Luciasih Agustini Morag Glen Heru Indrayadi Dono Wahyuno Fadjar Sagitarianto Bayo Alhusaeri

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1 Luciasih Agustini Morag Glen Heru Indrayadi Dono Wahyuno Fadjar Sagitarianto Bayo Alhusaeri The Second International Conference of Indonesia Forestry Researchers (INAFOR) August 2013 Root rot the most damaging disease to the majority of plantation resources (incl. Oil palms, rubber, plup wood : A.mangium & E.pellita) The mortality of A.mangium due to root rot: 5 25% after 10 years in Malaysia (Lee, 1997) 3 28% aged 3-5 years old in the 2 nd rotation, in Sumatra & Kalimantan (Irianto et al., 2006) E. Pellita is a promising species, an alternative to A.mangium: - Better & healthier growth (Hardiyanto, 2003) - Less susceptible to foliar disease (Harwood et al., 1997) - Less susceptible to Ganoderma root-rot in pot trial (Indrayadi, 2007 unpublished). 1

2 In a survey undertaken at the beginning of project FST/2003/048, the largest number of dead tress Phellinus sporocarps was observed. Phellinus is one of root-rot pathogens. It has a wide host range (± 150 host: mahogany, teak, rubber, tea, coffee, oil palm, cacao etc.) Examination of root-rot incidence in the current EP plantation is required in order to avoid a massive loss due to root-rot. 1) Characterize root disease in E. pellita by examining & describing the fungal signs 2) Determine disease spread in several monitored plots 3) Test some potential saprophytic fungi for controlling root-rot A. Root-rot Examination Surveying 12 sites affected by root-rot. Sporocarps Identification based on macro- & micro- scopic features; molecular ID Sampling of infected root fungal isolation molecular ID 2

3 B. Monitoring Disease Spread Establishment of 10 monitored plots in 4 different sites.(in total) 3 times monitoring assessments of crown health & signs of root-rot. Interval of assessments : 4 months C. Tests for Selecting Potential Biocontrol In vitro interaction between putative biocontrols & root-rot pathogens. Microscopic examination of hyphal interaction Re-growing test from the hyphal confrontation zones Pseudo-sclerotia A. Root-rot Examination Fungal Signs : Phellinus noxius Morphological ID: Basidiocarp: solitary to imbricate, effusoreflexed to resupinate, consistency hard Setal hyphae: abundant; projecting into the hymenium Basidiospores: hyaline, elliptical to ovoid, smooth & thin walled Molecular ID: 20 of 56 isolates match with GenBank accessions EF EF & EF from P. noxius. Honeycomb rot type 3

4 Fungal signs: Ganoderma spp. Morphological ID: Basidiocarps: single or in a small group; stipitate; light. Pileus: applannate broadly attached to dimidate /stipitate spatulate; laccate to dark red-brown. Basidia: four-spored; hyaline Basidiospores: pale yellow-brown, echinulate, truncate ellipsoid. Molecular ID: - G. philippii (predominant isolate 30 of 48) - G. mastoporum - G. australe group - G. subresinosum - Amauroderma sp. G. mastoporum Redish-Brown Rhizomorph Species Numbers of Isolate(s) Isolated from Isolated from Sites # Cerrena sp. 1 Eucalyptus root 11 Ganoderma australe 5 Eucalyptus root 3 and 10 Ganoderma mastoporum 8 Eucalyptus root, sporocarps 3 and 6 Ganoderma philippii * Eucalyptus root, 1,3,5,6,7,8, and 9 *Acacia mangium root Ganoderma subresinosum 1 Sporocarp 3 Ganoderma sp. 1 Eucalyptus root 9 Gymnopilus sp. 3 Eucalyptus root, sporocarp 3, 10 and 11 Lenzites aff. elegans 2 Eucalyptus root 11 Neonothopanus nambii 3 Eucalyptus root 3 Phellinus noxius 16 Eucalyptus root, sporocarp 2,6,10 and 11 Phlebiopsis sp. 11 Eucalyptus root 11 Rigidoporus sp. 1 Eucalyptus root 11 Tinctoporellis epimiltinus 8 Eucalyptus root 11 Root-rot pathogens Putative Biocontrols Why Cerrena & Phlebiopsis??? Fungal Isolates from Sporocarps & Eucalyptus pellita root samples Causal Agents of root-rot in the monitored plots: Phellinus sp. & G.philippii Sporocarps is not always indicate that root-rot causal agent(s). Gp sporocarps were not observed, but fungal isolates taken from infected roots were most frequently identified as Gp. 4

5 Number of dead trees (%) 28/08/2013 Cerr.& Phleb. were isolated from stumps that have vigourous coppice growth The site with had root-rot incidence in the previous planting. Proposed the possible role of Cerrena & Phlebiopsis in reducing root-rot & contributing to the healthy coppice growth. Spores of Phlebiopsis gigantea Apply to the stumps for reducing inocula of root pathogen. Suspended into 25L water Rotstop B. Monitoring Disease Spread % * 0.62% 0.08% % 1.55% 0.93% June 2008 Oct 2008 Feb 2009 Sept % * Numbers of treedeath per month 0 3A 3B 3C 6A 6B 6C 10A Plots Only 7 out of 10 monitored plots showed disease spreading; Plots 11A,B,C no obvious tree death caused by rootrot. Root-rot incidence is inversely related to infection rate. - Plots 6A,6B,6C had <25% incidence infection rate: up to 1.55% per month - Plot 10A had >50% incidence infection rate: up to 0.08% per month In the latest survey (Sept 2012): tree death caused by root-rot ranged 40 60% 5

6 C. In vitro Test for Putative Biocontrols Biocontrols vs G.philippii Mycoparasitic reaction at the confluence zones: Phlebiopsis & Cerrena showed a blocking response through condensed coiling around the G.philippii mycelia Biocontrol isolates (Cerrena & Phlebiopsis) grow faster, then overgrowth the G. philippii isolates. Re-growing Tests: Section 1,2,3,4 generate Cerrena/Phlebiopsis Section 5 generates G.philippii Gp seems possible to be controlled by Cerrena & Phlebiopsis. Biocontrols vs P.noxius Cerrena sp. vs P. noxius The pigmented lines indicate: antagonism of mycelia of two (more) different fungi occupying the same substratum (Darus et al., 1989) ability for prolonged-survival in unfavourable condition (Li, 1981) Phellinus has a good defense reaction Applying Cerrena or Phlebiopsis for controling P.noxius need to be considered carefully. Brown demarcation lines (pseudo-sclerotia) consist of pigmented-swollen hyphae Phlebiopsis sp. vs P. noxius 6

7 1. P. noxius & G. philippii appears to be the most prominent root-rot causing agents in E. pellita plantation surveyed. 2. Root-rot incidence in E. pellita plantation may increase by the time from one rotation to the next (~ experienced in A. mangium). survey across larger areas of E. pellita plantation are required to confirm this possibility. 3. Cerrena & Phlebiopsis seem to be potential biocontrol for G. philippii. 4. Controling Phellinus seems more difficult due to its defense reaction. 5. Possibility of appliying Cerrena or Phlebiopsis as root-rot biocontrols are need to be tested in field trials. ACIAR (project FST/2003/048) CSIRO Ecosystem Sciences PT. Arara Abadi FORDA Center for Biotechnology & Tree Improvement FORDA Center for Forest Conservation & Rehabilitation 7

8 Thank you 8