Ex Situ Research Articles

REBUILDING OF ORCHID GERMPLASM CONSERVATION VIA MOLECULAR MULTI-OMICS APPROACHES

The Orchidaceae family, the world’s second-largest flowering plant, dominates the International Union for Conservation of Nature (IUCN) red list. Threats to orchid survival include illegal harvesting, habitat destruction, temperature change, and unique association requirements in pollination systems and orchid mycorrhiza interaction. Some research breakthroughs in ex-situ and in-situ germplasm integration techniques for conservation application, including automated precision approaches in orchid research. Exsitu conservation approaches, including gene bank, seed bank, cryopreservation, field gene bank, and in-vitro culture, can play indispensable roles in the success of orchid conservation and preservation, especially for endangered orchids. Recently, multi-omics technology emerged as an important tool to bridge the knowledge gaps in orchid conservation so more effective measurement could take place. These provide a good conservation framework that confluences with technology and knowledge.

Population structure and diversification of Gymnospermium kiangnanense, a plant species with extremely small populations endemic to eastern China

Background Gymnospermium kiangnanense is the only species distributed in the subtropical region within the spring ephemeral genus Gymnospermium. Extensive human exploitation and habitat destruction have resulted in a rapid shrink of G. kiangnanense populations. This study utilizes microsatellite markers to analyze the genetic diversity and structure and to deduce historical population events of extant populations of G. kiangnanense. Methods A total of 143 individuals from eight extant populations of G. kiangnanense, including two populations from Anhui Province and six populations from Zhejiang Province, were analyzed with using 21 pairs of microsatellite markers. Genetic diversity indices were calculated using Cervus, GENEPOP, GenALEX. Population structure was assessed using genetic distance (UPGMA), principal coordinate analysis (PCoA), Bayesian clustering method (STRUCTURE), and molecular variation analysis of variance (AMOVA). Population history events were inferred using DIYABC. Results The studied populations of G. kiangnanense exhibited a low level of genetic diversity (He = 0.179, I = 0.286), but a high degree of genetic differentiation (FST = 0.521). The mean value of gene flow (Nm) among populations was 1.082, indicating prevalent gene exchange via pollen dispersal. Phylogeographic analyses suggested that the populations of G. kiangnanense were divided into two lineages, Zhejiang (ZJ) and Anhui (AH). These two lineages were separated by the Huangshan-Tianmu Mountain Range. AMOVA analysis revealed that 36.59% of total genetic variation occurred between the two groups. The ZJ lineage was further divided into the Hangzhou (ZJH) and Zhuji (ZJZ) lineages, separated by the Longmen Mountain and Fuchun River. DIYABC analyses suggested that the ZJ and AH lineages were separated at 5.592 ka, likely due to the impact of Holocene climate change and human activities. Subsequently, the ZJZ lineage diverged from the ZJH lineage around 2.112 ka. Given the limited distribution of G. kiangnanense and the significant genetic differentiation among its lineages, both in-situ and ex-situ conservation strategies should be implemented to protect the germplasm resources of G. kiangnanense.

Essential Omega-3 (n-3PUFA) Nutritional Content of Ptilinopus melanosphila and Chalcophaps indicia Meat

Ptilinopus melanosphila and Chalcophaps indica, known as Puna birds among local people of the Sula Islands, North Maluku, Indonesia, while Treron aromaticus, known as Puna Giu, belong to the Columbidae family. These birds are frequently hunted for consumption due to the delectable taste of their meat. This study aims to examine the essential Omega-3 (n-3PUFA) nutritional content, utilizing these two avian species (Ptilinopus melanosphila and Chalcophaps indicia) as samples. Local perceptions consider these birds as identical, hence this study also provides insights into the disparities between the two species based on meat quality analysis. The analysis encompasses proximate analysis, fatty acids, and amino acids, conducted on 6 samples, each encompassing 3 breast meats. These samples were mixed and blended, followed by chemical analysis at the integrated laboratory of Universitas IPB Baranangsiang, with proximate analysis executed at PAU IPB. The results reveal that Ptilinopus melanosphila meat contains crucial nutrients for human consumption, specifically long-chain omega-3 unsaturated fatty acids (n-3PUFA), which are more abundant compared to Chalcophaps indica and several other meat products. The myriad of significant benefits offered by this avian species underscores the importance of recommending its preservation to parties engaged in ex-situ conservation efforts. Furthermore, it is advocated to stimulate the establishment of regional culinary businesses specializing in nutrient-rich Black-Naped Fruit Dove meat.

