152 Plant Protection Quarterly Vol.13(4) 1998 • Is blackberry still a problem on pasture All these and other questions arose dur- L. agg. in south-eastern Victoria. Plant land because the message from re- ing the planning of the workshop. An- Protection Quarterly 13, 182-5. search is not reaching landholders? swers to such questions need not neces- McGregor, G. (1998). Relationships be- • What is the optimum combination of sarily follow, but I hope the definition of tween weedy and commercially grown existing control methods for use in dif- future research directions and the exten- Rubus species. Plant Protection Quarterly ferent situations? sion of those existing and new research 13, 157-9. • How can these existing methods be results will flow from the following pro- Milne, B.R. and Dellow, J.J. (1998). Herbi- adapted to areas of natural vegetation, ceedings. cide responses of blackberry (Rubus which often abut waterways or are in- fruticosus agg.) in the Central Table- accessible for conventional methods of References lands of New South Wales. Plant Protec- herbicide application to be used? Amor, R.L. and Miles, B.A. (1974). Tax- tion Quarterly 13, 180-1. • What novel methods of control may be onomy and distribution of Rubus Mueller, F. (1895). ‘Select extra-tropical developed for such ecological situa- fruticosus L. agg. (Rosaceae) naturalized plants, readily eligible for industrial tions, such as mycoherbicides? in Victoria. Muelleria 3, 37-62. culture or naturalization’, 9th edition, • Do we need to look anew in drier areas Amor, R.L., Richardson, R.G., Pritchard, p. 475. (Victorian Government Printer, of Europe for strains of rust better G.H. and Bruzzese, E. (1998). Rubus Melbourne). adapted to drier southern Australian fruticosus L. agg. In ‘The Biology of Parsons, W.T. and Cuthbertson, E.H. climates? Australian Weeds’, Volume 2, eds. F.D. (1992). ‘Noxious weeds of Australia’, • What native plant and animal species Panetta, R.H. Groves and R.C.H. Shep- 692 pp. (Inkata Press, Melbourne and are at risk from encroaching blackberry herd, pp. 225-46. (R.G. and F.J. Sydney). thickets? Richardson, Melbourne). Vere, D.T. and Holst, P.J. (1979a). Using • Which native plant species will be Evans, K.J., Symon, D.E. and Roush, R.T. goats to control blackberries and briars. strongly competitive with weakened (1998). Taxonomy and genotypes of the Agricultural Gazette of New South Wales blackberry plants as a result of imple- Rubus fruticosus L. aggregate in Aus- 90, 11-13. menting control? tralia. Plant Protection Quarterly 13, Vere, D.T. and Holst, P.J. (1979b). The eco- • How much will all this extra research 152-6. nomics of using goats to control Rubus cost? Mahr, F.A. and Bruzzese, E. (1998). The fruticosus. Proceedings of the 7th Asian- • Who will pay for this additional re- effect of Phragmidium violaceum (Schultz) Pacific Weed Science Society Confer- search? Winter (Uredinales) on Rubus fruticosus ence, pp. 207-9.

Taxonomy and genotypes of the Rubus fruticosus L. for improved biological control of black- berries. aggregate in Australia Introduction K.J. EvansA, D.E. SymonB and R.T. RoushA Blackberry (Rubus fruticosus L. agg.) is an A Department of Crop Protection and CRC for Weed Management Systems, important weed of natural and agricul- tural ecosystems in Australia and is University of Adelaide, Waite Campus, PMB 1, Glen Osmond, South Australia widely distributed in the high-rainfall re- 5064, Australia. gions of each State. Weed managers use B The Botanic Gardens of Adelaide and State Herbarium, North Terrace, the common name ‘blackberry’ to encom- Adelaide, South Australia 5000, Australia. pass all the Rubus taxa that comprise this aggregate. An ability to recognize the different Rubus taxa is a skill weed man- Summary agers could use to monitor the coloniza- Blackberry (Rubus fruticosus L. aggre- precise information regarding the tion and spread of individual taxa. New gate) is an important weed of agricultural amount and distribution of genetic vari- approaches for blackberry management, and natural ecosystems in Australia. ation within Rubus. We present the use such as biological control, require a know- Weed managers require accurate taxo- of DNA fingerprinting as a tool for (i) ledge of blackberry taxonomy because nomic keys for Rubus so that they can determining the genotype of an indi- biocontrol agents may provide more effec- identify which taxa are contributing