A Rapid Detection Method for Tomato Gray Mold Spores in Greenhouse Based on Microfluidic Chip Enrichment and Lens-Less Diffraction Image Processing
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It is of great significance to find tomato gray mold in time and take corresponding control measures to ensure the production of tomato crops. This study proposed a rapid detection method for spores of Botrytis cinerea in green-house based on microfluidic chip enrichment and lens-free diffraction image processing. Microfluidic chip with a regular triangular inner rib structure was designed to achieve the enrichment of Botrytis cinerea spores. In order to obtain the diffraction image of the diseased spores, a lens-less diffraction imaging system was built. Furthermore, the collected spore diffraction images were processed and counted. The simulation results showed that the collection efficiency of 16 μm particles was 79%, 100%, and 89% at the inlet flow rate of 12, 14 and 16 mL/min, respectively. The experimental verification results were observed under a microscope. The results showed that when the flow rate of the microfluidic chip was 12, 14 and 16 mL/min, the collection efficiency of Botrytis cinerea spores was 70.65%, 87.52% and 77.96%, respectively. The Botrytis cinerea spores collected in the experiment were placed under a microscope for manual counting and compared with the automatic counting results based on diffraction image processing. A total of 10 sets of experiments were carried out, with an error range of the experiment was 5.13~8.57%, and the average error of the experiment was 6.42%. The Bland-Altman method was used to analyze two methods based on diffraction image processing and manual counting under a microscope. All points are within the 95% consistency interval. Therefore, this study can provide a basis for the research on the real-time monitoring technology of tomato gray mold spores in the greenhouse.Keywords:
Microfluidic chip
Botrytis
The investigation forming the subject of this report was planned to study certain factors affecting Botrytis diseases on certain crop plants. Results are summarised in the following points: 1. The optimum temperature for growth of Botrytis cinerea Pers., Botrytis cinerea f. lini, Botrytis fabae Sard., Botrytis galanthina Berk. & Br., Botrytis narcissicola Kleb., Botrytis paeoniae Oudem. and Botrytia squamosa Walker was 21
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Botrytis calthae is a necrotrophic plant pathogen, closely related to the ubiquitous broad host range fungus Botrytis cinerea , but highly host specific. Botrytis isolates from lesions of Caltha palustris grown at different locations were classified with genetic markers as either B. calthae or Botrytis pseudocinerea , or less frequently as B. cinerea . A PCR‐based identification of B. calthae was developed. Seven haplotypes of B. calthae could be distinguished. Compared to B. cinerea , mycelium growth of B. calthae was similar, but conidiation less abundant, and sclerotia formation was only partially repressed by light. Conidia of B. calthae germinated more slowly, and showed a highly acidic optimum ( pH 2·5) compared to B. cinerea conidia ( pH 5·3). All B. calthae isolates were sensitive to common anti‐ Botrytis fungicides, but showed partial resistance to the succinate dehydrogenase inhibitors boscalid, fluopyram and carboxin. Infection experiments revealed a weak capability of B. calthae to induce necrotic lesions on plants that are hosts for B. cinerea . On C. palustris leaves, B. calthae induced similar lesions to B. cinerea . These data provide a basis for comparative molecular investigation of the physiology and host specificity of B. calthae and closely related Botrytis species.
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Gray mold, caused by Botrytis cinerea, is an important disease that causes world-wide extensive damage to a wide range of economically important crops. When it is necessary to determine the spore concentration of Botrytis cinerea in a certain area, it is important to develop a method that can capture the spores of Botrytis cinerea and that can identify them. For the identification and enumeration of the plant pathogen Botrytis cinerea in the environment the easiest method available for the moment is the use of a selective medium. Several selective media for the isolation of Botrytis spp. have been developed by other research groups. All these media contain fungicides that are usually non-toxic towards Botrytis species and tannic acid, which is oxidized to produce a brown pigment that visualises the growth of Botrytis cinerea on the selective media. It seemed that different isolates of Botrytis cinerea that are found in nature have different sensitivities towards the different fungicide concentrations that are used in the selective media. Making the "optimal" selective media for Botrytis cinerea, we have to take in consideration that so many as possible Botrytis cinerea isolates must be able to germinate and grow on this selective medium and that the contamination of other micro-organisms on the selective medium must be minimized. Before the final composition of our selective medium for Botrytis cinerea, different combinations of fungicide concentrations were tried out of the following three fungicides: Rubigan, maneb and PCNB (pentachloronitrobenzene). All these selective media with different fungicides concentrations were tested out for spore germination and mycelium growth of Botrytis cinerea. Because it was obvious that the percentage Botrytis cinerea that germinated on PDA (Potato Dextrose Agar) was higher than on the selective medium a few experiments were executed in which the percentage of spore germination on PDA was compared with the percentage of spore germination on selective media. From the results of these experiments a correction factor was calculated that will be used when spore concentrations have to be determined for air detections that are carried out in glasshouses.
