et al .

Australian Journal of Entomology (2005) 44, 68–70

Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), and huanglongbing disease do not exist in the Stapleton Station area of the Northern Territory of Australia

Glenn Bellis,1* David Hollis2 and Sarah Jacobson3

1Australian Quarantine and Inspection Service, Northern Australia Quarantine Strategy, GPO Box 3000, Darwin, NT 0801, Australia. 2The Natural History Museum, London (BMNH), London SW7 5BD, UK. 3Australian Quarantine and Inspection Service, Northern Australia Quarantine Strategy, PO Box 1054, Mareeba, Qld 4880, Australia.

Abstract A series of specimens of the Asian citrus psyllid, Diaphorina citri, collected from the Northern Territory (NT) in 1915 was recently rediscovered in the Natural History Museum, London. Surveys were conducted in 2002 on suitable hosts in the locality of the 1915 collections to see if the infestation had persisted. These failed to detect either D. citri or the bacterium that it transmits and that causes huanglongbing disease in citrus. It is presumed that D. citri was eradicated fortuitously by the removal of all citrus plants above latitude 19∞S during an eradication program for citrus canker in the NT from 1916 until 1922. Key words Asian citrus psyllid, citrus, Diaphorina citri, huanglongbing, Quarantine.

INTRODUCTION 95 citrus plants in this area in 1912. The source of the infection is believed to be plants imported into the Darwin Botanic The Asian citrus psyllid, Diaphorina citri Kuwayama, is a Gardens, probably from China or Japan, and it is possible that major pest of citrus in central and southern Asia extending as the infestation of D. citri arose from a similar importation. In far east as the island of New Guinea (Aubert 1990). Much of 1915 and 1916 prohibitions on the importation of citrus plants its pest status is due to its ability to transmit the bacterium and fruit into Australia from canker-affected countries were ‘Candidatus (Liberibacter) asiaticus’ that causes huanglong- implemented and in 1918 all trees on affected properties were bing (HLB) disease of citrus (Aubert 1990). Although it has destroyed, mostly by burning (Hill 1918). In 1922 a decision not officially been recorded from Australia, the Natural His- was made to eradicate all citrus trees in NT north of latitude tory Museum, London (BMNH) collection holds a sample of 19∞S, which lies about 700 km south of Darwin, and to pro- 20 pinned specimens of D. citri, in poor condition, bearing the hibit the replanting of citrus trees until 1925 (Anonymous labels: Australia: Stapleton. N.T. 16.vii.1915. G.F.Hill./Pres. 1922; Mertin 1952). by Imp.Bur.Ent. Brit.Mus. 1923-387./G. Heslop-Harrison All known host plants of D. citri belong to the family Det.195[2] [Diaphorina citri (Kuw)]/[] = handwritten. One of Rutaceae and incorporate a wide range of species including the specimens also bears a printed and handwritten label (? 10 from genera other than Citrus (Aubert 1990). Two of these, original): Stapleton [16.7.15.] N.T. G.F.Hill and at right angles Bergera koenigii (curry bush) and Murraya paniculata (jas- on the righthand margin: [352]/[] = handwritten. The relevant mine orange), are excellent hosts and are commonly grown in data in BMNH accession register states ‘120 Rhynchota. gardens in the NT. The latter species is also native to coastal Presented by Imperial Bureau of Entomology 16.viii.1923. and subcoastal vine thickets of the NT, but has not been Submitted by G.F. Hill (Australia)’. recorded from the area around Stapleton Station (Brock 1993). Stapleton Station 13∞10.53¢S 131∞01.38¢E was founded in The ability of D. citri to survive on other rutaceous plants the 19th century on the rail line between Pine Creek and native to the NT is unknown. It breeds on new flush with Batchelor, about 100 km south of Darwin, Northern Territory populations being higher and easier to detect on flushing trees (NT), Australia. An exotic disease of citrus, citrus canker Xan- and survival of D. citri is better during cool, rather than hot, thomonas axonopodis pv. citri, was detected in an orchard of humid weather (Aubert 1989). No records exist of subsequent collections of D. citri in the NT, nor have there been any reported cases of HLB, despite *Author to whom correspondence should be addressed (email: the presence of a viable citrus industry. Additionally, surveys [email protected]). for exotic pests and diseases, including D. citri and HLB,