Enhancing biotechnological approaches for the in vitro micropropagation: Protecting endangered wild tulip species in Samarkand, Uzbekistan

Tulipa, a genus of bulbous geophytes, boasts a globally recognized ornamental value. Anthropogenic influences have precipitated a decline in the wild populations of Tulipa, including those in Uzbekistan, necessitating the development of forward-looking conservation strategies. This study focuses on advancing biotechnological strategies for in vitro micropropagation to conserve endangered wild Tulipa species in Samarkand, Uzbekistan. Tulipa ingens and Tulipa fosteriana, listed as vulnerable and near threatened, respectively, on the IUCN Red List, face numerous anthropogenic threats including habitat loss and overexploitation. Our research demonstrates the effectiveness of sterilization protocols, successful micropropagation from seeds, and the importance of adapting to non-sterile conditions. Through meticulous experimentation, a novel sterilization approach involving a 5 percent Domestos solution coupled with brief ethanol exposure yielded the highest viability rates for plant materials. Micropropagation from seeds resulted in successful shoot and microbulb formation, providing a promising conservation strategy for these rare species. The study underscores the urgency of conservation efforts in Uzbekistan, where the extent of occurrence and area of occupancy of these species classify them as endangered. Our findings contribute valuable data for ex-situ conservation measures, offering hope for the preservation of red-listed Tulipa species in the region.

Ex-situ Conservation of Woody Plants within the Framework of the Millennium Seed Bank Partnership in Georgia

Ex-situ Conservation of Woody Plants within the Framework of the Millennium Seed Bank Partnership in Georgia

Application of 4A Concept in Bogor Botanical Garden Tourism Development Strategy

Indonesia is very rich in natural tourism potential (ecotourism), one of which is natural tourism in conservation areas. Bogor Botanical Garden is an ex-situ conservation area that has long been used by the wider community as a tourist attraction and in many countries, Bogor Botanical Garden is one of the most popular tourist destinations because it presents a panoramic view of natural landscape architecture. The research entitled "Application of the 4A Concept in the Development of Bogor Botanical Garden Tourism" focuses on the 4A concept, namely Attraction, Accessibility, Amenities, and Ancillary services. This type of research is qualitative, using qualitative descriptive analysis methods. Data collection is carried out by means of field observation, interviews, and literature studies. SWOT analysis was also used in this study, to identify the strengths and opportunities of Bogor Botanical Garden, and determine strategies to reduce weaknesses and challenges. The results showed changes in the management system of Bogor Botanical Garden. By analyzing the 4A concept, this study seeks to provide recommendations for the right development strategy for KRB, so that it can develop better and the vision of Bogor Botanical Garden can be achieved.

Conservation and Landscape Evaluation of Some Endangered Species in Türkiye

In addition to in-situ conservation activities related to endangered taxa, it has a great importance to provide ex-situ conservation activities to increase awareness and to determine their usage areas by examining reproduction methods. In the line with, the aim of the study is to prevent the extinction of some endangered endemic taxa in Ayaş, Beypazarı and Nallıhan regions of Ankara province in Türkiye, to evaluate the taxa within the framework of sustainable conservation and use principles by raising awareness, and thus to protect the biodiversity in the region. For these purposes, during their vegetation periods throughout the years 2020-2022, some observations were recorded in landscape point of view from the habitats of 9 Critically Endangered (CR) and 3 Vulnerable (VU) taxa. Moreover, all the taxa are photographed and have been archived in the Collections of National Botanical Garden of Türkiye (NBGT) as National Herbarium (TC), DNA and Tissue Banks. Also, samples have been kept in 70% EtOH for Microscobic Slide Collection of NBGT. In addition, the possibilities of using taxa in the landscape area were evaluated. The results of the habitat observations for years serve as a basis for future conservation and landscape gain efforts for the taxa.