to vidual plant, (ii) estimating the genetic tive control of some taxa when compared the weed problem. Blackberry comprises variation within and among Rubus taxa, with the level of control of other taxa. a few diploid sexual species (e.g. R. and (iii) clarifying some taxonomic prob- Taxonomic keys for Rubus are available in ulmifolius) and a large number of poly- lems in the genus Rubus. Twenty differ- each State, but there are inconsistencies ploid agamospecies (e.g. taxa in Aus- ent genotypes were identified among 13 among the States in the names provided. tralia named R. polyanthemus, R. different Rubus taxa. No genetic varia- The status of Rubus taxonomy in Aus- laciniatus and the widespread R. affin. tion was observed among 50 plants of R. tralia is explained partly by the general armeniacas (= R. discolor sensu auct. affin. armeniacas sampled from 29 loca- taxonomic problems encountered in the aust. non Weihe & Nees)). We review the tions throughout Australia, suggesting genus Rubus. In Europe, only a few dip- status of Rubus taxonomy in Australia that this common blackberry is probably loid blackberries, including R. ulmifolius, and present some new information re- a single clone. In contrast, seven differ- can be treated as true sexual species. The garding existing taxa based on collec- ent genotypes were observed among 26 remaining taxa in R. fruticosus L. agg. are a tions made in South Australia and exam- plants of R. ulmifolius sens. lat. sampled complex of polypoid and apomictic ined by Rubus specialists in Europe. The from six locations in Victoria. Two of biotypes (Asker and Jerling 1992, Nybom utility of Rubus taxonomy for research these genotypes were sampled from a 1995, Weber 1996). Apomixis, however, is workers and weed managers is also ex- single thicket of R. ulmifolius sens. lat. facultative, which means that a small pro- amined. Whereas the biological species We illustrate the utility of genotyping portion of seed may be produced sexually concept may be useful for weed manag- Rubus plants in studies to identify viru- as a result of hybridization between ers, research workers often require more lent strains of the European rust fungus apomicts or between apomicts and sexual Plant Protection Quarterly Vol.13(4) 1998 153 species (Nybom 1988, 1995). Spontaneous but is based on a study of Victorian taxa F15, was released in all Australian States as hybrids may have a limited local distribu- conducted in the 1960s (Amor and Miles a biological control agent in the summers tion or become a stabilized clone with a 1974). In all States, there are likely to be of 1991 and 1992. Although R. laciniatus is wide distribution. These biotypes, in ad- some Rubus taxa that cannot be given a classified as a resistant biotype when in- dition to forming large or small popula- species name because their morphology oculated with strain F15 (Bruzzese and tions, differ from one another in large or is different from that of taxa described Hasan 1986a) , it shows symptoms of rust small degrees (Weber 1996). The most re- in Europe. The distribution of the well- disease in the field (E. Bruzzese personal cent account of Rubus in Britain lists 307 defined taxa can be implied from her- communication). This taxon is probably species with even more in continental Eu- barium records, but the last detailed study being infected by the strain, or strains, of rope where taxonomic studies are incom- of taxon distribution (Amor 1971) again rust that were introduced illegally to plete (Edees and Newton 1988). Given that relates only to Victoria in the 1960s. A na- southern Victoria in 1984. In regions the taxonomic treatment of blackberry tional review of Rubus taxonomy and the where the rust disease is severe, continues to evolve, Weber (1996) pro- distribution of taxa is warranted so that landholders are advised to identify black- vides criteria for allocating apomictic inconsistencies among the States are mini- berry thickets showing no symptoms of biotypes to a ‘taxonomic species’, based mized and so that weed managers can de- rust disease and to note which species are mainly on the range of distribution of the fine more accurately which taxa are con- present in an attempt to identify taxa that biotype. tributing to the weed problem. may be resistant to the strain, or strains, In Australia, the different blackberry Accurate taxonomic keys would en- of rust fungus present in that location