during disease monitoring , a stem with small , irregular , brown lesions was sampled at a farm in cheonan , chungnam province , in september of 2011 .
the infected twig was rotted , and sunken cankers and reddish - brown needles were seen on the infected twig ( fig .
1 ) . after incubation in a humid condition , white mycelia , having dark brown necrosis , developed around the twig cavity . prior to further analysis
the ducc505 isolate was grown on pda and maintained at 25. a 5-mm diameter mycelial plug was cut from the margin of a 5-day - old culture of the isolate , and was placed centrally in an 85-mm petri dish containing pda .
the isolate was cultured at 25 , and colony characteristics such as color , shape and size were recorded .
the colony diameter was measured daily by scoring the average length for a period of 7 days .
colonies were whitish , having crenated edges , aerial mycelium on the surface , and with black gregarious fruiting bodies ( fig .
conidia were 17~22  3.5~4.2 m , fusiform , 4-septate , and straight to slightly curved .
the basal cell was of conical shape with an obtuse end , and pale brown in color .
the three median cells were brown : the second cell from base was pale brown , the third cell was a darker brown , and the fourth cell was brown .
there were 2~3 tubular , apical appendages arising from the apex of the apical cell .
the ducc505 isolate grew better on pda than oat meal agar and malt extract agar ( fig .
the optimum temperature for mycelial growth of the isolate ducc505 on pda was 25 ( fig .
the isolate ducc505 grew well in a broad range of ph , from 5 to 10 ( fig .
3c ) . these growth properties could be attributed to overcome the ph and low temperature stress in environment .
so far , none of fungicides have been registered for the disease control of acanthopanax sp . in the agrochemical use guide book in korea .
also , no fungicide has ever been tested for p. ellipsospora isolated from acanthopanax sp .
we therefore tested five kinds of fungicides for this study that are commercially available for ascomycete plant pathogens in korea . to understand the agrochemical sensitivity of the ducc505 isolate , we grew it with different concentration of fungicides , and the results are summarized in fig .
3d . in the benomyl and tebuconazol supplemented media , the mycelial growth was completely inhibited at 10 g / ml . this result is similar to pestalotiopsis microspora that is sensitive to tebuconazole . however , the isolate showed relative resistance in all media containing azoxystrobin , dimethomorph and triflumizole .
overall , it is suggested that among the five fungicides , benomyl and tebuconazol are the appropriate choice for the control of p. ellipsospora .
the ducc505 fungal isolate was grown on pda plates for 5 days at 25. mycelia were harvested by scraping the fungal colonies with a sterile blade .
genomic dna was extracted as described by kim et al . , with modifications . from the extracted genomic dna , partial translation elongation factor 1
polymerase chain reaction ( pcr ) was performed as described previously using the universal primers tef728 ( 5'-cat cga gaa gtt cga gaa gg-3 ' )   and tef1 ( 5'-gcc atc ctt gga gat acc agc-3 ' ) .
the pcr products were purified with a high pure pcr purification kit ( roche , basel , swiss ) and sequencing was carried out by macrogen inc .
the nucleotide sequence of partial tef1- gene of the fungal isolate shares 99% ( 531/536 ) sequence identity with that of known pestalotiopsis ellipsospora .
the tef1- gene sequence of the ducc505 was deposited in genbank dna database under accession number kc534872 .
4 ) . thus , based on the molecular and morphological data generated , the isolate was identified as p. ellipsospora .
the young twigs were inoculated with a droplet of the ducc505 conidia suspension ( 1.3  10 conidia / ml ) , which was prepared from the fungal cultures grown on pda .
the inoculated young twigs were incubated in a humid chamber at 25 for 7 days . as rotting progressed , superficial white mycelium and small black acervuli were observed ( fig . 5 ) .
p. ellipsospora was reisolated from the rotted twig lesion , thus fulfilling koch 's postulates .
these results demonstrated that p. ellipsospora ducc505 was able to infect a twig of a. divaricatus .
this is the first detailed report describing p. ellipsospora isolated from acanthopanax in korea , and it appears to be the first confirmation proving its pathogenicity on acanthopanax twigs .
 acanthopanax divaricatus , a member of the araliaceae family , has been used as an invigorant in traditional korean medicine . during disease monitoring , a stem with small , irregular 
 , brown lesions was sampled at a farm in cheonan in 2011 . 
 the symptoms seen were sunken cankers and reddish - brown needles on the infected twig . 
 the isolated fungal colonies were whitish , having crenated edges and aerial mycelium on the surface , and with black gregarious fruiting bodies . 
 the reverse plate was creamy white . 
 conidia were 17~22  3.5~4.2 m , fusiform , 4-septate , and straight to slightly curved . 
 the nucleotide sequence of the partial translation elongation factor 1 alpha gene of the fungal isolate , shares 99% sequence identity with that of known pestalotiopsis ellipsospora . based on the results of the morphological and molecular analyses , 
 the fungal isolate was identified as p. ellipsospora . in korea , this is the first report of canker on a. divaricatus . 
Disease symptoms and isolation of fungi
Mycological characterization
Molecular identification of 
Pathogenecity test