Guizhou Institute of Plant Protection

Guiyang, China

Guizhou Institute of Plant Protection

Guiyang, China
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Chen R.Y.,Guizhou Institute of Upland Food Crops | Zhang J.H.,Guizhou Institute of Animal Husbandry and Veterinary Science | Cheng J.,Guizhou Institute of Upland Food Crops | Zheng C.X.,Guizhou Institute of Upland Food Crops | And 2 more authors.
Plant Disease | Year: 2017

Tobacco (Nicotiana tabacum L.) is a leafy, annual, solanaceous plant grown commercially for its leaves. China accounts for around 40% of the total global tobacco production (U.S. Census Bureau 2005) and consumption (British American Tobacco 2013). In August 2016, leaf spots developed on N. tabacum cv. Yunyan 85 growing in a 2 ha commercial tobacco field in Guizhou, China (26.68°N, 107.52°E). Approximately 30% of the leaves on 3% of the plants were affected. The symptoms began as chlorotic areas and subsequently turned into dark colored, necrotic spots. The lesions expanded and became gray, grayish-brown, or light brown, circular to oblong with dark brown borders that often were surrounded by a yellow halo. Severely infected plants had poor leaf quality and at least 5% yield loss on one plant. Necrotic tissues from symptomatic leaf pieces were surface-sterilized, placed on potato dextrose agar (PDA) medium, and incubated at 25°C in the dark. After 6 days, blackish-brown colonies with a black reverse side grew, and dark brown septate conidiophores developed within the colony. One isolate was examined in detail. Conidia of the pathogen were short (26 to 28 × 11 to 12 μm), with predominantly 3-septa, straight or often curved, with end cells frequently paler than intermediate cells. Based on these characteristics, the fungus was identified as Curvularia trifolii (Falloon 1976). Species verification of this pathogen was completed by PCR amplification using universal primers ITS1/4 and BLAST analysis of the partial internal transcribed spacer (ITS) region of ribosomal DNA (rDNA). The sequence of this region of rDNA from the isolate (GenBank accession no. KX865088) was identical to the sequence of several C. trifolii accessions (99% similarity) (KM979920.1, GQ241277.1, and KJ88716.1). Pathogenicity of the fungus was confirmed by performing Koch’s postulates as follows. Five detached tobacco leaves (cv. K326) were surface disinfested with 5% NaOCl and then wound-inoculated. Plugs of the fungus (5 mm in diameter) obtained from the colony margins were transferred onto 3 × 3 mm wounds taken with a needle on the surface of five sterilized leaves. Inoculated leaves were incubated at 25°C, 100 to 120 μEm–2s–1, relative humidity (RH) >80%, and 16 h light per day to encourage disease development. Symptoms consistent with those described above developed on leaves within 7 days after inoculation. The pathogen was reisolated from affected leaves, but not from the noninoculated control leaves. C. trifolii causing leaf spot also has been reported from Berseem Clover in Nepal (Khadka 2016). However, to the best of our knowledge, this is the first report of C. trifolii causing leaf spot on tobacco in Guizhou Province of China. The disease needs to be considered in relation to existing disease management practices for spot of tobacco, and the biology of the pathogen. © 2017, American Phytopathological Society. All rights reserved.

Sun X.,CAS Institute of Microbiology | Kang S.,CAS Institute of Microbiology | Zhang Y.,Shanxi University | Tan X.,Hunan Academy of Agricultural Science | And 8 more authors.
PLoS ONE | Year: 2013

Rice false smut caused by the fungal pathogen Ustilaginoidea virens is becoming a destructive disease throughout major rice-growing countries. Information about its genetic diversity and population structure is essential for rice breeding and efficient control of the disease. This study compared the genome sequences of two U. virens isolates. Three SNP-rich genomic regions were identified as molecular markers that could be used to analyze the genetic diversity and population structure of U. virens in China. A total of 56 multilocus sequence types (haplotypes) were identified out of 162 representative isolates from 15 provinces covering five major rice-growing areas in China. However, the phylogeny, based on sequences at individual SNP-rich regions, strongly conflicted with each other and there were significant genetic differences between different geographical populations. Gene flow between the different geographical populations and genetic differentiation within each geographical population were also detected. In addition, genetic recombination and genetic isolation resulting from geographic separation was also found. © 2013 Sun et al.

