scholarly journals Seed Cryopreservation, Germination, and Micropropagation of Eastern Turkeybeard, Xerophyllum asphodeloides (L.) Nutt.: A Threatened Species from the Southeastern United States

Plants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1462
Author(s):  
Michelle Issac ◽  
Princy Kuriakose ◽  
Stacie Leung ◽  
Alex B. Costa ◽  
Shannon Johnson ◽  
...  

Xerophyllum asphodeloides (Xerophyllaceae), known as eastern turkeybeard, is an herbaceous perennial found in eastern North America. Due to decline and destruction of its habitat, several states rank X. asphodeloides as “Imperiled” to “Critically Imperiled”. Protocols for seed cryopreservation, in vitro germination, sustainable shoot micropropagation, shoot establishment in soil, and seed germination are presented. Seeds from two tested sources were viable after 20 months of cryopreservation. Germination of isolated embryos in vitro was necessary to overcome strong seed dormancy. Shoot multiplication and elongation occurred on ½ MS medium without PGRs. Shoots rooted in vitro without PGRs or with 0.5 mg/L NAA or after NAA rooting powder treatment and placement in potting mix. When planted in wet, peaty soil mixes, shoots grew for two months and then declined. When planted in a drier planting mix containing aged bark, most plants continued growth. In the field, plant survival was 73% after three growing seasons. Safeguarding this species both ex situ and in situ is possible and offers a successful approach to conservation. Whole seeds germinated after double dormancy was overcome by incubation under warm moist conditions for 12 weeks followed by 12 weeks cold at 4 °C and then warm.

Zygote ◽  
2021 ◽  
pp. 1-7
Author(s):  
Luciana Diniz Rola ◽  
Eveline dos Santos Zanetti ◽  
Maite del Collado ◽  
Ellen de Fátima Carvalho Peroni ◽  
José Maurício Barbanti Duarte

Summary In vitro production of embryos has gained prominence as a tool for use in wildlife conservation programmes in situ and ex situ. However, the development of this technique depends on steps that include ovarian stimulation, collection and oocyte maturation. The purpose of this study was to assess the feasibility of an ovarian stimulation protocol for follicular aspiration, the efficiency of videolaparoscopy for follicular aspiration and test a medium for in vitro oocyte maturation for the species Mazama gouazoubira. Five females were submitted to repeated ovarian stimulation (hormone protocol using controlled internal drug release), and estradiol benzoate on D0 and eight injections of follicle-stimulating hormone, once every 12 h, from D4 onwards at 30-day intervals. Fourteen surgical procedures were performed in superstimulated females, resulting in the collection of 94 oocytes and an average of 17.1 ± 9.1 follicles observed, 13.5 ± 6.6 follicles aspirated and 7.2 ± 3.7 oocytes collected per surgery. After collection, the oocytes were submitted to in vitro maturation for 24 h and stained with Hoechst 33342 dye to evaluate their nuclear status; 64.5% of the oocytes reached MII and 16.1% were spontaneously activated by parthenogenesis. The nuclear status of oocytes that did not undergo in vitro maturation was evaluated; 80.9% were found to be immature.


Genetika ◽  
2004 ◽  
Vol 36 (3) ◽  
pp. 221-227
Author(s):  
Jelena Aleksic ◽  
Sasa Orlovic

Principles of the conservation of genetic resources of elms (Ulmus spp) do not differ fundamentally from the general principles accepted for the conservation of genetic resources of other common Noble Hardwoods. Efficient conservation can best be achieved through appropriate combination of in situ and ex situ methods, which have distinct advantages. Besides that, ex situ conservation is employed when emergency measures are needed for rare endangered populations and when populations are too small to be managed in situ (e.g. risks of genetic drift and inbreeding). The aim of our research is ex situ conservation of genetic resources of field elm {Ulmus minor Mill) and European white elm (Ulmus laevis Pall) through establishment of field genebanks. Sampling was conducted in one population of field elm and one population of white elm. Plant material (buds) from 8 trees of field elm and 10 trees of white elm was used for in vitro production of clones. Obtained clones will be used for establishment of field genebanks on the experimental estate of the Institute of Lowland Forestry and Environment.


