Evaluation on the efficacy of microbial approaches in the control of Ganoderma boninense and their effects to soil microbial diversity

Basal Stem Rot (BSR) is the most destructive disease of oil palm (Elaeis guineensis Jacq.) which caused by bracket fungus; Ganoderma boninense. With no remedy to date, this disease has jeopardized the industry of oil palm critically. Several approaches including cultural, chemical, mechanical and biological measures have been reported in attempt to control this disease, but unfortunately, none have given satisfactory result. Therefore, a more reliable approach that could manage this disease, besides a continuous and sustainable oil palm production is worth to be further investigated. Introduction of microbes in disease management have been used for decades. An astute observation of low incidence of disease due to Ganoderma pathogen in natural ecosystem suggests that a healthy natural ecosystem can hinder the incidence of Ganoderma pathogen outbreak. In this respect, study on the potential of microbes in suppressing the growth of Ganoderma boninense and its effect to the soil microbial diversity were investigated in this project. Three microbes-based products which are Living Soil Microbes (LSM), Agrinos (AGN) and Agriorganica (AO) were tested in this study. LSM contains multi-strain of Bacillus spp. and Trichoderma spp., whereas AGN consist of Bacillus spp., Pseudomonas spp. and Aspergillus sp. Meanwhile, AO comprises of three living food microbes which are Lactobacillus, Nattobacillus and Saccharomyces cerevisiae developed from Japanese fermentation and brewing technology. To examine the ability of the treatments in reducing BSR disease incidence, assessments in nursery and field trial were conducted. The results from both trials showed that LSM, AGN and AO were able to reduce the colonization of G. boninense based on the reduction of ergosterol content compared to untreated control

In vitro seed germination of Coelogyne asperata Lindl. (Orchidaceae)

A protocol for in vitro seed germination of Coelogyne asperata Lindl. has been established successfully. Immature seeds from 182 days old capsule were cultured on three different basal media; Murashige and Skoog (MS), Knudson C (KC), and Vacin and Went (VW) and maintained under continuous light at 25 ± 2 ˚C. After 30 days of culture (DAC), more than 90% of seeds were germinated on KC and VW media and about 84% of seeds germinated on MS medium. The incorporation of organic additives, including coconut water, potato homogenate and tomato juice each at 10% (v/v) in KC medium was tested to determine their effect on seed germination of C. asperata. The result revealed that KC basal medium alone without addition of organic additives promoted over 90% of seed germination at 30 DAC. Therefore, the protocol of using standard KC basal medium for C. asperata seeds germination could be suggested for mass propagation and conservation of this wild scented orchid

Early detection and management of ganoderma basal stem rot disease: a special report from Sabah

Basal Stem Rot (BSR) disease caused by Ganoderma spp. is the most devastating disease of oil palm in Southeast Asia. This paper discusses sustainable approaches in managing BSR disease particularly on early detection and control of Ganoderma with some examples from oil palm estates in Sabah. New detection methods such as ergosterol analysis and Ganoderma signature via Fourier Transform Infrared Spectroscopy (FTIR) are emphasized. Latest disease control methods with great potentials such as combination of biological control agents (BCAs), enhancing defense mechanism of oil palm through enviro-friendly approach, potential biomarkers for selection of resistant breeding materials and utilization of eco-friendly fungicide were also discussed

Evaluation on the efficacy of different disruption method of oil palm roots and ganoderma boninense cell wall

The cell wall is the outermost layer of cell encountered by pathogens, determining the infection fate. However, the chemical composition involves during host-pathogen interaction has not been fully elucidated. Prior to determining the composition in the cell walls, the walls must be isolated first from the intercellular component. Selected methods for oil palm roots and G. boninense cell disruption were evaluated for their suitability for cell wall isolation and presented in this paper. The effect of selected disruption methods on the content of crude proteins and DNA absorbance under 260 and 280 nm wavelengths in the produced cell wall materials was established on the basis of their solubilized materials. The investigated methods were; i) homogenization with sonication; ii) grinding with liquid nitrogen and; iii) lyophilization with homogenization. The cell wall materials with the highest degree of cytoplasmic component released were produced using lyophilization with homogenization. This was confirmed with the highest DNA absorbance at 260/280 nm of 2.25/1.95 for G. boninense and 2.47/2.35 for oil palm roots in their solubilized materials. The thus-produced preparation also released 4.5 and 10.1 μg/g of crude proteins for G. boninense and oil palm roots respectively. The degree of cell wall purification from intracellular components was validated using Fourier Transform Infrared Spectroscopy (FTIR). Isolated cell wall can be further utilized in cell wall composition analysis

Some interpretations on FTIR results for the detection of Ganoderma boninense in oil palm tissue

The use of Fourier Transform Infrared (FTIR) spectroscopy has been employed for the past three decades in the study of biological samples and characterization of biomolecules. Objective: In this paper, the author have conducted the feasibility study of detecting Ganoderma boninense in infected oil palm tree using Fourier transform infrared spectroscopy (FTIR). Results: It was found that result from FTIR plot is capable of showing the presence of Ganoderma boninense with good sensitivity. Conclusion: In this paper, the mechanisms behind this detection are discussed. This is due to the unique functional groups that exist in the Ganoderma boninense which cannot be found in healthy oil palm tissue

