke d pe tals which is c ons idered normal for those cultivars . On indexing they were found to have RPF .
Ros e colour b reak was found only occas iona lly , mainly in glasshouse cutf lower varieties .
iv
PREFACE
Wi th increased unde rstanding and recognit ion o f the e f fects of virus infec tion , transmi s s i on and perpe t uat ion in veget ively propagated woody plant s it has become essential for the hort icultura l indus t ry
t o have access t o pro pagat ive mat erial as free o f vi ral pathogens as po s s ib le .
To achieve th i s obj ective a knowledge o f the virus and virus-like pa thogens infe c t ing a crop and rapid and reliab le methods for their detect ion are es sent ial .
Crops considered to b e o f s igni f icance t o the New Zealand economy have been given high priority by the Department o f S c ienti fi c and Indus t rial Re search for such inve s t i gat ions .
Although roses are an important nursery and cut flower crop within New Zea land they have a relat ively low export po tential and there fore despi t e an obvious need for inve s t igation of their viral s tatus the same priority c ould not be given to roses as to export fruit c rops .
However , in recognition of the urgent need for research in rose viruses the DS IR made funds availab le to Mas sey Univers i ty in 1 9 7 8 fo r a Ros e Research Contract .
The New Zealand rose industry and the ros e growing pub lic a t large are indeb ted t o D S IR for providing the funds whi ch made this
inve s t igat ion pos sib le . The cont inuing work by the New Zea land Nursery Research Centre , at Massey Universi ty , is now enab ling the pract i cal applicat ion of these resul t s .
I wish to express my thanks to Pro fessor K . S . Milne and the Department o f Hort icul t ure and Plant Health at Massey University for guidance , suppor t and pat i ence , and t o Mr H . F . Ne ilson for invaluab le t echnical assis tance . The o pportuni ty t o work with Dr J . B . Sweet (Massey Pos t-doct oral Fellow) , who had cons iderable
experience with viruses of rosaceous pl ants wa s also mo s t helpful .
V
Fo r the supply o f materials , anti sera and advice , thanks are due to Dr P . F . Fry , Plant Diseases Division , DSIR , Auckland ,
Professor R .W . Fulton , University o f Wi s conson , U . S . A . and
Dr B . D . Harr i son , Scottish Horti cultura l Research Ins ititute , U . K .
Members o f the nursery industry throughout New Zealand have always been supportive , allowing access to their crops and the opportuni ty to collect material for investigation . I trust these results wil l be o f benefit to them i n the future .
Last b ut not leas t, my thanks to my wife and fami ly for their patience and long-s uffering unders t anding.
P . C . Gardner June 1 9 8 3
vi
TABLE OF CONTENTS CHAPTER 1 . INTRODUCTION
S ource of Ros e Material Rose Sympt oms
Ros e Mosaic - Review
Rose Wilt and Prolife rat i on - Review
As sociation of Prunus Ne crotic Rings pot Vi rus with Ros e Mos ai c and Rose Wi lt Symptoms
CHAPTER 2 . INDEXING Bud Graft ing
Rosa multiflora 'ISU- 6 0' Rosa multiflora 'Burr'
Prunus serrulata 'Shiro fugen' App le Roo ts tocks
Peach 'Go lden Queen' Seedlings Me chanical Transmiss ion
Elec t ron Microscopy Sero logi cal Me thods
1 2 5 9 19 24
28 31 33 36 37 38 43 45
CHAPTER 3 . PRUNUS NECROTIC RINGSPOT VIRUS - Puri fication 4 6 Charac terist ics o f Parti c les
Ant iserum Produc tion and Tes ting
CHAPTER 4 . ENZYME-LINKED IMMUNOS ORBENT AS SAY Me thods and Results o f ELISA Indexing
" Di seases Co llection"
"High Health Mother Plant s "
S imultaneous ly for a Range o f Vi rus es
"Old Ros e Collec t ion"
Non Rose Hos t s and Rose Roots tocks
Dis t rib ut ion of Vi rus Wi thin the Rose Plant
CHAPTER 5 . VIRUS-LIKE FLOWER DIS ORDERS Rose Colour Break
Rose Pet al Fleck CHAPTER 6 . CONCLUSIONS BIBLIOGRAPHY
APPENDIX I
5 0 6 0 63 67 7 1 78 81 83 84 86 9 0 9 3 9 4 102 105 Back cover
vii
FIGURES
1 . Opt ical density pro files o f five isolates o f prunus necro t i c ringspot vi rus nucleoprote in
2 . Opt i cal density profile o f p runus necro t i c ringspot vi rus , ro se i solate ' 1 5 4 ' a fter polyacrylami de ge l electrophores i s
3 . ELISA p late showing react ions, agains t PNRSV-RA (Fulton) ant i serum, of isolates from the rose
"disease colle c t ion " , PNRSV-G (Ful ton) , ApMV ( Ful tons ' s serotype from rose) all ex cucumber
4 . ELISA plate showing reac t ions , aga inst ApMV-C ( Ful t on) ant i se rum , o f young leaf samp les from 46 roses in the "di sease collect ion" ( s amples ' 45 ' to
' 9 0 ' inclusive ) , healthy rose , C. quinoa infected wi th ApMV-C ( Fulton) , and cucumb er infected wi th PNRSV-G (Fulton)
5 . ELI SA p late showing PNRSV-RA (Fulton) , ApMV-C
(Fulton ) , and the comb inat ion o f these two ant isera with ArMV (Harrison ) aga ins t isolates o f rose ' 2 1 '
infected with PNRSV , Rosa multiflora gra fted wi th rose '72 ' (ApMV) , rose ' 10 1 ' in cucumb e r , and ApMV
( Fulton ) in c. quinoa
6 . Two flowers o f the rose cul tivar ' Bridal Pink ' showing severe symp toms o f rose co lour b reak
7 . Two fully open f lowers o f the rose cultivar ' Fragrant Cloud ' used as an indicator showing symptoms of rose petal fleck induced by an infe c t ed t e s t bud and a healthy negative t e s t
5 3
5 8
7 7a
7 7b
7 7 c
9 3a
94a
