Phylogenetic Relationships Among Global Populations of Pseudomonas syringae pv. actinidiae

ABSTRACT Pseudomonas syringae pv. actinidiae, the causal agent of canker in kiwifruit (Actinidia spp.) vines, was first detected in Japan in 1984, followed by detections in Korea and Italy in the early 1990s. Isolates causing more severe disease symptoms have recently been detected in several countries with a wide global distribution, including Italy, New Zealand, and China. In order to characterize P. syringae pv. actinidiae populations globally, a representative set of 40 isolates from New Zealand, Italy, Japan, South Korea, Australia, and Chile were selected for extensive genetic analysis. Multilocus sequence analysis (MLSA) of housekeeping, type III effector and phytotoxin genes was used to elucidate the phylogenetic relationships between P. syringae pv. actinidiae isolates worldwide. Four additional isolates, including one from China, for which shotgun sequence of the whole genome was available, were included in phylogenetic analyses. It is shown that at least four P. syringae pv. actinidiae MLSA groups are present globally, and that marker sets with differing evolutionary trajectories (conserved housekeeping and rapidly evolving effector genes) readily differentiate all four groups. The MLSA group designated here as Psa3 is the strain causing secondary symptoms such as formation of cankers, production of exudates, and cane and shoot dieback on some kiwifruit orchards in Italy and New Zealand. It is shown that isolates from Chile also belong to this MLSA group. MLSA group Psa4, detected in isolates collected in New Zealand and Australia, has not been previously described. P. syringae pv. actinidiae has an extensive global distribution yet the isolates causing widespread losses to the kiwifruit industry can all be traced to a single MLSA group, Psa3.     

Dutch elm disease in New Zealand: impacts from eradication and management programmes

Ophiostoma novo‐ulmi, the cause of the current pandemic of Dutch elm disease, is considered one of the 20 worst pests to have been introduced to New Zealand. An attempt to eradicate the pathogen came close to success, but was eventually stopped and replaced by management programmes. These programmes are slowing the spread of the pathogen. If left uncontrolled it is likely that New Zealand will lose 90% of its elm trees. Although elms are exotic trees in New Zealand, the loss of such a large number of popular amenity trees is likely to have numerous social, cultural, environmental and economic effects. It is estimated that this disease could cost public and councils over NZD $350 million in tree removal and replacement costs. Control of O. novo‐ulmi is limited, but biological control using d‐factor viruses has been proposed. Although O. novo‐ulmi was not eradicated from New Zealand, the eradication programme had many successes; in particular, the effective interaction of good management and science in the early years of the campaign. Also highlighted in this review are several tactics and steps that could be applied to eradication attempts for other species.     

Recent Invasions of Australia and New Zealand by Pathogenic Fungi and Counter Measures1

Despite the relative isolation of Australia and New Zealand and the strict quarantine regulations long imposed, many plant diseases have been introduced in recent years. The article analyzes 26 cases since 1960 which seem certain to be new arrivals, and have some economic significance. Wind-borne introduction from the islands to the north seems likely in several cases ( Cochliobolus heterostrophus, Peronosclerospora maydis, Puccinia arachidis, Mycosphaerella fijiensis var. difformis ) and air currents are certainly responsible for many arrivals in New Zealand from Australia (acacia rusts, Puccinia menthae, P. helianthi. Puccinia melanocephal (sugar cane rust) is one case of possible intercontinental movement west to east on prevailing air currents (from Africa to India and the Far East, then Australia and Central America). However, accidental introduction by air travellers, or illegally introduced planting material, seems most likely to explain the recent appearance in Australia of Puccinia striiformis and Melampsora spp. on poplar, even if these have since spread naturally to New Zealand. Dothistroma septospora , causing a needle blight of Pinus radiata , is unusual in that it apparently moved from New Zealand to Australia (around 1975). Internal restrictions can limit further spread in some cases. Otherwise, the « cost >> of these introductions has mainly arisen from the need for fungicide treatments and from the abondonment of valuable, but susceptible, cultivars. Introduced fungi may not behave as expected on the basis of earlier biological studies. Thus, P. striiformis survives the harsh Australian summers while D. septospora which necessitates costly fungicide treatments in New Zealand, has remained isolated and unimportant in Australia.     

