Assessing the Efficacy of Lemongrass (Cymbopogon citratus) and Sambong (Blumea balsamifera) Extracts in Combating Black Pod Disease: Sustainable Solutions for Controlling Phytophthora megakarya in Cameroon’s Cocoa Plantations

The use of plant extracts as antifungal agents is gaining increasing attention, particularly for the control of black pod disease in cocoa. Despite extensive research, current strategies haven’t been entirely effective. This study evaluated the effectiveness of Cymbopogon citratus and Blumea balsamifora leaf extracts, both individually and in combination, against Phytophthora megakarya. We assessed the efficacy of the most promising combination (75% B. balsamifera, 25% C. citratus) after storage at room temperature for up to 9 days. Agar microdilution and in vivo bioassays were conducted to determine antifungal susceptibility and effectiveness. Blumea extract exhibited the highest overall inhibitory activity, with the lowest minimum inhibitory concentration (117 µl mL−1) while C. citratus had a narrower range of MIC (146 to 233 µl mL−1). The combination of C. citratus and B. balsamifera demonstrated a synergistic effect against P. megakarya, achieving growth inhibition on V8 media (92.72 ± 4.20% to 100%) and on artificially infected detached pod cortex (92.24 ± 4.53% to 98.75 ± 1.25%), which was not significantly different from the positive control (Ridomil). Furthermore, this combination maintained its effectiveness for up to 9 days at room temperature. These findings suggest that combining plant extracts can enhance their antifungal properties.

Disease Analysis and Pathogen Identification of Black Spot Disease of Chinese Cabbage [Alternaria brassicae (Berk.) Sace.] in Bijie City

[Objective] Disease analysis and pathogen identification of black spot disease of Chinese cabbage [Alternaria brassicae（Berk.） Sace.] in Bijie City of Guizhou Province was carried out,so as to raise the corresponding prevention measures.[Method] Pathogen was isolated from diseased leaves using tissue surface separation method for cultivation,microscopic examination,spore size determination and pathogen identification.[Result] Microscopic examination showed that the disease was caused by Alternaria sp.,the size of its spores was（15.0-38.5） μm×（7.5-13.5） μm.Combining with the characteristics of host and pathogen,the pathogen caused black spot disease of Chinese cabbage was identified to be Alternaria brassicicola.[Conclusion] A.brassicicola was the pathogenic fungi of black spot disease of Chinese cabbage in Bijie City of Guizhou Province,and damage caused by the disease could be reduced through agricultural measures and chemical control.

Cloning and Sequencing Characteristics of rDNA ITS Region of the Pathogen Causing Black Spot Disease on Tumorous Stem Mustard

Taking tumorous stem mustard infected by black spot disease as the research material, the ribosomal 5.8S rDNA and its flanking ITS region were cloned, sequenced and aligned in the study. The results showed that the base sequence of pathogens collected from five different sites was almost identical with that of Alternaria brassicae, and the similarity degree reached 99.68%, without base difference greater than 3 bp. It had obvious differences with the sequences of A. brassicicola and A. japonica, and the base difference was greater than 3 bp with a lot of fragment deletions. It was preliminarily determined that the pathogen cau- sing black spot disease on tumorous stem mustard was A. brassicae.

16S rRNA gene sequencing analysis on changes in the intestinal flora of Procambarus clarkii with“Black May”disease

The morbidity and mortality peak of farmed Procambarus clarkii occurs around May every year,a phenomenon known as"Black May"disease(BMD).Increasing evidence shows that the intestinal flora is closely related to host health.We analyzed and compared the microbiota of healthy and BMDaffected P.clarkii intestines.The results show that there was no significant difference in bacterial a-diversity(richness P=0.59;evenness P=0.43;and diversity P=0.052)between the diseased group and the control group.Four dominant phyla in the intestines of crayfish in the control group,namely Tenericutes(30.86%),Bacteroidetes(29.99%),Firmicutes(22.23%),and Proteobacteria(15.23%),were identified.However,a striking shift in the microbial compo sition were found in the intestines of P.clarkii with BMD.Bacteroidetes was a dominant phylum in healthy P.clarkii,whereas the prevalence was low in diseased P.clarkii(1.87%).By contrast,the prevalence of Proteobacteria was significantly higher(P<0.05)in P.clarkii with BMD than in P.clarkii without BMD.Candidatus Bacilloplasma,Bacteroides,Vibrio,and Aeromonas showed significant differences(P<0.05)at the genus level.Tax4 Fun function prediction indicated that the relative abundance of genes involved in energy metabolism in the intestinal flora of P.clarkii with BMD was significantly reduced(P<0.05).Therefore,BMD can change the composition of the intestinal microbiota of P.clarkii.This study contributes to the understanding of the relationship between intestinal flora and host especially in aquatic animals.