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본 연구는 백합 잎마름병에 관여하는 B. elliptica와 B. cinerea의 형태적, 배양적 특성을 조사하고, 유전자 염기서열에 기초한 분자계통분석을 하며 병원성을 비교하기 위하여 실시하였다. 한국, 일본, 네덜란드의 주요 백합 재배지역에서 잎마름병에 걸린 백합으로부터 79개의 Botrytis속 균주를 분리하였다. 분생자경과 분생포자의 형태적 특징에 의하여 Botrytis 균들은 2개의 그룹으로 분리되었으며 이들은 각각 B. elliptica와 B. cinerea로 동정되었다. 균사생장 속도는 B. cinerea가 B. elliptica에 비해 빨랐으며 이러한 형태적, 배양적 특성에 기초하여 79개의 균주 중 54균주(68%)가 B. elliptica로, 25균주(32%)가 B. cinerea로 동정되었다. RPB2와 HSP60의 염기서열 분석을 통해, 공시균주는 두 그룹으로 구분되었으며 이들은 형태적으로 동정한 그룹과 일치하였다. 포자 현탁액을 이용한 B. elliptica와 B. cinerea의 병원성 검정 결과 B. elliptica 균주들은 백합 잎과 꽃잎 모두에 병원성을 나타내었다. 그러나 B. cinerea 분생포자 현탁액은 꽃잎에만 병원성을 나타내었으며 PDB가 첨가된 포자현탁액을 접종원으로 사용했을 때에만 백합 잎에 병원성을 나타내었다. The seventy nine isolates of Botrytis spp. were obtained from leaf blight lesions of lily in Korea, Japan and Netherlands. Morphological and cultural characteristics of them were investigated and molecular characteristics of them were determined using sequence analysis of RNA polymerase II (RPB2) and heat-shock protein 60 (HSP60) gene. A selection of Botrytis isolates were evaluated for their pathogenicity to lily. Based on morphological and cultural characteristics, the Botrytis isolates were divided into two groups, and identified as B. elliptica (n = 54) and B. cinerea (n = 25). Based on analysis of RPB2 and HSP60 sequences, the Botrytis isolates were also divided into two groups and well supported morphological groupings. Spore suspensions of B, elliptica showed significant pathogenicity on lily leaves and flowers, however those of B. cinerea showed pathogenicity only on flowers but not on leaves. The latter showed pathogenicity on lily leaves only when spore suspensions amended with PDB were used as inocula.
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ABSTRACT Two alternatives to chemical control of Botrytis in strawberries were assessed, post-harvest defoliation and finger harrowing, the latter being used primarily in weed control but also claimed to affect the occurrence of Botrytis. Post-harvest defoliation, though having some positive effect on marketable yield, did not reduce the incidence of Botrytis significantly. This was the case regardless of whether or not cut leaf material was removed from the plots. Finger harrowing, though in some instances having a negative effect on yield, did not control Botrytis significantly either. However, it was clearly demonstrated that, in organic strawberry growing, the incidence of Botrytis increases with plant age. Therefore, a short-term cropping system may be more important in controlling Botrytis than either post-harvest defoliation or finger harrowing. In conclusion, the combined use of several cultural methods may be needed when controlling Botrytis in organically grown strawberries.
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Botrytis blossom blight and fruit rot, caused by
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Iprodione
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The objective of the paper is to investigate the sulfur dioxide(SO2) to inhibit Botrytis cinera Pers to control the decay of grapes during storage to select suitable SO2 concentration inhibiting Botrytis cinera Pers to provide a bass for controlling decay of grapes during storage.The fuming treatment medium with different SO2 concentrations and Botrytis cinera Pers on the surface of grapes fruit were used to study inhibition effects on Botrytis cinera Pers and effects on grapes quality and to select suitable SO2 treatment concentration.Resuts showed that there was positive correlation between the inhibition effect of Botrytis cinera Pers in vitro and the inhibition and concentration of Botrytis cinera Pers.The significant effect of SO2 to control Botrytis cinerea Pers was beginning from 200 μL/L.Grapes inoculated Botrytis cinerea Pers bacteriostatic experiment showed,200 μL/L or 250 μl/L treatment effect was significantly better than the other concentrations.But 250 μl/L SO2 handling was harmful to grape quality.Suming up the inhibition of Botrytis cinerea Pers and the affection of grape fruit quality,SO2 concentration of 200 μL/L was suit for embalmed thompson seedless storage.
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