Bacteria were isolated from the crop and midgut of field collected Bactrocera cacuminata (Hering) and Bactrocera tryoni (Froggatt) (Diptera: Tephritidae). Two methods were used, firstly isolation onto two types of bacteriological culture media (PYEA and TSA) and identification using the API-20E diagnostic kit, and secondly, analysis of samples using the 16S rRNA gene molecular diagnostic method. Using the API-20E method, 10 genera and 17 species of bacteria in the family Enterobacteriaceae were identified from cultures growing on the nutrient agar. The dominant species in both the crop and midgut were Citrobacter freundii, Enterobacter cloacae and Klebsiella oxytoca. Providencia rettgeri, Klebsiella pneumoniae ssp ozaenae and Serratia marcescens were isolated from B. tryoni only. Using the molecular cloning technique that is based on 16S rRNA gene sequences, five bacteria classes were dignosed — Alpha-, Beta-, Gamma- and Delta- Proteobacteria and Firmicutes — including five families, Leuconostocaceae, Enterococcaceae, Acetobacteriaceae, Comamonadaceae and Enterobacteriaceae. The bacteria affiliated with Firmicutes were found mainly in the crop while the Gammaproteobacteria, especially the family Enterobacteriaceae, was dominant in the midgut. This paper presents results from the first known application of molecular cloning techniques to study bacteria within tephritid species and the first record of Firmicutes bacteria in these flies.
A novel leaf fall disease of rubber trees (Hevea brasiliensis) has been recently noted in Thailand. The fungal pathogens of this disease were identified based on both morphological and molecular characteristics as Neopestalotiopsis cubana and N. formicarum. Portions of internal transcribed spacer (ITS) region and the large subunit (LSU), translation elongation factor 1‐α (TEF1‐α) and β‐tubulin (TUB) genes were PCR amplified with the primer pairs ITS1/ITS4, LR0R/LR5, EF1‐728F/EF2 and T1/Bt2b, respectively. Sequencing of the PCR products and a phylogenetic tree based on the combined ITS, TEF1‐α and TUB confirmed these pathogens as N. cubana and N. formicarum. Pathogenicity test results showed that the pathogens cause leaf spot and leaf fall similar to that observed in natural infections. This is the first report on the novel leaf fall disease of rubber trees in Thailand, with the results demonstrating that it is associated with N. cubana and N. formicarum.
Biocontrol fungi (BFs) play a key role in regulation of pest populations. BFs produce multiple non-ribosomal peptides (NRPs) and other secondary metabolites that interact with pests, plants and microorganisms. NRPs—including linear and cyclic peptides (L-NRPs and C-NRPs)—are small peptides frequently containing special amino acids and other organic acids. They are biosynthesized in fungi through non-ribosomal peptide synthases (NRPSs). Compared with C-NRPs, L-NRPs have simpler structures, with only a linear chain and biosynthesis without cyclization. BFs mainly include entomopathogenic and mycoparasitic fungi, that are used to control insect pests and phytopathogens in fields, respectively. NRPs play an important role of in the interactions of BFs with insects or phytopathogens. On the other hand, the residues of NRPs may contaminate food through BFs activities in the environment. In recent decades, C-NRPs in BFs have been thoroughly reviewed. However, L-NRPs are rarely investigated. In order to better understand the species and potential problems of L-NRPs in BFs, this review lists the L-NRPs from entomopathogenic and mycoparasitic fungi, summarizes their sources, structures, activities and biosynthesis, and details risks and utilization prospects.
A review of female mutillids in southern Thailand is presented, based on literature, loaned material, and two field expeditions. Thirty-three species in 23 genera have been previously recorded from Thailand (Lelej 2005; Lelej et al. 2016, 2017; Okayasu et al. 2018). In March and April 2017, 337 female specimens were collected in the provinces of Krabi, Nakhon Si Thammarat, Phatthalung, Songkhla, and Trang. In May 2018, 255 female specimens were collected in the provinces of Kanchanaburi, Phetchaburi, and Surat Thani. These collected specimens were sorted into 38 morphospecies: 13 already known from Thailand, nine previously described species newly recorded from Thailand, and 16 new to science and described here. Based on loaned material, three more described species were newly recorded from southern Thailand and one more undescribed species was found. Keys to females of the 31 mutillid genera found in southeast Asia and 42 mutillid species studied from southern Thailand are provided. Illustrations and diagnoses are provided for each species studied in southern Thailand, including the following 17 new species: Bischoffitilla tokay Williams, Bischoffitilla cambrai Williams, Mickelomyrme puttasoki Williams, Mickelomyrme kinguri Williams, Nordeniella maleeae Williams, Smicromyrme helarctos Williams, Smicromyrme borkenti Williams, Smicromyrme thaochani Williams, Krombeinidia agricola Williams, Krombeinidia chang Williams, Krombeinidia baanmaka Williams, Orientidia manleyi Williams, Orientidia thoawanae Williams, Petersenidia modkhong Williams, Petersenidia leleji Williams, Trogaspidia pittsi Williams, and Trogaspidia wilsoni Williams. Three new synonyms and one new combination are recognized: Bischoffitilla lamellata (Mickel, 1933) = Squamulotilla arundinacea Pagden, 1934, syn. nov., Mickelomyrme pusillaeformis (Hammer, 1962), comb. nov. = Mickelomyrme yunnanensis Lelej, 1996, syn. nov., Trogaspidia pagdeni (Mickel, 1933) = Timulla (Trogaspidia) pagdeni nodoa Mickel, 1933, syn. nov. Finally, two other taxa are raised to full species from subspecies status, Eotrogaspidia oryzae (Pagden, 1934), status resurr., and Trogaspidia lingnani (Mickel, 1933), status nov. New combination is also proposed for Krombeinidia meeungensis (Cockerell, 1928), comb. nov. Twenty-three new country records are presented: 12 from Thailand, four from Vietnam, four from Laos, two from Cambodia, and one from Indonesia.