Screening and Identification of Biocontrol Strains against Potato Black Shank Disease,Pectobacterium atrosepticum

[Objective]The paper was to screen and identify biocontrol strains against potato black shank disease in Inner Mongolia.[Method]Totally 58 strains of Bacillus spp.preserved in the laboratory were used to screen and identify biocontrol agents against potato black shank disease,and the antagonistic activities of strains were determined by plate confrontation culture method.[Result]A total of 17 strains showed antagonistic activity against Pectobacterium atrosepticum in potato,accounting for 29.31%of the tested strains.Five strains with better antagonistic effect were obtained by further screening,namely Chi8-7,Genyun4-4,Gen7-4,Jing6-7,and S-7.Genyun4-4 had the most obvious bacteriostatic effect,and the average inhibition zone diameter reached 20.7 mm,significantly higher than those of other tested strains.Results of bioactivity determination also demonstrated that the strain had better bacteriostatic effect.Through morphological observation,molecular identification and physiological and biochemical characteristics determination,the strain Chi8-7 was identified as Bacillus amyloliquefaciens;the strain Genyun4-4 was identified as B.subtilis;and strains Gen7-4,Jing6-7 and S-7 were identified as B.methylotrophicus.[Conclusion]The results will lay a theoretical basis for biological control against potato black shank disease in Inner Mongolia.

Pathogenesis, Prevention and Control of Coffee Black Fruit Disease ( Fusarium solani )

Coffee black fruit disease causes great harm to the yield and quality of coffee. Multiple factors, such as climate factors, insect pests, pathogen infection, physiological disorders, improper planting density, etc. , can cause coffee black fruit disease, which can be categorized into three types: pathogen infection type, physiological disorder type and insect pest type. Through the analysis of pathogenesis, the prevention and control methods and techniques of the disease corresponding to different types are put forward.

Field Efficacy of a Biopesticide Based on <i>Tithonia diversifolia</i>against Black Sigatoka Disease of Plantain (<i>Musa</i>spp., AAB)

Black Sigatoka disease (BSD) is a foliar disease caused by Mycosphaerella fijiensis, responsible of reduction of the photosynthetic area of banana plant and yield at harvest since it has an influence on fruit physiology. The control of BSD relies on the use of chemicals which are not affordable for the small holder farmers and increase the cost of production. Moreover, this chemical control is ineffective, negatively impacting the environment and human health, and is at the origin of strain resistance. Tithonia diversifolia is known as rich in many compounds such as mineral elements, defense metabolites, some phytochemicals;and it is increasingly used in agriculture. Recently, the protective effect of Tithonia diversifolia liquid extract against BSD development on plantain vivoplants in the nursery was highlighted. The aim of our study was to evaluate the efficacy of a biopesticide base on Tithonia diversifolia on the BSD development in a plantain field under high disease pressure. The effect of Tithonia diversifolia biopesticide on Mycosphaerella fijiensis mycelial growth in vitro was evaluated. An experimental field at the flowering stage was selected and treated with the biopesticide base on Tithonia diversifolia at three different concentrations: undiluted (100%), diluted at 1/2 (50%) and diluted at 1/4 (25%) for 17 weeks. The disease severity, the number of functional leaves, the youngest spotted leaf (YSL) and the youngest necrotic leaf (YNL) were evaluated in course of time. The biopesticide treatments significantly (P < 0.001) reduce the BSD severity in course of time, but it is more effective for the most diluted concentration (25%). The number of leaves increases in course of time as well as the rank of the YSL and the YNL confirming the efficiency of BSD control. The efficacy of this biopesticide base on Tithonia diversifolia could be a hopeful ecoresponsible solution for the plantain sector in general and in particular for poor small farmers.

