Wicaksono A, Mursidawati S. 2020. Short Communication: Lugol’s iodine test on Rafflesia patma-Tetrastigma leucostaphylum intersection tissue for preliminary starch visualization. Nusantara Bioscience ...12: 91-96. As holoparasitic plant, Rafflesia has no recognizable plastid genome, but it has plastid-like organelle. Despite the fact that it obtains nutrients from host plant, it is unknown if Rafflesia stores primary metabolites, such as carbohydrates, from its host. A study was performed to visualize the starch in Rafflesia patma Blume proximal tissue which was intersected to its host root, Tetrastigma leucostaphylum (Dennst.) Alston, using modified Sachs’ test with Lugol’s iodine. The result revealed the absence of blackening in the R. patma tissue caused by starch reaction with the iodine, but occurred in the root cortical tissue of T. leucostaphylum. The absence of starch in R. patma tissue indicated that possibly the plastid-like organ has no similar function to amyloplast, and starch is not used for storage in the flower. It is likely that R. patma relies completely on the host’s photosynthate to maintain the flower metabolism during anthesis period. However, detailed histochemical analysis for starch or carbohydrate is needed for confirmation whether the starch is existing even in small quantity, and molecular genetic observation on sucrose intake and flowering (anthesis) regulatory genes will also be required to confirm if Rafflesia takes the photosynthate directly from its host and possibility if Rafflesia coordinates the formation of flower bud and anthesis between the endophytic tissues to prevent overexploitation of nutrients from its host.
Plants in the genus Rafflesia (Rafflesiaceae) bear the largest flowers in the world. Almost all members of this holoparasitic plant family have limited ranges and are rare or threatened. This genus ...is therefore important from a scientific and conservation perspective. An ex situ conservation collection of a population of Rafflesia patma Blume has been monitored at Bogor Botanic Garden (BBG) since 2004, the first time that this has ever been done. Studies have been made, including propagation trials on both seed and vegetative material at BBG. Of all the propagation methods tested, grafting on to the host plant Tetrastigma scariosum (Blume) Planch proved to be the most successful for growing R. patma, resulting in blooming having occurred seven times since 2010. Grafting is the quickest way to propagate Rafflesia in cultivation. This cultivation process provides a new hope for the conservation of this endangered and charismatic genus. This paper describes the methods trialled including both unsuccessful techniques as well as those which resulted in blooming events.
The early life history of Rafflesia R.Br. is hidden by its cryptic nature. Being a holoparasitic plant, early growth and development of species in this genus have been suggested to occur inside the ...host plant. In this study, using microscopic analyses, we examined how Rafflesia patma Blume grows from a simple clump of cells or protocorm inside the cambium tissue of Tetrastigma leucostaphylum (Dennst.) Alston, its host plant. We further show how the protocorm expands in about 2 months into an early form of a flower knob or bud, just before the flower begins to emerge from the host plant vine. Our study suggests that the endophyte, R. patma, spreads within T. leucostaphylum vascular cambium tissue in a linear manner, but not as a continuous strand. We suspect that the parasitic endophyte spreads inside the host vascular cambium and is pushed farther away from its point of origin to another part of the host as the host vine cambium fusiform initial cells divide and enlarge over months. Moreover, the protocorm matures in the cambium region of the T. leucostaphylum vine which lies close to the xylem, and grow outwards, bursting through the host phloem, then the host cortex, and finally towards the host periderm as it begins to emerge. Although we believe this might be the closest prediction on how Rafflesia develops at an early stage and is distributed within its host body, a future analysis of endophytic tissue growth in a controlled environment, such as the use of tissue culture, is required to strengthen our findings.
