Sumatra disease in cloves induced by a xylem-limited bacterium following mechanical inoculation

Suspensions of a Gram negative, non motile nutritionally fastidious bacterium, cultured from clove trees affected by Sumatra disease and inoculated by needle puncture into the xylem of seedling cloves and young trees, caused the death of over 60% of the plants withing 2 year. Bacteria, confined to zylem vesssels, occurred throughout the vascular systems of dying plants and were also still active in some of the plants which survived for 3 years after inoculation. Bactria resembling those used as inoculum were readily reisolated form dying plants but not from any water-treated controls. Symptoms in experimental plants and the appearance and distribution of vessels occluded by bacteria were indistinguishable from those resulting from natural infection. All clove types tested were susceptible, including progeny from trees which had survived past disease attack. Result suggest the importance of using vigorously growing plants and providing protection against full sunlight during incubation for the successful establishment of beacterial infection in juvenile clove plants follwing mechanical inoculation.     

Isolation, culture and ultrastructure of a xylem-limited bacterium associated with Sumatra disease of cloves

A xylem-limited bacterium (XLB) previously shown to be associated with Sumatra disease of clove trees was consistently isolated from diseased but not from healthy trees. Isolation of the XLB was most successful using freshly expressed bacterial ooze obtained from root and branch tissues with low to moderate levels of xylem vessel invasion in trees at early stages of disease. XLB were commonly isolated from trees with the rapid form of disease but infrequently from those with slow decline. Bacterial colonies grew slowly on nutrient agar, reaching a maximum diameter of 1 mm in 5-7 days at 28^oC. Cells of the bacterium were 0.5-0.6 μm in width and 1.0-1.5 μm in length and did not possess flagella. The bacterial envelope was strongly rippled and Gram negative.     

Suppression of Sumatra disease symptoms in cloves treated with antibiotics

Repeated infusions of a mixture of oxytetracycline and streptomycin into the wood of initially symptomless clove trees exposed to natural infection by Sumatra disease resulted in a delay in symptom onset and slower rates of symptom development compared with untreated trees. The numbers of bacterially occluded vessels in root samples of treated trees were significantly fewer than those from controls. These results strongly support the hypothesis that a xylem-limited. Gram-negative bacterium, known to be consistently associated with Sumatra disease symptoms, is the causal agent. The productive life of treated trees was usefully extended but some phytotoxicity was observed, resulting in leaf scorching and stunted flower buds. In view of the high value of the crop, antibiotic therapy merits further investigation as a short-term control measure.     

Occurrence of blood disease of banana in Sumatra,Indonesia

Since 1991, the sudden death of cultivated banana plants has been widely observed in the southern region of Sumatra Island, Indonesia. Wilting from loss of petiole and midrib turgidity, yellowing, and necrosis of leaves was followed by death of the whole plant. Reddish brown bacterial ooze exuded from the cut surface of infected pseudostems and fruits. The colony appearance of the isolated bacterium was similar to that of Ralstonia solanacearum. The bacterium was pathogenic to banana plants but not to tomato. Its bacteriological properties agreed with those of blood disease bacterium (BDB) of banana described previously. The 16S rDNA sequence of strain Banana E had conserved bases characteristic of BDB. Based on these results, the causal agent was identified as BDB, which is a close relative of Ralstonia. The isolates have resistance against antibiotics, such as chloramphenicol and tetracycline.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number AB095535     

Characteristics of the transmission of Sumatra disease of cloves by tube-building cercopoids, Hindola spp.

Repeated transmission tests showed that the tube-building cercopoids Hindolafulva (in Sumatra) and H. striata (in Java) were natural vectors of Sumatra disease of cloves, caused by Pseudomonas syzygii . In tests on experimental transmission of P , syzygii, H , striata collected from a disease-free area acquired bacteria within 4 h of access to infected plants and transmitted them within 4 h of access to healthy test plants. Transmission was persistent and commenced within 24 h of acquisition feeding, but there was evidence for a short latent period between acquisition and transmission of bacteria. After acquisition feeding, P . syzygii was isolated from 67% of insects that transmitted the disease and also from 3% of those that did not. Times to death of test plants following first exposure to both naturally and experimentally infective insects showed log-normal distributions with similar slopes. Younger (1-year) seedlings died slightly sooner (median response time = 200 days) than older (2- to 3-year) test plants (median response time = 280 days). The bacterial content of test plants was positively correlated with time to death.     

