Morpho-cultural, Pathogenicity and Molecular Characterization of Phyllosticta capitalensis Inciting Cavendish Banana Freckle Disease in Hainan

[Objectives]The study was to identify the casual agent of freckle disease on Cavendish banana in Hainan Province,China.[Methods]Fungal isolates were isolated from affected leaf tissues and identified by the morphological features,molecular identification and pathogenicity test.[Results]The fungus isolated from affected leaf tissues was identified as Phyllosticta capitalensis based on the morphological properties of the colony and spore,coupled with sequence analyses of the internal transcribed spacer(ITS)region and the large subunit(LSU)rDNA gene.Koch s postulates were fulfilled by successfully re-isolating the pathogen from the artificial inoculated leaves.[Conclusions]P.capitalensis is a new pathogen responsible for Cavendish banana freckle disease in Hainan.

Biological Characteristics of the Pathogen Phyllosticta capitalensis Causing Banana Freckle Disease in Hainan Province

[Objectives]This study was conducted to clarify the biological characteristics of the pathogen Phyllosticta capitalensis,the causal agent of freckle disease on Cavendish banana in Hainan Province,China.[Methods]The impact of various nutritional and environmental factors,including media,carbon sources,nitrogen sources,temperature,pH and light on the growth and sporulation of P.capitalensis was assessed using two distinct methods:mycelium growth rate and blood counting chamber.[Results]The mycelial growth and sporulation of P.capitalensis on different media exhibited notable differences.The use of banana leaf extract dextrose agar(BLEAD)and carrot agar(CA)was observed to facilitate rapid mycelial growth.The potato dextrose agar(PDA)and potato sucrose agar(PSA)were conducive to the production of conidia.The utilization of distinct carbon and nitrogen sources exerted a pronounced influence on the growth of P.capitalensis.Maltose,dextrose,fructose,and casein acid hydrolysate were the preferred substrates for mycelial growth.The tested carbon and nitrogen sources did not significantly stimulate conidial production,whereas dextrose and NaNO 3 were found to favor sporulation.The optimal temperature for mycelial growth and conidial production was determined to be 28 and 32℃,respectively.No mycelial growth was observed at 5℃.Active mycelial growth was observed at pH 6-10,with pH 6-7 being particularly conducive to sporulation.Complete darkness was conducive to mycelial growth and sporulation.[Conclusions]It is recommended that BLEDA and PDA should be incubated at 28℃for 14 d in the dark for the purpose of mycelial growth and sporulation of P.capitalensis,respectively.

Molecular Characterization,Morphology and Pathogenicity of Curvularia pseudobrachyspora,A New Causal Agent of Leaf Spot on Banana

[Objectives]The paper was to study molecular characterization,morphology and pathogenicity of Curvularia pseudobrachyspora,a new causal agent of leaf spot on banana.[Methods]Banana(Musa acuminate)leaves with streaks or long ellipse-shaped lesions were sampled in an orchard of Danzhou City,Hainan Province,China in 2021.Fungus isolates were isolated from the diseased tissues and further identified as C.pseudobrachyspora based on morphological characteristics of colony,conidiophore and spore,phylogenetic analyses of the ITS region,GAPDH and TEF-1αgenes.[Results]In the pathogenicity test,the fungus re-isolated from inoculated leaves with necrotic lesions was identified morphologically and molecularly,fulfilling Koch's postulates.[Conclusions]C.pseudobrachyspora is a new pathogen causing leaf spot of banana in China and the world.

Morphological and Molecular Identification of Alternaria alternata Causing Leaf Spot Disease on Huangdi Banana in China

[Objectives]The paper was to identify Alternaria alternata causing leaf spot disease on Huangdi banana in China.[Methods]Fungal isolates were isolated and identified by morphological features,molecular identification and pathogenicity test.[Results]There were light to dark brown,tiny oval spots on leaves.The causal agent isolated from affected leaves was identified as A.alternata based on the morphological properties,coupled with sequence analyses of the internal transcribed spacer(ITS)region,large subunit ribosomal DNA(LSU rDNA)and the translation elongation factor 1-alpha(TEF-1α)gene.Koch s postulates were fulfilled by successful re-isolation of pathogen from the artificial inoculated leaves.[Conclusions]To our knowledge,this is the first report of leaf spot caused by A.alternata on Huangdi banana in China.The identification of A.alternata as the causal agent of the observed leaf spot disease on Huangdi banana is critical to the prevention and control of this disease in the future.

