Freshwater biodiversity conservation has received substantial attention in the scientific literature and is finally being recognized in policy frameworks such as the Global Biodiversity Framework and its associated targets for 2030. This is important progress. Nonetheless, freshwater species continue to be confronted with high levels of imperilment and widespread ecosystem degradation. An Emergency Recovery Plan (ERP) proposed in 2020 comprises six measures intended to \"bend the curve\" of freshwater biodiversity loss, if they are widely adopted and adequately supported. We review evidence suggesting that the combined intensity of persistent and emerging threats to freshwater biodiversity has become so serious that current and projected efforts to preserve, protect and restore inland-water ecosystems may be insufficient to avert substantial biodiversity losses in the coming decades. In particular, climate change, with its complex and harmful impacts, will frustrate attempts to prevent biodiversity losses from freshwater ecosystems already affected by multiple threats. Interactions among these threats will limit recovery of populations and exacerbate declines resulting in local or even global extinctions, especially among low-viability populations in degraded or fragmented ecosystems. In addition to impediments represented by climate change, we identify several other areas where the absolute scarcity of fresh water, inadequate scientific information or predictive capacity, and a widespread failure to mitigate anthropogenic stressors, are liable to set limits on the recovery of freshwater biodiversity. Implementation of the ERP rapidly and at scale through many widely dispersed local actions focused on regions of high freshwater biodiversity and intense threat, together with an intensification of ex-situ conservation efforts, will be necessary to preserve native freshwater biodiversity during an increasingly uncertain climatic future in which poorly understood, emergent and interacting threats have become more influential. But implementation of the ERP must be accompanied by measures that will improve water, energy and food security for humans - without further compromising the condition of freshwater ecosystems. Unfortunately, the inadequate political implementation of policies to arrest widely recognized environmental challenges such as climate change do not inspire confidence about the possible success of the ERP. In many parts of the world, the Anthropocene future seems certain to include extended periods with an absolute scarcity of uncontaminated surface runoff that will inevitably be appropriated by humans. Unless there is a step-change in societal awareness of - and commitment to - the conservation of freshwater biodiversity, together with necessary actions to arrest climate change, implementation of established methods for protecting freshwater biodiversity may not bend the curve enough to prevent continued ecosystem degradation and species loss.

Myrteae is the most species-rich tribe in the Myrtaceae family, represented by a range of socioeconomically and ecologically significant species. Many of these species, including commercially relevant ones, have become increasingly threatened in the wild, and now require conservation actions. Tissue culture presents an appropriate in vitro tool to facilitate medium-term and long-term wild germplasm conservation, as well as for commercial propagation to maintain desirable traits of commercial cultivars. So far, tissue culture has not been extensively achieved for Myrteae. Here, tissue culture for Eugenia, one of the most species-rich genera in Myrteae, is reviewed, giving directions for other related Myrteae. This review also focuses on ex situ conservation of Australian Myrteae, including using seed banking and field banking. Despite some progress, challenges to conserve these species remain, mostly due to the increasing threats in the wild and limited research. Research into in vitro methods (tissue culture and cryopreservation) is paramount given that at least some of the species are \'non-orthodox\'. There is an urgent need to develop long-term in vitro conservation for capturing the remaining germplasm of threatened Myrteae.

The gut microbiome plays important roles in metabolic and immune system related to the health of host. This study applied non-invasive sampling and 16S rDNA high-throughput sequencing to study the gut microbiota structure of red pandas (Ailurus fulgens) for the first time under different geographical latitudes in captivity. The results showed that the two predominant phyla Firmicutes (59.30%) and Proteobacteria (38.58%) constituted 97.88% of the total microbiota in all the fecal samples from north group (red pandas from Tianjin Zoo and Jinan Zoo) and south group (red pandas from Nanjing Hongshan Forest Zoo). The relative abundance of Cyanobacteria in north group was significantly higher than that in south group. At the genus level, Escherichia-Shigella (24.82%) and Clostridium_sensu_stricto_1 (23.00%) were common dominant genera. The relative abundance of norank_f__norank_o__Chloroplast, Terrisporobacter and Anaeroplasma from south group was significantly higher than that of north group. Alpha and Beta analysis consistently showed significant differences between north group and south group, however, the main functions of intestinal microbiota were basically the same, which play an important role in metabolic pathways, biosynthesis of secondary metabolites, microbial metabolism in different environments, and amino acid biosynthesis. The variations in gut microbiota between the northern and southern populations of the same species, both kept in captivity, which are primarily driven by significant differences in climate and diet. These findings provide a deeper understanding of the gut microbiota in red pandas and have important implications for their conservation, particularly in optimizing diet and environmental conditions in captivity.

