Pest categorisation of Witches' broom disease of lime (Citrus aurantifolia) phytoplasma

The EFSA Panelon Plant Health performed a pest categorisation for the Witches' broom disease of lime (Citrusaurantifolia) phytoplasma for the EU territory. The pest has been reported in a few countries in the Middle East and is not known to occur in the EU. The disease is caused by a well-defined phytoplasma strain in the Candidatus Phytoplasma aurantifolia' species, for which efficient molecular detection assays are available. The most important known natural host is Citrusaurantifolia, which is only grown for ornamental purposes in the EU. Sweet limes, rough lemon and trifoliate orange are also naturally infected by that phytoplasma. The latter can be transmitted by grafting also to some citrus species. Other citrus species were reported to be resistant; however, their susceptibility has been assessed only by symptom observations, and the possible presence of phytoplasmas in symptomless plants cannot be ruled out. The phytoplasma is transmitted by the leafhopper Hishimonusphycitis, which is not known to occur in the EU. There is no information on the vector status of other phloem feeding insects of citrus present in the EU. The pest is listed in Annex IIAI of Directive 2000/29/EC. The main pathways for entry, plants for planting and the vector insect, are closed by existing legislation on import of citrus plants. Nevertheless, should the pest enter, it could establish and spread. In countries where Witches' broom disease of lime (WBDL) is present, it has significant impact. The main knowledge gaps concern (1) and vertical transmission of the phytoplasma to H.phycitis eggs (2) lack of information regarding susceptibility of citrus crops grown in the EU (3) status of potential insect vectors in the EU. Therefore, the WBDL phytoplasma meets the criteria assessed by EFSA for consideration as a potential Union quarantine pest

Pest categorisation of Palm lethal yellowing phytoplasmas

The EFSA Panelon Plant Health performed a pest categorisation of Palm lethal yellowing phytoplasmas for the EU territory. This name is used to describe diseases that share the same succession of symptoms in palms that are caused by a number of strains of phytoplasma, for which efficient molecular detection assays are available. The pest is not known to occur in the EU and therefore does not meet one of the criteria for being a Union regulated non-quarantine pest. For Candidatus Phytoplasma palmae', the planthopper Haplaxius crudus, which is not known to be present in the EU, is the confirmed vector, but for the other strains, the vectors are unknown. The host range of the pest is restricted to Arecaceae species, in particular coconut. The pest is regulated on all known hosts in Annex IIAI of Directive 2000/29/EC. It could potentially enter the EU via plants for planting or through infected vectors. The phytoplasmas could become established in the EU as host plants are present. It is unknown whether arthropods present in the EU could be vectors. The potential impact of the pest if introduced into the EU is difficult to assess given this uncertainty but is estimated to be limited. The main knowledge gaps concern the status of potential vector insects in the EU; the possibility for seed transmission of the phytoplasmas; the origin and volume of the trade in palm seeds and plants for planting; the host status and susceptibility of many palm species grown in the EU and the potential new assignments of phytoplasmas to this categorisation that might have associated alternate hosts. Palm lethal yellowing phytoplasmas meet the criteria assessed by EFSA for consideration as Union quarantine pest

Construction of an interactive online phytoplasma classification tool, iPhyClassifier, and its application in analysis of the peach X-disease phytoplasma group (16SrIII)

Phytoplasmas, the causal agents of numerous plant diseases, are insect-vector-transmitted, cell-wall-less bacteria descended from ancestral low-G+C-content Gram-positive bacteria in the Bacillus–Clostridium group. Despite their monophyletic origin, widely divergent phytoplasma lineages have evolved in adaptation to specific ecological niches. Classification and taxonomic assignment of phytoplasmas have been based primarily on molecular analysis of 16S rRNA gene sequences because of the inaccessibility of measurable phenotypic characters suitable for conventional microbial characterization. In the present study, an interactive online tool, iPhyClassifier, was developed to expand the efficacy and capacity of the current 16S rRNA gene sequence-based phytoplasma classification system. iPhyClassifier performs sequence similarity analysis, simulates laboratory restriction enzyme digestions and subsequent gel electrophoresis and generates virtual restriction fragment length polymorphism (RFLP) profiles. Based on calculated RFLP pattern similarity coefficients and overall sequence similarity scores, iPhyClassifier makes instant suggestions on tentative phytoplasma 16Sr group/subgroup classification status and ‘Candidatus Phytoplasma’ species assignment. Using iPhyClassifier, we revised and updated the classification of strains affiliated with the peach X-disease phytoplasma group. The online tool can be accessed at http://www.ba.ars.usda.gov/data/mppl/iPhyClassifier.html