•Rafflesia patma Blume is a holoparasitic plant.•Rafflesia patma parasitizes the cambium tissue of Tetrastigma leucostaphylum.•We use microscopic analyses to assess protocorm growth and flower knob formation.•Rafflesia patma spreads within T. leucostaphylum vascular cambium linearly.•The protocorm matures in the cambium region of the T. leucostaphylum vine.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Sukarno N, Mursidawati S, Listiyowati S, Nugraha NH, Fadillah WN, Waite M. 2023. Root associated Fusarium solani Species Complex (FSSC) in epiphytic and terrestrial orchids. Biodiversitas 24: ...2577-2586. Members of the Fusarium solani Species Complex (FSSC) are ecologically diverse, covering human and plant pathogens, saprobes, and endophytic fungi of economically important plants. The role of FSSC on tropical orchids, however, has received little attention. This research aimed to isolate and identify the FSSC found as endophytes in roots of the epiphytic orchids Cymbidium finlaysonianum Lindl. and Vanda tricolor Lindl., and the terrestrial orchids Calanthe triplicata (Willemet) Ames and Phaius tankervilleae (Banks) Blume. Fungal isolation was obtained from peloton structures within the root. Fungal identification was done using combined morphological and molecular characteristics using ITS rDNA sequences. Four isolates of Fusarium were identified based on morphological characteristics. The BLAST analysis showed that all the isolates were in the Fusarium solani Species Complex (FSSC). Further phylogenetic relationship analysis indicated that all the FSSC isolates belonged to FSSC5 lineage or Fusarium solani sensu stricto, which is nested in FSSC clade 3 as a subgroup. The fungi F. solani sensu stricto V2 and CF34 were isolated from the epiphytic orchids, and F. solani sensu stricto C5 and P44 were isolated from the terrestrial orchids. It is interesting that the FSSC5 isolated from the epiphytic orchids form different groups than those of the terrestrial orchids. This is the first report showing the tropical orchids are the host of the FSSC5 as endophytes and this broadens the known host range and ecological behavior of the FSSC5.
Rafflesia spp Wicaksono, Adhityo; Mursidawati, Sofi; Sukamto, Lazarus A. ...
Planta,
08/2016, Volume:
244, Issue:
2
Journal Article, Book Review
Peer reviewed
Members of the genus Rafflesia (Rafflesiaceae), which are holoparasitic plants known to grow on a host vine, Tetrastigma sp., are widely spread from the Malayan Peninsula to various islands ...throughout Indonesia. The plant’s geographical distribution as well as many other aspects pertaining to the basic biology of this genus have still not been studied. The young flower buds and flowers of wild Rafflesia hasseltii Suringar, Rafflesia keithii Meijer and Rafflesia cantleyi Solms-Laubach are used in local (Malaysia and Indonesia) traditional ethnomedicine as wound-healing agents, but currently no formal published research exists to validate this property. To maintain a balance between its ethnomedicinal and ornamental use, and conservation, Rafflesia spp. must be artificially cultivated to prevent overexploitation. A successful method of vegetative propagation is by host grafting using Rafflesia impregnated Tetrastigma onto the stem of a normal Tetrastigma plant. Due to difficulties with culture contamination in vitro, callus induction was only accomplished in 2010 for the first time when picloram and 2,4-D were added to a basal Murashige and Skoog medium, and the tissue culture of holoparasitic plants continues to be extremely difficult. Seeds harvested from fertile fruit may serve as a possible method to propagate Rafflesia spp. This paper provides a brief synthesis on what is known about research related to Rafflesia spp. The objective is to further stimulate researchers to examine, through rigorous scientific discovery, the mechanisms underlying the ethnomedicinal properties, the flowering mechanisms, and suitable in vitro regeneration protocols that would allow for the fortification of germplasm conservation.
Tetrastigma (Vitaceae) are a genus of tropical lianas that are hosts for parasitic plants in the family Rafflesiaceae. Since the stem of Tetrastigma is creeping and often devoid of leaves, it is ...often mistakenly visually identified as a root. Rafflesia flowers at the ground level (i.e. terrestrial flowers) have been documented to grow on Tetrastigma roots, but it is unclear whether those that arise aerially emerge from host root or stem organs. In this study, we present a new, lessdestructive means of sampling Tetrastigma leucostaphylum, which is a host of many Rafflesia species. Histological observation of two 250 cm above-ground aerial samples confirmed that both were stems, as shown by distinct anatomical features (compactly arranged vascular bundles with simple phloem bundles) in contrast to roots, which had loosely arranged vascular bundles with ramified phloem bundles. Clearer identification of Tetrastigma vine infected with aerial Rafflesia is needed to confirm whether the parasitic flower grows on a stem or on an erect aerial root to better understand the biology of this enigmatic Rafflesia species that have evaded cultivation and ex situ conservation efforts. This new sampling method may be a useful way to identify the Tetrastigma vine in the future due to its less invasive nature compared to severing the whole vine.