The long-tailed goral is close to extinction, and ex situ conservation is essential to prevent this phenomenon. Studies on the gut microbiome of the long-tailed goral are important for understanding the ecology of this species. We amplified DNA from the 16S rRNA regions and compared the microbiomes of wild long-tailed gorals and two types of captive long-tailed gorals. Our findings revealed that the gut microbiome diversity of wild long-tailed gorals is greatly reduced when they are reared in captivity. A comparison of the two types of captive long-tailed gorals confirmed that animals with a more diverse diet exhibit greater gut microbiome diversity. Redundancy analysis confirmed that wild long-tailed gorals are distributed throughout the highlands, midlands, and lowlands. For the first time, it was revealed that the long-tailed goral are divided into three groups depending on the height of their habitat, and that the gut bacterial community changes significantly when long-tailed gorals are raised through ex situ conservation. This provides for the first time a perspective on the diversity of food plants associated with mountain height that will be available to long-tailed goral in the future.

UNASSIGNED: In this study, we use simulations to determine how pollen flow and sampling constraints can influence the genetic conservation found in seed collections.
UNASSIGNED: We simulated genotypes of parental individuals and crossed the parentals based on three different ranges of pollen flow (panmictic, limited, and highly limited) to create new seed sets for sampling. We tested a variety of sampling scenarios modeled on those occurring in nature and calculated the proportion of alleles conserved in each scenario.
UNASSIGNED: We found that pollen flow greatly influences collection outcomes, with panmictic pollen flow resulting in seed sets containing 21.6% more alleles than limited pollen flow and 48.6% more alleles than highly limited pollen flow, although this impact diminishes when large numbers of maternal plants are sampled. Simulations of realistic seed sampling (sampling more seed from some plants and fewer from others) showed a relatively minor impact (<2.5%) on genetic diversity conserved compared to ideal sampling (uniform sampling across all maternal plants).
UNASSIGNED: We conclude that future work must consider limited pollen flow, but collectors can be flexible with their sampling in the field as long as many unique maternal plants are sampled. Simulations remain a fruitful method to advance ex situ sampling guidelines.

Worldwide, oak species are threatened with extinction due to habitat loss, pathogens, and changing fire regimes. Ex situ conservation through tissue culture may protect the remaining genetic diversity of Quercus dumosa, or the coastal sage scrub oak, from further loss. We designed three basal salt formulations based on the mineral composition of shoot tips and first leaves from mature Q. dumosa and explored carbohydrate source, stress-mitigating compounds, and plant growth regulator concentrations to develop a method of cultivating many Q. dumosa culture lines in vitro. All three novel basal salt formulations led to decreased necrosis compared with commercial basal salt formulas WPM, MS, and DKW. Substitution of 30 g L-1 sucrose with glucose and adding 250 mg L-1 ascorbic acid, 5.2 mg L-1 SNP sodium nitroprusside, and 103 mg L-1 y-aminobutyric acid improved culture health overall. In an experiment involving 115 culture lines, 0.66 mg L-1 6-benzylaminopurine produced the highest average shoots per explant, but 0.33 mg L-1 produced the greatest proportion of shoots 2 cm or greater. Incubation for 24 h in 20 mg L-1 indole-3-butyric acid led to the most rooting. These methods show promise for the ex situ conservation of many genotypes of endangered Q. dumosa.

This paper focuses on the creation of an in vitro collection of grapevine hybrids from the breeding program of the Kazakh Scientific Research Institute of Fruit Growing and Viticulture and investigates the presence of Plasmopara viticola resistance mediated by Rpv3 and Rpv12 loci. We looked at the optimization of in vitro establishment using either shoots taken directly from field-grown plants or from budwood cuttings forced indoors. We further screened for the presence of endophyte contamination in the initiated explants and optimized the multiplication stage. Finally, the presence of the resistance loci against P. viticola was studied. The shoots initiated from the field-sourced explants were the more effective method of providing plant sources for in vitro initiation once all plant accessions met the goal of in vitro establishment. The concentration of phytohormones and the acidity of the culture medium have a great effect on the multiplication rate and the quality of in vitro stock cultures. Out of 17 grapevine accessions, 16 showed the presence of single or combined resistance loci against P. viticola. The grapevine accessions identified as carrying Rpv3 and Rpv12 alleles represent important genetic resources for disease resistance breeding programs. These accessions may further contribute to the creation of new elite cultivars of economic interest.

