A case of rotenone poisoning from ingesting Derris trifoliata Lour. (Tuba fruit/pod) in Malaysia.

Derris trifoliata is mainly found in mangrove area in tropical regions and the plant extract is traditionally used for fishing by poisoning. This is the first case report of rotenone poisoning in a child from ingesting Derris trifoliata seed. The child developed altered consciousness, vomiting, hypotension, metabolic acidosis, and acute kidney injury. Species identification of this case requires the collaborative efforts of various agencies. She survived from the poisoning with no neurological sequelae.

In Vitro Efficacy of Aqueous and Methanol Extract of Cassia siamea Against the Motility of Caenorhabditis elegans.

Gastrointestinal nematode infections can cause great losses in revenue due to decrease livestock production and animal death. The use of anthelmintic to control gastrointestinal nematode put a selection pressure on nematode populations which led to emergence of anthelmintic resistance. Because of that, this study was carried out to investigate the efficacy of aqueous and methanol extract of Cassia siamea against the motility of C. elegans Bristol N2 and C. elegans DA1316. Caenorhabditis elegans Bristol N2 is a susceptible strain and C. elegans DA1316 is an ivermectin resistant strain. In vitro bioassay of various concentrations of (0.2, 0.6, 0.8, 1.0 and 2.0 mg mL-1) aqueous and methanol extracts of C. siamea was conducted against the motility of L4 larvae of C. elegans Bristol N2 and C. elegans DA1316. The L4 larvae were treated with 0.02 µg mL-1 of ivermectin served as positive control while those in M9 solution served as negative control. The activity of the extracts was observed after 24 h and 48 h. A significant difference was recorded in the extract performance compared to control at (P < 0.001) after 48 h against the motility of the larvae of both strains. The methanol extracts inhibited the motility of C. elegans Bristol N2 by 86.7% as well as DA1316 up to 84.9% at 2.0 mg mL-1 after 48 h. The methanol extract was more efficient than aqueous extract (P < 0.05) against the motility of both strains of C. elegans. Cassia siamea may be used as a natural source of lead compounds for the development of alternative anthelmintic against parasitic nematodes as well ivermectin resistant strains of nematodes.

Droplet-vitrification of Aranda Broga Blue orchid: Role of ascorbic acid on the antioxidant system and genetic fidelity assessments via RAPD and SCoT markers.

A droplet-vitrification cryopreservation protocol has been successfully developed for Aranda Broga Blue orchid hybrid using protocorm-like bodies (PLBs). However, maximum growth regeneration percentage was recorded at 5% only based on previous report. Thus, to improve growth recovery of cryopreserved PLBs, cryopreservation stages were supplemented with ascorbic acid, tested at 50, 100 and 150 mg/L. However, results demonstrated that exogenous ascorbic acid was not favorable in regeneration of cryopreserved explants (maximum value of 1.67 % with 50 mg/L ascorbic acid supplementation). Total soluble protein and various antioxidant enzyme activities such as catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX) were evaluated after each cryopreservation stages in conjunction with the application of exogenous ascorbic acid. Addition of antioxidant must be carefully evaluated and its application may not guarantee successful growth recovery. RAPD and SCoT molecular analysis confirmed the genetic stability of regenerated cryopreserved PLBs as no polymorphism was detected compared to control PLBs culture.

Biochemical analyses of Dendrobium Sabin Blue PLBs during cryopreservation by vitrification.

Dendrobium Sabin Blue is an important orchid hybrid that has been grown extensively as cut flower, potted plant and is also popular for its deep purplish blue flowers.  The most efficient long term conservation method of this hybrid is through cryopreservation. Cryopreservation involving the vitrification method consists of explants exposure to highly concentrated cryoprotective solution followed by freezing rapidly in liquid nitrogen. However, these treatments involved highly concentrated cryoprotectant that could incur toxicity to the explants. Hence, cryopreservation protocol requires biochemical analyses in understanding the damages or injuries occurred during cryopreservation treatments. In this study, biochemical analyses revealed a general reduction in chlorophyll, carotenoid and porphyrin content to 0.40 µg/g F W (thawing stage), 31.50 µg/g F W unloading stage and 2230.41 µg/g F W (thawing stage), respectively in comparison to the control treatments. In addition, increased level in proline content were obtained at different cryopreservation stages with highest level (5.42 µmole/g F W) recorded at the PVS2 dehydration stage. Fluctuated outcomes were obtained in catalase (CAT), ascorbate peroxidase (APX) and peroxidase (POX) enzyme activities in PLBs exposed to different cryopreservation stages. Lowest values recorded for CAT enzyme activity were obtained at the dehydration stage (3.94 U/g). Lowest POX enzyme activities were obtained at the dehydration (122.36 U/g) and growth recovery (106.40 U/g) stages. Additionally, lowest APX enzyme activities values were recorded at the thawing (7.47 U/g) and unloading (7.28 U/g) stages. These have contributed to low regeneration of Dendrobium Sabin Blue protocorm like bodies (PLBs) following cryopreservation. Hence, in the future experimental design, exogenous antioxidant could be included in the cryopreservation procedures to improve the existing protocol.