taxa are grouped as the R. fruticosus L. ag- hance research efforts to improve the bio- (Bruzzese and Lane 1996). If these thickets gregate. Because of the subtle differences logical control of blackberry using the Eu- are allowed to expand, then the black- between these taxa and the inadequate ropean rust fungus. Blackberry leaf rust is berry infestation is maintained by replac- amount of reference material that has been a disease of blackberry caused by Phrag- ing one taxon with another. This phenom- authenticated, the names of the biotypes midium violaceum (Bruzzese and Lane enon occurred in the biological control introduced in this country are debatable. 1996). The susceptibility of different Rubus program for skeleton weed (Chondrilla The Rubus section of the Flora of Victoria taxa to individual strains of the European juncea) where the narrow-leaf form was (Walsh and Entwisle 1996) includes many rust fungus varies (Bruzzese and Hasan controlled successfully by a strain of the of the names applied in Australia to date 1986a,b,c). One strain of the rust fungus, rust fungus Puccinia chondrillina. The other

Table 1. Taxa, genotypes and localities of blackberry taxa collected in the 1996/1997 flowering season. No. of Rubus taxon GenotypeB StateC LocationsD samples R. affin. armeniacas Weber = 8 SA Scott Creek (7; -), Mt. Lofty (4; 15380, 15708, -, -), Stirling (2; 15378), 18 R. discolor sensu auct. aust. non Crafers West (1; -), Belair (1; 15712), Penwortham (1; 15376), Weihe & Nees Wilpena (1; 15348) Vic Murrongowan (3; EB2, -,-), Buenba (2; -), Somerville (5; -) 10 Tas New Norfolk (1; CG1), Hamilton (1; CG2), Maydena (2; CG5, CG6), 9 Mt. Field (1; CG3), Hesterway (1; CG4), Launceston (1; SW3), Newnham (1; SW4), Longford (1; SW5) ACT Black Mountain (3; LM1, -, -). 3 NSW Towomba (1; LM4), Harolds Cross (3; -) 4 WA Bunbury (1; EO1), Denmark (1, SL2), Manjimup (1; SL3), 6 Nannup (1; SL4), Grimwade (1; SL5), Quininup (1; SL6) R. polyanthemus Lindeb. 3 Vic Goongarah (5; -), Callignee (5; 15714, -, -, -, -) 10 R. polyanthemus Lindeb. 2 Vic Goongarah (1; -) 1 R. laciniatusA Willd. 5 SA Lenswood (1; 15406) 1 R. rubritinctusA W.C.R. Watson 7 SA Lenswood (1; 15362), Charleston (1; 15710) 2 R. erythropsA Edees & Newton 11 SA Mt. Lofty (3; 15457, 15457, 15711) 3 R. leucostachysA Schleich. ex Sm. 15 SA Slapes Gully (1; 15367), Seven Hill (1; 15709) 2 R. ulmifolius sens. lat. 14 Vic Frankston (5; -), Shellys Long Camp (4; -) 9 R. ulmifolius sens. lat. 1 Vic Buffalo River (2; -) 2 R. ulmifolius sens. lat. 9 Vic Foster (2; -) 2 R. ulmifolius sens lat. 10 Vic Foster (6; -) 6 R. ulmifolius sens lat. 13 Vic Benambra (1; -) 1 R. ulmifolius sens. lat. ? 6 Vic Frankston (5; -) 5 R. ulmifolius sens. lat. ? 4 Vic Callignee (1; -) 1 R. ulmifoliusA Schott 16 SA Mt. Osmond (1; 15421) 1 R. ulmifolius var. anoplothyrsusA Sud. 18 SA Waterfall Gully (1; 15370) 1 R. thibetanus Franch. 12 SA Mt. Lofty (1; 15713) 1 R. ? 17 SA Penwortham (1; 15377), Seven Hill (1; 15377) 2 R. parvifolius L. 19 SA Waterfall Gully (1; 15707) 1 R. parvifolius L. 20 NSW Harolds Cross (1; -) 1 A Taxa named by A. Newton and H. Weber. B Illustrated in Figure 1. C SA = South Australia, Vic = Victoria, Tas = Tasmania, ACT = Australian Capital Territory, NSW = New South Wales, WA = Western Australia. D The figures in parentheses refer to the number of plants genotyped followed by the reference number of the State Herbarium specimen, South Australia. A dash (-) means that the plant is not represented as a herbarium specimen. 154 Plant Protection Quarterly Vol.13(4) 1998 two apomictic forms of C. juncea are now Zimmerman et al. 1989, Meyer kbp spreading into areas formerly dominated et al. 1991). M13 DNA, when 9.4 by the narrow-leaf form (Cullen and hybridized to genomic DNA – Delfosse 1990). that has been digested with re- The Co-operative Research Centre for striction enzymes, detects Weed Management Systems has estab- length variants in minisatellite 6.6 lished a project to determine the suscepti- sequences among different ge- – bility of the different blackberry taxa to nomic DNA samples. Indi- strains of the European rust fungus vidual plants that have differ- present in Australia. The first objective is ent restriction-fragment pat- to define the blackberry taxa to be used in terns (DNA fingerprints) are 4.4 the pathogenicity and virulence studies. genetically different. A prob- – To overcome some of the current taxo- ability