On the possibility of using FTIR for detection of Ganoderma Boninense in infected oil palm tree

Ganoderma boninense is a basidiomycetes fungus that causes basal stem rot disease (BSR) in oil palm trees. In Malaysia alone, the loss caused by this disease was estimated between RM 225 Million to RM 1.5 Billion in 2011 by Malaysian Palm Oil Board. Unfortunately, many planters do not realize that their fields were infected with BSR until it is too late. Several methods have been proposed for early detection of Ganoderma boninense infection. In this paper, Fourier transform infrared spectroscopy (FTIR) is investigated as a tool to detect the presence of Ganoderma boninense in oil palm tree. It is shown that there are differences in the FTIR result from the infected and healthy oil palm tree that resembles the FTIR characteristics from pure Ganoderma boninense. The result presented in this paper shows the possibility of FTIR as a tool for detecting the infection of the fungi in oil palm tree

Evaluation on the effectiveness of organic acids combination against Ganoderma boninense, the causal pathogen of basal stem rot in oil palm

Basal Stem Rot (BSR) disease mainly caused by Ganoderma boninense has become a serious threat to the South East Asia oil palm industry. With no conclusive remedy to date, the oil palm industry is still in search of effective ways to manage this disease. The present work reports the effectiveness of organic acids combination (OAC) in managing Ganoderma infection in oil palm. In this study, the pre-formulated organic acids combination from a product to control BSR caused by Ganoderma was carried out both in the field and nursery. The trial was conducted for a duration of approximately 18 months. The field trial was carried out at Bode Estate of Kretam Plantations Sabah in Sandakan. The possibility of the OAC in preventing the infection from spreading to newly planted seedlings in the area with Ganoderma history was also assessed via nursery trial at Mile 25, estate of Kam Cheong Sdn Bhd. In the field trial, three different sets of protocols i.e.: A (0.4% v/v with 5 rounds of application), B (0.4% v/v with 3 rounds of application), and C (0.5% v/v with 3 rounds of application) of the OAC treatment were applied along with Ganoderma Selective Medium (GSM) analysis, ergosterol content analysis, in vitro antagonistic evaluation and Scanning Electron Microscope (SEM) observation to comprehensively investigate the efficacy of the combination. Protocols A, B and C had significantly reduced the colonisation / amount of ergosterol content (8.832-9.095 μg/g of trunk tissues) in the infected palms in comparison to those Ganoderma infected but left untreated palms (48.956 μg/g of trunk tissues). However, there was no significant difference between the effectiveness among the three protocols in reduction of Ganoderma colonisation till month-12, in which protocol C proved to perform better compared to the other two protocols. There was slight ergosterol content increment in oil palm tissues treated with various protocols of the OAC at month-18, but were much lesser compared to untreated palms. Nonetheless, none of the protocols in application of OAC gave an absolute control of Ganoderma till the end of the trial, as the treated palms remained infected but with much lower ergosterol content compared to untreated palms. Application of the OAC as soil treatment for prevention of Ganoderma infection to seedlings replanted in the area with Ganoderma history in Kam Cheong Estate showed lesser disease incidences compared to those untreated ones. The infected seedlings which were treated by this product also showed lesser amount of ergosterol content which represents lesser colonisation of the pathogenic fungi. However, OAC-treated seedlings still recorded the presence of ergosterol from low to moderate in some of the tested samples. In vitro experiment of OAC and Ganoderma mycelia further elaborates the possible interaction between these organic acids with Ganoderma when in contact with either the tissues or soil. The in vitro results suggest OAC has destructive effect against the mycelia of Ganoderma with SEM evidences of massive damaging effects of the product to the mycelia of the fungi. Based on the GC-MS analysis, the OAC were identified from the products propanoic acid, acetic acid, benzoic acid, sorbic acid and besylic acid

Optimisation of cell wall lipids extraction from oil palm using mixture of chloroform and methanol via response surface methodology

Understanding oil palm’s cell wall lipid during pathogenic invasion, especially from Ganoderma fungus helps us to devise better strategies in managing Basal Stem Rot caused by the fungus effectively. However, as the lipids of plant’s cell wall is a delicate subject, while acquiring them in ample amount with decent quality for research purpose is an excruciating work, therefore it is crucial to develops an optimised method for acquisition. In this study, the effects of solvent ratio, extraction time, extraction temperature and biomass to solvent ratio pertaining effective lipids extraction from oil palm roots cell wall were investigated, aided with Central Composite design (CCD) of Response Surface Methodology (RSM) analysis to obtain optimum oil palm cell wall lipid extraction conditions. Using the projection of RSM model and validation of empirical evaluation, the results revealed that the optimum cell wall lipids yield was obtained under the following extraction conditions: 10 mL of solvent mixture (chloroform: methanol (2:1, v/v)) per gram tissue for 120 min with gentle agitation at 30˚C of extraction temperature. Ultimately, Fourier Transform Spectroscopy (FTIR) analysis was carried out to validate the successful and complete extraction were achieved using the optimised condition