Autofluorescence study and selected cyanidin quantification in the Jewel orchids Anoectochilus sp. and Ludisia discolor.

Anoectochilus sp. and Ludisia discolor are known as Jewel orchids. Both species are terrestrial wild orchids that grow in shaded areas of forests. The Jewel orchids are renowned for the beauty of their leaves, which are dark-green laced with silvery or golden veins. The orchids are used as a cure in various parts of Asia. Overharvesting and anthropogenic disturbances threaten the existence of the Jewel orchids in the wild, necessitating human intervention in their survival. An understanding of the structure and adaptations of a plant may assist in its survival when propagated outside of its habitat. In this study, ex vitro leaves of Anoectochilus sp. and L. discolor were subjected to freehand sectioning, and then inspected through brightfield and fluorescence microscopy. The study indicated that all parts of both plants presented typical monocotyledonous characteristics except the leaves. The leaves displayed dorsiventrality with distinct palisade and spongy mesophyll layers. The spongy mesophyll layer contained cells which fluoresced a bright red when exposed to ultraviolet, blue, and green light wavelengths, hinting at the presence of anthocyanins for photoprotection. Cyanidin was detected in the leaves of L. discolor, as enumerated through high performance liquid chromatography (HPLC). The observations indicated that Anoectochilus sp. and L. discolor are well-adapted to live under shaded conditions with minimal exposure to light.

The dual defensive strategy of Amorphophallus throughout its ontogeny.

Amorphophallus bufo is a rarely studied plant in Malaysian tropical rainforests. We measured the spectral reflectance of different developmental stages of A. bufo (seedlings, juveniles and adults), background soil/ debris and leaves from other neighboring plant species. Results show that the leaves of A. bufo seedling have a similar reflectance curve as the background soil and debris. Adults and juveniles of A. bufo are similar to other neighboring plants' leaf colors. We hypothesize that the cryptic coloration of A. bufo seedlings plays an important role in camouflage and that the numerous black spots on the surface of the petioles and rachises, may serve as a defensive mimicry against herbivores.

Early Studies on Protoplast Isolation of Ludisia discolor, A Wild Orchid.

The terrestrial Ludisia discolor, also referred to as the jewel orchid is prized for the quality of its leaves. L. discolor is known as a medicinal herb and is touted for its heat- and pathogen-resisting qualities. L. discolor is valuable in the production of both flavonoids and anthocyanins, antioxidants that are exalted in the health industry. Plant cell cultures have emerged as alternative sources of anthocyanin production. Plant protoplast cultures are used frequently in transient gene expression studies and in the establishment of callus and cell suspension cultures. Benefits of plant protoplast system include similarity to cells found in plant tissues, reproduction under controlled conditions, and prevention of masking of stress responses to previous handling techniques. A study was conducted to assess the amenability of the stem and leaves of L. discolor to protoplast isolation. The stem and leaf segments were weighed, sliced into thin layers, immersed in a digestion medium, washed and then cultured onto a recovery medium. Results indicated that the production of plant protoplasts from L. discolor may be viewed as an alternative in the generation of cell cultures and ultimately in the production of anthocyanins from the cell cultures.

Ascorbate Peroxidase Activity of Aranda Broga Blue Orchid Protocorm-like Bodies (PLBs) In Response to PVS2 Cryopreservation Method.