value, however, must be nomic difficulties in assigning names attached to the inference that 8 6 14 14 9 9 10 10 10 18 8 3 4 19 20 to some Rubus biotypes, we are determin- two plants are clones based on Genotype ing genotypes of the plant material to be the fact that their DNA finger- used in bioassays with the rust fungus. prints are identical (Jeffreys et Figure 1. Southern hybridization of M13mp18RF The genotype of an individual plant can al. 1985). DNA to HaeIII digested total Rubus DNA be determined by DNA fingerprinting Total DNA was extracted (approximately 3 µg per lane) from various (Jeffreys et al. 1985, Nybom 1996). The from 1.0 g samples of fresh Ru- genotypes as listed in Table 1. Each lane range of Rubus genotypes for which a rust bus leaflets or from 0.4 g sam- contains DNA from a different plant. strain is compatible will then be deter- ples of dried Rubus leaflets us- mined. In a separate project, conducted by ing a CTAB (cetyl trimethyl ammonium Results D. Symon at the State Herbarium, South bromide) extraction buffer as described by Of the fifty collections sent to A. Newton Australia, the taxonomy of Rubus is being Doyle and Doyle (1990). DNA samples and H.E. Weber, eight were identified reviewed as part of a larger project on Ro- were digested with the restriction enzyme to Series only as follows: Sylvatici, 5 saceae for the Flora of Australia. We have HaeIII and fractionated by electrophoresis collections; Vestiti, 1 collection; Hystrix, 1 linked the two projects to clarify some in a 1% agarose gel using Tris-acetate collection; and Micantes, 1 collection (Ta- taxonomic problems. Here we report the (TAE) buffer, pH 7.8, containing 40 mM ble 1). This may have been due to inad- correlation between the taxonomy, based Tris, 20 mM sodium acetate and 1 mM equacies of the specimens or to variants on morphological characters, and the EDTA. A sample of digested DNA from R. arising in Australia. Newton and Weber genotypes of various Rubus taxa as esti- affin. armeniacas was run in lanes two and provided the following names not previ- mated by DNA markers. eleven of each 20-well gel. The DNA was ously used in Australia: R. erythrops, R. transferred to Hybond-N (Amersham) ny- leucostachys, R. rubritinctus, R. ulmifolius Materials and methods lon membranes according to the method var. anoplothyrsus plus R. armeniacas and R. Plant collection of Southern (Southern 1975) and cross- anglocandicans for 17 specimens we named In December, 1996 and January, 1997, linked to the membranes using UV light R. discolor. Weber re-examined the mate- voucher specimens were prepared from from a GS Gene Linker (Bio-Rad). The rial we named R. discolor and considered Rubus plants collected from fifty locations DNA of bacteriophage M13 (M13mp18 none of them to be typical of R. armeniacas. in South Australia and sent to taxonomists RF, Biolabs, New England) was hybrid- Until this matter is resolved, we use R. in the United Kingdom (A. Newton, ized to the target DNA at 60°C according affin. armeniacas to cover the two (or one) Leamington Spa, Warwickshire) and in to the two-step procedure of Weihe et al. taxa. Germany (H.E. Weber, University of (1990). The probe was radio-labelled with Well-defined genotypes were deter- Vechta, Vechta). Eight voucher specimens a-32P-dCTP by random oligo-labelling us- mined using the DNA fingerprinting tech- were prepared at each location. Newton ing DNA labelling beads (Pharmacia nique. For each plant tested, this tech- and Weber each received one specimen Biotech). The labelled DNA was separated nique detected up to twelve distinct HaeIII per location, assessed the material inde- from unincorporated nucleotides in a restriction-fragments in the size range pendently then discussed the results be- MicroSpin™ (Pharmacia Biotech) column, 4–10 kbp (Figures 1 and 2). Restriction- tween themselves before concurring on denatured, then added to 10 mL of hy- fragments outside this size range were not the names of the taxa. The remaining bridization solution. Membranes were scored because they were either absent specimens are deposited in the State Her- washed at 65°C (Weihe et al. 1990) and ex- (>10 kbp) or too difficult to resolve (<4 barium, South Australia, and some of posed to X-ray film (X-Omat AR, Kodak) kbp). All Rubus taxa tested could be differ- these will be sent to herbaria located in at -80°C. entiated using this technique and 20 