The Relationship between Future Wilderness and in Situ and Ex Situ Plant Conservation

Extrapolation of current (1993) rates of wilderness destruction indicate that no significant areas are likely to remain by the year 2300. This likely trend is reinforced by the current continued growth of the human population. The concepts of in situ and ex situ plant species conservation are examined in the context of wilderness loss and practical consequences are discussed including the concept of ad valorem species conservation.

Evaluation of the initial growth of Shorea platyclados wildlings through the application of mycorrhizal biofertilizer under different levels of shading

The obstacles to the propagation of Shorea platyclados involve the uncertain flowering and fruiting seasons, the recalcitrant nature of the seeds, and the dependence on shade for growth. Furthermore, a mutualistic symbiosis, called mycorrhizae, usually exists between the roots and fungi, similar to other species of Dipterocarpaceae. This consequently hinders the regeneration process, which in turn may threaten the sustainability of this species in the wild. Ex-situ conservation using wildings is one of the measures that can be taken to overcome this issue. This study aimed to assess the initial growth of S. platyclados seedlings by administering different amountsof mycorrhizal biofertilizers under varying shade conditions at four and eight weeks of age in the nursery. A randomised complete block design was employed with the dose of biofertilizer as the treatment (0, 2.5, 5, 7 and 9 g/polybag) and shade as the group (0, 50, 60 and 70%). Observations were taken in the nursery at four and eight weeks of age. The observation parameters included the survival percentage, height growth, diameter growth, and vigour of the seedlings. There were significant differences in percentage of survival. Still, significant differences in height growth were observed with different doses of biofertilizer and shading at both four and eight weeks of age. Significant differences in diameter growth were observed with different doses of biofertilizer at both four and eight weeks of age, but not with different shades. No significant differences were observed for seedling vigour with different doses of biofertilizer and shade at both four and eight weeks of age. The optimal amount of biofertilizer was 5 g/polybag, resulting in maximum growth. This was achieved in a shade of 60%.

Workforce Motivation at Malaysian Orangutan Conservation Centres

Most conservation research and efforts focus on a targeted animal taxon and we often tend to forget about the human component in conservation work. It is often alleged that conservation work is in itself rewarding, rather than motivated by monetary gain. However, this remains anecdotal, and this study approaches this topic through qualitative analysis of the motivation of the workforce in orangutan conservation. The objective of this paper was to assess the driving motivations for conservation workers at three Malaysian ex-situ conservation facilities for orangutans. Using structured interviews and a thematic approach, we classified five main types of motivation amongst the workers. The “Environment/Surrounding” factor describing positive team work experiences emerged as the primary motivator for conducting this job, whereas “Monetary” and “Conservation” were similarly often mentioned as the second motivators. “Personal Development” and “Humanistic” factors were the third and least mentioned motivators, respectively. Our results indicate the importance of providing and maintaining a good work environment and relationship with co-workers, as well as adequate salaries to motivate the conservation workforce. The findings from this study could help the management of any conservation areas/centers to effectively manage and encourage their workforce towards a better care of captive animals

A Study on the Ex-Situ Conservation and Promotion of Medicinal Plants in Dr. Sushila Tiwari Herbal Garden, Rishikesh, Uttarakhand, India

A study was conducted at the Dr. Sushila Tiwari herbal garden in Rishikesh, Uttarakhand, with the aim of scientific documentation and data collection of medicinal plants. The study found there are approximately 126 species present in the herbal garden, trees being the most abundant species followed by shrubs, herbs, climbers and grasses. The major families found in the herbal garden are Fabaceae, Lamiaceae, Apocynaceae, Moraceae, Poaceae, and Solanaceae etc. Critically endangered species such as Chlorophytum borivilianum, Endangered species such as Oroxylum indicum, Ginkgo biloba, Cammiphora wightii, Rauvolfia serpentina, and Strychnos lucida. Near threatened species such as Aegle mermelos, were also found in the garden. Herbal gardens are crucial for conserving rare and endangered medicinal plants, while also promoting the utility of commonly available and frequently used medicinal plants among people, beneficiaries and other stakeholders.