Support for a stepwise mutation model for pathogen evolution in Australasian Puccinia striiformis f.sp. tritici by use of molecular markers

Since its initial detection in Australia in 1979, wheat yellow (stripe) rust ( Puccinia striiformis f.sp. tritici ) has evolved in Australia and New Zealand into more than 20 pathotypes with assorted virulence characteristics. This evolution is believed to have occurred in a stepwise fashion from an original single pathotype, with no subsequent new introductions. A combination of random amplified polymorphic DNA (RAPDs) and amplified fragment length polymorphisms (AFLPs) was used to examine the level of molecular variation in Australian and New Zealand isolates, and to compare this with variation amongst other isolates of P. striiformis . Using 60 RAPD primers on seven Australian isolates representing seven different pathotypes collected between 1979 and 1991, more than 300 potentially polymorphic loci were analysed and no polymorphisms were detected. Using the same primers on two UK isolates, 3% of loci showed a polymorphism. A similar level of polymorphism was found between UK isolates using AFLP primers, and between 5 and 15% of fragments were polymorphic between an isolate from the UK, an isolate from Denmark, and one from Colombia. However, no AFLP polymorphisms were found amongst 14 Australian and New Zealand isolates tested, at over 100 potentially polymorphic loci. The lack of molecular variation in the Australian and New Zealand collection is consistent with the stepwise mutation theory of pathotype evolution from a single introduction.     

Epidemiology of sharka disease in France

Plum pox virus was first detected in France in the 1960s. Both PPV-D and PPV-M strains are present but epidemics related to the PPV-M strain detected in the late 1980s are the most problematic. The two PPV strains have unequal distributions in peach and apricot orchards and different prevalences. More than 20 different aphid species have been identified as vectors of PPV but most of them do not colonize Prunus species. Thus, aphids involved in the spread of PPV in orchards are essentially visiting aphids. The main sources of inoculum for the vectors are leaves and fruits of infected stone-fruit trees. Spontaneous, wild and ornamental Prunus species such as Prunus dulcis , P. spinosa or P. pissardii are susceptible to PPV isolates found in France but their role as a reservoir in sharka epidemics is probably negligible. The disease spreads rapidly in orchards but the rate of progression may vary according to the identity of the PPV strain and the Prunus species. Analysis of spatial patterns of disease has shown that secondary spread by aphids frequently occurs over short distances in the orchards (aggregated patterns) but also that dissemination at longer distances (of several hundred metres) is a common event.     

Reclassification of an isolate of Guignardia citricarpa from New Zealand as Guignardia mangiferae by sequence analysis

Citrus black spot was recorded as present in New Zealand in international databases on the basis of one isolate (ICMP 8336) identified by morphological features as Guignardia citricarpa . This isolate was from a soft rot, not a typical symptom of citrus black spot, on a Seminole tangerine fruit grown in the northern region of New Zealand. Sequence analysis of the internal transcribed spacer (ITS) region (ITS1, 5·8S and ITS2) showed that this isolate was 99% identical to the ITS region of G. mangiferae , a closely related saprotroph. Despite climatic conditions being suitable for this disease, citrus black spot symptoms have never been seen on citrus fruits grown in New Zealand. Thus the absence of symptoms on citrus in New Zealand is probably because G. citricarpa is not present. On the basis of these results, the record of citrus black spot occurring in New Zealand should be re-examined.     

Hosts and symptoms of Plum pox virus: Herbaceous hosts

Plum pox virus (PPV) is polyphagous and epidemic. Apart from cultivated and wild Prunus species, a large number of herbaceous plants can be hosts of the virus. New herbaceous host species are continuously being reported following artificial inoculation studies. Some of these herbaceous hosts, Chenopodium foetidum , Nicotiana clevelandii , N. benthamiana and Pisum sativum are very useful for concentrating and purifying the virus. The list of plants that have been found to be infected with PPV in their natural environment is shorter than the list of plants which can be experimentally infected. The role of weed species in PPV survival and spread in orchards is poorly understood. It is widely accepted that annual plants or weeds are not important in the epidemiology of PPV.     

