EFIKASI SUHU PROSES PRODUKSI ASAP CAIR DAN KONSENTRASI ASAP CAIR DARI KAYU LABAN (VITEX PUBESCENS) TERHADAP JAMUR OPHIOSTOMA PILIFERUM

Efficacy wood vinegar produced from wood laban (Vitex pubescens) against Ophiostoma piliferum fungus was evaluated. The objectives of this research to perform in vitro antifungal of wood vinegar from laban wood against O. piliferum. The source of lignocelluloses biomass was carbonization process used three temperature i.e. 350, 400 and 450 °C. Efficacy of wood vinegar to the fungus carried on PDA (potato dextrose agar) in a Petri dish that has been mixed with the wood vinegar with a concentration of 0, 0.5, 1.0, 1.5, and 2.0 % (v/v). Antifungal test was a factorial 3 by 5 in a completely randomized design. The means were seperated using using Tukey’s test at p = 0.05. All data were analyzed using the SAS softwere (version 8.2, SAS Institute Inc., NC. USA). The test results showed that the pyrolysis suhue effect and concentration of wood vinegar on the inhibition of fungal growth. The higher pyrolysis temperature and concentration the inhibition of fungal growth was increased.Efikasi asap cair dari kayu laban (Vitex pubescens)terhadap Ophiostoma piliferum telah dilakukan.Penelitian ini bertujuan mengevaluasi kemampuan asap cair sebagai antijamur (O. piliferum) secara in vitro. Asap cair yang digunakan diperoleh dengan cara pirolisis pada suhu produksi asap cair dan konsentrasi asap cair yang berbeda. Efikasi asap cair terhadap jamur dilakukan pada media PDA (potato dextrose agar) dalam cawan Petri dengan campuran asap cair pada konsentrasi 0, 0,5, 1,5, dan 2,0 (v/v).Pengujian aktivitas antijamur dilakukan menggunakan rancangan acak lengkap tipe faktorial.Data yang diperoleh dianalisis dengan sidik ragam.Pengaruh antarperlakuan dilakukan uji BNJ pada taraf nyata 5%. Analisis data menggunakan SAS versi 9.13. Hasil pengujian menunjukkan bahwa suhu pirolisis produksi asap cair dan konsentrasi asap cair berpengaruh terhadap daya penghambatan pertumbuhan jamur. Makin tinggi suhu pirolisis produksi asap cair dan konsentrasinya dan makin tinggi daya penghambatan terhadap pertumbuhan jamur, O. piliferum.

The Mitogenomes of Ophiostoma minus and Ophiostoma piliferum and Comparisons With Other Members of the Ophiostomatales

Fungi assigned to the Ophiostomatales are of economic concern as many are blue-stain fungi and some are plant pathogens. The mitogenomes of two blue-stain fungi, Ophiostoma minus and Ophiostoma piliferum, were sequenced and compared with currently available mitogenomes for other members of the Ophiostomatales. Species representing various genera within the Ophiostomatales have been examined for gene content, gene order, phylogenetic relationships, and the distribution of mobile elements. Gene synteny is conserved among the Ophiostomatales but some members were missing the atp9 gene. A genome wide intron landscape has been prepared to demonstrate the distribution of the mobile genetic elements (group I and II introns and homing endonucleases) and to provide insight into the evolutionary dynamics of introns among members of this group of fungi. Examples of complex introns or nested introns composed of two or three intron modules have been observed in some species. The size variation among the mitogenomes (from 23.7 kb to about 150 kb) is mostly due to the presence and absence of introns. Members of the genus Sporothrix sensu stricto appear to have the smallest mitogenomes due to loss of introns. The taxonomy of the Ophiostomatales has recently undergone considerable revisions; however, some lineages remain unresolved. The data showed that genera such as Raffaelea appear to be polyphyletic and the separation of Sporothrix sensu stricto from Ophiostoma is justified.