Throughout the cryopreservation process, plants were exposed to a series of abiotic stresses such as desiccation and osmotic pressure due to highly concentrated vitrification solution. Abiotic stress stimulates the production of reactive oxygen species (ROS) which include hydrogen peroxide, superoxide radicals, and singlet oxygen. Higher production of ROS may lead to oxidative stress which contributes to the major injuries in cryopreserved explants. Antioxidant enzymes in plant such as ascorbate peroxidase (APX) can protect plants from cell damage by scavenging the free radicals. This study was determined based on APX enzyme activity of Aranda Broga Blue orchid's protocorm-like bodies (PLBs) in response to PVS2 (Plant Vitrification Solution 2) cryopreservation treatments at different stages. PLBs that were precultured at 0.25 M sucrose for 3 days were subjected to vitrification cryopreservation method. Results obtained showed that the highest APX activity was achieved at PVS2 cryoprotectant treatment prior liquid nitrogen (LN) storage. This phenomenon indicating that accumulation of osmotic and dehydrating stress throughout the cryopreservation treatment resulted in oxidative burst which in turn leads to higher APX activity in order to control the excess production of ROS. To conclude, PVS2 treatment was revealed as the most detrimental step throughout cryopreservation treatment. Thus, this research also suggested that exogenous antioxidant such as ascorbic acid can be added throughout cryopreservation procedure especially at PVS2 treatment in the future experiments to aid in regrowth of cryopreserved explants by reducing oxidative stress.

Cryptic coloration of Macaranga bancana seedlings: A unique strategy for a pioneer species.

Macaranga bancana is considered as a successful pioneer plant species. Usually found in disturbed and open areas, most of the current research focused on its relations with ants. One of the unique feature of the plants is that the seedling leaves are red, resembling and almost matching the background. Using a portable spectrometer, we measured the color reflectance of M. bancana seedlings (less than 20 cm in height). We also measured the leaf litter reflectance, adult M. bancana leaves and also seedlings of several other species found in the vicinity of M. bancana seedlings. The reflectances of M. bancana seedlings are very similar to that of the leaf litter background. We suggest that this cryptic coloration is crucial during the early stages of the plant when it still cannot rely on the protection of ants.

The distribution of anacardiaceae in teluk bahang forest reserve, pulau pinang.

To investigate the distribution of Anacardiaceae in Teluk Bahang Permanent Forest Reserve (TBPFR) in Pulau Pinang, all trees with a diameter at breast high (DBH) ≥ 5 cm were enumerated in a study site constituting 0.4 ha of the reserve. Seventy five individuals of Anacardiaceae (14% of all trees) are recorded. These individuals represent 4 genera and 5 species, namely, Mangifera pentandra, Mangifera macrocarpa, Gluta elegans, Campnosperma auriculatum and Swintonia floribunda. The mean density of Anacardiaceae within the study plots is 7.50±8.14 (mean±S.D.) per ha whereas the basal area (BA) calculated is 0.97 m(2)/0.40 ha. The importance value (IVi) for Anacardiaceae is 81%. The estimated total aboveground biomass (TAGB) for Anacardiaceae is 24.24 ton/0.40 ha. A total of 333 Anacardiaceae saplings with a DBH < 5 cm are recorded. These saplings have been identified as juveniles of the genera Gluta (9.99%), Swintonia (84.90%) and Mangifera (5.11%).

Soils on exposed Sunda shelf shaped biogeographic patterns in the equatorial forests of Southeast Asia.

The marked biogeographic difference between western (Malay Peninsula and Sumatra) and eastern (Borneo) Sundaland is surprising given the long time that these areas have formed a single landmass. A dispersal barrier in the form of a dry savanna corridor during glacial maxima has been proposed to explain this disparity. However, the short duration of these dry savanna conditions make it an unlikely sole cause for the biogeographic pattern. An additional explanation might be related to the coarse sandy soils of central Sundaland. To test these two nonexclusive hypotheses, we performed a floristic cluster analysis based on 111 tree inventories from Peninsular Malaysia, Sumatra, and Borneo. We then identified the indicator genera for clusters that crossed the central Sundaland biogeographic boundary and those that did not cross and tested whether drought and coarse-soil tolerance of the indicator genera differed between them. We found 11 terminal floristic clusters, 10 occurring in Borneo, 5 in Sumatra, and 3 in Peninsular Malaysia. Indicator taxa of clusters that occurred across Sundaland had significantly higher coarse-soil tolerance than did those from clusters that occurred east or west of central Sundaland. For drought tolerance, no such pattern was detected. These results strongly suggest that exposed sandy sea-bed soils acted as a dispersal barrier in central Sundaland. However, we could not confirm the presence of a savanna corridor. This finding makes it clear that proposed biogeographic explanations for plant and animal distributions within Sundaland, including possible migration routes for early humans, need to be reevaluated.