2020 ◽  
Vol 21 (20) ◽  
pp. 7459
Author(s):  
María Elena González-Benito ◽  
Miguel Ángel Ibáñez ◽  
Michela Pirredda ◽  
Sara Mira ◽  
Carmen Martín

Epigenetic variation, and particularly DNA methylation, is involved in plasticity and responses to changes in the environment. Conservation biology studies have focused on the measurement of this variation to establish demographic parameters, diversity levels and population structure to design the appropriate conservation strategies. However, in ex situ conservation approaches, the main objective is to guarantee the characteristics of the conserved material (phenotype and epi-genetic). We review the use of the Methylation Sensitive Amplified Polymorphism (MSAP) technique to detect changes in the DNA methylation patterns of plant material conserved by the main ex situ plant conservation methods: seed banks, in vitro slow growth and cryopreservation. Comparison of DNA methylation patterns before and after conservation is a useful tool to check the fidelity of the regenerated plants, and, at the same time, may be related with other genetic variations that might appear during the conservation process (i.e., somaclonal variation). Analyses of MSAP profiles can be useful in the management of ex situ plant conservation but differs in the approach used in the in situ conservation. Likewise, an easy-to-use methodology is necessary for a rapid interpretation of data, in order to be readily implemented by conservation managers.


Author(s):  
Marine Deshors ◽  
Olivier Guais ◽  
Virginie Neugnot-Roux ◽  
Xavier Cameleyre ◽  
Luc Fillaudeau ◽  
...  
Keyword(s):  
Ex Situ ◽  

Author(s):  
Jane Muthoni ◽  
Hussein Shimelis ◽  
Rob Melis

Plant genetic resources (PGRs) play an important role in agriculture, environment protection, cultural property and trade; they need to be conserved. There are two fundamental approaches for the conservation of PGRs: in situ and ex situ. In situ conservation is the conservation of ecosystems and natural habitats and the maintenance and recovery of viable populations of species in their natural surroundings. Ex situ preservation is the storage of seeds or plant materials under artificial conditions to maintain their long term viability and availability for use. Genebanks employ seed storage, field collections of living plants and in vitro storage (tissue culture or cryopreservation) for ex situ preservation of PGR. Storage of orthodox seeds, which are tolerant to low moisture content and low temperatures at appropriate temperature and humidity, is the most convenient ex situ conservation method. Plants that produce recalcitrant seeds or non-viable seeds are conserved in field genebanks as well as in-vitro in slow growth media for short-to-medium term and cryopreservation in liquid nitrogen at -1960C for long-term periods. Cryopreservation is very expensive and needs trained personnel; this could explain why this method is rarely used for conservation of plant genetic resources in most developing countries. Potato tubers are bulky and highly perishable; the crop is generally conserved as clones either in field genebanks (with annual replanting), in-vitro conservation in slow growth media for short-to-medium term and cryopreservation for long term. Field genebanks are expensive to maintain and the crop is exposed to many dangers; hence, cryopreservation is the only feasible method for long term conservation. However, given the high cost of cryopreservation, long-term conservation of potato genetic resources is poorly developed in most resource-poor countries leading to high rates of genetic erosion. This paper looks into the various methods that that can be applied to conserve potato genetic resources and the status of conservation of potatoes in major genebanks and some countries.


2021 ◽  
Vol 273 ◽  
pp. 01007
Author(s):  
Valentina Puzirnova ◽  
Natalia Doroshenko

The paper is devoted to the problem of plant biodiversity conservation. This problem is acute all over the world. Lower Don Region has a centuries old history of viticulture and winemaking. There are many valuable vine varieties which are worthy of preservation for future generations. Classical methods no longer cope with this task. Applying advances of biotechnology in addition to traditional methods of ex situ and in situ biodiversity conservation allows sustainable management of genetic resources. This article summarizes the study of methods for creation slow growing collection for grapevine variety Fioletoviy Ranniy. Keeping plants in a slow-growing collection is one of the best ways to preserve biodiversity. This study analyzed the effect of various media compounds on vigor of vine in order to elongate the time between replantings.


2020 ◽  
Vol 60 (10) ◽  
pp. 1227
Author(s):  
P. Comizzoli

More than 50 deer species live in diverse ecosystems around the world. Unfortunately, most of them are threatened or endangered because of over-hunting, poaching or habitat destruction. Protection of wild populations (in situ) and management of animal collections in zoos and breeding centres (ex situ) are complementary conservation efforts relying on multidisciplinary approaches. Reproductive biology of deer species is one of the critical areas that still needs to be thoroughly studied to ensure the success of in situ or ex situ programs. Interestingly, there is a vast diversity in reproductive traits within the deer family (from anatomy to breeding-season patterns). On the basis of this fundamental knowledge, adapted reproductive biotechnologies have been developed to enhance reproduction and preserve fertility of individuals. Early works on artificial insemination (AI), in vitro fertilisation (IVF), and germplasm freezing in the more common red deer, sika deer and white-tailed deer have been highly inspiring to projects aiming at saving endangered deer species. A few fawn births following AI or IVF using frozen semen have been reported in wild species (e.g. Eld’s deer, Rucervus eldii thamin); however, assisted reproductive techniques and cryo-banking are currently not integrated into the management of rare and endangered populations. Knowing that many deer populations are rapidly declining in situ and ex situ, there is now an urgent need for better strategies and more fertility preservation options. The objectives of the present article are to review (1) existing reproductive biotechnologies to preserve fertility of different deer species and (2) how to integrate these approaches into the management of rare and endangered populations to address conservation issues.


