Characteristics of sugarcane white leaf phytoplasma transmission by the leafhopper Matsumuratettix hiroglyphicus

The leafhopper Matsumuratettix hiroglyphicus (Matsumura) (Hemiptera: Cicadellidae) is the most important vector of sugarcane white leaf (SCWL) phytoplasma that significantly affects the sugarcane crop in Asia. Here, we aimed to study the characteristics of SCWL phytoplasma transmission by M. hiroglyphicus. To this end, the stylet penetration activities performed during the acquisition access period (AAP) and inoculation access period (IAP) were investigated by the direct current electrical penetration graph technique and confirmed by quantitative polymerase chain reaction (qPCR). Additionally, the latent period (LP) of SCWL phytoplasma in the vector was determined by qPCR and localised by fluorescent in situ hybridisation. The results indicated that the acquisition of SCWL phytoplasma occurred during phloem ingestion (waveform D), whereas its inoculation was associated with salivation into the phloem sieve element (waveform C). The minimum AAP was 15 min and the minimum duration of phloem ingestion was 2.35 min. The minimum LP of SCWL phytoplasma in the vector was at least 14 days; then, SCWL phytoplasma moved to the salivary glands of the insect, enabling the transmission of the pathogen to the host plants. The minimum IAP for a successful transmission of SCWL phytoplasma to the host plants was 11–12 min, with a minimum duration of salivation into phloem of 1.35 min. The female vectors had higher SCWL phytoplasma copy numbers than the male vectors, and displayed faster AAP, IAP, and LP. Overall, our findings provide important information related to the feeding behaviour of M. hiroglyphicus and its effect on the transmission of SCWL phytoplasma.     

Transmission of sugarcane white leaf phytoplasma by Yamatotettix flavovittatus, a new leafhopper vector

Sugarcane white leaf disease is caused by plant pathogenic phytoplasmas that are transmitted to the plant by the leafhopper Matsumuratettix hiroglyphicus (Matsumura). To determine whether there are other insect vectors that transmit this disease pathogen, leafhopper species in sugarcane, Saccharum officinarum L., fields in northeastern Thailand were monitored by using light traps. Sixty-nine leafhopper species from family Cicadellidae were found. Using nested polymerase chain reaction (PCR) with specific primers, a 210-bp amplified DNA fragment corresponding to phytoplasma associated with sugarcane white leaf disease was detected from 12 species of leafhoppers [Balclutha rubrostriata (Melichar), Balclutha sp., Bhatia olivacea (Melichar), Exitianus indicus Distant, Macrosteles striifrons Anufriew, Matsumuratettix hiroglyphicus (Matsumura), Recilia distincta (Motschulsky), Recilia dorsalis (Motschulsky), Recilia sp., Thaia oryzivora Ghauri, Yamatotettix flavovittatus Matsumura, and Xestocephalus sp.]. The percentage of individual infection with phytoplasma varied from 5% in B. olivacea to 35% in Xestocephalus sp. The most abundant leafhopper species, i.e., E. indicus, Y. flavovittatus, and M. hiroglyphicus were used in transmission tests to determine their vector status for the sugarcane white leaf phytoplasma transmission. Infected insects were reared on healthy plants and specific PCR followed by sequencing of the amplicons was used to determine whether the phytoplasma was transmitted to the plants. The results showed that both Y. flavovittatus and M. hiroglyphicus, but not E. indicus, can transmit sugarcane white leaf phytoplasma to healthy sugarcane plants. The transmission efficiency of M. hiroglyphicus (55%) was higher than that of Y. flavovittatus (45%). We conclude that Y. flavovittatus is a newly discovered vector for sugarcane white leaf disease, in addition to M. hiroglyphicus. These two species peak at different times of the year and therefore complement each other in the transmission of the phytoplasma. Because there are no known alternative host plants for the sugarcane white leaf, management of the disease will necessarily require the control of both Y. flavovittatus and M. hiroglyphicus.     