Wang H.,Guizhou Tobacco Science Institute | Yang S.,Guizhou Province Tobacco Company | Yang S.,China Agricultural University | Wang M.,Guizhou Tobacco Science Institute | And 8 more authors.
Crop Protection | Year: 2013

Tobacco black shank caused by Phytophthora parasitica is a devastating soil-borne disease of tobacco in China. Control of tobacco black shank relies on numerous fungicide applications. A new carboxylic acid amide (CAA) fungicide, mandipropamid, was examined for its effects on various asexual developmental stages of P. parasitica in vitro and in tobacco seedlings. Although mandipropamid did not affect discharge of zoospores from sporangia, it strongly inhibited mycelial growth (mean EC50 value of 0.0112 μg ml-1), sporangia production (mean EC50 value of 0.009 μg ml-1), germination of encysted zoospores (mean EC50 value of 0.014 μg ml-1), and germination of sporangia (mean EC50 value of 0.017 μg ml-1). For protective activity in tobacco seedlings, various applications of mandipropamid were superior in reducing black shank compared to that of metalaxyl and of azoxystrobin; while for curative activity assay, 100 μg ml-1 of mandipropamid exhibited the same efficacy as that of metalaxyl, and presented superior activity than that of azoxystrobin. In 2010 and 2011, 119 isolates of P. parasitica from Guizhou Province of China were characterized for the baseline sensitivity to mandipropamid by measuring mycelial growth. Values of effective concentrations for 50% mycelia growth inhibition varied from 0.0068 to 0.0285 μg ml-1 and averaged 0.013 (±0.0045) μg ml-1, with a unimodal distribution. This information will serve as a baseline for tracking future changes in sensitivities of P. parasitica populations to mandipropamid in China. © 2012 Elsevier Ltd.

Li W.-H.,Guizhou University | Li W.-H.,Guizhou Institute of Plant Protection | Jin D.-C.,Guizhou University | Li F.-L.,Guizhou Institute of Plant Protection | And 2 more authors.
Symbiosis | Year: 2016

Gut bacterium Pantoea sp. is one of the predominant bacterial species in the larval gut of the diamondback moth, Plutella xylostella. The phenotypic characters of Pantoea sp. were investigated with BIOLOG phenotype MicroArray (PM) in this study. Totally 950 different metabolic phenotypes were tested using the PM plates 1–10. Results exhibited that Pantoea sp. was able to metabolize 37.37 % of the tested carbon sources, 91.32 % of nitrogen sources, 100 % of sulfur sources, and 98.31 % of phosphorus sources. Most informative utilization patterns for carbon sources of Pantoea sp. were organic acids and carbohydrates, and for nitrogen were various amino acids. The bacterium had 94 different biosynthetic pathways. It had a wide range of adaptabilities, and could still metabolize in osmolytes with up to 9 % sodium chloride, 6 % potassium chloride, 5 % sodium sulfate, 20 % ethylene glycol, 4 % sodium formate, 4 % urea, 5 % sodium lactate, 200 mmol/L sodium phosphate (pH 7.0), 100 mmol/L ammonium sulfate (pH 8.0), 100 mmol/L sodium nitrate, and 100 mmol/L sodium nitrite, respectively. It also exhibited active metabolism under pH values between 4.5 and 10. Pantoea sp. showed active decarboxylase activities while poor deaminase activities in the presence of various amino acids. The phenotypic characterization of Pantoea sp. increased our knowledge of the bacterium, in particular its interactions with insect hosts and the adaptability in gut environments, and showed us some possible approaches to controlling diamondback moth through decreasing Pantoea sp. density. © 2016 Springer Science+Business Media Dordrecht

Tao G.,CAS Institute of Microbiology | Tao G.,Guizhou Institute of Plant Protection | Tao G.,Guizhou Key Laboratory of Agricultural Biotechnology | Hyde K.D.,Mae Fah Luang University | Cai L.,CAS Institute of Microbiology
Journal of Applied Microbiology | Year: 2013

Colletotrichum kahawae is a strongly aggressive pathogen causing coffee berry disease and is specific to Arabica coffee (Coffea arabica) in Africa. In this article, we developed a real-time PCR assay for the species-specific diagnosis of C. kahawae by designing the primers and a TaqMan probe derived from the single nucleotide polymorphism-rich region of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene. Methods and Results: DNA markers from rDNA internal transcribed spacer, actin, β-tubulin and GAPDH genes of the ex-type culture of C. kahawae and 10 reference strains of Colletotrichum species were analysed for intra- and interspecific variations. The GAPDH gene was selected to develop a species-specific DNA marker. A TaqMan real-time PCR assay for species-specific detection of C. kahawae was developed, and its accuracy was tested against type strains of other phylogenetically closely related species in the C. gloeosporioides species complex, with the detection sensitivity of 80fgμl-1 of genomic DNA. Conclusions: This real-time PCR assay is highly specific and sensitive for the diagnosis of C. kahawae and can be applied in qualitative and quantitative tests. Significance and Impact of the Study: This protocol allows for a rapid and sensitive detection of C. kahawae and will be useful in disease management and pest detection to prevent further spread of this pathogen. © 2012 The Society for Applied Microbiology.

