Virulence Pattern of Pyricularia oryzae Pathotypes Towards Blast Monogenic Lines

Rice blast caused by Pyricularia oryzae (P. oryzae) is one of the most serious diseases infecting rice worldwide. In the present study, virulence pattern of six P. oryzae pathotypes (P0.0, P0.2, P1.0, P3.0, P7.0 and P9.0) identified from the blast pathogen collected in Peninsular Malaysia, were evaluated using a set of 22 IRRI-bred blast resistance lines (IRBL) as well as to determine the resistance genes involved. The information on the virulence of the blast pathotypes and the resistance genes involved is important for breeding of new rice variety for durable resistance against blast disease. The IRBL was established from 22 monogenic lines, harbouring 22 resistance genes [Pia, Pib, Pii, Pit, Pi3, Pi5(t), Pish, Pi1, Pik, Pik-s, Pik-m, Pik-h, Pik-p, Pi7(t), Pi9, Piz, Piz-5, Piz-t, Pi19, Pi20(t), Pita-2, and Pita=Pi4(t)]. Based on the disease severity patterns, the tested pathotypes were avirulence towards seven IRBLs [IRBLi-F5, IRBLk-Ka, IRBLkh-K3, IRBLz-Fu, IRBLsh-S, IRBLPi7 (t) and IRBL9-W] of which these IRBLs harbouring Pii, Pik, Pik-h, Piz, Pish, Pi7(t) and Pi9 resistance genes, respectively. Therefore, the results suggested that the seven IRBLs carrying seven resistance genes [Pii, Pik, Pik-h, Piz, Pish, Pi7(t) and Pi9] would be suitable candidates of resistance genes to be incorporated in new breeding lines to combat the current blast pathotypes in the field.

Vegetative Propagation of Hoya imperialis and Hoya coronaria by Stem Cutting and Micropropagation

Hoya imperialis (H. imperialis) and H. coronaria (Apocynaceae) are known to have ornamental value due to their beautiful flowers; however, the feasibility of propagating these plants have not been reported despite the wild populations in Brunei Darussalam being highly threatened due to habitat loss and overcollection. Thus, the present study aimed to conduct a preliminary study of the feasibility of two alternative propagation methods, stem cutting and micropropagation, as a potential approach for their ex situ conservation. Hoya stem cuttings were treated with either indole-3-butyric acid (IBA) or 1-naphthaleneacetic acid (NAA) (0–2000 mg/L), and then propagated onto a mixture of peat moss and perlite. For micropropagation, Hoya leaf explants were cultured onto Murashige and Skoog (MS) agar media that were supplemented with IBA and/or kinetin (KN) (0–10.0 mg/L). This present study shows that both Hoya species were successfully propagated by stem cutting even without hormone treatment. However, interestingly, in H. imperialis, when compared with control, the mean number of new leaves (6.3 ± 1.0) and the mean relative growth rate (RGR) based on stem diameter (0.004 ± 0.0007 cm cm−1 day−1) significantly increased when treated with 500 mg/L NAA and 2000 mg/L IBA, respectively. Meanwhile, in H. coronaria, significantly higher mean number of roots was achieved by treating with 1000 mg/L NAA (16.6 ± 1.4) or 2000 mg/L IBA (17.5 ± 2.7) compared with control. For micropropagation, callus induction was not promising and could only be observed at specific concentrations of both IBA and KN, with H. imperialis appearing to be more responsive towards these hormones in comparison to H. coronaria. The present study showed that stem cutting appeared more feasible in propagating both Hoya species.

Preliminary Study on the Effect of Nitrogen Fertilisation on Phytochemical Content Quality of Gynura procumbens

The study was carried out to examine the effects of nitrogen (N) fertilisation on phytochemical content quality, to determine types of phytochemicals compositions, and to establish phytochemicals marker compounds in Gynura procumbens. In this two factors study, three stages of harvesting (H) time and three rates of N were laid out according to Randomised Complete Block Design (RCBD). Physiological and biochemical attributes were recorded to exhibit the trend for plant quality. The results showed that, N has affected phytochemical content significantly (p < 0.05) with stronger effect on physiological and biochemical attributes (p < 0.01). The results suggested that 0 and 90 kg/ha N, respectively are highly and least effecting the Photo, Cond, TChlC, TCC, TPrc, TLiC, TPC and TFC.

Chitinolytic and Antagonistic Activity of Streptomyces Isolated from Fadama Soil against Phytopathogenic Fungi

Chitinases which degrade chitin have attracted attention as biological antifungal agents. The purpose of this study is to isolate Streptomyces from Fadama soil and assess its chitinolytic and antagonist potential against phytopathogenic fungi for application as biocontrol agent. Streptomyces were isolated from Fadama soil. The selected isolate CT02 exhibited chitinolytic characteristics. Chitinase production was performed under different temperatures, pH and varying incubation period. The highest chitinase production by CT02 isolate was observed after five days of cultivation. The highest chitinase activity was observed at 35°C and pH 7. The crude extracellular enzyme exhibited a specific activity of 4.20 U/μg whereas partially purified extracellular enzyme exhibited a specific activity of 6.19 U/μg with purification fold of 1.47. The selected isolate CT02 and its extracellular crude chitinase showed in vitro antifungal antagonist potential by inhibiting the growth of Aspergillus niger and Aspergillus oryzae. This indicates that Streptomyces derived chitinases are potential biocontrol agents against phytopathogenic fungi.