Molecular biology and pathogenicity of phytoplasmas

Phytoplasmas are a large group of plant-pathogenic wall-less, non-helical, bacteria associated with diseases, collectively referred to as yellows diseases, in more than a thousand plant species worldwide. Many of these diseases are of great economic importance. Phytoplasmas are difficult to study, in particular because all attempts at culturing these plant pathogens under axenic conditions have failed. With the introduction of molecular methods into phytoplasmology about two decades ago, the genetic diversity of phytoplasmas could be elucidated and a system for their taxonomic classification based on phylogenetic traits established. In addition, a wealth of information was generated on phytoplasma ecology and genomics, phytoplasma–plant host interactions and phytoplasma–insect vector relationships. Taxonomically, phytoplasmas are placed in the class Mollicutes, closely related to acholeplasmas, and are currently classified within the provisional genus ‘Candidatus Phytoplasma’ based primarily on 16S rDNA sequence analysis. Phytoplasmas are characterised by a small genome. The sizes vary considerably, ranging from 530 to 1350 kilobases (kb), with overlapping values between the various taxonomic groups and subgroups, resembling in this respect the culturable mollicutes. The smallest chromosome, about 530 kb, is known to occur in the Bermuda grass white leaf agent ‘Ca. Phytoplasma cynodontis’. This value represents the smallest mollicute chromosome reported to date. In diseased plants, phytoplasmas reside almost exclusively in the phloem sieve tube elements and are transmitted from plant to plant by phloem-feeding homopteran insects, mainly leafhoppers and planthoppers, and less frequently psyllids. Most of the phytoplasma host plants are angiosperms in which a wide range of specific and non-specific symptoms are induced. Phytoplasmas have a unique and complex life cycle that involves colonisation of different environments, the plant phloem and various organs of the insect vectors. Furthermore, many phytoplasmas have an extremely wide plant host range. The dynamic architecture of phytoplasma genomes, due to the occurrence of repetitive elements of various types, may account for variation in their genome size and adaptation of phytoplasmas to the diverse environments of their plant and insect hosts. The availability of five complete phytoplasma genome sequences has made it possible to identify a considerable number of genes that are likely to play major roles in phytoplasma–host interactions. Among these, there are genes encoding surface membrane proteins and effector proteins. Also, it has been shown that phytoplasmas dramatically alter their gene expression upon switching between plant and insect hosts

Coconut lethal yellowing diseases: a phytoplasma threat to palms of global economic and social significance

The recent discovery of Bogia coconut syndrome in Papua New Guinea (PNG) is the first report of a lethal yellowing disease (LYD) in Oceania. Numerous outbreaks of LYDs of coconut have been recorded in the Caribbean and Africa since the late Nineteenth century and have caused the death of millions of palms across several continents during the Twentieth century. Despite the severity of economic losses, it was only in the 1970s that the causes of LYDs were identified as phytoplasmas, a group of insect-transmitted bacteria associated with diseases in many other economically important crop species. Since the development of polymerase chain reaction (PCR) technology, knowledge of LYDs epidemiology, ecology and vectors has grown rapidly. There is no economically viable treatment for LYDs and vector-based management is hampered by the fact that vectors have been positively identified in very few cases despite many attempted transmission trials. Some varieties and hybrids of coconut palm are known to be less susceptible to LYD but none are completely resistant. Optimal and current management of LYD is through strict quarantine, prompt detection and destruction of symptomatic palms, and replanting with less susceptible varieties or crop species. Advances in technology such as loop mediated isothermal amplification (LAMP) for detection and tracking of phytoplasma DNA in plants and insects, remote sensing for identifying symptomatic palms, and the advent of clustered regularly interspaced short palindromic repeats (CRISPR)-based tools for gene editing and plant breeding are likely to allow rapid progress in taxonomy as well as understanding and managing LYD phytoplasma pathosystems

Description of the genus \u2018Candidatus Phytoplasma\u2019, a taxon for the wall-less non-helical prokaryotes that colonize plant phloem and insects

The trivial name 'phytoplasma' has been adopted to collectively name wall-less, non-helical prokaryotes that colonize plant phloem and insects, which were formerly known as mycoplasma-like organisms. Although phytoplasmas have not yet been cultivated in vitro, phylogenetic analyses based on various conserved genes have shown that they represent a distinct, monophyletic clade within the class Mollicutes. It is proposed here to accommodate phytoplasmas within the novel genus 'Candidatus (Ca.) Phytoplasma'. Given the diversity within 'Ca. Phytoplasma', several subtaxa are needed to accommodate organisms that share 97.5% similar to those of other 'Ca. Phytoplasma' species, are characterized by distinctive biological, phytopathological and genetic properties. These include 'Ca. Phytoplasma pyri' (associated with pear decline), 'Ca. Phytoplasma prunorum' (associated with European stone fruit yellows), 'Ca. Phytoplasma spartii' (associated with spartium witches'-broom), 'Ca. Phytoplasma rhamni' (associated with buckthorn witches'-broom), 'Ca. Phytoplasma allocasuarinae' (associated with allocasuarina yellows), 'Ca. Phytoplasma ulmi' (associated with elm yellows) and an additional taxon for the stolbur phytoplasma. Conversely, some organisms, despite their 16S rRNA gene sequence being < 97-5% similar to that of any other 'Ca. Phytoplasma' species, are not presently described as Candidatus species, due to their poor overall characterization