dif- Sydney, Canberra, Melbourne, Hobart ferent genotypes were observed among and Perth. In the same flowering season, Data analysis the 13 putative taxa (Table 1, Figure 2). No single voucher specimens were collected For each lane on the autoradiograph, genetic variation was observed among 50 at various locations in the other States as the relative front (RF) of each restriction plants of R. affin. armeniacas sampled from specified in Table 1. fragment, within the approximate size 29 locations throughout Australia. range of 4–10 kilobase pairs (kbp) and Genetic variation was observed within DNA fingerprinting producing a strong hybridization signal, R. ulmifolius sens. lat., R. polyanthemus and Young, healthy leaflets from the plant was calculated. In order to compare re- R. parvifolius. Seven different genotypes from which a voucher specimen was pre- striction-fragment patterns among auto- were identified among 26 plants of R. pared were processed for the extraction of radiographs, an adjusted RF was calcu- ulmifolius sens. lat. sampled from six loca- DNA. The genotypes of Rubus plants were lated as follows: tions in Victoria. Two of these genotypes determined by generating Restriction adjusted RF = 80 - [RFarm. - RF] were sampled from a single blackberry Fragment Length Polymorphisms (RFLPs) where RFarm. is the RF for the largest re- thicket at Foster. Genotype 14 of R. ulmi- using DNA from bacteriophage M13 as a striction fragment for R. affin. armeniacas folius sens. lat. had a very similar DNA fin- probe (Van Wezenbeek et al. 1980, observed on the same autoradiograph as gerprint to genotype 15 of R. leucostachys. Rogstad et al. 1988, Vassart et al. 1987, the sample. The DNA extracted from genotypes 14 Plant Protection Quarterly Vol.13(4) 1998 155 and 15 was separated on different gels. If, Nybom 1995). The two types of however, the DNA had been separated on data were well correlated, al- 1 the same gel, then it could be determined though interspecific variation 2 3 whether or not the smallest restriction- was sometimes more significant 4 fragment observed for both genotypes using morphological charac- 5 6 was a co-migrating fragment. Two geno- ters. 7 types of R. polyanthemus were detected This preliminary survey of 8 among six plants collected at the one loca- the genotypes of Rubus present 9 10 tion (Goongarah, Victoria) and two plants in Australia will be continued to 11 12 of R. parvifolius collected from SA or NSW include other taxa listed in New Genotype 13 were different genotypes. South Wales, Victoria and other 14 States, such as R. chloocladus, R. 15 Discussion cissburiensis, R. laciniatus subsp. 16 17 This study represents the first report of selmeri, R. leightonii, R. radula, R. 18 Rubus genotypes occurring in Australia. rosaceus and R. vestitus. In con- 19 20 We present the correlation of 20 genotypes junction with studies of mor- to 13 putative taxa of Rubus in Australia phological characters, we shall and provide six taxon names not previ- test the hypotheses that (a) 40 60 80 ously used in South Australia. A compari- some R. ulmifolius sens. lat. are Adjusted RF son of the voucher specimens from South R. leucostachys, (b) R. rubritinctus Figure 2. DNA fingerprints of various Rubus Australia with material collected in other = R. pyramidalis, and (c) R. genotypes as listed in Table 1. States has yet to be undertaken. leightonii and R. rosaceus are DNA fingerprinting, using the M13 closely related to R. erythrops. Once the would separate this genetically-distant probe, was found to be a useful tool for material from all States is compared, there species from the other Rubus taxa. The re- determining Rubus genotypes. Using this may be additional cases of synonomy. striction-fragment patterns for R. parvi- method, Antonius and Nybom (1994) de- R. laciniatus and R. ulmifolius are likely to folius were different from the patterns ob- tected 24 genotypes among 24 plants sam- be well-defined in all States. served for the other Rubus taxa (Figure 2). pled from a population of a wild and sexu- The results of the DNA fingerprinting Genetic variation is to be expected in this ally reproducing R. idaeus and estimated conducted in this study provide a prelimi- diploid species which probably repro- that the probability for the occurrence, by nary indication of the amount of genetic duces sexually (Thompson 1997). Another chance, of identical DNA fingerprints was variation occurring within