Nest Characteristics of the Sumatran Orangutan (Pongo abelii) in the Wildlife Sanctuary Soraya Station in Aceh Province, Indonesia

Orangutans (Pongo spp.) populations used to be widely distributed throughout Southeast Asia, from Java in the south to the Southern China in the north during the Pleistocene. Their populations have declined up to 75% of their original size and are now distributed only in parts of the tropical rainforests of Borneo and Sumatra. Pongo pygmaeus, Pongo tapanuliensis and Pongo abelii are the three most representative species, in this study, here we discussed the latter. Sumatran forests are generally suffering from deforestation with rates ranging from 3.74% to 49.85% between 2000 and 2012. Thus, human wildlife conflict intensity has escalated and gained more traction. Orangutans are known as arboreal great apes and need to build nests for resting. We applied the transect line method (three transects; each 1,000 m long) at different elevations in Soraya Research Station, Gelombang Village, Sultan Daulat sub-district, Subulussalam district, and assessed the nest characteristics. The characteristics are: (1) nesting position referring to the position of nest on a tree; (2) nest successional stages defining the age and leaf decay used in constructing a nest indicated with I (new) until V (almost gone); and (3) nest density to predict the density of nest per square km. Afterwards, the identified nesting trees along the transect were further identified based on their species, and assessed based on three characteristics (the tree height, diameter and the height of a nest measured from the soil). A total of 27 nests were found, and 44% were located in transect III or at the riparian. Out of 27, four orangutans’ nests were found on Moraceae (Streblus elongatus) and Myrtaceae (Syzigium spp.), while three nests were found on Dipterocarpus sp. The tree height, tree diameter and nest height were 10 m–25 m (mean = 17.5 m; SD = ± 0.25), 10 cm–30 cm (mean = 20 cm; SD = ± 0.4) and 16 m–20 m (mean = 18 m; SD = ± 0.35), respectively. Meanwhile, nest density calculated based on the form: d = [N/ (L* 2w)], and the values obtained were 8.4, 13.45, 26.9 nests/km2 located on transect I, II and III or at the riparian. The most commonly found nest successional stages and position were stage III and position 3, respectively. This study could serve as a baseline research in primate conservation and nest characterisation could be used as guidance for any future activity planning (e.g. tree reforestation) in a particular region and the existence of various tree species diversity are indispensable for maintaining orangutan habitats’ quality.

Effects of Recreational Camping on the Environmental Values of National Parks in Sri Lanka

Camping is a popular activity in the contemporary nature-based tourism domain and rapidly gaining momentum as a key recreational activity in Sri Lanka’s national parks (NPs). Recreational uses such as camping in natural areas can induce significant and often localised resource impacts that can affect soil, vegetation, wildlife and water, with the severity of such impacts varying according to the intensity of use. Hence, monitoring of the biophysical conditions of campsites has become an important component in the reserve management agenda in many places, especially in developed countries. To the best of our knowledge, this is the first study to examine the biophysical impacts associated with the recreation ecology of camping in Sri Lanka. Ten campsites from three dry zone NPs were selected to assess biophysical impacts of camping activities. Field measurements were based on the fixed radial transect method. Gathered data included the total area of the campsite, erosion potential measured as the area of exposed soil (devoid of vegetation or organic litter), number of exposed roots and human damage to trees, number of fireplaces/ fire scars on the ground, visual counts of litter, soil compaction measured by penetrometer, loss of woody debris. This study reports significant levels of environmental degradation related to all the indictors of biophysical impacts at both high and low use campsites. There was no evidence for any difference in the level of environmental degradation associated with high and low use campsites. The loss of natural values associated with campsites negatively impacted visitors’ nature-based experience. These findings highlight the importance of managing biophysical impacts in campsites to provide a high-quality visitor experience, while sustainably managing tourism activities in NPs.