The epigeal predatory arthropods in aubergine fields at South Sumatra (Indonesia) with regard to the plant cover and the use of insecticides

Journal of Plant Diseases and Protection - For the first time, the predatory arthropods of aubergine fields in South Sumatra, Indonesia, were studied by pitfall trapping and by square flooding in...     

欧文氏杆菌引起梨果腐的一种新病害研究初报

近年来,在山东泰安发现梨果实软腐的新病害。从病部分离到一种细菌。根据个体形态,培养性状,生理化反应,致病性郑州 研究结果,确认致病菌为大黄颈腐欧文氏杆菌Ｅｒｗｉｎｉａｒｈａｐｏｎｔｉｃｉ（Ｍｉｌｌａｒｄ）Ｂｒｕｋｈｏｌｄｅｒ,非梨火疫菌Ｅ．ａｍｙｌｏｖｏｒａ。     

Association of mycoplasma-like organisms with tomato big bud disease in Greece

The big bud disease of tomato is described in Greece for the first time. Its identity and aetiology were ascertained by electron microscopy, fluorescence and bright-field microscopy, transmission tests and antibiotic treatments. Pleomorphic mycoplasma-like organisms (MLOs) were detected in the sieve tubes of infected leaf tissues by thin-section electron microscopy and also by fluorescence and bright-field microscopy after staining with the DNA-binding fluorescent stain Hoechst 33258, and with toluidine blue. Infected tomato plants showed remission of symptoms when treated with tetracycline-HCl, but not with benzylpenicillin, while healthy plants developed big bud symptoms when graft inoculated with diseased tomato shoots. The occurrence of natural spread of big bud disease in Greece indicates that one or more vector species are present in the country.     

Association of Rhizoctonia strains with bare patch disease of wheat in Western Australia

Rhizoctonia -like fungi were isolated from the roots of diseased wheat plants sampled from the centre and periphery of three bare patches, and from apparently healthy plants from outside the patches. Of the isolates recovered, 81% were multinucleate and belonged to R. solani anastomosis group 8, and pectic zymogram group 1-1; the remaining isolates were binucleate Rhizoctonia spp. The multinucleate isolates could be grouped into highly virulent, intermediately virulent, and weakly virulent types. The binucleate isolates were all non-pathogenic. The multinucleate isolates were obtained at a significantly higher frequency from plants within the patches compared with outside the patches, and with the exception of a single isolate, the highly virulent isolates were not found outside the patches. The weakly virulent isolates were present at much lower frequencies than the highly virulent and intermediately virulent forms within the patches. The frequency of occurrence of binucleate isolates did not vary significantly among the locations sampled. None of the multinucleate isolates contained plasmids. Some of the isolates contained a prominent single dsRNA species and one or more minor dsRNA species. The distribution of these dsRNAs was not correlated with pathogenicity.     

Association of Tomato leaf curl Joydebpur virus and a betasatellite with leaf curl disease of eggplant

Leaf samples (five) from brinjal/eggplant fields showing upward leaf curling symptoms were collected from Varanasi, Uttar Pradesh state, India. The full length genome of begomovirus and associated betasatellite were amplified by PCR, cloned and sequenced. Sequences of homologous DNA-A and its betasatellite in all samples were the same. The samples failed to amplify DNA-B, suggesting that the begomovirus associated with leaf curl disease of eggplant was monopartite. The complete genome (homologous of DNA-A) consists of 2758 nts, whereas the betasatellite has 1352 nts and the genome organization is typical of Old World begomoviruses. The sequence analysis showed high levels of nucleotide sequence identity (79.8–91.7%) of virus with Tomato leaf curl Joydebpur virus (ToLCJoV) infecting chilli in India, suggesting it as a strain of ToLCJoV based on the current ICTV taxonomic criteria for begomovirus strain demarcation. However, the betasatellite associated was identified as a variant of Tomato leaf curl Bangladesh betasatellite (ToLCBDB), with which it shared highest sequence identity of 84.7–94.8%. Phylogenetic analyses of the genome further supported the above results. The recombination analyses of both genome and betasatellite showed that a major part of genome sequences are derived from begomoviruses (ToLCJoV, ChiLCuV, AEV) infecting chilli, tomato, ageratum and betasatellite from PaLCuB as the foremost parents in evolution, suggesting this as a new recombinant virus strain. This is the first report of a monopartite begomovirus and a betasatellite molecule associated with the leaf curl disease of eggplant.     