and among explanation for the genetic variation in R. 1.4 × 10-3. In our study, DNA extracted taxa. The fact that no genetic variation was parvifolius is that the plant collected in from the same genotype at different times observed among the large sample of R. South Australia is from a population that and from different locations produced the affin. armeniacas plants suggests that this is isolated from the one occurring in New same restriction-fragment pattern, dem- common blackberry is probably a single South Wales (Bean 1997). onstrating the reproducibility of this tech- clone. Very low levels of genetic variabil- DNA fingerprinting of Rubus taxa was nique. DNA fingerprinting is particularly ity occur within apomictic taxa in Europe initiated in order to provide a record of the useful in clarifying the taxonomy of such as R. plicatus, R. scissus, R. gracilis, R. genotypes of the plant material to be used closely related taxa, where the separation infestus, R. grabowski, R. pedemontanus and in studies of the genetic interaction of the of two taxa based on morphological char- R. polyanthemus (Kraft and Nybom 1995, European rust fungus (Phragmidium vio- acters is questionable. When presented Kraft et al. 1996). Variation in DNA finger- laceum) with its blackberry host. This with two names for R. affin. armeniacas, we prints was not detected among five plants project could now be expanded to deter- concluded that these taxa were probably a of R. polyanthemus in Europe (Kraft and mine the amount and distribution of single genotype. Nybom 1995) whereas we identified one genetic variation within and among Rubus Similar problems to those described plant that had a slightly different geno- taxa in Australia. The collection of earlier have been investigated in Europe type (genotype 2) from that of the other 10 voucher specimens is essential in these where populations of Rubus with different plants (genotype 3) that were analysed. studies and provides the means for corre- names in different countries yielded simi- More samples of R. polyanthemus should lating genotypes with taxa. The informa- lar DNA fingerprints. Kraft and Nybom be tested to confirm the existence of this tion generated could be used by research (1995) present two cases where Swedish- DNA polymorphism. workers in a number of ways; for exam- named taxa and German-named taxa had Considerable genetic variation was ob- ple, to determine the most efficient strate- identical DNA fingerprints; that is, R. served among plants of R. ulmifolius sens. gies for introducing agents for biological scheutzii = R. muenteri and R. hartmanii = R. lat. from different locations in this study. control. Given that characterized and fuscus. In the latter case, there were signifi- The genotypes observed within this group uncharacterized strains of the European cant biometrical differences in several covered a large spectrum of the genotypes rust fungus are already present in this characters between the taxa which might observed in the study. Given that voucher country, the next step is to identify taxa be due to environmental effects and/or specimens have not yet been prepared for and genotypes that exhibit disease resist- the accumulation of somatic mutations. these plants, we make no conclusions re- ance when inoculated with these strains. Phenotypic plasticity may be common in garding the taxonomy of this group. A Additional strains could then be intro- Rubus. Populations of genetically identical more detailed study of this taxon is war- duced to target those resistant taxa. Not R. nessensis plants in Sweden (Antonius ranted. If some of these plants were the only should the resistant taxa be identi- and Nybom 1994) exhibited significant result of hybridization between taxa, then fied, but their genetic variability and bio- variation in parameters related to plant experiments to determine the origin of geography should be estimated. demography and flowering phenology these hybrids could be conducted. Differential susceptibility of Rubus (Nybom 1987a,b). DNA fingerprint data Rubus parvifolius is a widespread native genotypes to strains of the rust fungus is have been compared with the results from species (Bean 1997) that is not closely re- likely to occur and individual strains are a biometrical analysis of morphological lated to R. fruticosus L. agg. This species likely to cause more damage in popula- characters measured on herbarium