Black gill disease in Litopenaeus vannamei made by various agents

Melanization of shrimp gills is a general symptom in shrimp black bill diseases(sBG);therefore,the cause is not immediately apparent.This study aimed to determine the primary and secondary agents associated with a black gill outbreak in the Delvar complex in the Bushehr Province of Iran.During July and August of 2020,450 healthy and affected shrimps were collected based on brown to black gill color from three farms of the Bushehr Province in Iran.Characterization of pathogens was performed by gross signs,wet mounts,microbiological tests,histopathology methods,and sequencing of molecular markers.The mean weight and length of infected shrimps was significantly(P<0.05)lower than the healthy shrimps.The histological examination of sBG indicated the adhesion of secondary lamellae,accumulation of hemocyte cells at the end of the secondary lamellae,and necrosis of epithelial cells with pyknosis and karyorrhexis.According to our results,Aspergillus sydowii as a primary agent correlates with sBG disease in Litopenaeus vannamei from Iran,and other biotic factors,including bacterial(Vibrio owensii and V.parahaemolyticus)probably are secondary agents associated with sBG disease.

Stimulatory Effect of Tithonya diversifolia-by Products on Plantain Banana Vivoplants in Nursery (A Review)

Plantain banana is an important cash crop that serves as stable food for millions of people around the world and contributes to income generation. Indeed, they provide a major staple food crop for millions of people and play an important role in the social fabric of many rural communities. Plantain banana cultivation encounters major problem of seedlings unavailability that are essential for the creation of new plantations, as well as parasitic constraints. Mycosphaerella fijiensis is the main pathogen attack constraints of banana plant responsible of black Sigatoka disease, and viruses, which can severely reduce the photosynthetic leaf area, leading to banana production losses of more than 80% in plantations with soil fertility problems. The repeated use of synthetic input is the origin of contamination to the environment, different pollution sources of plants and human health, as well as resistance to some strains of pathogens and plant fertilization problems over time. Recent works carried out in nursery have shown that vivoplants of plantains treated with biostimulants based on natural products notably Tithonia diversifolia biopromote good growth and less susceptibility to M. fijiensis. Indeed, an increase in agromorphological characteristics, good accumulation of growth and defense biomarkers was also observed. In this context, Tithonia diversifolia is shown to be involved in the stimulatory effect mechanism of growth promotion and defensive reaction of plantain vivoplants against various pathogens and it is suggested to be acting as a vital stimulator. This article reviews the current state of knowledge on plantain banana cultivation constraints and on the potential of Tithonia diversifolia in relation with its different stimulatory effects on plantain vivoplants.

Identification of differentially expressed proteins in poplar leaves in-duced by Marssonina brunnea f. sp. Multigermtubi

Black spot disease in poplar is a disease of the leaf caused by fungus. The major pathogen is Marssonina brunnea f. sp. multigermtubi. To date, little is known about the molecular mechanism of poplar （M. brunnea） interaction. In order to identify the proteins related to disease resistance and understand its molecular basis, the clone ＂NL895＂ （P. euramericana CL＂NL895＂）, which is highly resistant to M. brunnea f. sp. multigermtubi, was used in this study. We used two-dimensional gel electrophoresis （2-DE） and mass spectrometry （MS） to identify the proteins in poplar leaves that were differentially expressed in response to black spot disease pathogen, M. brunnea f. sp. multigermtubi. Proteins extracted from poplar leaves at 0, 12, 24, 48, and 72 h after pathogen-inoculation were separated by 2-DE, About 500 reproducible protein spots were detected, of which 40 protein spots displayed differential expression in levels and were subjected to Matrix assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF MS） followed by database searching. According to the function, the identified proteins were sorted into five categories, that is, protein synthesis, metabolism, defense response and unclassified proteins.