中国蔷薇科归化植物新纪录——尖齿黑莓

安徽省滁州市发现中国蔷薇科新记录植物——尖齿黑莓(Rubus argutus Link)。该种原产北美洲,是许多栽培树莓的亲本,目前已在日本、新西兰、夏威夷等地归化,并造成一定的生态危害。尖齿黑莓具有掌状复叶、茎具棱、皮刺极为发达,易与国产悬钩子属植物显著区别。文章调查了其分布面积、提供详细的照片与形态描述,介绍了其生物学特性、危害、价值及国外的防治手段,并就其入侵风险进行讨论。凭证标本保存于安徽师范大学生命科学学院植物标本馆(ANUB)。     

Susceptibility of native New Zealand Myrtaceae to the South African strain of Austropuccinia psidii: A biosecurity threat

Austropuccinia psidii, cause of myrtle rust, has spread globally where Myrtaceae occur. Multiple strains of A. psidii have been identified, including a unique strain found only in South Africa. The South African strain is a biosecurity concern for species of Myrtaceae worldwide. This is because preliminary testing of South African Myrtaceae suggests it could have a wide host range, and thus has the potential to be invasive. In this study, we assessed the ability of the South African strain to infect other species of Myrtaceae by testing the susceptibility of New Zealand provenance Myrtaceae. Seedlings of four native New Zealand Myrtaceae species (Metrosideros excelsa, Leptospermum scoparium, Kunzea robusta, and Kunzea linearis) were artificially inoculated in South Africa with a single-uredinium isolate of the South African strain. Fourteen days after inoculation, uredinia, and in many cases telia, had developed on the young leaves and stems of all four host species, which led to shoot tip dieback in the more severe cases. When comparisons were made between the species, K. robusta was the least susceptible to the South African strain of A. psidii, while L. scoparium and M. excelsa were the most susceptible. While only a limited number of seed families were tested, only a small proportion of the seedlings showed resistance to infection by the South African strain. This preliminary testing highlights the potential invasive risk the South African strain poses to global Myrtaceae communities, including New Zealand Myrtaceae.     

Characterization of a strain of Apple stem grooving virus in Actinidia chinensis from China

A new strain of Apple stem grooving virus (ASGV) has been identified in Actinidia chinensis imported from China. The leaves of these plants exhibited a variety of symptoms including interveinal mottling, chlorotic mosaics and ringspots. Capillovirus-like particles were observed under the electron microscope, and the virus could be mechanically transmitted to a range of herbaceous indicators. The virus was detected using ELISA with antisera raised against ASGV. Sequencing of the virus revealed that it had more than 95% amino acid identity with ASGV in the putative coat and movement proteins. From the morphological, transmission, serological and molecular evidence, it was concluded that the virus is a strain of ASGV. It is not known how this strain of ASGV is transmitted, other than by grafting, nor is it known what effect the virus has on the growth of infected vines. The Actinidia -infecting strain of ASGV does not occur in New Zealand, and infected plants will not be released from quarantine. The detection methods used during the research will assist quarantine and the safe movement of breeding material.     

The current status of pesticide resistance in Australian populations of the redlegged earth mite (Halotydeus destructor)

BACKGROUND: The redlegged earth mite, Halotydeus destructor Tucker, is an important pest of broad‐acre farming systems in Australia, New Zealand and South Africa. In 2006, high levels of resistance to synthetic pyrethroids were discovered in this species in Western Australia. RESULTS: Between 2007 and 2010, the authors monitored resistance in field populations and found it had spread considerably within the state of Western Australia. Twenty‐six paddocks from 15 individual properties were identified with resistance, and these paddocks ranged over 480 km. To date, resistance has not been detected in any other Australian state. Resistance in H. destructor appears to be found across the entire pyrethroid group, but not to other chemical classes such as organophosphates and carbamates, or other chemistries with novel modes of action. CONCLUSION: The high levels of resistance occurring in Western Australia have caused considerable economic losses due to ineffective chemical applications and mortality of crop plants at seedling establishment. These findings highlight the need for a comprehensive resistance surveillance programme to be developed for H. destructor within Australia. Growers need to consider non‐chemical approaches for pest control and should be encouraged to implement pesticide resistance management programmes for H. destructor. Copyright © 2011 Society of Chemical Industry     

新西兰出口苹果的有害生物监控及检疫监管体系

新西兰苹果出口到欧盟、亚洲、美洲和中东等65个国家和地区,其国内水果生产建立了较完善的出口苹果有害生物监控及检疫监管体系,通过介绍新西兰苹果出口的监管体系、有害生物监控防治、出口检验检疫等,供国内检验检疫部门和农业生产部门借鉴和参考。     

A further record of the New Zealand flatworm Artioposthia triangulata (Dendy) in England

Studies have shown the New Zealand flatworm to be a predator of earthworms and to be widely distributed throughout most of Ireland and Scotland. Except for one record in 1965, it had not been found in England until, in December 1992, it was positively identified from a garden centre near Manchester. Viable and empty egg capsules were also found suggesting that it had reproduced and become established in that nursery. During 1993 other confirmed reports were received from Yorkshire and East Anglia, indicating that it may be able to survive and multiply across large areas of England.     

