In vitro seed germination of paphiopedilum lowii, an endangered slipper orchid in North Borneo

Paphiopedilum lowii (Lindl.) Stein is alarmingly decreasing in numbers due to habitat destruction and over-collection. Propagation through seed is one effort to conserve the species. In vitro method offers better results for seed germination in orchids, but the success of germination is species-specific. Hence, this research aimed to evaluate different types of media [(½ strength Murashige & Skoog and Orchimax Medium (Duchefa Biochemie)], and to determine the effects of sucrose [0% 1%, 2% and 3% (w/v)], peptone [0%, 0.1% and 0.2% (w/v)], coconut water [0%, 10%, 20% and 30% (v/v)] and fertilizer [Orchid Focus-0%, 0.1% and 0.2% (v/v)] on seed germination. The protocorm size was also determined after 12 weeks of observation. Half-strength MS media consistently gave higher seed germination, ½ MS supplemented with 1% (w/v) sucrose, 0.1% (w/v) peptone, 0% (v/v) coconut water and 0.1 % (v/v) fertilizer resulted in the highest seed germination at 5.32% ± 5.9, 19.27% ± 9.34, 11.33% ± 3.80 and 19.31% ± 9.03 respectively. Despite being 12 weeks in culture, the sizes of the protocorms are small (0.222±0.089 mm diameter, 0.703±0.280 mm circumference, and 0.045±0.043 mm2 area); this requires future investigation. These findings can serve as base information for further enhancement of seed germination and development of propagation methods of P. lowii for use in a conservation program. © 2022, Society for Indonesian Biodiversity. All rights reserved

A Species Checklist of Wild Orchids in Selected Sites in Kadamaian, Kota Belud, Sabah

A brief orchid diversity study was conducted from 14th to 19th October 2019 in Kadamaian, located northwest of Kinabalu area, through a scientific geographic expedition. A convenience sampling method was employed with only the flowering individuals collected and preserved as herbarium specimens. A total of 58 species and 32 genera were identified during the field visits to the Mount Nopungguk, Melangkap Noriou, Ulu Sungai Melawa, and basecamp site. The finding comprises of 43 epiphytes, 13 terrestrials, and two mycoheterotrophs. Appendicula congesta, Bulbophyllum disjunctum, Dendrobium kiauense, and Goodyera rostellata are species endemic to Borneo found during the expedition along with Crepidium multiflorum, a hyper-endemic species to Kinabalu area. Several rare species which narrowly distributed to primary forest habitat were discovered, including jewel orchids; Cystorchis variegata var. variegata and Anoectochilus geniculatus, and two mycoheterotrophs; Aphyllorchis pallida and Lecanorchis multiflora var. multiflora

A preliminary survey and chemical profiling of wild ginger species in Kadamaian, Kota Belud, Sabah

A preliminary survey of the diversity of gingers (Zingiberaceae) was conducted in Kadamaian, Kota Belud from 14th to 19th October, 2019. Wild ginger species is utilized widely as one of the most important material in traditional medicine among indigenous people of Sabah. However, few of these plant species have been studied for their chemical constituents and beneficial properties. In order to investigate the compound composition, the essential oil from Etlingera brevilabrum, Alpinia nieuwenhuizii and Hornstedtia havilandii were screened. The essential oil was obtained from leaves, stems and rhizomes of the plant through hydro-distillation and analysed for their chemical composition through Gas Chromatography-Mass Spectrometry (GC-MS). The result of this study indicated that the chemical constituents of all three parts for all species are similar; all have terpenoids (monoterpene and sesquiterpene), aldehyde, hydrocarbon, ketone and alcohol in the essential oil extracts. GC-MS analyses of the oils led to the identification of 35 compound constituents from the leaves, stems and rhizomes of E. brevilabrum, which is the highest. Meanwhile, A. nieuwenhuizii displayed 34 chemical compositions from all parts (leaf, stem and rhizome) of the plant. H. havilandii showed the lowest number of volatiles from all plant parts (24 compounds). Monoterpene is dominant in all wild ginger studied, except for rhizome of E. brevilabrum. On the contrary, E. brevilabrum showed sesquiterpene as the most abundant compound in its composition. This shows that the volatile oil composition of wild ginger species is extremely variable. This study provides preliminary key chemical information for evaluating the quality of local wild gingers in Kadamaian, Kota Belud, Sabah