Black band disease as a possible factor of the coral decline at the northern reef-flat of Yongxing Island,South China Sea

Black band disease （BBD）, characterized by the Cyanobacterial dominated pathogenic consortium, is thought to play a key role in the global decline of the coral reef ecosystems. The present paper originally documents a case of BBD from Yongxing Island （Xisha Islands, South China Sea）, and further probes the reasons of this abnormal phenomenon. Prior to 2007, corals at northern reef-flat of Yongxing Isand were in healthy growth. Catastrophic coral mortality occurred between 2007 and 2008. The 16S rRNA gene sequencing and PCR amplification, with universally conserved primers, were applied to detect the conta- gious bacterial community of the microbial mat. The results demonstrated that six bacterial divisions constituted the clone libraries derived from the BBD mat, and that Cyanobacteria are the most diversely represented group that inhabit BBD bacteri- al mats, despite the fact that species in five others divisions （a-Proteobacteria, y-Proteobacteria, Bacteroidetes, Verrucomi- crobia and Actinobacteria） are also consistently diverse within the BBD mats of diseased coral. Other factors such as coral bleaching, typhoons, ocean acidification and crown-of-thorns starfish outbreaks, are not primarily responsible for the coral mortality within such a short time interval. The disaster expansion of BBD associated with Cyanobacterial blooms is a more likely mechanism impacting these coral reefs. Excessive human activity enhances the eutrophication of the marine water of the reefal region and may result in occurrence of the BBD.

Nonphytotoxic copper oxide nanoparticles are powerful“nanoweapons”that trigger resistance in tobacco against the soil-borne fungal pathogen Phytophthora nicotianae

Investigations into the potential application of nanoparticles acting as nanofungicides in sustainable agriculture are rapidly expanding due to the high antimicrobial properties of these compounds,which do not risk inducing pathogen resistance to fungicides.A detailed understanding of the impact of copper oxide nanoparticles(CuO NPs)on soil-borne phytopathogenic fungi is yet to be obtained.This study aimed to explore the in vitro antifungal activity and control efficacy of CuO NPs applied via irrigation with respect to tobacco black shank(TBS)disease caused by Phytophthora nicotianae.The results revealed that CuO NPs greatly interfered with the reproductive growth process of this fungus,repressing hyphal growth,spore germination and sporangium production.Additionally,morphological damage,intracellular ROS accumulation and increased SOD enzyme activity in hyphae were the antifungicidal mechanisms of these NPs.In pot experiments,treatment with CuO NPs at 100 mg L^(–1)significantly suppressed TBS development,compared with the effect on control plants,and the control efficacy reached 33.69%without inducing phytotoxicity.Exposure to CuO NPs significantly activated a series of defense enzymes,and resistance genes in tobacco can further explain the mechanisms by which CuO NPs suppressed fungal infection.The Cu content in both the leaves and roots of P.nicotianae-infested plants increased by 50.03 and 27.25%,respectively,after treatment with 100 mg L^(–1)CuO NPs,compared with that of healthy plants.In particular,a higher Cu content was observed in infected roots than in leaves.Therefore,this study showed the potential of CuO NPs applied as nanofungicides and as nanoinducers of fungus resistance genes for the management of TBS through inhibition of pathogen infection and stimulation of plant defenses.

Black gill disease of Paciifc white leg shrimp(Litopenaeus vannamei)byAspergillus lfavus

Objective:To study the epidemiology of black gill disease in white leg shrimp which is a major problem being faced by the commercial shrimp farmers who are culturing Litopenaeus vannamei(L.vannamei)in India.Methods:The normal and infected shrimps were collected from shrimp pond and the gill was preserved in appropriate preservative for histopathological examination and scanning electron microscope analysis.Pathogenic fungus was isolated from black gill of L.vannamei in potato dextrose agar medium.Morphological study and fungal strain identification were done by using light microscopy and scanning electron microscope.Fungal DNA was amplified by ITS4 and ITS5 primers and gene sequencing was done by Macrogen Inc.,Korea.Phylogenetic tree was prepared by using MEGA 6 software.Results:Fungal spores and hyphae were observed both in internal and external gill surface of infected shrimps.Fungal spores were round in shape and mature sporangium was observed.The histopathology study showed clearly that infected gill was damaged by the fungi.Scanning electron microscopic study showed adherence of fungi in infected gill.Internal transcribed spacer gene sequencing revealed that it was caused by Aspergillus flavus.Conclusions:The outcome of the present study would help to know the cause of black gill disease and to understand the effect of pathogenic fungi in shrimp culture.This study will initiate researchers for work in field of treatment or prevention of black gill disease in commercial L.vannamei culture.

