In vitro culture and conservation of microalgae: Applications for aquaculture, biotechnology and environmental research

Summary Microalgae are a highly diverse group of unicellular organisms comprising the eukaryotic protists and the prokaryotic cyanobacteria or blue-green algae. The microalgae have a unique environmental status; being virtually ubiquitous in euphotic aquatic niches, they can occupy extreme habitats ranging from tropical coral reefs to the polar regions, and they contribute to half of the globe’s photosynthetic activity. Furthermore, they form the basis of the food chain for more than 70% of the world’s biomass. Microalgae are a valuable environmental and biotechnological resource, and the aim of this review is to explore the use of in vitro technologies in the conservation and sustainable exploitation of this remarkable group of organisms. The first part of the review evaluates the importance of in vitro methods in the maintenance and conservation of microalgae and describes the central role of culture collections in applied algal research. The second part explores the application of microalgal in vitro technologies, particularly in the context of the aquaculture and biotechnology industries. Emphasis is placed upon the exploitation of economically important algal products including aquaculture feed, biomass production for the health care sector, green fertilizers, pigments, vitamins, antioxidants, and antimicrobial agents. The contribution that microalgae can make to environmental research is also appraised; for example, they have an important role as indicator organisms in environmental impact assessments. Similarly, designated culture collection strains of microalgae are used for ecotoxicity testing. Throughout the review, emphasis is placed on the application of in vitro techniques for the continued advancement of microalgal research. The paper concludes by assessing future perspectives for the novel application of microalgae and their products.     

一种采集水稻二化螟卵(块)的高效简便新方法

比较了二化螟Chilosuppressalis卵块的塑料袋收集法和稻株法。结果表明 ,塑料袋法和稻株法收集的卵粒总数及卵孵化率都没有显著的差异 ,均为 2 79粒卵 雌蛾和 95 %。塑料袋法收集的卵块数较稻株法的多 ,但小卵块比例较高 ( <1 0 0粒卵 卵块 )。还讨论了采用塑料袋法收集二化螟及其它昆虫卵的优点。     

A method to generate multilocus barcodes of pinned insect specimens using MiSeq

For molecular insect identification, amplicon sequencing methods are recommended because they offer a cost‐effective approach for targeting small sets of informative genes from multiple samples. In this context, high‐throughput multilocus amplicon sequencing has been achieved using the MiSeq Illumina sequencing platform. However, this approach generates short gene fragments of <500 bp, which then have to be overlapped using bioinformatics to achieve longer sequence lengths. This increases the risk of generating chimeric sequences or leads to the formation of incomplete loci. Here, we propose a modified nested amplicon sequencing method for targeting multiple loci from pinned insect specimens using the MiSeq Illumina platform. The modification exists in using a three‐step nested PCR approach targeting near full‐length loci in the initial PCR and subsequently amplifying short fragments of between 300 and 350 bp for high‐throughput sequencing using Illumina chemistry. Using this method, we generated 407 sequences of three loci from 86% of all the specimens sequenced. Out of 103 pinned bee specimens of replicated species, 71% passed the 95% sequence similarity threshold between species replicates. This method worked best for pinned specimens aged between 0 and 5 years, with a limit of 10 years for pinned and 14 years for ethanol‐preserved specimens. Hence, our method overcomes some of the challenges of amplicon sequencing using short read next generation sequencing and improves the possibility of creating high‐quality multilocus barcodes from insect collections.     

南京中山植物园活植物信息管理子系统

以植物园植物记录国际传输格式（ITF）为标准,采用以数据库为中心的模块化与结构化的方法,开发研制了南京中山植物园活植物信息管理子系统,并于1994年正式投入使用。该系统界面友好,操作方便,修改灵活,响应速度快。文章对系统的实现、数据处理、软件结构以及软件的功能进行了讨论。     

Records of ocean sunfish Mola mola along the Norwegian coast spanning two centuries, 1801–2015

Records of the ocean sunfish Mola mola along the Norwegian coast were compiled from all possible sources: literature, media, databases and museums. A total of 216 records were found between 1801 and 2015. They were distributed along the whole coast, except for the most north‐eastern part. Nearly all years with more than five records were after 2000, with 1985 as the only exception and with a maximum of 23 records in 2014. Most, 92·4%, were from July to December. Records from before 1979 were more incidental and random and no sunfish were recorded in 54 separate years between 1879 and 2015. The northernmost record was from 70° 44′ N in December 1881. No relationship between year and latitude was found. Forty‐four per cent of the records were of sunfish caught in fishing gear, 27% were found stranded and 30% were observed alive at sea. A trend of a decrease in numbers of observations of living individuals and an increase in numbers of strandings as dead individuals as autumn progressed was found. Geographical differences in the three categories of observation were also found. Among the specimens whose size was recorded, most were small, <30 kg and 70 cm in total length. Only 10 were heavier than 100 kg, all these were from before 1960. Annual numbers correlated weakly with sea temperatures. This was supported by a weak negative correlation with the North Atlantic Oscillation (NAO) index, because the majority of years since 2000 had a negative NAO.     