In Vitro Multiplication and Ontogeny of Ferns Tectaria zeylanica (Houtt.) (Dryoptridaceae) from South India under Current Climate Conditions

The oak leaf fern, Tectaria zeylanica (Houtt.) Sledge, is an endangered fern in India, native to the Southeast Asian region. For the ex-situ conservation of this endangered fern, spores were collected and cultured aseptically on full-strength KC Medium with sucrose at pH 5.8. The spores are monolete with a prominent winged perispore. The exine ruptures at the laesura, and three or four germ filaments grow out, following a Vittaria-type germination pattern. The prothallial development was of the Aspidium type. Adult gametophytes are cordate and much broader than their length. Antheridia develop between 120 and 140 days. They are characteristically 3-celled, with a basal cell, a median cell, and an opercular cell. The female sex organs, archegonia, develop between 150 and 170 days. They are superficial and arranged on the lower side of the prothallus near the midrib and the meristematic region. Fertilization and subsequent development of the sporophytes were observed after four months. The highest percentage of sporophyte formation was observed on KC medium fortified with Kin 0.5 mg/l. Sporophytes had uniseriate, unicellular, and multicellular hairs, similar to those found on adult prothalli. The highest percentage of root formation was observed in the medium with IBA + NAA (1.0 mg/l). The in vitro raised plants showed a 54% establishment during hardening in the field. The present study has established a reproducible protocol for the ex-situ conservation of this endangered oak leaf fern.

Will Dalbergia species survive climate change? Predicting the potential future distribution of threatened species in Madagascar

Many species of Dalbergia, the source of highly prized rosewood, are threatened with extinction, a situation compounded by climate change. Accurate estimates of how suitable habitat will be impacted by global change are needed to inform their sustainable management. We selected five commercially exploited species endemic to humid eastern Madagascar (D. baronii, D. louvelii, D. maritima, D. normandii, and D. orientalis) to explore the potential value of species distribution modeling for understanding factors influencing their current distributions and estimating future patterns of suitable habitat, using MaxEnt and locality information for 715 taxonomically and spatially validated occurrences. Eight bioclimatic variables were used for modeling; projections were made under three scenarios for climate change and two for CO2 emissions. The distribution of each of the five species is primarily influenced by precipitation seasonality. Their potential geographical range is predicted to decrease by 2100 (-5% to -33%) under the optimistic CO2 scenario (RCP 2.6) except for a slight expansion of D. normandii (+2%), and by -14% to -58% under the pessimistic scenario (RCP 8.5). Our results show that suitable habitat will likely decrease with climate change, even within protected areas. They provide valuable information for future application of IUCN Red List criterion A to exploited species with fragmented distributions, targeting ex-situ conservation and management, and guiding restoration, reforestation, and other actions to establish and reinforce Dalbergia populations in areas where they are more likely to persist during climate change over the present century and beyond.

Creation of traditional Jain Vatika in Teerthanker Mahaveer University campus, Moradabad, Uttar Pradesh, India - An ex-situ conservation approach

The creation of a traditional Jain Vatika in Teerthanker Mahaveer University (TMU) campus in Uttar Pradesh, India is a holistic and innovative approach to ex-situ conservation approach. This initiative serves a dual purpose: preserving plant species with cultural and religious significance within Jainism and contributing to the broader biodiversity conservation mission. The traditional Jain Vatika has become a spiritual and educational sanctuary, fostering a deeper understanding of Jain culture and values among students and visitors. Simultaneously, it plays a vital role in the conservation of plant diversity, providing a secure harbor for a diverse range of species. The twenty-four plants are planted in the traditional Jain Vatika which represent the twenty-four Lord Teerthankers of Jain religion.

Human factors and health status of the highland forest ecosystems in North Shewa Zone, Central Ethiopia

The main purpose of this study was to evaluate anthropogenic factors and ecological health status of the forest ecosystems in the north Shewa zone (NSZ), Ethiopia. Tree cutting, grazing, cultivation, and fires were the major anthropogenic factors that led to deforestation and loss of biodiversity. The ecological analysis showed that the sites of Mehal-Wonz, Mehal-Tsebel, and Abogedam forests were very heavily disturbed, while Debdebo and Zego forests were heavily and moderately disturbed, respectively, implying the severe degradation of most forests in NSZ. Therefore, to restore the degraded forest areas of the zone, in-situ and ex-situ community-based conservation strategies were recommended.

