Integrating drivers influencing the detection of plant pests carried in the international cut flower trade.

This paper analyses the cut flower market as an example of an invasion pathway along which species of non-indigenous plant pests can travel to reach new areas. The paper examines the probability of pest detection by assessing information on pest detection and detection effort associated with the import of cut flowers. We test the link between the probability of plant pest arrivals, as a precursor to potential invasion, and volume of traded flowers using count data regression models. The analysis is applied to the UK import of specific genera of cut flowers from Kenya between 1996 and 2004. There is a link between pest detection and the Genus of cut flower imported. Hence, pest detection efforts should focus on identifying and targeting those imported plants with a high risk of carrying pest species. For most of the plants studied, efforts allocated to inspection have a significant influence on the probability of pest detection. However, by better targeting inspection efforts, it is shown that plant inspection effort could be reduced without increasing the risk of pest entry. Similarly, for most of the plants analysed, an increase in volume traded will not necessarily lead to an increase in the number of pests entering the UK. For some species, such as Carthamus and Veronica, the volume of flowers traded has a significant and positive impact on the likelihood of pest detection. We conclude that analysis at the rank of plant Genus is important both to understand the effectiveness of plant pest detection efforts and consequently to manage the risk of introduction of non-indigenous species.

Molecular evidence that the aphid-transmitted Tomato mild mottle virus belongs to the Potyviridae family but not the Potyvirus genus.

Molecular studies have been carried out on Tomato mild mottle virus (TMMV) and 2322 nucleotides of the aphid-transmitted virus have been sequenced. This translates to a 2187 nucleotide open reading frame and a 135 nucleotide untranslated region (UTR) followed by a poly A tail. The sequence is consistent with the NIb and coat protein regions of a member of the Potyviridae family of viruses. Sequence comparisons show this virus to have no close relationship with any previously sequenced aphid-transmitted virus; the highest identity was found with the whitefly-transmitted Ipomoviruses. The vector of transmission and the low level of sequence identity with other aphid-transmitted viruses suggest TMMV does not belong to any existing genus of the Potyviridae family.

Subgroup Determination of Bulgarian Isolates of Cucumber Mosaic Virus and the Presence of Satellite RNAs.

Cucumber mosaic virus (CMV) is one of the most important viruses in Bulgaria, causing severe losses to agriculture, but little is known about the occurrence and distribution of subgroups within the country or the presence of satellite RNAs (satRNAs). Samples showing typical symptoms (mild to severe mosaic, vein clearing, vein necrosis, leaf deformation, stunting, and fruit necrosis) on several important crops (tomato, cucumber, pepper, bean, courgette, and tobacco) were collected from the main agricultural regions of the country. Isolates were maintained by sap inoculation in tobacco plants. Total RNAs were isolated from 38 samples (including two from bean) and used in reverse transcription-polymerase chain reaction (RT-PCR) assay with primers corresponding to the coat protein (CP) gene of RNA3 (3). A single strong band, 870 bp in length, was produced from all these samples. Amplified products were analyzed for subgroup differentiation by digestion with the restriction endonucleases MspI (3), PvuII, and EcoRI. The MspI subgroups 2 and 1 designated by Rizos et al. (3) according to their restriction endonuclease digest data correspond to the subgroups I and II widely used in the literature and based on serology, sequence data, and other properties. In this report, the subgroups are referred to as I and II for the sake of clarity. Isolates in both subgroups were found in all the main regions of Bulgaria. A few variations in MspI and EcoRI digestion patterns were seen, indicating some variability between isolates within subgroups. Only five samples, three from tomato and two from pepper, were found to be subgroup II. Subgroup I isolates were found in all the crops sampled. The PCR product from one representative isolate of each subgroup was cloned and sequenced by standard procedures. Alignment of the nucleotide and predicted amino acid sequences with published sequences of the CMV CP gene confirmed that the amplified products were derived from CMV. A further eight samples from bean gave only weak amplification and digestion of the products suggested they were likely to be subgroup II. However, these samples were unusual in not inducing symptoms in inoculated tobacco and in being difficult to propagate. The nature of these virus isolates is therefore unclear. Only a single occurrence in Bulgaria of satRNA of CMV has been reported (4) but in this study satRNAs were detected by RT-PCR (1) in total plant RNA extracts of 21 of the 38 samples tested. Amplified products of two of them, NB and 146D, were sequenced; comparison with published sequences confirmed that they were derived from CMV satellite. As expected from the symptoms induced by these isolates, a sequence homologous to the domain of satRNA Y responsible for bright yellow mosaic on tobacco (2) was identified in satRNA NB but not in satRNA 146D. satRNAs were not detected in the eight bean samples that had given only weak amplification with the CMV CP gene primers. The results presented here clearly demonstrate the presence of both subgroups of CMV in Bulgaria. Although CMV in Bulgaria has been studied previously by serological methods, no evidence had been found for the presence of subgroup II. References: (1) F. Grieco et al. Virology 229:166, 1997. (2) C. Masuta and Y. Takanami. Plant Cell 1:1165, 1989. (3) H. Rizos et al. J. Gen. Virol. 73:2099, 1992. (4) E. Stoimenova. J. Cult. Collect. 1:45, 1995.

Rapid differentiation of closely related isolates of two plant viruses by polymerase chain reaction and restriction fragment length polymorphism analysis.

Immunocapture reverse transcriptase-polymerase chain reaction (RT-PCR) followed by restriction fragment length polymorphism (RFLP) analysis of the product has been shown to be an effective procedure for discriminating serologically indistinguishable isolates of two plant viruses, raspberry bushy dwarf (RBDV) and zucchini yellow mosaic (ZYMV). For both viruses, only limited sequence information was available at the time of primer design, but most of the isolates which were tested could be amplified (the one exception being a serologically quite distinct isolate of ZYMV). Restriction endonucleases revealing diagnostic RFLPs were readily identified. Each of two isolates of ZYMV could be detected in the presence of the other and the relative proportions approximately quantified by visual estimation of the relative intensity of the appropriate bands. A range of isolates of different RBDV pathotypes were compared; isolates were grouped in ways that accorded with their known history. Computer analysis of the published sequence from which the primers had been derived showed the sequenced isolate to be identical with an isolate imported from the USSR. The PCR/RFLP procedure is rapid (it can be completed in less than 2 days), effective and will probably be generally applicable to distinguishing closely related virus isolates, even where little sequence information is available.