State of the art and perspectives on neotropical fern and lycophyte systematics

For ferns and lycophytes, the Neotropics is a hotspot of diversity (3000–4500 species), and second only to Southeastern Asia in richness and endemism. This paper presents the current state of knowledge on fern and lycophyte systematics in the Neotropics, and emphasizes sampling sufficiency and current taxonomic and phylogenetic knowledge. Plant systematics plays an important role in documenting diversity and geographic distribution patterns that are needed to understand relationships and evolutionary patterns, and a vital role in species conservation. Although in recent decades this field of science has undergone a revolution because of new approaches and techniques, data presented in this work shows that large gaps remain and there is still a long path towards fully understanding fern and lycophyte systematics in Neotropics. Approaches and how to choose areas that should be targeted in order to try to fulfill these knowledge gaps are discussed.     

Long‐term preservation of a collection of Rhizoctonia solani using cryogenic storage

Rhizoctonia solani is an important plant pathogen for a number of crops and maintaining an extensive collection of reference isolates is important in understanding relationships of this pathogen with multiple hosts. Current long‐term storage methods typically call for frequent transfer increasing the risk of changes in morphological, physiological or virulence characteristics. Cryopreservation using storage in liquid nitrogen (LN) was evaluated to examine the potential for storage of a R. solani culture collection containing 106 isolates (primarily from sugar beet). Cultures were stored on autoclaved barley grains in the vapour phase of LN. After 60 days, 5 years and 10 years in storage, all isolates were tested for viability by calculating the percentage of barley grains from which R. solani mycelia grew. Five years after initial storage, all isolates except one had no change in viability. After 10 years in storage, 67 of 106 isolates had no significant decrease in viability, 39 of 106 isolates had a significant decrease in viability but only 9 isolates had less than 10% growth, with 4 having no growth. A subset of isolates stored for 10 years were tested for pathogenicity on a susceptible (FC901) and resistant (FC703) sugar beet germplasm. All isolates tested maintained approximately the same level of virulence that they had prior to storage on both germplasms. This indicates that cryogenic methods are suitable for the preservation or storage of R. solani culture collections, although efficacy may vary with individual isolates.     

Museomics Dissects the Genetic Basis for Adaptive Seasonal Coloration in the Least Weasel

Dissecting the link between genetic variation and adaptive phenotypes provides outstanding opportunities to understand fundamental evolutionary processes. Here, we use a museomics approach to investigate the genetic basis and evolution of winter coat coloration morphs in least weasels (Mustela nivalis), a repeated adaptation for camouflage in mammals with seasonal pelage color moults across regions with varying winter snow. Whole-genome sequence data were obtained from biological collections and mapped onto a newly assembled reference genome for the species. Sampling represented two replicate transition zones between nivalis and vulgaris coloration morphs in Europe, which typically develop white or brown winter coats, respectively. Population analyses showed that the morph distribution across transition zones is not a by-product of historical structure. Association scans linked a 200-kb genomic region to coloration morph, which was validated by genotyping museum specimens from intermorph experimental crosses. Genotyping the wild populations narrowed down the association to pigmentation gene MC1R and pinpointed a candidate amino acid change cosegregating with coloration morph. This polymorphism replaces an ancestral leucine residue by lysine at the start of the first extracellular loop of the protein in the vulgaris morph. A selective sweep signature overlapped the association region in vulgaris, suggesting that past adaptation favored winter-brown morphs and can anchor future adaptive responses to decreasing winter snow. Using biological collections as valuable resources to study natural adaptations, our study showed a new evolutionary route generating winter color variation in mammals and that seasonal camouflage can be modulated by changes at single key genes.     

Deficiency in African plant distribution data – missing pieces of the puzzle

Biodiversity is spatially unevenly distributed and so is the information on its spatial patterns. This uneven distribution of information on species occurrences is an important impediment to the conservation of biodiversity. Based on 185 427 collection records of 5873 plant species in sub-Saharan Africa, we analyse the availability of distribution data suitable for the GIS-based mapping of plant diversity patterns at a one-degree resolution. Using the bioclimatic model GARP, distribution ranges for each species were modelled. In order to identify data-deficient areas, the documented and modelled diversity patterns were compared. Only for a few, well-known centres of plant diversity are there comparatively many data collection records available. For several of the areas with very few collection records, such as the Guinean montane forests, the north-western Congolian lowland forests, and the southern Albertine Rift montane forests, the model predicts a species richness much higher than currently documented. In many of the data deficient areas, difficult conditions for scientific research appear to have limited collection activities for decades. Only strategic field collections can fill these gaps. Another cause of data deficiency is that data collected and digitized do not match the quality requirements for GIS-based work at the super-regional scale. In particular, regional databases documenting partial ranges of species are rarely connected. One challenge for the Global Strategy for Plant Conservation is therefore to establish international collaborative structures and technical standards that will allow analysis of biogeographical patterns across political boundaries.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 150 , 355–368.