Tao G.,CAS Institute of Microbiology | Tao G.,Guizhou Institute of Plant Protection | Tao G.,Guizhou Key Laboratory of Agricultural Biotechnology | Liu Z.-Y.,Guizhou Key Laboratory of Agricultural Biotechnology | And 3 more authors.
Fungal Diversity | Year: 2013

Thirty-six strains of endophytic Colletotrichum species were isolated from leaves of Bletilla ochracea Schltr. (Orchidaceae) collected from 5 sites in Guizhou, China. Seventeen different species, including 7 new species (namely C. bletillum, C. caudasporum, C. duyunensis, C. endophytum, C. excelsum-altitudum and C. guizhouensis and C. ochracea), 8 previously described species (C. boninense, C. cereale, C. destructivum, C. karstii, C. liriopes, C. miscanthi, C. parsonsiae and C. tofieldiae) and 2 sterile mycelia were identified. All of the taxa were identified based on morphology and phylogeny inferred from multi-locus sequences, including the nuclear ribosomal internal transcribed spacer (ITS) region, partial genes of β-tubulin (TUB2), actin (ACT) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Comprehensive morphological descriptions and illustrations are provided for new species. Our investigation indicates a high diversity of Colletotrichum species in B. ochracea. © 2013 Mushroom Research Foundation.

Wang H.,Guizhou Tobacco Science Institute | Li W.,Guizhou Institute of Plant Protection | Chen Q.,Guizhou Tobacco Science Institute | Huang Y.,Yangtze University | And 6 more authors.
Phytopathology | Year: 2012

A simple, rapid, small-scale microbioassay for infection of tobacco seedlings by Phytophthora parasitica var. nicotianae was developed here. This assay uses tobacco seedlings cultivated in petri dishes for a standardized method for quantitation of initial zoospore inocula and highthroughput screening of antagonistic bacteria. Zoospore inocula between 102 to 105 spores per petri dish were inoculated on 14-day-old tobacco seedlings for the susceptibility test. The optimum inocula was established to be ten thousand zoospores. One hundred and fifty pure culture bacteria with different pigments, growth rates, and morphologies were isolated from rhizosphere soil of tobacco and screened for protective ability against tobacco black shank. Fifteen bacteria presented high activity against P. parasitica on tobacco seedlings. They were identified by Biolog GEN III MicroPlate and distributed as Bacillus amyloliquefaciens, B. licheniformis, Paenibacillus pabuli, B. atrophaeus, B. subtilis, B. pumilus, and B. endophyticus, respectively. Four antagonists chosen randomly from the 15 bacteria all exhibited the same 100% protective activity in planta as that in the petri dishes. This microassay proved to be a rapid, reproducible, and efficient method for screening of potential biological agents or microorganisms and may be useful for studying mechanisms of infection and control of Phytophthora spp. Under hydroponic conditions. © 2012 The American Phytopathological Society.

Li H.-B.,Yangzhou University | Li H.-B.,Guizhou Institute of Plant Protection | Zheng Y.-T.,Yangzhou University | Sun D.-D.,Yangzhou University | And 2 more authors.
Pesticide Biochemistry and Physiology | Year: 2014

Temperature and pesticide are two important factors that affect survival, reproduction and other physiological processes of insects. To determine interactions of elevated temperature and avermectins treatment on the western flower thrips, Frankliniella occidentalis, newly emerged adults were exposed to combinations of three temperatures (21, 26 and 33. °C) and two avermectins concentrations (0, 45. ppm), and survival rate, reproduction, longevity, antioxidant enzymes activities and heat shock proteins (hsps) induction were analyzed. The results showed that the survival, longevity and reproduction of F. occidentalis decreased with increased temperature and avermectins treatment. While elevated temperature and avermectins treatment significantly decreased activity of SOD, activities of POD and GST significantly increased after exposure to elevated temperature, avermectins or their combination. Elevated temperature had no effect on activity of CAT, but it was obviously improved by the combination of temperature and avermectins treatment. Expression analysis of hsps showed that four heat shock proteins (hsp90, hsc702, hsp60 and hop) were up-regulated by the induction of elevated temperature with small fold changes. After treatment with avermectins, expression levels of hsp90, hsc701, hsc702 and hop were significantly up-regulated with increased temperature and higher than those of their respective control at higher temperature. Surprisingly, expression level of hps60 was down-regulated with increased temperature, but the expression level at 21 or 26. °C remained higher than that of control. Overall, our studies suggest that elevated temperature enhance toxicity of avermectins and their combination induced acute oxidative damage to F. occidentalis. Therefore, consideration of temperature in evaluating avermectins toxicity is necessary to make accurate prediction of its effect on F. occidentalis and other insects. © 2014 Elsevier Inc.