speci- was included in this study to assess how tions that are genetically homogeneous. mens in Sweden and Germany (Kraft and well the DNA fingerprinting technique Burdon and Marshall (1981) argue that the 156 Plant Protection Quarterly Vol.13(4) 1998 genetic structure of a target species has violaceum, a potential biological control Rogstad, S.H., Patton, J.C.I. and Schaal, important implications with respect to the agent of European blackberry. Annals of B.A. (1988). M13 repeat probe detects likelihood of success of biological control. Applied Biology 108, 585-96. DNA minisatellite-like sequences in In their survey and analysis of data in the Bruzzese, E. and Hasan, S. (1986c). Infec- gymnosperms and angiosperms. Pro- literature, biological control programs at- tion of Australian and New Zealand ceedings of the National Academy of Sci- tempted with 44 asexually-reproducing Rubus subgenera Dalibarda and Lampo- ence, USA 85, 9176-8. plant species were successful in 21 cases batus by the European blackberry rust Southern, E.M. (1975). Detection of spe- (48%), while in sexually-reproducing fungus Phragmidium violaceum. Plant cific sequences among DNA fragments plant species biological control was suc- Pathology 35, 413-16. separated by gel electrophoresis. Jour- cessful in only four of 29 (14%) cases. In Bruzzese, E. and Lane, M. (1996). ‘The nal of Molecular Biology 98, 503-17. those Rubus populations that are geneti- blackberry management handbook’. Thompson, J.N. and Burdon, J.J. (1992). cally heterogenous, the release strategy (Keith Turnbull Research Institute, Gene-for-gene coevolution between for the European rust fungus might be to Frankston). plants and parasites. Nature 360, 121-5. deploy mixtures of strains. Before and af- Burdon, J.J. and Marshall, D.R. (1981). Bio- Thompson, M.M. (1997). Survey of chro- ter strains are released, however, genetic logical control and the reproductive mosome numbers in Rubus (Rosaceae: structures of rust and blackberry popula- mode of weeds. Journal of Applied Ecol- Rosoideae). Annals of the Missouri Bo- tions should be characterized in order to ogy 18, 649-58. tanical Gardens 84, 128-64. monitor co-evolutionary events (Thomp- Cullen, J.M. and Delfosse, E.S. (1990). Van Wezenbeek, P.M.G.F., Hulsebos, son and Burdon 1992). We demonstrate Progress and prospects in biological T.J.M. and Schoenmakers, J.G.G. (1980). the utility of genotyping Rubus plants for control of weeds. Proceedings of the Nucleotide sequence of the filamentous clarifying taxonomic problems and rec- Ninth Australian Weeds Conference, bacteriophage M13 DNA genome: com- ommend that DNA markers be used to Adelaide, pp. 452-76. parison with phage fd. Gene 11, 129-48. characterize rust and blackberry popula- Doyle, J.J. and Doyle, J.L. (1990). Isolation Vassart, G., Georges, M., Monsieur, R., tions and thus elucidate the genetic factors of plant DNA from fresh tissue. Focus Brocas, H., Sophie, A. and Christophe, required for successful biological control. 12, 13-15. D. (1987). A sequence in M13 phage de- Edees, E.S. and Newton, A. (1988). ‘Bram- tects hypervariable minisatellites in Acknowledgments bles of the British Isles’. (Ray Society, human and animal DNA. Science 235, We thank A. Newton (UK) and H.E. We- London). 683-4. ber (Germany) for their assistance in nam- Jeffreys, A.J., Wilson, V. and Thein, Walsh, N.G. and Entwisle, T.J. (1996). ing the Rubus collections. We thank Franz S.L. (1985). Individual-specific ‘finger- ‘Flora of Victoria’, Volume 3. (Inkata Mahr and El Bruzzese (Department of prints’ of human DNA. Nature 316, Press, Melbourne, Victoria). Natural Resources and Environment, Vic- 76-9. Weber, H.E. (1996). Former and modern toria) for supplying all the plant material Kraft, T. and Nybom, H. (1995). DNA fin- taxonomic treatment of the apomictic from Victoria. We also thank the other gerprinting and biometry can solve Rubus complex. Folia Geobotanica et Rubus collectors and advisors; namely, some taxonomic problems in apomictic Phytotaxonomica 31, 373-80. John Hosking, Louise Morin, Andy blackberries (Rubus subgen. Rubus). Weihe, A., Niemann, C., Lieckfield, D., Sheppard, Sandy Lloyd, Eric Orchard, Watsonia 20, 329-43. Meyer, W. and Borner, T. (1990). An im- Gwen Mayo, Andrew Bishop, Stephen Kraft, T., Nybom, H. and Werlemark, G. proved hybridization procedure for Welsh and Christian Goninon. (1996). 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