Candidates for symbiotic control of sugarcane white leaf disease

The leafhopper Matsumuratettix hiroglyphicus (Matsumura) is the most important vector of a phytoplasma pathogen causing sugarcane white leaf (SCWL) disease. The purpose of this study was to evaluate candidate bacterial symbionts for possible use as vehicles in the control of the disease. 16S rRNA bacterial genes were amplified from whole bodies of M. hiroglyphicus leafhoppers and analyzed by cloning and sequencing. Two dominant groups were found: one belonged to the Betaproteobacteria that did not closely match any sequences in the database and was named bacterium associated with M. hiroglyphicus (BAMH). Another one found to be abundant in this leafhopper is "Candidatus Sulcia muelleri" in the order Bacteroidetes, which was previously reported in the insect members of the Auchenorrhyncha. Most M. hiroglyphicus leafhoppers carry both BAMH and "Ca. Sulcia muelleri." Fluorescent in situ hybridization showed that BAMH and "Ca. Sulcia muelleri" colocalized in the same bacteriomes. BAMH was present in the midgut and ovaries of the leafhopper and was found in all developmental stages, including eggs, nymphs, and adults. Because BAMH appears to be specific for the SCWL vector, we evaluated it as a candidate for symbiotic control of sugarcane white leaf disease.     

Metabolomics reveal alterations in arachidonic acid metabolism in Schistosoma mekongi after exposure to praziquantel

BackgroundMekong schistosomiasis is a parasitic disease caused by the blood-dwelling fluke Schistosoma mekongi. This disease contributes to human morbidity and mortality in the Mekong region, posing a public health threat to people in the area. Currently, praziquantel (PZQ) is the drug of choice for the treatment of Mekong schistosomiasis. However, the molecular mechanisms of PZQ action remain unclear, and Schistosoma PZQ resistance has been reported occasionally. Through this research, we aimed to use a metabolomic approach to identify the potentially altered metabolic pathways in S. mekongi associated with PZQ treatment.Methodology/Principal findingsAdult stage S. mekongi were treated with 0, 20, 40, or 100 μg/mL PZQ in vitro. After an hour of exposure to PZQ, schistosome metabolites were extracted and studied with mass spectrometry. The metabolomic data for the treatment groups were analyzed with the XCMS online platform and compared with data for the no treatment group. After low, medium (IC₅₀), and high doses of PZQ, we found changes in 1,007 metabolites, of which phosphatidylserine and anandamide were the major differential metabolites by multivariate and pairwise analysis. In the pathway analysis, arachidonic acid metabolism was found to be altered following PZQ treatment, indicating that this pathway may be affected by the drug and potentially considered as a novel target for anti-schistosomiasis drug development.Conclusions/SignificanceOur findings suggest that arachidonic acid metabolism is a possible target in the parasiticidal effects of PZQ against S. mekongi. Identifying potential targets of the effective drug PZQ provides an interesting viewpoint for the discovery and development of new agents that could enhance the prevention and treatment of schistosomiasis.     

Differentiation of phytoplasmas associated with sugarcane and gramineous weed white leaf disease and sugarcane grassy shoot disease by RFLP and sequencing

 Restriction fragment length polymorphism (RFLP) and sequencing were used to elucidate the genetic relationship between phytoplasmas that cause white lead disease and grassy shoot disease in sugarcane and white leaf disease in gramineous weeds found in the cane-growing areas (Crowfoot grass, Bermuda grass and Brachiaria grass). A 1.35-kb DNA fragment encoding for the 16s rRNA was amplified by PCR using universal primers and analysed by digestion with nine restriction endonucleases. A DNA fragment containing the 3′ end of the 16s rRNA and the spacer region between the 16s rRNA and the tRNA(Ile) was amplified by PCR and sequenced. Analysis of the RFLP patterns and of the sequence showed that grassy shoot and white leaf diseases in sugarcane are caused by two different phytoplasmas. Sequence analysis of phytoplasma DNA obtained from three species of weeds showing symptoms of white leaf disease failed to detect any organism that is identical to those infecting the sugarcane. Moreover the phytoplasma species that infect the three types of gramineous weeds, although closely related, are nevertheless different Received: 15 April 1997 / Accepted: 18 April 1997     