A preliminary survey of the flora and vegetation of an ex-situ conservation area (Hyde Park Zoo Sanctuary and Tropical Gardens Inc., Guyana)

Hyde Park Zoo Sanctuary and Tropical Gardens Inc. is a local ex-situ conservation area located at Land of Canaan, East Bank Demerara, Guyana. It serves to manage and conserve species of wild flora and fauna, boasts several diverse native and exotic species, and hosts a rescue centre for wild captures. Over a six (6) week period, flora & vegetative visual surveys were conducted there to i) identify and classify the species of flora found at accessible parts of the site (according to the official established map of the area), and ii) report these findings in a comprehensive report in order to establish a database of the preliminary findings of the study. Based on the preliminary findings of the study, a total of ninety-nine (99) plant species from twenty (20) families were identified. The largest family group was found to be Arecaceae with 15 species. Leguminosae and Bromeliaceae both contained 6 species each. Other representative families were Apocynaceae, Orchidaceae, Rosaceae and Aracaceae. Seventy-nine species (80.8%) were exotic and nineteen (19.4%) species were found to be native. None of the identified species were found to be under threat, or near extinction. Based on these preliminary findings, it is established that the area has great taxonomic and conservation potential, and with effective management along with strategic planning, the area can serve as an ex-situ conservation model, where both native and exotic species thrive together.

Potential Vulnerability of Natural Populations of Camu-camu (Myrciaria dubia) to Anthropogenic Stressors in Southwestern Amazon

One economically promising Amazonian species is camu-camu (Myrciaria dubia) which fruits have the highest content of vitamin C recorded, with a high potential for applications on the food, supplement and cosmetic industry and which can be a cornerstone for agroforestry initiatives for Amazonian peasants. Camu-camu has an Amazon wide distribution although most studies and collections have occurred on the Central and Western Amazon. The aim of this study is to assess the distribution and potential vulnerability of camu-camu populations in the state of Rondônia, Brazil, the southern limit of its distribution, which exhibit a drier climate. We use data recorded in herbaria and a bioprospecting expedition and data on anthopogenic stressors, estratified at the municipal level, like fire events, deforestation and dams. Camu-camu records occur preferentially in the Central and Northern part of the state, however most of the collections are around cities, major rivers and roads, showing a strong collector bias. Municipalities with camu-camu records suffered higher rates of deforestation and show higher occurrence of fire events and planned dams than municipalities without camu-camu records. Camu-camu populations in Rondônia are thriving on drier conditions and may be more adapted for future prevalent conditions for the Amazon under climate change. These adaptations may be useful for future domestication efforts. We call for an increasing effort of collecting and salvaging camu-camu populations on human dominated landscapes and to encourage participatory conservation with local farmers, for an ex-situ conservation on rural landscapes, especially under the prospect of climate change.

Effect of Pre-treatments on Germination of Pterocarpus marsupium Roxb. for Developing Field Gene-Banks in Eastern India

Aims: Pterocarpus marsupium Roxb. is an important timber species with high medicinal properties belonging to the Fabaceae family. The species is severely threatened by anthropogenic and ecological pressures coupled with less natural regeneration. No results in vegetative propagation methods and very less germination is reported for the species under nursery conditions. This study investigates the germination potential of P. marsupium seeds under the prevailing climatic conditions of regions in eastern India. Place and Duration of Study: The study was carried out at the ICFRE - Institute of Forest Productivity (IFP), Lalgutwa, Ranchi, during the 24th to the 32nd Meteorological Standard Week (MSW) of 2022, spanning from June to August. Methodology: Seeds were collected from three CPTs selected from natural forests of Jharkhand, Bihar and West Bengal and the experiment was carried out following split plot design. Observations on several germination parameters and seed vigour indices were recorded until eight weeks after sowing and subjected to factor multivariate analysis of variance (MANOVA) in SPSS 25.0. Results: The experiment revealed that alternate wetting and drying of seeds significantly (P <. 001) outperformed all other treatments with a highest germination percentage of 60±1.196%. Accessions from Jharkhand and West Bengal exhibited significantly (P < .001) better vigour indices (1959.8, 1855.35 for SVI-I and 538.715, 527.742 for SVI-II, respectively) compared to the accession from Bihar (1796.49 and 532.618 for SVI-I and SVI-II). Although the study did not find any significant interaction between the treatments and accession, there was a statistically significant impact on the aggregate dependent variables (F (126,503.279) = 1.579, P <. 001; Wilks' Λ = .077). Conclusion: The results of this study report an economically feasible and rapid seed treatment method for establishing field gene banks of P. marsupium in eastern India to aid its ex-situ conservation practices.