Kesan kepelbagaian media kultur, pengawalatur tumbuhan dan aditif organik ke atas mikropropagasi orkid selipar terancam Sabah

Paphiopedilum hookerae var volonteanum dan Paphiopedilum dayanum adalah spesies orkid selipar yang endemik di Sabah yang terkenal dengan keunikan morfologi bunga. Ini telah mengakibatkan orkid ini sering diambil dan diperdagangkan secara haram. Kini, orkid ini telah dilaporkan sebagai spesies yang terancam oleh International Union for Conservation of Nature (IUCN) dan tersenarai dalam Appendix 1: Convention on International Trade in Endangered Species (CITES). Ini mendorong kepada kegiatan pemuliharaan bagi mengekalkan populasi orkid selipar. Namun, kadar pertumbuhan orkid selipar adalah sangat rendah dan mengambil masa yang lama. Teknik kultur tisu dapat meningkatkan jumlah individu spesies dalam masa yang singkat. Oleh itu, kajian ini bertujuan untuk membangunkan protokol kultur tisu untuk meningkatkan bilangan individu orkid selipar P. hookerae var volonteanum dan P. dayanum dengan menggunakan biji benih sebagai eksplan. Percambahan biji benih kedua-dua spesies dilakukan ke atas media yang ditambahkan dengan sukrosa (0, 1, 2 dan 3%, w/v), pepton (0, 0.5, 1.0, 1.5 dan 2.0 g/L), dan air kelapa (0, 5, 10, 15 dan 20%, v/v). Seterusnya, jenis dan kepekatan pengawalatur tumbuhan tunggal iaitu 6-benzylaminopurina (BAP; 0, 0.2, 0.4, 0.6, 0.8, 1.0 mg/L), asid alpha-naftalenasetik (NAA; 0, 0.1, 0.2, 0.3, 0.4, 0.5 mg/L) dan thidiazuron (TDZ 0, 0.2, 0.4, 0.6, 0.8, 1.0 mg/L) telah diuji sebagai kajian awal. Kemudian, pengawalatur atur tunggal yang terbaik daripada kajian awal iaitu 1.0 mg/L TDZ digabungkan dengan asid 2,4-diklorofenoksiasetik (2,4-D; 0, 0.2, 0.4, 0.6, 0.8, 1.0 mg/L). Kajian percambahan diteruskan bagi P. hookerae var volonteanum untuk meningkatkan percambahan pada media Murashige & Skoog (MS), ½MS, Robert Ernst (RE), dan New Dogashima Medium (NDM). Seterusnya, jus tomato (0, 0.05, 0.2, 0.3 dan 0.5%, v/v) dan homogenat pisang (0, 0.05, 0.2, 0.3 dan 0.5%, w/v) serta faktor kematangan biji benih berumur 240 HSP dan 360 HSP telah diuji terhadap biji benih orkid P. hookerae var volonteanum . Selepas 80 hari pengkulturan, percambahan tertinggi bagi P. hookerae var volonteanum ialah pada media ½MS dengan 2% (w/v) sukrosa (57.11±1.1%), 2.0 g/L pepton (69.76±7.7%) dan 10% (v/v) air kelapa (55.76±1.9%). Bagi P. dayanum pula, percambahan tertinggi adalah pada 2% (w/v) sukrosa (82.47±8.7%), 2.0 g/L pepton (91.93±7.4%) dan 10% (v/v) air kelapa (90.26±7.7%). Pengawalatur tumbuhan BAP, NAA dan TDZ mencatatkan peratusan percambahan yang rendah berbanding kawalan. Ini menunjukkan kesan perencatan terhadap percambahan biji benih bagi kedua-dua spesies. Kombinasi pengawalatur tumbuhan 1 mg/L TDZ + 1 mg/L 2,4-D memberikan percambahan yang tinggi bagi kedua-dua P. hookerae var volonteanum (59.00±7.2%) dan P. dayanum (71.25±8.2%). Kesan jus tomato 0.5% (v/v) dan homogenat pisang 0.3% (w/v) memberikan percambahan yang rendah terhadap biji benih P. hookerae var volonteanum masing-masing dengan 19.90±3.6% dan 18.08±7.8%. Biji benih P. hookerae var volonteanum yang berumur 240 HSP adalah yang terbaik untuk percambahan biji benih secara in vitro . Untuk kajian proliferasi protokorm terhadap P. dayanum , pengawalatur tumbuhan kinetin (0, 1.0, 2.0, 3.0, 4.0, 5.0 mg/L), BAP (0, 1.0, 