Survey for Asian citrus psyllid in NT 69 which are regularly conducted in Darwin by the Northern Young foliage of all rutaceous plants growing on these Territory Department of Business, Industry and Resource properties was inspected visually for D. citri adults and lar- Development (NTDBIRD) and throughout coastal NT by the vae and sampled using a net. Native vegetation growing on Northern Australia Quarantine Strategy (NAQS) of the these properties, particularly along Stapleton Creek, was Australian Quarantine and Inspection Service, have not thus examined and any rutaceous plants were inspected similarly far detected either organism (ESC Smith, pers. comm. 2003; for D. citri. GA Bellis unpubl. data 2003; Weinert et al. 2004). Despite Citrus plants exhibiting interveinal leaf chlorosis consistent this, it was possible that a population of D. citri had managed with their having HLB were tested for the presence of the to survive undetected on native rutaceous plants in the area causative agent, ‘Candidatus Liberibacter asiaticus’. The mid around Stapleton Station. The threat posed by D. citri and veins of symptomatic leaves were excised, dried over calcium HLB to the Australian citrus industry was sufficiently serious chloride and screened at the NAQS Mareeba laboratory (spec- to warrant investigation to determine if this infestation still imen #GB22), using DNA extraction techniques described by existed. This paper outlines surveys of the area around Staple- Ahrens and Seemüller (1992) and polymerase chain reaction ton Station for D. citri and HLB. primers described by Jagoueix et al. (1996).

METHODS AND MATERIALS RESULTS

The exact site of the orchard of 95 trees at Stapleton from The rutaceous plants growing at the six sites are listed in which the specimens of D. citri were probably collected could Table 1. No D. citri were found on any of the plants inspected not be located accurately. Hill (1918) notes that it was close at any of these sites in either survey. During the September to a permanent creek from which it was irrigated. Five prop- survey, the majority of Citrus sp. trees were flushing, but few erties in the vicinity of the original Stapleton Homestead, were flushing in March. including those containing the springs which supply Stapleton The only rutaceous plants growing in native vegetation in Creek, the only permanent creek in the area, were included in the area were Melicope elleryana, which is not a recognised the surveys. Two surveys were conducted during different host of D. citri, and no psyllids were found on the 13 plants seasons to ensure that flushing citrus were surveyed, thereby inspected. ensuring maximal opportunity of detecting D. citri. The first Only one citrus plant exhibited symptoms similar to that of occurred on 24 March 2002 at the end of the hot, wet season HLB. Leaves from this plant tested negative for the causative and included the four properties Mabuhay Farm, Milton organism. Springs, Litchfield Springs and Camp Creek Station. The sec- ond survey occurred on 7 September 2002 at the end of the cool, dry season and included the three properties Mabuhay DISCUSSION Farm, Jim Roddy’s property (the site of the original Stapleton Homestead) and Milton Springs. Property owners indicated There is no evidence that D. citri has persisted in the Stapleton that no citrus or related plantings existed beyond the confines Station area. No native plants known to be capable of support- of their homesteads and searches for native rutaceous plants ing this insect were present and a number of attractive host were confined to creek lines and springs that provided the only plants were not infested either during the wet season or in the suitable habitat. cool dry season when many of these plants were flushing.