The Reproductive Biology and Reproductive Success of Pterocarpus macrocarpus Kurz1

The reproductive biology of Pterocarpus macrocarpus Kurz (pradu) was studied in 37‐year‐old plantation trees in Thailand to determine the causes of seed and fruit loss. Trees flowered at the end of March or early in April at the end of the hot dry season and start of the rainy season. Flowering occurred over about a one‐month period. Fruits developed over the next six months during the rainy season and matured at the start of the cool dry season in October and November. Phenology was similar in the four trees that were studied in detail. Racemes averaged 30 flowers each and each raceme was receptive for several days, although each flower was only receptive for one day. After pollination, floral parts were shed over several days and fruits began to develop. Pradu is entomopholous but its insect pollinators were not identified. The stigma is covered by hairs and a secretion is produced. A high proportion of flowers were pollinated. Then, there was a rapid loss of flowers and young fruits. These observations and earlier genetic studies indicate the probability of a high level of self‐incompatibility in this predominantly outbreeding species. Pradu may have a very late‐acting self‐incompatibility mechanism found in many other hardwoods. The zygote remains quiescent for six weeks as the endosperm develops. During this time most of the ovules and fruits abort, suggesting resource allocation preferentially to cross‐pollinated ovules. Pradu has a high reproductive potential but a low preemergence reproductive success (0.8), which is common for many hardwood species. The major cause of the low reproductive success was fruit loss during early development. Fruit production may be enhanced by increased cross‐pollination among unrelated parent trees. This may be accomplished in seed orchards and seed production areas by the introduction of additional insect pollinators that travel greater distances between trees and by the relatively close spacing of unrelated parent trees.     

Dispersal of the leafhoppers <Emphasis Type="Italic">Matsumuratettix hiroglyphicus</Emphasis> and <Emphasis Type="Italic">Yamatotettix flavovittatus</Emphasis> (Homoptera: Cicadellidae), vectors of sugarcane white leaf disease

Sugarcane white leaf disease is a serious problem in many Asian Countries. The leafhoppers Matsumuratettix hiroglyphicus (Matsumura) and Yamatotettix flavovittatus Matsumura are the main vectors of sugarcane white leaf disease phytoplasma. Gaining a better understanding of the dispersal behavior of such insect vectors is essential for both disease epidemiology and vector control. The dispersal distances of M. hiroglyphicus and Y. flavovittatus in a sugarcane field were estimated by means of mark–release–recapture experiments. Adult leafhopper vectors collected from the fields were marked using fluorescent dye powder and released at a central release point in a sugarcane field. The marking method did not have a significant effect on the survival or flight activity of the leafhopper vectors. The overall release–recapture rates of M. hiroglyphicus and Y. flavovittatus within 50 m were 10.1 and 13.4 %, respectively. The estimated natural mean dispersal distances for M. hiroglyphicus and Y. flavovittatus were 162.1 and 387.5 m, respectively. Wind appears to be the main factor influencing the leafhopper dispersal direction.     

Annotated catalogue of the types of Indo-Burmese non-marine Gastropoda deposited in the Faculty of Tropical Medicine,Mahidol University,Thailand

ABSTRACT

Four decades after its publication, Rolf A.M. Brandt’s 1974 monograph on the non-marine molluscs of Thailand remains the main authority on freshwater and estuarine species for Southeast Asia and includes up to 165 new species of snails and bivalves described by Brandt and colleagues in the same book and preceding publications. All the holotypes are lodged at the Forschungsinstitut und Naturmuseum Senckenberg in Germany, and are largely inaccessible to Thai and other Southeast Asian researchers, who rely heavily on the Brandt collection as a key reference. Paratypes were, however, donated to various other collections, including some in Thailand. We present the first catalogue of 45 paratypes of gastropods of the Brandt collection, described from Thailand, Laos and Cambodia, which are lodged at the Faculty of Tropical Medicine, Mahidol University, Thailand.