Assessing the virulence of ophiostomatoid fungi associated with the pine-infesting weevils to scots pine Pinus sylvestris L. seedlings

The pine-infesting weevils are known to be effective vectors of ophiostomatoid fungi. To understand more about fungal virulence of these fungi, inoculation studies were conducted on Scots pine (<em>Pinus sylvestris</em> L.). Two-year-old seedlings were wound-inoculated with one of eleven ophiostomatoid fungi associated with pine-infesting weevils. After 11 weeks, a darkened lesion, extending from the point of inoculation, was observed in all species, except for <em>Ophiostoma</em> cf. <em>abietinum</em> Marm. &amp; Butin, <em>Ophiostoma quercus</em> (Georgev.) Nannf., and <em>Sporothrix inflata </em>de Hoog. Seedling mortality was observed in seedlings inoculated with <em>Leptographium truncatum</em> (M.J. Wingf. &amp; Marasas) M.J. Wingf., <em>Leptographium lundbergii</em> Lagerb. &amp; Melin<em>,</em> <em>Leptographium procerum</em> (W.B. Kendr.) M.J. Wingf., <em>Grosmannia radiaticola</em> (J.J. Kim, Seifert &amp; G.H. Kim) Zipfel, Z.W. de Beer &amp; M.J. Wingf., <em>Ophiostoma floccosum</em> Math.-Käärik,<em> Ophiostoma minus</em> (Hedgc.) Syd. &amp; P. Syd., and <em>Ophiostoma piliferum </em>(Fr.) Syd. &amp; P. Syd. <em>Ophiostoma minus</em> and <em>L. truncatum</em> caused the largest lesions and sapwood blue-stain in Scots pine. <em>Grosmannia radiaticola, Ophiostoma piceae </em>(Münch) Syd. &amp; P. Syd., <em>O. floccosum</em>,<em> O. piliferum,</em> <em>L. lundbergii</em>,and<em> L. procerum </em>produced significantly smaller lesions and sapwood blue-stain than <em>O. minus</em> and <em>L. truncatum</em>, while<em> O</em>. cf. <em>abietinum</em>, <em>O. quercus</em> and <em>S. inflata</em> did not cause any lesions.

Fungal pretreatment of pine wood to reduce the emission of volatile organic compounds

Sterilized pine wood strands were treated with the ascomycete Ophiostoma piliferum (Cartapip 97™) in an attempt to reduce aldehyde emissions through degradation of aldehyde-forming precursors. Wooden boards were then produced from the treated strands by means of a laboratory press. VOC emissions of these boards were characterized and the concentrations of seven major substances were followed for a period of 28 days in Markes μ-CTE micro-chambers employing Tenax-TDAS/GC/MS analytics. Boards made from treated strands showed a highly significant (P<0.001) reduction in aldehyde emissions by 70% compared to the control boards, while differences in the monoterpene emissions were not significant (P>0.05). Wood extractives from treated and untreated strands were also analyzed by GC/MS after milling and acetone/water extraction. A marked decrease of C18 unsaturated fatty acids was detected in agreement with the reduction of aldehyde emissions, and the monosaccharides as indicators of fungal growth were depleted and stilbenes and lignans were partially degraded.

Ophiostomatoid fungi isolated from Pinus radiata logs imported from New Zealand to Korea

Ophiostomatoid fungi discolor lumber, logs, and tree sapwood. Stained wood has a lower market value and can be refused by importing customers because such products can potentially carry pathogenic fungi. Little information is available on the ophiostomatoid fungi that colonize Pinus radiata D. Don (radiata pine) logs imported from New Zealand into Korea. In this work, we attempted to identify the native and non-native fungi colonizing wood imported into Korea. At least 12 species of ophiostomatoid fungi, including an unknown Ophiostoma sp. A, were identified among the fungi-staining radiata pine. They were Leptographium procerum (Kendr.) Wingf., Leptographium bistatum Kim & Kim, Ophiostoma floccosum Mathiesen, Ophiostoma huntii (Rob.) Hoog & Scheff., Ophiostoma ips (Rumbold) Nannf., Ophiostoma nigrocarpum (David.) Hoog, Ophiostoma piceae (Münch) H. & P. Sydow, Ophiostoma piliferum (Fries) H. & P. Sydow, Ophiostoma quercus (Georgév.) Nannf., Ophiostoma radiaticola Kim et al., and Ophiostoma setosum Uzunovic et al. Of these species, O. floccosum was the dominant species in both logs and boards. We confirmed that many of the sapstain species that we isolated have been previously reported in Korea. However, four species, O. radiatcola, O. setosum, O. huntii, and O. nigrocarpum have not been reported previously in Korea. We also found the new species, L. bistatum, along with an unknown Ophiostoma sp. A.Key words: radiata pine, ophiostomatoid fungi, New Zealand, non-native organisms, β-tubulin gene.