The HPLC determination of residues of maleic hydrazide in cloves of garlic bulbs following foliar application

A method was developed for the extraction and HPLC analysis of the plant growth regulator maleic hydrazide (MH) from garlic bulbs. Recovery of MH from fortified garlic tissue was 75.8(±6.1)% at the 1 and 2 mg kg^?1 fortification levels. MH residues were determined by HPLC in cloves of garlic bulbs following treatment at three sites in southern Ontario with foliar applications at 3.75 kg ha^?1. MH residues in the cloves were in the order of 3 mg kg^?1 at two sites, and 11 mg kg^?1 at the third side.     

Association of aster yellows subgroup 16SrI-B phytoplasmas with a disease of Rehmannia glutinosa var. purpurea

A new phytoplasma disease of Rehmannia glutinosa var. purpurea was observed in the Czech Republic in 1998. Infected plants showing severely proliferating shoots, leaves reduced in size with vein clearing and chlorosis, shortened internodes and virescent petals died in advanced stages of the disease. Electron microscopy examination of the ultra-thin sections revealed the presence of numerous polymorphic bodies in phloem tissue of leaf midribs and petioles. The disease was successfully transmitted from infected plant via a dodder bridge into periwinkle ( Catharanthus roseus ). The phytoplasma aetiology of this disease was further confirmed by polymerase chain reaction (PCR) using universal primers R16F2/R16R2. Restriction fragment length polymorphism (RFLP) analysis of amplification products indicated the presence of aster yellows related phytoplasmas (16SrI-B) in naturally infected samples of R. glutinosa var . purpurea and in symptomatic periwinkle after dodder transmission of the agent. A comparison of the amplified sequence with 17 sequences available in the GenBank confirmed the classification of the phytoplasma in the subgroup 16SrI-B. This is the first report of natural occurrence of phytoplasma-associated disease in R. glutinosa var. purpurea.     

Activation of systemic disease resistance in pea by an avirulent bacterium or a benzothiadiazole, but not by a fungal leaf spot pathogen

Inoculation of first expanded leaves of pea seedlings with an avirulent strain of Pseudomonas syringae pv. pisi , or treatment with sprays of a benzothiadiazole (20 or 100  μ g a.i. mL⁻¹), decreased the susceptibility of subsequent leaves 7 or 14 days later to challenge inoculation with Mycosphaerella pinodes . Inoculation of first leaves with a virulent strain of P. syringae pv. pisi or with M. pinodes did not decrease the susceptibility of plants to M. pinodes . Treatments effective in decreasing susceptibility to M. pinodes were similarly active against Uromyces viciae-fabae and virulent P. syringae pv. pisi . Effective treatments also enhanced the activities of the enzymes β-1,3-glucanase and chitinase in untreated upper leaves 6 days later. Ineffective treatments for decreased susceptibility had no effect on the activity of the enzymes. None of the treatments enhanced peroxidase activities. The results are discussed in relation to the reported signalling effects of the benzothiadiazole and in relation to a suggested high activity of the avirulent P. syringae pv. pisi strain and inactivity of M. pinodes in enhancing natural signalling.     