Transcriptome data analysis provides insights into the conservation of Michelia lacei, a plant species with extremely small populations distributed in Yunnan province, China.

Michelia lacei W.W.Smith (Magnoliaceae), was classified as a Plant Species with Extremely Small Populations (PSESP) by the Yunnan Provincial Government in both action plans of 2012 and 2021. This evergreen tree is known for its high ornamental and scientific value, but it faces significant threats due to its extremely small population size and narrow geographical distribution. The study aims to understand the genetic structure, diversity, and demographic history of this species to inform its conservation strategies. The analysis of transcriptome data from 64 individuals across seven populations of M. lacei identified three distinct genetic clusters and generated 104,616 single-nucleotide polymorphisms (SNPs). The KM ex-situ population, originating from Longling County, exhibited unique genetic features, suggesting limited gene flow. The genetic diversity was substantial, with significant differences between populations, particularly between the KM lineage and the OTHER lineage. Demographic history inferred from the data indicated population experienced three significant population declines during glaciations, followed by periods of recovery. We estimated the effective population size (Ne) of the KM and OTHER lineages 1,000 years ago were 85,851 and 416,622, respectively. Gene flow analysis suggested past gene flow between populations, but the KM ex-situ population showed no recent gene flow. A total of 805 outlier SNPs, associated with four environmental factors, suggest potential local adaptation and showcase the species' adaptive potential. Particularly, the BZ displayed 515 adaptive loci, highlighting its strong potential for adaptation within this group. The comprehensive genomic analysis of M. lacei provides valuable insights into its genetic background and highlights the urgent need for conservation efforts. The study underscores the importance of ex-situ conservation methods, such as seed collection and vegetative propagation, to safeguard genetic diversity and promote population restoration. The preservation of populations like MC and BZ is crucial for maintaining the species' genetic diversity. In-situ conservation measures, including the establishment of in-situ conservation sites and community engagement, are essential to enhance protection awareness and ensure the long-term survival of this threatened plant species.

Sex identification of birds in Taipei Zoo.

As a conservation and breeding institution for birds, Taipei Zoo plays an important role in restoring endangered species. As approximately half of all bird species are monomorphic, precisely confirming the sex of individuals is critical for the management of ex-situ conservation breeding populations, as well as for understanding the sex ratio of those in the wild. Generally, PCR is used more reliably for sex determination versus traditional methods such as plumage, behavior or hormone levels. Nevertheless, the various primer sets and annealing temperatures vary between species, and so inaccurate sexing can occasionally happen due to inadequate PCR conditions. To reduce the probability of misidentification, and to establish a PCR condition database for sex determination across the diverse range of avian taxa, we tested multiple primer sets and annealing temperatures for amplification of the bird sex-specific gene fragments (CHD1) for each captive or rescued avian species held at Taipei Zoo since 2014. A total of 162 species across 22 orders were tested using one or two primer sets. One hundred and fifty-five species were successfully sexed by the primer set 2550F/2718R and the success rate of sex typing reached over 90% of species tested in each order. Most species have suitable PCR annealing temperatures between 45°C and 55°C, and the species in the same avian taxa showed similar results in temperature. This indicates that it is possible to select the annealing temperature of other species in the same family when the species had not been tested before. We expect this study will improve the success rate of identifying sex by using applicable PCR conditions and reduce the time for searching references every time before attempts to PCR sex birds.