Disruption of the Subtilase Gene, albin1, in Ophiostoma piliferum

ABSTRACT Wood sapstaining fungi produce multiple proteases that break down wood protein. Three groups of subtilases have been identified in sapstaining fungi; however, it is not known if these groups have distinct physiological roles (B. Hoffman and C. Breuil, Curr. Genet. 41:168-175, 2002). In this work we examined the role of the subtilase Albin1 from Ophiostoma piliferum. Reamplification of cDNA ends PCR was used to obtain the albin1 gene sequence. The encoded subtilase is probably extracellular and involved in nutrient acquisition. This gene was disrupted with an Agrobacterium tumefaciens-mediated transformation system. Two of the disruptants obtained had significantly lower levels of proteolytic activity, slower growth in bovine serum albumin, and significantly reduced growth on wood. Thus, albin1 plays an important role in O. piliferum's ability to acquire nitrogen from wood proteins.

Notes on Ceratocystis brunnea and some other Ophiostoma species based on partial ribosomal DNA sequence analysis

Ribosomal gene sequence data were obtained from a nonfruiting culture originally identified as Ceratocystis brunnea R.W. Davidson; this species was considered a nomen dubium by Upadhyay (1981) due to a presumptive lack of teleomorph material. The data showed that C. brunnea is a valid species that should be transferred to Ophiostoma, demonstrating that DNA data can compensate for the presumed or actual lack of morphological features lost during either long-term culturing or disintegration of holotype specimens. Use of partial large ribosomal sequence data to assess the relationship of C. brunnea to other Ophiostoma spp. showed that it is not a synonym of Ophiostoma piliferum (Fr.:Fr.) Syd. & P. Syd. as has been suggested; instead, it appears to be distantly related to Ophiostoma piceae (Münch) Syd. & P. Syd. The data obtained for the Ophiostoma piliferum strains included in this study suggest that hardwood-derived isolates may be distinct from those obtained from conifers. In addition, molecular characters support transferring Ceratocystis pseudonigra Olchow. & Reid, Ceratocystiopsis concentrica (Olchow. & Reid) Upadhyay, Ceratocystiopsis pallidobrunnea (Olchow. & Reid) Upadhyay, and Ceratocystiopsis crenulata (Olchow. & Reid) Upadhyay to Ophiostoma.Key words: rDNA, phylogenetics, Ophiostoma piliferum, Ophiostoma, Ceratocystsis.

Albino Strains of Ophiostoma Species for Biological Control of Sapstaining Fungi

SummaryAlbino strains ofOphiostoma floccosum,O. piceaeandO. pluriannulatumwere selected and screened for biological control of sapstaining fungi on New Zealand radiata pine (Pinus radiata). Albino strains were obtained through matings and single ascospore isolations from cultures of prevalent species in New Zealand. These strains do not synthesize the melanin-like hyphal pigments of common sapstaining fungi. Additional mating studies were also carried out to obtain isolates ofO. piceaeandO. floccosumthat lacked pigmented fruiting structures. Several albino isolates ofO. piceaewith colorless synnemata and isolates ofO. floccosumwith little to no synnemata were obtained. Biological control potential of the albino isolates was evaluated in the laboratory by challenging them on wood chips with fungi that cause extensive sapstain in pine,Leptographium procerum,Ophiostoma piliferumandSphaeropsis sapinea. Many albino isolates ofO. floccosum,O. piceaeandO. pluriannulatumwere effective in stopping the challenge fungi from staining wood chips and were fast growing and colorless when grown unchallenged on wood chips. Selected albino isolates ofO. pluriannulatumwere used in two field trials in New Zealand to control sapstain. Several strains were found to significantly reduce dark sapstain as compared to the untreated control logs.