As wildlife protection continue to strengthen, research on the gut microbiota of wildlife is increasing. Carrying out conservation and research on endangered species in the Qinghai Tibet Plateau plays an important role in global biodiversity conservation. This study utilized 16S rRNA sequencing of fecal samples to investigate the composition, function, and changes of the gut microbiota of bharal in different environments, seasons, and genders. The results showed that Firmicutes and Bacteroidota were the dominant phyla and UCG-005, Bacteroides, UCG-010 were the dominant genera of bharal. In the wild, the abundance of Firmicutes increased which was conducive to the decomposition and utilization of cellulose, hemicellulose, and carbohydrate. Due to the variety of food types and nutrition in different seasons, the composition and function of gut microbiota were obviously different between genders. Compared with zoo, higher alpha diversity, a more complex gut microbiota network structure, and stronger metabolic function were conducive bharal to adapting to the wild environment. In the zoo, captive bharals were fed foods rich in high fat and protein, which increased the abundance of Bacteroidota and reduced the alpha diversity of gut microbiota. A fixed diet unified the gut microbiota between genders of bharal. It is very important to pay attention to the impact of captive environments and maintain the native gut microbiota of wildlife.

Genetic resources serve as the foundation of our food supply and are building blocks for the development of new crop varieties that support sustainable crop production in the face of climate change, as well as for the delivery of healthy diets to a continuously growing global population. With the encouragement of the FAO and with technical guidance and assistance from the International Board for Plant Genetic Resources (IBPGR), almost 2000 genebanks have been established worldwide for the ex situ conservation of genetic resources since the middle of the last century. The global genetic resources\' conservation and use system has evolved over several decades and presents apparent weaknesses, without a clear blueprint. Therefore, a Special Issue (SI) of Plants on \'A Critical Review of the Current Approaches and Procedures of Plant Genetic Resources Conservation and Facilitating Use: Theory and Practice\' was initiated. This SI comprises 13 review and research papers that shed light on the history and the political dimensions of the global system; its current strengths, weaknesses, and limitations; and how the effectiveness and efficiency of the system could be improved to satisfy the germplasm users (plant breeders, researchers) and benefit consumers and society at large. This SI provides insight into new approaches and technical developments that have revolutionised ex situ conservation and the use of germplasm and related information. It also reflects on complementary conservation approaches (in situ, on-farm, home gardens) to ex situ genebanks, as well as how-through new forms of collaboration at national, regional, and global levels and through stronger links between public genebanks-synergies between the private breeding sector and botanic garden community could be achieved to strengthen the global conservation and use system. Special attention has also been given to the governance of genetic resources and access and benefit-sharing issues that increasingly hamper the needed access to a wide range of genetic resources that is essential for plant breeders to fulfil their mission.

The Louisiana pinesnake (Pituophis ruthveni) is considered one of the rarest snakes in North America and was federally listed under the Endangered Species Act in 2018. Captive breeding and reintroduction of zoo-bred hatchlings has been successful, however, limited founders in the captive population and the inability to bring new, wild genes into the captive colony presents a major concern for the conservation of this species. The use of artificial insemination (AI) was first applied to snakes in the 1980s but further development of the technique has since received little attention. Our goal was to develop a method of AI for use in breeding Louisiana pinesnakes to facilitate gene flow from wild to captive populations. We inseminated two captive Louisiana pinesnakes with semen collected from one donor male, novel to both females. Timing of AI occurred following the emergence of females from brumation, and when large, distinct follicles were detected using digital palpation. Females were inseminated four and five times over a period of 14 and 19 days, respectively, using fresh and 2-day refrigerator stored semen. One female laid seven eggs, which resulted in four fertile eggs and two viable hatchlings, while the second female produced three fertile of seven eggs laid but no viable hatchlings. Genetic analyses confirmed the donor male was the sire of hatchlings. This is the first successful AI of an endangered snake species and provides a framework for the use and optimization of assisted reproductive technologies for use in conservation breeding programs.