Societal Impact Statement
Rafflesia
is the genus that contains the world's largest flowers. Despite their global appeal, most of the 42 known species are now at risk of extinction. Urgent action is ...needed to protect these remarkable flowers. A combined approach to conservation is recommended, including a greater level of habitat protection and support for local community action groups.
Rafflesia
is a suitable new icon for conservation in the Asian tropics.
Summary
The genus
Rafflesia
, which includes the world's largest flowers, has aroused curiosity among scientists for centuries and features prominently in local culture across Southeast Asia. The plant has long been used in ethnobotanical medicine and, more recently, as a source of revenue from ecotourism. But despite its acclaim,
Rafflesia
remains poorly understood in many respects. Taxonomy is disputed, new species are described each year, and the plant has proven recalcitrant to cultivation. This has hindered conservation, and most of the 42 known species are now severely threatened, yet only one is listed by the International Union for Conservation of Nature (IUCN). We estimate that 60% of
Rafflesia
species face a severe risk of extinction (equivalent to Critically Endangered CR). Moreover, we predict that at least 67% of known habitats fall outside protected areas, exacerbating their vulnerability. Alarmingly, recent observations suggest taxa are still being eradicated before they are even known to science. We present recent scientific discoveries and probable extinctions and highlight case studies of conservation success, with a focus on the role of local people. We propose a multi‐pronged conservation approach combining strengthened taxonomy, ex situ propagation, ecotourism, and an extension of protected areas. We suggest action devolved to local communities and awareness campaigns linked to social media networks will be crucial outside of protected jurisdictions. Finally, we propose to establish
Rafflesia
as a new icon for plant conservation in the Asian tropics. A combined approach might just save some of the world's most remarkable flowers, most of which are now on the brink of being lost.
Ang
Rafflesia
ay isang urihay ng halaman na kilalang namumukadkad ng pinakamalalaking bulaklak sa buong mundo. Bagama't nakabibighani at kahalihalina, karamihan sa apatnapu't dalawang sarihay ay nanganganib pa ring malipol sa kanilang likas na pinamumugaran. Dahil rito, kailangan ng agarang pagkilos upang maproteksyunan ang kahanga‐hanga at kakaibang mga bulaklak na ito. Matitiyak ang pangangalaga ng mga ito sa pamamagitan ng kolektibong pamamaraan tulad ng masidhing pangangalaga sa kagubatan, at pagpapaigting ng suporta sa lokal na komunidad. Ang
Rafflesia
ay simbolo ng konserbasyon sa tropikal na kagubatan sa Asya.
Rafflesia
adalah genus kepada bunga terbesar di dunia. Walaupun menjadi tarikan diperingkat global, dengan kebanyakan spesies daripada sejumlah 42 spesies telah kenalpasti, namun masih mempunyai risiko untuk mengalami kepupusan. Oleh itu, tindakan bagi melindungi bunga yang istimewa ini perlu dilaksanakan dengan kadar segera. Gabungan pendekatan untuk pemuliharaan adalah disyorkan, iaitu merangkumi usaha yang lebih tinggi bagi meningkatkan tahap perlindungan habitat, dan memberi pelbagai jenis sokongan dan dokongan untuk kumpulan tindakan dikalangan masyarakat setempat. Ini dapat menjadikan
Rafflesia
sebagai ikon baharu yang sesuai untuk usaha pemuliharaan di kawasan tropika Asia.
