Spring needle cast (SNC) in Tasmania is defined by the early casting of pine needles and appears in 6-to 7-year-old pine plantations after canopy closure. The severity of SNC can vary significantly among trees growing within the same plantation. Fungal communities were surveyed in a Pinus radiata family trial, using DNA extraction from needles, PCR with fungal-specific primers and sequencing to detect the presence of fungal species. Samples were taken at the same time as the trees were scored for disease severity. Trees with contrasting levels of SNC disease severity have significantly different needle fungal communities, but family pedigree and different ages of needle are not clearly distinguished by their fungal communities. All common fungal pathogens previously implicated in causing SNC were identified from the study, but of these, only Lophodermium pinastri was correlated with high levels of disease. Several species of Teratosphaeriaceae were detected, and one of these was also strongly associated with needle cast.

Summary Cyclaneusma Needle Cast is associated with the fungal pathogen Cyclaneusma minus. The disease has led to significant losses of Pinus radiata production in New Zealand, but the disease appears less serious in Australia. Two cultural morphotypes of C. minus have been discriminated in New Zealand, and isolates from Australia also correspond to these two morphotypes. To examine if the two morphotypes of Cyclaneusma are different species, phylogenetic analysis using sequences from five gene regions (rDNA ITS, mt‐LSU, n‐LSU, tef–1 and βT‐1) was conducted. This study revealed that C. minus consists of two distinct clades which correspond to the two morphotypes. One morphotype (‘verum’) was more closely related to Cyclaneusma niveum than to the other morphotype (‘simile’). A morphological description is required to formalise the new species represented by the ‘simile’ clade; an investigation of the individual pathogenic behaviours of both morphotypes is also required.

Background: Needle fungi in conifers have been extensively studied to explore their diversity, but environmental factors influencing the composition of fungal communities in Pinus radiata D.Don needles have received little attention. This study was conducted to examine the influence of the environment as defined by rainfall, elevation and temperature on the composition of fungal communities in pine needles at an age prior to that at which spring needle cast (SNC) is generally observed. Elucidating the entire fungal community in the needles is a first step towards understanding the cause of the disease. Methods: Needle samples were collected from 5-year-old P. radiata trees, their age predating the onset of SNC, from 12 plantations in Tasmania. Interpolated data for the climate variables, including seasonal components for rainfall and temperature, were obtained from an enhanced climate data bank. Pooled needle samples were examined for the fungi they contained using DNA sequencing of cloned polymerase chain reaction (PCR) products. Clones were grouped into operational taxonomic units (OTUs) and identified to their lowest possible taxonomic level by comparison with reference isolates and public DNA databases. Results: DNA sequencing revealed that needle fungal communities differed greatly, depending upon the total annual rainfall and needle age. Needle fungi that have been previously associated with pathogenic behaviour, such as Cyclaneusma minus, Dothistroma septosporum, Lophodermium pinastri, Strasseria geniculata and Sydowia polyspora, were all found in the needles in this study. Also, of 70 ascomycetous OTUs that were discriminated by their DNA sequences, 21 were identified as belonging to the Teratosphaeriaceae. Conclusions: An overview of the endophytic fungi present in the needles of P. radiata has been obtained from trees whose age precedes the onset of SNC. All of the previously known pathogenic species for needle cast diseases were present, in addition to a range of unidentified OTUs belonging to the Teratosphaeriaceae.

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Root rot caused by Ganoderma philippii (Bres. & Henn. ex Sacc.) Bres., is an important disease in Acacia plantation. A strategy to control this disease is currently being developed, particularly on the application of biological control agent (BCA). Species specific primers for rapid identification of potential fungi as BCA were developed. This study aimed to obtain the best DNA condition and the best primers for species specific identification. DNA with 20× dilution is the best condition for amplification of the ITS fragment thus used for rapid species identification. The best primer set to detect Cerrena sp. is CrF1/CrR1, while PbF2/PbR2 is the best primer set for Phlebiopsis sp. 1. There was no specific primer suitable to detect Phlebia sp. 1 and Phlebia sp. 2 only, but Pl-2F1/Pl-2R4 is the best primer set for Phlebia spp.