Serological characterization of Italian isolates of plum pox potyvirus1

An Italian isolate of plum pox potyvirus (PPV) from apricot, Ispave 17, was used as antigen for production of monoclonal antibodies. Six clones secreting specific antibodies to PPV were obtained. All these monoclonal antibodies were used to test a collection of different Italian PPV isolates, collected from plum, apricot and peach orchards, and other European isolates (including PPV-D and PPV-M serotypes), using DAS-ELISA, SDS-PAGE, western blot and GIEM. In western blot analysis, the PPV-M and PPV-D coat protein, detected directly from crude peach GF305 extracts, showed different electrophoretic mobility, the coat protein of PPV-M being slightly larger than that of PPV-D. ELISA tests, performed with fixed dilutions of antibodies and limiting dilutions of clarified samples, showed with some monoclonal antibodies a marked difference between PPV-M and PPV-D strains, at ratios greater than 1:40 (w/v). Also in GIEM some monoclonal antibodies gave a good labelling reaction only with PPV-D serotype. With the help of this differentiation, it was found that all Italian isolates tested were of the D serotype and none of the severe M strain of PPV, which has not been reported in Italy.     

Control and monitoring: eradication of Plum pox virus in Switzerland

In 1967, sharka disease ( Plum pox virus ; PPV) was detected for the first time in Switzerland. By the end of the 1970s, the disease was considered as eradicated after a rigorous campaign of survey and eradication. Since then, periodical inspections and random testing of imported plants and young orchards have been performed by Swiss plant protection services. This permitted the detection of outbreaks of PPV in 2004. Consequently, a new survey and eradication campaign began.     

Detection and characterization of Plum pox virus: serological methods

Tremendous progress has been made in the research and development of Plum pox virus (PPV) serological reagents and methods in recent years. Two facts have revolutionised the serological detection and characterization of the virus: the development of the ELISA method in 1977, and the later emergence of specific monoclonal antibody technology. The availability of commercial kits has popularised PPV diagnosis, now making diagnosis possible at large scale for quarantine purposes, eradication programmes and control of the disease in nurseries. The use of the universal monoclonal antibody 5B-IVIA, used in DASI-ELISA, is the most accurate system for routine PPV detection. Likewise, the use of typing monoclonal antibodies gives exact characterization of the main PPV types described: 4DG5 for PPV-D, AL for PPV-M, EA24 for PPV-EA, and TUV and AC for PPV-C. There is, in general, an excellent correlation between serological data obtained with PPV specific monoclonal antibodies and data obtained by molecular PCR based methods. ELISA using a single or a mixture of monoclonal antibodies will remain the preferred method for universal detection and routine screening of PPV for years to come. Today, other serological methods and reagents are also recommended in the EPPO Diagnostic Protocol, increasing the number of reliable tests available for PPV detection. These developments have helped to control sharka disease in recent years. International co-operation in this field has been crucial to the improvement and validation of serological tools for PPV detection and characterization.     

Plum pox virus detection in dormant plum trees by PCR and ELISA

An immunocapture polymerase chain reaction (IC-PCR) protocol and ELISA were compared for their effectiveness in detecting plum pox virus (PPV) in dormant plum material. Although the IC-PCR was about one thousand times more sensitive than ELISA, PPV was detected by ELISA in 71–80% of bark samples collected in December, January and March 1996/97 from pot-grown rootstock trees inoculated with PPV the previous March, compared with 85–86% detection in the same samples by IC-PCR. In similar samples from one-year-old shoots taken from infected branches of orchard trees, 66–81% were positive by ELISA compared with 81–87% by IC-PCR. With bulked samples taken from the fibrous roots of the pot-grown trees, PPV was detected in 92–100% of samples by IC-PCR in winter compared with only 38–65% by ELISA. These results were confirmed in samples from the roots and shoots of the same trees in 1997/98. Three samples per shoot would have been sufficient to detect PPV by ELISA in 87 of the 88 infected shoots tested during the two winters. However, infected shoots are irregularly distributed in diseased trees and PCR assays of root samples offer the potential for improving the reliability of identifying trees infected with PPV.