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DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, UILJ, UKNU, UL, UM, UPUK
Orchids form a symbiotic relationship with endophytic fungi throughout their life cycle, including their seed germination. Research on orchid endophytic fungi in Indonesia is still limited. This ...study aimed to isolate and identify endophytic fungi of Phalaenopsis amabilis, Dipodium brevilabium, Vanilla planifolia, and Aerides odorata orchids. Endophytic fungi were isolated from surface-sterilized orchid root pelotons. Fungal identification was based on morphological and molecular characteristics using ITS rDNA sequences, followed by BLAST and phylogenetic tree analysis. All orchids studied had peloton structures in root cortex tissues. From the four orchids studied, 22 fungal isolates were obtained. Five fungal isolates were obtained from the P. amabilis orchid, consisting of 1 Penicillium and 4 sterile mycelium isolates. The D. brevilabium had 7 fungal isolates, namely 1 Penicillium and 6 Fusarium isolates. The V. planifolia had 6 Fusarium isolates, whereas the A. odorata had 4 isolates, such as 1 Penicillium, 2 sterile mycelium, and 1 unidentified isolate. The genus Penicillium was found in 3 out of the 4 orchid species studied. One Penicillium isolate was molecularly identified as Penicillium citrinum. Endophytic fungi in P. amabilis were mostly sterile mycelium, while D. brevilabium, V. planifolia, and A. odorata were dominated by Fusarium.
Many orchids require mycorrhizal symbioses with fungi for their development and survival. Rhizanthella gardneri the Western Australian underground orchid is associated with the companion plant ...Melaleuca uncinata and its ectomycorrhizal fungus symbiont. Much less is known about the habitat requirements of its sister species, R. slateri, which occurs in Eastern Australia. The absence of chlorophyll from Rhizanthella gardneri and R. slateri results in total dependency on associations with fungal symbionts. Many ecological and biological aspects of these fascinating orchids remained poorly known, including the identity of the fungal associates and the nature of their tripartite associations with Rhizanthella and Melaleuca. Extremely high specificity of these mycorrhizal relationships is likely to be the most important factor explaining the highly specific habitat requirements of underground orchids. The purpose of this study was to conduct further investigations of the role of the mycorrhizal associations of Australian underground orchids by identifying the fungi involved in these associations, optimising their growth in sterile culture and devising efficient means for synthesising their tripartite associations with R. gardneri and M. uncinata. In total, 16 isolates of fungi were successfully obtained from the two underground orchids and used in a series of experiments to understand both the nature of the fungi and their relationship with orchids. The identity of these fungi was established by using conventional morphological and molecular methods. Cultural and morphological studies revealed that all isolates from R. gardneri and R. slateri were binucleate rhizoctonias with affinities to members of the genus Ceratobasidium. However, the teleomorph state that was observed from the R. slateri symbiont during this study more closely resembled a Thanatephorus species. Further identification using ITS sequence comparisons confirmed that mycorrhizal fungi of Rhizanthella belonged to the Rhizoctonia alliance with relatives that include Thanatephorus, Ceratobasidium, or Rhizoctonia from other continents with over 90% similarity. Most of these related fungi are known as plant pathogens, but some were orchid mycorrhizal fungi. However, the isolates from the two underground orchids were most closely related to each other and formed a discrete group relative to other known members of the Rhizoctonia alliance. Sterile culture experiments determined culture media preferences for mycorrhizal fungi from Rhizanthella and other orchids. A fully defined sterile culture medium designed to more closely resemble Australian soil conditions was formulated. This new medium was compared to undefined media containing oats or yeast extract and recommendations for growth of these fungi are provided. The undefined media based on oats provided the best growth of most fungi, but the new Australian soil media was also effective at growing most orchid mycorrhizal fungi and this fully defined media was less prone to contamination and should provide more reproducible results. A comparison of three methods for inoculating M. uncinata with the underground orchid fungi resulted in the production and characterisation of ectomycorrhizal roots and hyphae formed by fungi isolated from R. gardneri and R. slateri. These underground orchid fungi could easily be distinguished from other mycorrhizal fungi (caused by airborne contamination) by the characteristic appearance of these roots and hyphae. A new system for growing and observing tripartite mycorrhizal associations was devised using pots with side viewing windows and the use of transparent seed packets to contain Rhizanthella seeds. This method allowed all the stages of seed germination to be observed in the glasshouse, culminating in the production of underground orchid rhizomes. Seed germination was only successful when seed was placed directly over active M. uncinata ectomycorrhizas confirmed to belong to the correct fungus by microscopic observations through the side of window pots. The importance of these new scientific discoveries concerning the biology and ecology of the underground orchids and their associated fungi for the recovery of these critically endangered orchids are discussed.
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