Zheng Y.-T.,Yangzhou University | Li H.-B.,Yangzhou University | Li H.-B.,Guizhou Institute of Plant Protection | Lu M.-X.,Yangzhou University | Du Y.-Z.,Yangzhou University
PLoS ONE | Year: 2014

Quantitative real time PCR (qRT-PCR) has emerged as a reliable and reproducible technique for studying gene expression analysis. For accurate results, the normalization of data with reference genes is particularly essential. Once the transcriptome sequencing of Frankliniella occidentalis was completed, numerous unigenes were identified and annotated. Unfortunately, there are no studies on the stability of reference genes used in F. occidentalis. In this work, seven candidate reference genes, including actin, 18S rRNA, H3, tubulin, GAPDH, EF-1 and RPL32, were evaluated for their suitability as normalization genes under different experimental conditions using the statistical software programs BestKeeper, geNorm, Normfinder and the comparative DCt method. Because the rankings of the reference genes provided by each of the four programs were different, we chose a user-friendly web-based comprehensive tool RefFinder to get the final ranking. The result demonstrated that EF-1 and RPL32 displayed the most stable expression in different developmental stages; RPL32 and GAPDH showed the most stable expression at high temperatures, while 18S and EF-1 exhibited the most stable expression at low temperatures. In this study, we validated the suitable reference genes in F. occidentalis for gene expression profiling under different experimental conditions. The choice of internal standard is very important in the normalization of the target gene expression levels, thus validating and selecting the best genes will help improve the quality of gene expression data of F. occidentalis. What is more, these validated reference genes could serve as the basis for the selection of candidate reference genes in other insects. Copyright © 2014 Zheng et al.

Lu M.-X.,Yangzhou University | Li H.-B.,Yangzhou University | Li H.-B.,Guizhou Institute of Plant Protection | Zheng Y.-T.,Yangzhou University | And 2 more authors.
Journal of Thermal Biology | Year: 2016

The western flower thrips, Frankliniella occidentalis, is an important invasive pest with a strong tolerance for extreme temperatures; however, the molecular mechanisms that regulate thermotolerance in this insect remain unclear. In this study, four heat shock protein genes were cloned from F. occidentalis and named Fohsp90, Fohsc701, Fohsc702 and Fohsp60. These four Hsps exhibited typical characteristics of heat shock proteins. Subcellular localization signals and phylogenetic analysis indicated that FoHsp90 and FoHsc701 localize to the cytosol, whereas FoHsc702 and FoHsp60 were located in the endoplasmic reticulum and mitochondria, respectively. Analysis of genomic sequences revealed the presence of introns in the four genes (three, four, seven, and five introns for Fohsp90, Fohsc701, Fohsc702 and Fohsp60, respectively). Both the number and position of introns in these four genes were quite different from analogous genes in other species. qRT-PCR indicated that the four Fohsps were detected in second-stage larvae, one-day-old pupae, and one-day-old adults, and mRNA expression levels were lowest in larvae and highest in pupae. Fohsc701 and Fohsc702 possessed similar expression patterns and were not induced by cold or heat stress. Expression of Fohsp60 was significantly elevated by heat, and Fohsp90 was rapidly up-regulated after exposure to both cold and heat stress. Exposure to -8 °C had no effect on expression of the four Fohsps; however, expression of Fohsp90 and Fohsp60 was highest after a 2-h incubation at 39 °C. Furthermore, cold and heat hardening led to significant up-regulation of the four Fohsps compared to their respective controls. Collectively, our results indicate that the four FoHsps contribute to insect development and also function in rapid cold or heat hardening; furthermore, FoHsp90 and FoHsp60 contribute to thermotolerance in F. occidentalis. © 2016 Elsevier Ltd.

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