Mechanism of disease development caused by a multihost plant bacterium, Pseudomonas cichorii, and its virulence diversity

Pseudomonas cichorii causes rot on lettuce leaves, distinct from the necrotic spots of infected eggplant leaves. On lettuce leaves, P. cichorii invades intercellular spaces through stomata and grows vigorously, causing rot on the leaves. Surprisingly, P. cichorii does not produce pectate lyase, the most important enzyme for degrading plant cell walls. Alternatively, infection with P. cichorii causes heterochromatin condensation and DNA laddering in lettuce cells, followed by induced cell death, which results in disease symptoms. Thus, apoptotic programmed cell death (PCD) is associated with the symptoms on lettuce leaves. Although PCD in P. cichorii-infected eggplant leaves is also associated with necrotic spots caused by the bacteria, there are differences in the responses of the two hosts. P. cichorii harbors the N-acetyltransferase family gene (pat) and the hrp genes (hrp) encoding a type III secretion system, adjacent to an aldehyde dehydrogenase gene (aldH). The virulence of P. cichorii is hrp-dependent in eggplant, but not in lettuce. Furthermore, hrp, pat and aldH are implicated in the diversity of P. cichorii virulence on susceptible Asteraceae species. The involvement of hrp, aldH and pat in bacterial virulence on the respective species has no relationship with the phylogeny of the plants species. When these results are considered together, P. cichorii has multiple virulence determinants. The involvement of not only hrp but also aldH and pat in P. cichorii virulence arose after species diversification of host plants. Host responses implicated in symptom development have been responsible for the development of virulence determinants of P. cichorii.     

Association of Pythium coloratum and Pythium sulcatum with cavity spot disease of carrots in Western Australia

All 170 Pythium isolates from carrot cavity spot lesions from a field in Western Australia were found to belong to either P. coloratum or P. sulcatum . All isolates of P. coloratum produced large, brownish-black, water-soaked and depressed lesions on mature carrots inoculated with agar plugs colonized by the pathogen. In comparison, only a few isolates of P. sulcatum produced lesions and these were small. In glasshouse trials, P. coloratum produced substantial and numerous lesions at an inoculum density of 0.5% (weight of millet seed-based inoculum/weight of soil), whilst P. sulcatum produced few and small lesions at inoculum densities of 0.8 and 1% and none at 0.5%. This is the first record of P. coloratum as a causal agent of cavity spot of carrots.     

The Natural Occurrence of Two Distinct Begomoviruses Associated with DNAbeta and a Recombinant DNA in a Tomato Plant from Indonesia

ABSTRACT Two begomoviruses (Java virus-1 and Java virus-2), two satellite DNAs (DNAbeta01 and DNAbeta02), and a recombinant DNA (recDNA) were cloned from a single tomato plant from Indonesia with leaf curl symptoms, and the role of these satellite DNAs in the etiology of begomovirus disease was investigated. The genome organizations of the two viruses were similar to those of other Old World monopartite begomoviruses. Comparison of the sequences with other begomoviruses revealed that Java virus-1 was a newly described virus for which the name Tomato leaf curl Java virus (ToLCJAV) is proposed. Java virus-2 was a strain of Ageratum yellow vein virus (AYVV) (AYVV-[Java]). ToLCJAV or AYVV-[Java] alone did not induce leaf curl symptoms in tomato plants. However, in the presence of DNAbeta02, both ToLCJAV and AYVV-[Java] induced leaf curl symptoms in tomato plants. In the presence of DNAbeta01, these viruses induced mild leaf curl symptoms in tomato plants. The recDNA had a chimeric sequence, which arose from recombination among ToLCJAV, AYVV-[Java], DNAbeta01, and DNAbeta02; it was replicated only in the presence of AYVV-[Java] in tomato plants.     

Association between genetic variability and titre of Grapevine Pinot gris virus with disease symptoms

The present work was carried out in order to verify the possible association between a new grapevine disease, characterized by leaf mottling and deformation, and the genetic variability and concentration of Grapevine Pinot gris virus (GPGV), a recently identified virus tentatively associated with the pathology. After vineyard surveys and the establishment of real‐time qPCR assays, characterization of GPGV isolates and evaluation of GPGV titre were assessed in more than 100 samples of grapevine Glera, collected from plants regardless of whether or not they showed the symptomatology. Results showed that there was an important association between the GPGV variants and manifestation of the symptoms, and that grapevines with symptoms harboured significantly higher GPGV titre than symptomless vines. Moreover, an interesting relationship among the phylogenetic clustering of the isolates originating from plants with symptoms and some epidemiological characteristics of the disease was found. The current study confirmed the role of GPGV in the emergent disease characterized by grapevine leaf mottling and deformation.