2.0, 3.0, 4.0, 5.0 mg/L), TDZ (0, 1.0, 2.0, 3.0, 4.0, 5.0 mg/L), NAA (0, 0.5, 1.0, 1.5, 2.0 mg/L), dan 2,4-D (0, 0.5, 1.0, 1.5, 2.0 mg/L) secara tunggal dan kombinasi auksin-sitokinin telah diuji. Selepas 250 hari pengkulturan, tiada proliferasi protokom dapat dilihat pada media kawalan, kinetin dan BAP. Proliferasi protokom dapat dilihat pada 2.0 mg/L TDZ (10.6±1.5) protokorm baru, 1.5 mg/L 2,4-D (8.3±1.1) protokorm baru dan 1.0 mg/L NAA (6.6±2.5) protokorm baru. Proliferasi protokorm yang tertinggi dicatatkan pada media kombinasi 1.0 mg/L NAA+3.0 mg/L TDZ (3.7±2.6) protokorm baru dan 1.5 mg/L 2,4-D+1.0 mg/L TDZ (4.9±0.9) protokorm baru. Untuk perkembangan protokorm P. dayanum, baja berasaskan ekstrak rumpai laut dan baja Orchid Focus pada kepekatan 0.25% (v/v) telah diuji untuk kajian awal. Kemudian, baja ekstrak rumpai laut diuji pada kepekatan yang berbeza (0, 0.01, 0.25, 0.5, dan 1%, v/v). Kompleks aditif keladi, ubi keledek putih dan ubi keledek ungu mentah dan dimasak pada kepekatan 0.25% (w/v) juga diuji untuk perkembangan protokorm. Selepas 180 hari pengkulturan, perkembangan protokorm yang baik adalah pada media rawatan baja berasaskan ekstrak rumpai laut (0.25%, v/v) dengan panjang daun diukur (0.78±0.3 cm) dan akar (0.84±0.6 cm). Baja berasaskan rumpai laut pada kepekatan 0.01% (v/v) menghasilkan peratus pokok hidup sebanyak 96% serta panjang pucuk 1.1±0.3 cm dan akar 2.3±0.8 cm. Ubi keledek ungu dimasak memberikan daun yang tinggi (2.1±0.7 cm) dan ubi keladi dimasak memberikan akar yang panjang (2.9±0.2 cm). Protokol kultur tisu ini berjaya bagi orkid selipar P. dayanum. Namun, percambahan biji benih P. hookerae var volonteanum adalah perlahan dan tiada protokorm berproliferasi. Protokol yang telah dibangunkan ini boleh diaplikasikan dalam program konservasi seperti mewujudkan hasil-hasil tisu kultur yang telah diaklimatisasi pada pasaran awam agar pengambilan secara haram dapat dikurangkan atau dikembalikan semula ke habitat asal agar spesies orkid ini tidak lagi terancam kepupusan. Walau bagaimanapun bagi spesies P. hookerae var volonteanum, kajian lanjut masih diperlukan untuk memendekkan jangka masa percambahan biji benih seterusnya membangunkan protokol yang bersesuaian

A species checklist of wild orchids in selected sites in Kadamaian, Kota Belud, Sabah

A brief orchid diversity study was conducted from 14th to 19th October 2019 in Kadamaian, located northwest of Kinabalu area, through a scientific geographic expedition. A convenience sampling method was employed with only the flowering individuals collected and preserved as herbarium specimens. A total of 58 species and 32 genera were identified during the field visits to the Mount Nopungguk, Melangkap Noriou, Ulu Sungai Melawa, and basecamp site. The finding comprises of 43 epiphytes, 13 terrestrials, and two mycoheterotrophs. Appendicula congesta, Bulbophyllum disjunctum, Dendrobium kiauense, and Goodyera rostellata are species endemic to Borneo found during the expedition along with Crepidium multiflorum, a hyper-endemic species to Kinabalu area. Several rare species which narrowly distributed to primary forest habitat were discovered, including jewel orchids; Cystorchis variegata var. variegata and Anoectochilus geniculatus, and two mycoheterotrophs; Aphyllorchis pallida and Lecanorchis multiflora var. multiflora