Characterisation of Plant Growth-Promoting Endophytic Bacteria from Sugarcane and Their Antagonistic Activity against Fusarium moniliforme

The use of endophytic bacteria in agriculture provides an effective way of improving crop yield and significantly reducing chemical usage, such as fungicides. This research was conducted to explore endophytic bacteria with plant growth promotion (PGP) and antifungal activities against Fusarium moniliforme AIT01. In this study, we obtained 52 isolates of endophytic bacteria associated with the roots and stems of sugarcane from Nakhon Ratchasima province, Thailand. In vitro antagonistic activity test showed that 14 out of 52 isolates had antagonistic activity against the fungal pathogen F. moniliforme AIT01. These antagonistic endophytic bacteria were identified as belonging to six different species as follows: Nguyenibacter vanlangensis, Acidomonas methanolica, Asaia bogorensis, Tanticharoenia aidae, Burkholderia gladioli and Bacillus altitudinis based on phenotypic characteristics, along with phylogenetic analysis of their 16S rRNA gene sequences. Seven isolates effectively inhibited F. moniliforme AIT01 mycelial growth by up to 40%. The volatile compounds of six isolates reduced the growth of F. moniliforme AIT01 by over 23%. Moreover, riceberry rice seedlings previously treated with B. gladioli CP28 were found to strongly reduce infection with phytopathogen by 80% in comparison to the non-treated control. Furthermore, the isolates also showed relevant PGP features, including ammonia production, zinc and phosphate solubilisation, auxin and siderophore biosynthesis. These results demonstrated that the tested endophytic bacteria could be successfully utilised as a source of PGP and biocontrol agent to manage diseases caused by F. moniliforme.

Isolation and Identification of Plant Growth Promoting Rhizobacteria from Sago Palm (Metroxylon sagu, Rottb.)

Plant growth promoting rhizobacteria (PGPR) are strains of naturally occurring soil bacteria that live in close vicinity to the plant’s rhizosphere region which possess the capability to augment host growth. This study was conducted to isolate and identify potential PGPR isolates indigenous to Metroxylon sagu, Rottb. rhizosphere. These potential isolates were characterised based on their beneficial plant growth promoting (PGP) properties and identified by molecular analysis via 16S rDNA sequencing. A total of 18 isolates were successfully isolated, out of which five isolates were tested, and designated as (S1A, S2B, S3A, S3C and S42). Among the five isolates, two isolates (S2B and S3C) were found to produce high levels of indole-3-acetic acid (2.96 μg/mL and 10.31 μg/mL), able to fix nitrogen and show significant activity in phosphate solubilisation. The analysis of their sequences via National Center for Biotechnology Information (NCBI) suggested their close identity towards Lysinibacillus sphaericus and Bacillus thuringiensis. It can be concluded that the isolated PGPR possesses beneficial PGP attributes. It can be implied that the isolated PGPR are potential to be used as inoculant biofertilisers, beneficial for Metroxylon sagu, Rottb. growth. Hence, further studies need to be done to evaluate the effectiveness of the beneficial microbes towards sago seedlings growth, under pot experiment.

Characterisation and Pathogenicity of Aspergillus tamarii Causing Banana Fruit Rot

Banana fruit rot is a common postharvest disease of the banana fruit. The appearance of rot symptoms on the surface of the fruits reduces the quality and marketability of banana. From rot lesions on banana fruits, three Aspergillus isolates were isolated. Based on morphological characteristics and sequences of Internal Transcribed Spacer, β-tubulin and calmodulin, the isolates were identified as A. tamarii. Pathogenicity tests of the isolates, conducted using mycelial plugs with wounded and unwounded treatments, showed A. tamarii as the pathogen of banana fruit rot. Rot symptoms were highly severe on wounded banana fruits compared to unwounded fruits, and therefore, wounded banana fruits are more susceptible to A. tamarii infection. To the best of our knowledge, this is the first report of A. tamarii as a causal pathogen of banana fruit rot. This study indicated A. tamarii is one of postharvest rot pathogens of banana.

Evaluation of Cellulolytic Endo-1,4-β-D-Glucanase Activity in the Digestive Fluid of Adult Phytophagous Beetle Hoplasoma unicolor

Insects of the taxonomic order Coleoptera are recognised for considerable cellulolytic activity in their digestive fluid. The cellulolytic activity of the gut fluid in Hoplasoma unicolor, a member of Coleoptera, however, remains unexplored. In this study, we, for the first time, report the qualitative and quantitative analysis of cellulolytic activity in the digestive fluid of this insect. The cellulolytic endo-1,4-β-D-glucanase activity was confirmed in the supernatant of the insect’s digestive fluid by agar plate assay using carboxymethyl cellulose as the substrate. To determine the optimum pH, enzyme activity was further assessed in an acidic pH range of 5 to 6, and the highest activity was observed at pH 5.3. For quantitative analysis, endoglucanase activity was measured using 3,5-dinitrosalicylic acid method which revealed that the specific activity of the gut sample was 0.69 (±0.01) units per mg of protein. For further characterisation of the cellulases in the sample, SDS-PAGE and zymogram analysis were carried out. Two active cellulolytic bands were detected on the zymogram suggesting the presence of two distinct endoglucanases which completely disappeared upon heating the sample at 55°C. Our study, therefore, highlights prospect of the gut fluid of H. unicolor as an important source of cellulase enzymes that merits further investigations into their extensive characterisation for potential industrial applications.