Characterization of Neopestalotiopsis clavispora , A New Etiological Agent of Leaf Spot Isolated from Banana

[Objectives]The study was to confirm the etiological agent of leaf spot on banana in Yunnan Province of China.[Methods]Fungus isolates were isolated from the diseased tissues,and cultured to observe the morphological characteristics of colony and spore.Furthermore,phylogenetic analyses and pathogenicity test were also conducted to confirm the pathogen.[Results]The fungus isolated from the diseased tissues was identified as Neopestalotiopsis clavispora.[Conclusions]N.clavispora is a new pathogen causing leaf spot of banana.This research provides the first description of N.clavispora as a causal agent on banana in China,and adds new insights related to the host range of N.clavispora.

Morphological and Molecular Identification of Curvularia geniculata Associated with Leaf Spot Disease of Banana in China

[Objectives]The paper was to identify the causal agent of leaf spot of banana in Hainnan,China.[Methods]Fungus isolates were isolated from the diseased tissues,and morphological characteristics of colony and spore were observed.Furthermore,phylogenetic analyses and pathogenicity test were conducted to confirm the pathogen.[Results]The fungus isolates from the diseased tissues was identified as Curvularia geniculata.[Conclusions]C.geniculata is a new pathogen causing leaf spot of banana in Hainan,China.This finding will help to broaden the distribution and host range of C.geniculata,indicating that it poses potential damage to banana in China.

Inhibitory Effects of Six Fungicides against Neocordana musae

[Objective]The paper was to screen effective fungicides to control Cordana leaf spot caused by Neocordana musae.[Method]The toxicities of six fungicides against mycelial growth of N.musae were measured by the method of mycelium growth inhibition.[Result]The average EC50 values of six fungicides against mycelial growth ranged from 0.1029 to 504.065μg/mL.Among them,22.5%picoxystrobin SC and 12.5%epoxiconazole SC showed strong inhibitory activity on mycelial growth,with the average EC50 values of 0.1029 and 0.6851μg/mL,respectively;followed by 50%thiophanate-methyl SC,with an average EC50 value of 1.993.80%Azoxystrobin WG showed a relatively low sensitivity against N.musae,with an average EC50 value of 504.046.[Conclusion]The six fungicides tested have great value in preventing and controlling Cordana leaf spot.22.5%Picoxystrobin SC,12.5%epoxiconazole SC and 50%thiophanate-methyl SC have better inhibitory activities on mycelial growth of N.musae,and can be further used in field trials.

PCR-based Assay for the Detection of Xanthomonas campestris pv. mangiferaeindicae Causing Bacterial Black Spot in Mango

[Objective] This study aimed to develop a PCR assay for detecting Xanthomonas campestris pv. mangiferaeindicae(Xcm) in culture and in planta. [Method] Primers(Xcm HF and Xcm HR) were designed based on the partial sequence of hrp B gene from xanthomonads to develop a PCR assay for Xcm. Furthermore, specificity and sensitivity of the primer pairs were analyzed in detection of genomic DNA and cell from Xcm. [Result] Amplication was positive only with genomic DNA from positive control ATCC11637 and 12 Xcm strains; no PCR products were amplified with genomic DNA from ten other xanthomonads and seven other bacterial species. The sensitivity of detection was 2.4 pg/μl genomic DNA, and 1.8 × 104CFU/ml cells. The primers also worked well for pathogen detection in direct PCR assays of Xcm colonies grown on liquid medium and in PCR assays of total DNA from leaf, branch and fruit lesions. [Conclusion] A PCR assay was successfully established for rapid detection of Xcm in culture and in planta.