Soil microorganisms are well studied for their beneficial effects on plant growth and their impact on biocontrol agents. The production of volatile antifungal compounds emitted from soil fungi is considered to be an effective ability that can be applied in biofumigants in the control of plant diseases. A soil fungus, Trichoderma asperelloides TSU1, was isolated from flamingo flower cultivated soil and identified on the basis of the morphology and molecular analysis of the internal transcribed spacer (ITS), rpb2, and tef1-α genes. To test T. asperelloides TSU1-produced volatile organic compounds (VOCs) with antifungal activity, the sealed plate method was used. The VOCs of T. asperelloides TSU1 inhibited the mycelial growth of fungal pathogens that were recently reported as emerging diseases in Thailand, namely, Corynespora cassiicola, Fusarium incarnatum, Neopestalotiopsis clavispora, N. cubana, and Sclerotium rolfsii, with a percentage inhibition range of 38.88–68.33%. Solid-phase microextraction (SPME) was applied to trap VOCs from T. asperelloides TSU1 and tentatively identify them through gas chromatography–mass spectrometry (GC/MS). A total of 17 compounds were detected in the VOCs of T. asperelloides TSU1, and the dominant compounds were identified as fluoro(trinitro)methane (18.192% peak area) and 2-phenylethanol (9.803% peak area). Interestingly, the commercial 2-phenyethanol showed antifungal activity against fungal pathogens that were similar to the VOCs of T. asperelloides TSU1 by bioassay. On the basis of our study’s results, T. asperelloides TSU1 isolated from soil displayed antifungal abilities via the production of VOCs responsible for restricting pathogen growth.
Members of the Bactrocera dorsalis Hendel (Diptera: Tephritidae) complex constitute well-recognized destructive pests of fruits in peninsular Thailand. The development and survival of immature stages of the carambola fruit fly, Bactrocera carambolae Drew & Hancock, and the Asian papaya fruit fly, Bactrocera papayae Drew & Hancock , were compared at six constant temperatures of 15, 20, 25, 27, 30, and 35°C, 70 ± 5% relative humidity, and a photoperiod of 12:12 (L:D). The objectives were to determine the effect of temperature on the developmental stages for optimizing rearing and to understand the geographical pattern of occurrence of these fruit fly species. A strong and positive linear relationship was observed between temperature and developmental rate of immature stages of B. carambolae. Similarly, a strong and positive linear relationship was observed between temperature and developmental rate of B. papayae. A temperature summation model was used to estimate the lower threshold temperature and the thermal constant. Bactrocera papayae was significantly faster in development and higher in survival and appeared to be better adapted to low temperatures than B. carambolae, as it exhibited the lowest threshold temperatures at all immature stages. The observed differences in response to various temperatures revealed to some extent the impact of temperature on these species’ distribution in peninsular Thailand and other parts of the world.
Entomopathogenic fungi (EF), who represent active agents to control insect natural populations, usually persist in terrestrial habitats. Southwest area in China has various climate conditions and abundant plant biodiversity (crop, forest, grassy, orchard and arable areas). Nevertheless, the potential of soil-inhabitant EF as insect pest biocontrol agents, is unknown. In this study, first the EF biodiversity from soil of three provinces (Guizhou, Sichuan, and Yunnan) was surveyed. Then, the virulence of 29 isolated strains against Bemesia tabaci and Spodoptera litura was assessed. After analyzing 212 soil samples, 497 isolated fungi were identified. Out of them, 490 isolates were classified in 45 species of 24 genera, whereas the other seven isolates, belonging to Paecilomyces and Purpureocillium genera, were not identified under species level. Furthermore, the EF biodiversity from soil of Sichuan, Yunan, and Guizhou areas, analyzed by Shannon Wiener Index (SWI) was rated at 2.98, 1.89, and 2.14, while SWIs-biodiversity in crop, forest, grassy, orchard and arable areas was rated at 2.88, 2.74, 3.05, 2.39, and 2.47. SWI data suggested that soil from Sichuan area and grassy had higher EF biodiversity compared with other analyzed provinces and areas. Virulence bioassay results indicated that, out of the 29 isolates tested, 24 were pathogenic against B. tabaci and S. litura, resulting in mortality rates >10%. In conclusion, this study reports the EF distribution and biodiversity in soil from three provinces located at Southwest China, whereas their potential use as a tool for the B. tabaci and S. litura biocontrol must be further investigated.
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