Flower endophytic fungi of Geodorum densiflorum endangered orchid

Geodorum densiflorum is an ornamental and medicinally important orchid. The medicinal metabolites are produced by endophytic fungi associated with orchid tissue, particularly flowers. However, there is no report of the endophytic fungi from the G. densiflorum flowers. This research aimed to study the endophytic fungi from different parts of G. densiflorum flower. Fungal isolation was carried out from sepal, petal, labium, stigma, and anther using surface sterilization method. The fungi were identified by combining morphological and molecular characteristics of ITS rDNA region. The results showed that each flower organ had different species of endophytic fungi. A total of seven species of endophytic fungi were obtained; four species were successfully identified by molecular identification and three species based on morphology. The four species, namely Hypomontagnella barbarensis, Aspergillus oryzae, Curvularia pseudobranchyspora, and Nigrospora chinensis, while the three species, namely Gonatobotrys sp., Humicola sp., Aspergillus section Nigri. The labium inhabited by Curvularia pseudobranchyspora, Nigrospora chinensis, Aspergillus section Nigri. Three species isolated from petals, namely Hypomontagnella barbarensis, Gonatobotrys sp., and Aspergillus oryzae. The sepal is inhabited by Humicola sp. There were no endophytic fungi in stigma and anther. This indicates that each flower part is a unique habitat of endophytic fungi.

Multiple shoot induction and regeneration of Vanilla borneensis Rolfe - a critically endangered orchid of Assam, India

In the present investigation, a micropropagation protocol has been developed for Vanilla borneensis Rolfe – a critically endangered orchid through multiple shoot regeneration. Through in vitro multiple shoot regeneration from both nodal and shoot tip explants, maximum (100%) shoot induction was observed. The minimum time required for shoot bud induction was observed from the shoot tip (5–7 days) on medium supplemented with BAP (4.44 mM) + KIN (2.32 mM) as compared to the nodal explants. Maximum multiple shoot regeneration was observed from nodal explants on the medium supplemented with BAP (4.44 mM) + TDZ (6.82 mM). However, maximum shoot length was observed on the medium supplemented with BAP (4.44 mM) + 15% CW and the number of nodes (5.27±0.33) per shoot after 90 days. Maximum (80-100%) of root initiation was observed in almost all the concentrations of NAA. The shortest time of root initiation was found on the medium supplemented with NAA (5.37 mM). Further, acclimatization period was found to be 15 days with 70% acclimatization while 60% of survivability was observed in the field condition. This efficient micropropagation method of V. borneensis could be successfully used for mass propagation as well as conservation of the critically endangered wild orchid.

Floral Trait and Mycorrhizal Similarity between an Endangered Orchid and Its Natural Hybrid

Hybridization can often lead to the formation of novel taxa which can have traits that resemble either or both parental species. Determining the similarity of hybrid traits to parental taxa is particularly important in plant conservation, as hybrids that form between rare and common taxa may more closely resemble a rare parental species, thereby putting the rare parental taxon at further risk of extinction via increased backcrossing and introgression. We investigated the floral (morphological and chemical) traits and orchid mycorrhizal (OrM) fungal associations of the endangered orchid Orchis patens, its more common sister species O. provincialis, and their natural hybrid O. × fallax in natural sympatric populations. We found that both morphological and chemical floral traits of O. × fallax are shared by the parents but are more similar to O. patens than O. provincialis. OrM fungi were shared among all three taxa, indicating that the availability of OrM fungi should not represent a barrier to establishment of individuals of any of these taxa. These results suggest that O. × fallax may be able to expand its distribution within a similar niche to O. patens. This highlights the importance of quantifying differences between hybrids and parental taxon in species conservation planning.

Determination of Sites of Special Importance for the Conservation of Threatened Orchid Species in Colombia

Colombia is the country with the highest number of orchid species (4270), whose optimal habitat is cold and humid forests. However, the outlook for conservation is alarming, considering that deforestation is causing the loss of millions of hectares of forests. This situation has led to the existence of 206 endangered orchid species. Therefore, this research was conducted to determine Sites of Special Importance for the Conservation of Threatened Orchid Species in Colombia (SSICO), through an analysis of their spatial and altitudinal distribution using various databases, to make a selection of nature reserves on a municipality scale, using Marxan software, and employing relevant parameters (richness, rarity, and IUCN category). Furthermore, the results were later compared with the Protected Areas System, determining their coverage to propose SSICOs. 674 records of the presence of threatened orchids in 277 municipalities were obtained. Urrao, Abrego, and Frontino were the areas with the greatest richness and rarity. Marxan selected 47 municipalities located mostly in the Andes region, and four SSICOs were prioritized, which are located in the Antioquia, Norte de Santander, Nariño and Putumayo provinces. These SSICOs, in addition to being points of great biodiversity, are areas with special socio-economic characteristics that influence the management of natural resources. These areas require timely attention, research, and intervention by environmental authorities because of their importance for conserving orchids and Andes Forests.

Development of species-specific SCAR markers for identification and authentication of three rare Peninsular Malaysian endemic Coelogyne (Orchidaceae) orchids

Background: Coelogyne kaliana, Coelogyne stenochila and Coelogyne tiomanensis are three valuable rare orchid species endemic to Peninsular Malaysia, currently rampantly traded illegally via the internet and through local nurseries, which label them as hybrids to avoid enforcement detection. Drastic measures to ensure the continued existence of their populations in the wild should be introduced as they are rapidly diminishing into extinction, including the development of rapid and accurate species-specific identification tools. These three orchid species are highly similar morphologically and currently it is impossible to distinguish among them without their reproductive structures. Methods: RAPD-based species-specific SCAR markers were developed to distinguish and authenticate the identity of these three endemic Peninsular Malaysian Coelogyne species. Results: Three SCAR markers were successfully developed in this study. SCAR marker primer pair, CKL_f / CKL_r was specific to C. kaliana as it produced a unique single band of 271 bp but not in C. stenochila and C. tiomanensis. SCAR marker primer pair CST_f / CST_r amplified a single band of 854 bp in C. stenochila and two bands of different sizes (372 bp and 858 bp) in C. tiomanensis, but no amplification in C. kaliana. The third SCAR marker primer pair, CTI_f / CTI_r produced a single band (about 500 bp) for both C. stenochila and C. tiomanensis, but showed no amplification in C. kaliana. Conclusions: Although not all these SCAR markers were species amplification specific, they could be used to discriminate among the three Coelogyne species effectively. Accurate species identification is one of the most important steps to allow a proper management plan to be established in the effort to conserve these three endangered orchid species of Peninsular Malaysia. Besides, it could effectively put a stop to the illegal trading of these rare endangered orchid species worldwide.