2013 ◽  
Vol 41 (1) ◽  
pp. 73 ◽  
Author(s):  
Victoria CRISTEA ◽  
Liliana JARDA ◽  
Irina HOLOBIUC

Within the current context of declining biodiversity, the botanical gardens play an essential role in its conservation. Dianthus callizonus, D. glacialis ssp. gelidus and D. spiculifolius are the species that we seek to preserve in "Alexandru Borza" Botanical Garden of Cluj-Napoca (Romania). Several replicates were collected for each taxon from different populations in order to avoid the genetic uniformity. The material collected from the natural sites, was planted on a rockery, specially designed for this collection in the Botanical Garden. At the time of planting, each individual was sampled for setting up an in vitro collection and further biochemical and molecular analyses. In case of ex situ outdoor conservation of the three Dianthus species, 80.6% of the individuals collected in the field survived during the first year but the percentage decreased drastically after four years. In the case of in situ collected individuals, as well as in the case of in vitro individuals, D. spiculifolius had the best ability to acclimatize in the Botanical Garden, and D. callizonus presented the lowest number of surviving individuals. The ex vitro acclimatization of the plantlets had 80% efficiency at 10ºC, using three different substrates: soil and pearl stone mix 1/1, soil and sand mix 1/1 and pearl stone. All the three species are preserved in vitro, whereas the plantlets are acclimatized outdoors. Ex situ conservation of these species will have a positive impact on the biodiversity conservation.


2005 ◽  
Vol 17 (7) ◽  
pp. 683 ◽  
Author(s):  
W. V. Holt ◽  
R. M. A. Elliott ◽  
A. Fazeli ◽  
N. Satake ◽  
P. F. Watson

Previous experiments have shown that boar sperm survival in vitro is enhanced when co-incubated with a solubilised protein extract of porcine oviducal apical plasma membrane proteins. Here, we examine the hypothesis that the effects are mediated by direct oviduct–sperm contact and use in situ biotinylation of the oviducal epithelial surface to trace the surface-exposed biotinylated proteins through purification and solubilisation steps. We have also examined the effectiveness of mechanical scraping as a method of recovering oviducal epithelial proteins. We show that a subset of proteins originally exposed at the oviducal surface eventually bind to spermatozoa during incubation in vitro, but also show that a different protein subset is implicated if the sperm incubation is performed with proteins that had been biotinylated after (ex situ) extraction from the oviduct. Apical plasma membrane fractions biotinylated after purification contained many more biotinylated protein bands than preparations labelled before purification and multiple protein bands were eventually found to associate with spermatozoa. Although the evidence presented here supports the hypothesis that protein(s) anchored to the oviducal epithelium bind populations of spermatozoa directly and may have a role in the enhancement of sperm viability, it also shows that the choice of investigative technique exerts a major influence on experimental outcomes.


2019 ◽  
Vol 20 (17) ◽  
pp. 4080 ◽  
Author(s):  
Giuseppe Pezzotti ◽  
Tetsuya Adachi ◽  
Francesco Boschetto ◽  
Wenliang Zhu ◽  
Matteo Zanocco ◽  
...  

The availability of osteoinductive biomaterials has encouraged new therapies in bone regeneration and has potentially triggered paradigmatic shifts in the development of new implants in orthopedics and dentistry. Among several available synthetic biomaterials, bioceramics have gained attention for their ability to induce mesenchymal cell differentiation and successive bone formation when implanted in the human body. However, there is currently a lack of understanding regarding the fundamental biochemical mechanisms by which these materials can induce bone formation. Phenomenological studies of retrievals have clarified the final effect of bone formation, but have left the chemical interactions at the cell–material interface uncharted. Accordingly, the knowledge of the intrinsic material properties relevant for osteoblastogenesis and osteoinduction remains incomplete. Here, we systematically monitored in vitro the chemistry of mesenchymal cell metabolism and the ionic exchanges during osteoblastogenesis on selected substrates through conventional biological assays as well as via in situ and ex situ spectroscopic techniques. Accordingly, the chemical behavior of different bioceramic substrates during their interactions with mesenchymal cells could be unfolded and compared with that of biomedical titanium alloy. Our goal was to clarify the cascade of chemical equations behind the biological processes that govern osteoblastogenic effects on different biomaterial substrates.


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