Table 1 Properties and plants inspected for Diaphorina citri and symptoms of huanglongbing disease in the area around Stapleton Station, Northern Territory of Australia

Site Plants inspected Notes Mabuhay Farm 1 lemonade (Citrus limon ¥ C. reticulata); 1 pink grapefruit No native Rutaceae (C. paradisi); 1 kaffir lime (C. hystrix); 1 orange (C. sinensis); 2 Eureka lemons (C. limon); 1 Lisbon lemon (C. limon); 2 pomello (C. grandis); 3 lime (C. aurantifolia); 7 kumquat (C. japonica); Jim Roddy’s Property 2 Poncirus trifoliata; 1 lime; 1 lemonade; 1 old kumquat; 1 Melicope elleryana was the only native rutaceous plant small ornamental Murraya paniculata; 12 Melicope elleryana growing around the natural spring at the back of the old homestead site. Milton Springs 4 ornamental Murraya paniculata and 1 pommelo, all at least Melicope elleryana was the only native rutaceous plant 20 years old; 1 Melicope elleryana growing around the natural spring at the back of the homestead. Litchfield Springs 12 Citrus sp. trees No native Rutaceae Camp Creek 5 Citrus sp. trees No native Rutaceae

70 G Bellis et al.

The presence of HLB-like symptoms in the absence of the amplifies a sequence of the 16S rRNA gene. Phytopathology 82, 828Ð causative agent is not unusual, as the symptoms are similar to 832. Anonymous. 1922. Annual Report, pp. 13Ð14. Administrator of the those caused by nutrient deficiency (Davis et al. 2000). Northern Territory, Darwin. The results of the present survey coupled with those con- Aubert B. 1989. Management of the citrus greening disease in Asian ducted by NTDBIRD and NAQS indicate that D. citri is not countries. In: Proceedings of the 2nd FAO-UNDP Regional Work- present in the NT. It is likely that D. citri was eradicated shop on Asian/Pacific Citrus Greening, Lipa, Philippines, 20Ð26 November 1988 (eds B Aubert, K Chung & CI Gonzales), pp. 51Ð fortuitously along with citrus canker in 1922. Stapleton Station 52. was well within the eradication area and had been specifically Aubert B. 1990. Integrated activities for the control of huanglongbing- targeted during another eradication program initiated 4 years greening and its vector Diaphorina citri Kuwayama in Asia. In: earlier. Monitoring by NTDBIRD and NAQS for D. citri will Proceedings of the Fourth FAO-UNDP International Asia Pacific Conference on Citrus Rehabilitation, Chiang Mai, Thailand, 4Ð10 continue as part of a national quarantine initiative. February 1990 (eds B Aubert, S Tontyaporn & D Buangsuwon), pp. 133Ð144. Brock J. 1993. Native Plants of Northern Australia. Reed Books Australia, ACKNOWLEDGEMENTS Kew, Australia. Davis RI, Jacobson SJ, Rahamma S & Gunua TG. 2000. Surveillance for citrus huanglongbing (greening) disease in New Guinea and north We thank the owners and caretakers of these properties for Queensland. Australasian Plant Pathology 29, 226. access to their plants and for information on the history of the Hill GF. 1918. History of citrus canker in the Northern Territory (with notes of its occurrence elsewhere). Bulletin of the Northern Territory Stapleton area. In particular we thank Bill Lane of Mabuhay of Australia 18, 1Ð8. Farm for his local knowledge. We are grateful to Gary Lambert Jagoueix S, Bové JM & Garnier M. 1996. PCR detection of the two formerly of Milton Springs for information on the history of ‘Candidatus’ Liberobacter species associated with greening disease the area and to Andrew Mitchell NAQS, for botanical support of citrus. Molecular and Cellular Probes 10, 43Ð50. Mertin JV. 1952. Plant quarantine survey in the Northern Territory. Jour- during the second survey. nal of the Australian Institute of Agricultural Science 18, 27Ð32. Weinert MP, Jacobson SC, Grimshaw JF et al. 2004. Detection of huanglongbing (citrus greening disease) in Timor-Leste (East REFERENCES Timor) and in Papua New Guinea. Australasian Plant Pathology 33, 135Ð136.

Ahrens U & Seemüller E. 1992. Detection of DNA of plant pathogenic mycoplasma-like organisms by a polymerase chain reaction that Accepted for publication 3 March 2004.