Ganoderma philippii is a root pathogen of many woody plants in tropical regions and is particularly aggressive to Acacia mangium, which is grown on a 6‐year rotation for pulpwood in Indonesia. The disease becomes progressively worse over each rotation and control measures have met with limited success. We studied the population genetics of G. philippii to evaluate the role of sexual and asexual reproduction in its mode of spread. Populations were genetically distinct with high levels of inbreeding, and clonal spread to adjacent trees increased after the first rotation. Despite the high levels of genetic diversity seen at all sampling scales, migration rates appear low. Measures to reduce the underground spread of the pathogen as well as methods to prevent the initiation of new infections from basidiospores will be needed to reduce the incidence of root rot in A. mangium plantations.

Identification of wood decay fungi based on morphological character of isolates is very helpful where identification of fruit body morphology is ambiguous. Nine isolates of Phlebiopsis sp.1 obtained from the isolation root of Eucalyptus pellita in permanent plots ABSTRAKPada jamur penghuni kayu, identifikasi berdasarkan karakter morfologi isolat sangat membantu apabila terdapat keraguan dalam identifikasi morfologi tubuh buah. Sembilan isolat Phlebiopsis sp.1 yang diperoleh dari isolasi akar Eucalyptus pellita pada plot permanen penyakit busuk akar mempunyai potensi sebagai pengendali hayati untuk G. philippii. Verifikasi berdasarkan sekuen ITS dari rDNA menunjukkan kedekatan isolat tersebut dengan P. gigantea dan P. flavidoalba. Karakterisasi morfologi dilakukan pada isolat yang ditumbuhkan pada media Malt Extract Agar dengan penambahan serbuk kayu. Berdasarkan pengamatan secara makroskopis isolat-isolat tersebut dapat dikategorikan ke dalam tiga tipe morfologi yang berbeda: (1). Cottony dari awal sampai pertumbuhan lanjut, satu isolat (Pb5); (2). Cottony pada awal kemudian berubah menjadi farinaceous to floccose pada pertumbuhan lanjut, 7 isolat (Pb1, Pb2, Pb4, Pb6, Pb8, Pb9, Pb10); (3). Absent pada awal kemudian berubah menjadi zonate pada pertumbuhan lanjut, satu isolat (Pb11). Tipe morfologi 2 merupakan karakter morfologi yang dominan dari isolat Phlebiopsis sp.1, dengan gumpalan miselium kecil halus berwarna putih yang berubah kecoklatan pada permukaan koloni. Tipe morfologi 3 mempunyai ciri karakter morfologi yang jauh berbeda dengan 8 isolat lainnya, dengan ciri khas adanya lingkaran konsentrisitas dengan perbedaan warna dan tekstur pada setiap lingkarannya. Laju pertumbuhan miselium berkisar antara 10,70 -10,85 cm²/hari untuk tipe morfologi 1 dan 2; 1,95 cm²/hari untuk tipe morfologi 3.

ABSTRAKPinus radiata merupakan jenis pohon berkayu lunak yang banyak ditanam di Selandia Baru dan Australia. Jamur endofit telah banyak dipelajari pada daun jarum beberapa jenis konifer melalui teknik isolasi jamur namun belum banyak dilakukan pada jenis Pinus radiata. Penelitian ini dilakukan untuk mengetahui kondisi daun jarum P. radiata yang optimal bagi proses ektraksi DNA jamur endofit serta mengetahui jenis-jenis jamur endofit yang berasosiasi dengan daun jarum P. radiata di hutan tanaman Plenty melalui metode PCR secara langsung. Daun jarum yang telah dikeringkan pada suhu ruangan memberikan hasil amplifikasi DNA yang lebih jelas dan konsisten dibandingkan dengan daun yang masih segar serta daun yang telah dikeringkan pada ruang pengering maupun pada alat pengering. Pada penelitian ini, Cap-nodiales merupakan kelompok jamur yang paling banyak dijumpai serta tidak ditemukan jenis jamur patogen. Data sekuen DNA yang diperoleh melalui ektraksi DNA secara langsung dari daun jarum P. radiata mampu memberikan perbedaan serta identifikasi bagi jenis-jenis jamur.Kata kunci: Pinus radiata, jamur endofit, ekstraksi DNA secara langsung, identifikasi jamur