Exploring the Biocontrol Efficacy of Trichoderma spp. against Rigidoporus microporus, the Causal Agent of White Root Rot Disease in Rubber Trees (Hevea brasiliensis).

Rigidoporus microporus, which causes white root rot disease (WRD) in Hevea brasiliensis, is a looming threat to rubber plantation in Malaysia. The current study was conducted to determine and evaluate the efficiency of fungal antagonists (Ascomycota) against R. microporus in rubber trees under laboratory and nursery conditions. A total of 35 fungal isolates established from the rubber tree rhizosphere soil were assessed for their antagonism against R. microporus by the dual culture technique. Trichoderma isolates can inhibit the radial growth of R. microporus by 75% or more in the dual culture test. Strains of T. asperellum, T. koningiopsis, T. spirale, and T. reesei were selected to assess the metabolites involved in their antifungal activity. Results indicated that T. asperellum exhibited an inhibitory effect against R. microporus in both volatile and non-volatile metabolite tests. All Trichoderma isolates were then tested for their ability in producing hydrolytic enzymes such as chitinase, cellulase and glucanase, indole acetic acid (IAA), siderophores production, and phosphate solubilization. From the positive results of the biochemical assays, T. asperellum and T. spirale were selected as the biocontrol candidates to be further tested in vivo against R. microporus. The nursery assessments revealed that rubber tree clone RRIM600 pretreated with only T. asperellum or with the combination of T. asperellum and T. spirale was able to reduce the disease severity index (DSI) and exert higher suppression of R. microporus compared to other pretreated samples, with the average DSI below 30%. Collectively, the present study demonstrates that T. asperellum represents a potential biocontrol agent that should be further explored to control R. microporus infection on rubber trees.

Virulence of Rigidoporus microporus Isolates Causing White Root Rot Disease on Rubber Trees (Hevea brasiliensis) in Malaysia.

Latex production from Hevea brasiliensis rubber tree is the second most important commodity in Malaysia, but this industry is threatened by the white root rot disease (WRD) caused by Rigidoporus microporus that leads to considerable latex yield loss and tree death. This study aimed to characterize and compare the virulence of five R. microporus isolates obtained from infected rubber trees located at different states in Malaysia. These isolates were subjected to morphological and molecular characterization for species confirmation and pathogenicity test for the determination of virulence level. BLAST search showed that the ITS sequences of all the pathogen isolates were 99% identical to R. microporus isolate SEG (accession number: MG199553) from Malaysia. The pathogenicity test of R. microporus isolates conducted in a nursery with 24 seedlings per isolate showed that isolate RL21 from Sarawak has developed the most severe above- and below-ground symptoms of WRD on the rubber clone RRIM600 as host. Six months after being infected with R. microporus, RL21 was evaluated with the highest average of disease severity index of 80.52% for above- and below-ground symptoms, followed by RL22 (68.65%), RL20 (66.04%), RL26 (54.38%), and RL25 (43.13%). The in vitro growth condition tests showed that isolate RL21 of R. microporus has optimum growth at 25-30 °C, with the preference of weakly acidic to neutral environments (pH 6-7). This study revealed that different virulence levels are possessed among different R. microporus isolates even though they were isolated from the same host species under the same climate region. Taken together, field evaluation through visual observation and laboratory assays have led to screening of the most virulent isolate. Determination of the most virulent isolate in the present study is vital and shall be taken into consideration for the selection of suitable pathogen isolate in the development of more effective control measures in combating tenacious R. microporus.

Application strategies by selective medium treated with entomopathogenic bacteria Serratia marcescens and Pseudomonas aeruginosa as potential biocontrol against Coptotermes curvignathus.

The success of microbial termiticides in controlling termites depends on the ability of microbes to grow in different media and the functionality of the microbes as a resistant barrier or toxic bait. This study was conducted to understand the mortality rate and behaviour changes of the subterranean termite Coptotermes curvignathus Holmgren introduced with different concentrations of Serratia marcescens strain LGMS 1 and Pseudomonas aeruginosa strain LGMS 3 using wood and soil as bacterial transfer medium. In general, higher concentration of bacteria in soil caused a reduction in tunnelling activity and wood consumption and an increase in mortality. However, application on wood revealed a different outcome. Wood treated with S. marcescens of 106 CFU ml-1 concentration proved to be more efficient as bait than higher concentration applications as it caused a high mortality rate while still highly palatable for termites. Wood or soil treated with S. marcescens concentration higher than 109 CFU ml-1 creates a high toxicity and repellent barrier for termites. Pseudomonas aeruginosa of 109 CFU ml-1 concentrations applied on wood served as a slow-acting toxic bait. However, the ability for S. marcescens and P. aeruginosa to survive on wood is low, which made the bait unable to retain a useful level of toxicity for a long period of time and frequent reapplication is needed.

Characterization of bioadsorbent produced using incorporated treatment of chemical and carbonization procedures.

The production of bioadsorbent from palm kernel shell (PKS) and coconut shell (CS) pretreated with 30% phosphoric acid (H3PO4) was optimized using the response surface methodology (RSM). Iodine adsorption for both bioadsorbents was optimized by central composite design. Two parameters including the H3PO4 pretreatment temperature and carbonization temperature were determined as significant factors to improve the iodine adsorption of the bioadsorbent. Statistical analysis results divulge that both factors had significant effect on the iodine adsorption for the bioadsorbent. From the RSM analysis, it was suggested that using 80 and 79°C as H3PO4 pretreatment temperature and 714 and 715°C as carbonization temperature would enhance the iodine adsorption of the CS and PKS bioadsobent, respectively. These results indicated that H3PO4 is a good pretreatment for preparing PKS and CS prior to carbonization process to produce bioadsorbent with well-developed microporous and mesoporous volume. The effort to produce alternative high grade and inexpensive adsorbent derived from lignocellulosic biomass, particularly in the nut shell form was implied in this research.

Production of bioadsorbent from phosphoric acid pretreated palm kernel shell and coconut shell by two-stage continuous physical activation via N2 and air.

In the present study, agricultural biomass-palm kernel shell (PKS) and coconut shell (CS)-was used to produce high porosity bioadsorbent using two-stage continuous physical activation method with different gas carrier (air and N2) in each stage. The activation temperature was set constant at 600, 700, 800 or 900°C for both activation stages with the heating rate of 3°C min-1. Two parameters, the gas carrier and activation temperature, were determined as the significant factors on the adsorption properties of bioadsorbent. BET, SEM, FTIR, TGA, CHNS/O and ash content were used to elucidate the developed bioadsorbent prepared from PKS and CS and its capacity towards the adsorption of methylene blue and iodine. The novel process of two-stage continuous physical activation method was able to expose mesopores and micropores that were previously covered/clogged in nature, and simultaneously create new pores. The synthesized bioadsorbents showed that the surface area (PKS: 456.47 m2 g-1, CS: 479.17 m2 g-1), pore size (PKS: 0.63 nm, CS: 0.62 nm) and pore volume (PKS: 0.13 cm3 g-1, CS: 0.15 cm3 g-1) were significantly higher than that of non-treated bioadsorbent. The surface morphology of the raw materials and synthesized bioadsorbent were accessed by SEM. Furthermore, the novel process meets the recent industrial adsorbent requirements such as low activation temperature, high fixed carbon content, high yield, high adsorption properties and high surface area, which are the key factors for large-scale production of bioadsorbent and its usage.