Phyllosticta species associated with citrus diseases in China

Phyllosticta species associated with diseases of four commercial Citrus species grown in China are reported.Totally,496 Phyllosticta strains were isolated from mandarins(Citrus reticulata),pomeloes(C.maxima),oranges(C.sinensis)and lemons(C.limon)in the main citrus producing regions across China,and 74 strains were selected for phylogenetic analysis.Analyses inferred from the sequences of internal transcribed spacer region(ITS1,5.8S nrDNA and ITS2),partial translation elongation factor 1-alpha(TEF1)and partial actin gene(ACT),showed these representative Phyllosticta isolates clustered in four distinct clades corresponding to three known,and one undescribed species.The newly resolved taxon,Phyllosticta citrichinaensis was isolated from leaves and fruits of all four Citrus species and is introduced in this paper.This taxon caused minor damage,showing irregular spots or freckles.Phyllosticta citriasiana,associated with tan spot of pomeloes,was isolated only from pomeloes,and never from lemons,mandarins and oranges.Phyllosticta citricarpa,the citrus black spot pathogen,which is presently subjected to phytosanitary legislation in the EU and United States,was isolated from lemons,mandarins and oranges,but never from pomeloes.The isolates of P.citricarpa clustered in two subclades,one from mandarins,the other from oranges and lemons.P.capitalensis was isolated from all four Citrus species as an endophyte,causing false melanose,or together with P.citricarpa or P.citriasiana.Morphological,cultural and biochemical characters were consistent with the results of phylogenetic analysis.In addition,a specific primer pair Pca8/ITS4 was designed and selected,and its corresponding PCR procedure was developed for the detection of P.citriasiana in this study.

引起我国葡萄黑根病的Dactylonectria novozelandica的鉴定

葡萄黑根病(Black foot disease)是重要的葡萄枝干病害之一,全球葡萄主要种植区均有发生[1],近两年我国有该病害发生的报道[1-2]。田间,黑根病主要危害8年树龄以下的幼树,症状主要表现为发芽异常,枝条节间缩短,叶片失绿黄化,根部出现褐色至黑色坏死等[3-5]。引起葡萄黑根病的病原菌复杂多样,已报道8个属34种病原菌[1]。

Synthesis of selenium-doped carbon from glucose:An efficient antibacterial material against Xcc

Se/C as a novel carbon-based biomaterial was prepared by using cheap and abundant glucose as the carbon source.It was highly active and could well restrain Xanthomonas campestris pv.campestris(EC50=4.7403 μg/mL),a very harmful germ causing the devastating cabbage black rot disease and resulting in huge economic losses.As a type of carbon material insoluble in wate r,Se/C is bio-compatible and can adhere onto the leaves of the plants to allow a slow and sustained release of its efficacy,affording an efficient method for the cabbage black rot disease prevention and cure.This work as the first report on the bioactivity studies of Se/C significantly expands the application scopes of the selenium-containing materials and may draw continuous attentions from a broad field.

Rapid and non-destructive decay detection of Yali pears using hyperspectral imaging coupled with 2D correlation spectroscopy

The black spot disease caused by Alternaria alternata on Yali pears is a great concern as it compromises their edible quality and commercial value.To realize rapid and non-destructive classification of this disease,hyperspectral imaging(HSI)technology was combined with two-dimensional correlation spectroscopy(2DCOS)analysis.A total of 150 pear samples at different decay grades were prepared.After obtaining the HSI images,the whole sample was demarcated as the region of interest,and the spectral information was extracted.Seven preprocessing methods were applied and compared to build the classification models.Thereafter,using the inoculation day as an external perturbation,2DCOS was used to select the feature-related wavebands for black spot disease identification,and the result was compared to those obtained using competitive adaptive reweighting sampling and the successive projections algorithm.Results demonstrated that the simplified least squares support vector model based on 2DCOS-identified feature wavebands yielded the best performance with the identification accuracy,precision,sensitivity,and specificity of 97.30%,94.60%,96.16%,and 98.21%,respectively.Therefore,2DCOS can effectively interpret the feature-related wavebands,and its combination with HSI is an effective tool to predict black spot disease on Yali pears.