Offspring production from cryopreserved primordial germ cells in Drosophila

AbstractThere is an urgent need to cryopreserve Drosophila stocks that have been maintained as living cultures for a long time. Long-term culture increases the risk of accidental loss and of unwanted genetic alteration. Here, we report that cryopreserved primordial germ cells (PGCs) can produce F1 progeny when transplanted into hosts. The cryopreserved donor PGCs could form germline stem cells in host gonads and contributed to continuous offspring production. Furthermore, the ability to produce offspring did not appear to vary with either differences between donor strains or cryopreservation duration. Therefore, we propose that our cryopreservation method is feasible for long-term storage of various Drosophila strains. These results underscore the potential usefulness of our cryopreservation method for backing up living stocks to avoid either accidental loss or genetic alteration.

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Rediscovering Zygorhizidium affluens Canter: Molecular Taxonomy, Infectious Cycle, and Cryopreservation of a Chytrid Infecting the Bloom-Forming Diatom Asterionella formosa

Applied and Environmental Microbiology ◽

10.1128/aem.01826-18 ◽

2018 ◽

Vol 84 (23) ◽

Cited By ~ 9

Author(s):

Cecilia Rad-Menéndez ◽

Mélanie Gerphagnon ◽

Andrea Garvetto ◽

Paola Arce ◽

Yacine Badis ◽

...

Keyword(s):

Molecular Taxonomy ◽

Cost Effective ◽

Trophic Levels ◽

Content Type ◽

Long Term Storage ◽

Asterionella Formosa ◽

Infectious Cycle ◽

Cryopreservation Method ◽

Term Storage

ABSTRACT Parasitic Chytridiomycota (chytrids) are ecologically significant in various aquatic ecosystems, notably through their roles in controlling bloom-forming phytoplankton populations and in facilitating the transfer of nutrients from inedible algae to higher trophic levels. The diversity and study of these obligate parasites, while critical to understand the interactions between pathogens and their hosts in the environment, have been hindered by challenges inherent to their isolation and stable long-term maintenance under laboratory conditions. Here, we isolated an obligate chytrid parasite (CCAP 4086/1) on the freshwater bloom-forming diatom Asterionella formosa and characterized its infectious cycle under controlled conditions. Phylogenetic analyses based on 18S, 5.8S, and 28S ribosomal DNAs (rDNAs) revealed that this strain belongs to the recently described clade SW-I within the Lobulomycetales. All morphological features observed agree with the description of the known Asterionella parasite Zygorhizidium affluens Canter. We thus provide a phylogenetic placement for this chytrid and present a robust and simple assay that assesses both the infection success and the viability of the host. We also validate a cryopreservation method for stable and cost-effective long-term storage and demonstrate its recovery after thawing. All the above-mentioned tools establish a new gold standard for the isolation and long-term preservation of parasitic aquatic chytrids, thus opening new perspectives to investigate the diversity of these organisms and their physiology in a controlled laboratory environment. IMPORTANCE Despite their ecological relevance, parasitic aquatic chytrids are understudied, especially due to the challenges associated with their isolation and maintenance in culture. Here we isolated and established a culture of a chytrid parasite infecting the bloom-forming freshwater diatom Asterionella formosa. The chytrid morphology suggests that it corresponds to the Asterionella parasite known as Zygorhizidium affluens. The phylogenetic reconstruction in the present study supports the hypothesis that our Z. affluens isolate belongs to the order Lobulomycetales and clusters within the novel clade SW-I. We also validate a cryopreservation method for stable and cost-effective long-term storage of parasitic chytrids of phytoplankton. The establishment of a monoclonal pathosystem in culture and its successful cryopreservation opens the way to further investigate this ecologically relevant parasitic interaction.

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The genes encoding the major 42S storage particle proteins are expressed in male and female germ cells of Xenopus laevis

Development ◽

10.1242/dev.113.3.851 ◽

1991 ◽

Vol 113 (3) ◽

pp. 851-856

Author(s):

B. Abdallah ◽

J. Hourdry ◽

S. Deschamps ◽

H. Denis ◽

A. Mazabraud

Keyword(s):

Xenopus Laevis ◽

Germ Cells ◽

5S Rna ◽

Male And Female ◽

Long Term Storage ◽

Genes Encoding ◽

Rna Accumulation ◽

Storage Particle ◽

Term Storage

As components of the 42S storage particles (thesaurisomes), thesaurin a and thesaurin b are involved in the long-term storage of tRNA and 5S RNA in previtellogenic oocytes of Xenopus laevis. Thesaurin a and thesaurin b are among the most abundant proteins in previtellogenic oocytes. We show here that the mRNAs encoding thesaurin a and thesaurin b are present not only in previtellogenic oocytes but also in pre-meiotic germ cells (oogonia). These mRNAs can also be detected in spermatogonia and early spermatocytes, and are translated into protein in testis, as they are in ovary. We conclude that male germ cells mimic female germ cells in several aspects of gene activity related to RNA accumulation and metabolism.

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Cryopreservation of papaya seeds cv. Sukma, Callina, and Caliso: Effect of loading treatment and immersion time in plant vitrification solution-2

Nusantara Bioscience ◽

10.13057/nusbiosci/n110112 ◽

2019 ◽

Vol 11 (1) ◽

pp. 71-78

Author(s):

FITRI FATMA WARDANI ◽

DARDA EFENDI ◽

DINY DINARTI ◽

Joko Ridho Witono

Keyword(s):

Immersion Time ◽

Storage System ◽

Long Term Storage ◽

Randomized Design ◽

Long Time ◽

Papaya Seeds ◽

Vitrification Solution ◽

Different Characteristics ◽

Term Storage

Abstract. Wardani FF, Efendi D, Dinarti D, Witono JR. 2019. Cryopreservation of papaya seeds cv. Sukma, Callina, and Caliso: Effect of loading treatment and immersion time in plant vitrification solution-2. Nusantara Bioscience 11: 71-78. Sukma, Callina, and Caliso are papaya cultivars released by the Center for Tropical Horticulture Studies, IPB University, Bogor, West Java, Indonesia. In general, papaya seeds cannot be stored for a long time with a conventional GenBank storage system, even though some of them consider as “orthodox” class of seed. Cryopreservation, storage at an ultra-low temperature (-196 0C) of liquid nitrogen, could be possible for long-term storage of papaya seeds. The experiment conducted to get the necessary of loading treatment and the best immersion time in PVS2 for cryopreservation of papaya seeds cv. Sukma, Callina, and Caliso, so they still had considerable viability. We conducted experiment as factorial in completely randomized design, with the first factor was loading treatment with two levels (with and without loading treatment), and the second factor was immersion time in PVS2 with three levels (15, 30 and 45 min). So, there was six treatment and we used 50 seeds for each treatment. Results showed that the three papaya cultivars gave different responses to the treatment before cryopreservation proved that papayas seed had different characteristics, depending on genotype. For papaya seeds cv. Sukma, loading treatment was not needed and the best immersion time in PVS2 was 15 min. The viability of papaya seeds cv. Callina was low, so we should try another treatment before cryopreservation. For papaya seeds cv. Caliso, loading treatment was needed and the best immersion time in PVS2 was 30 min.

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Development and Optimization of a Germination Assay and Long-Term Storage for Cannabis sativa Pollen

Plants ◽

10.3390/plants9050665 ◽

2020 ◽

Vol 9 (5) ◽

pp. 665 ◽

Cited By ~ 2

Author(s):

Daniel Gaudet ◽

Narendra Singh Yadav ◽

Aleksei Sorokin ◽

Andriy Bilichak ◽

Igor Kovalchuk

Keyword(s):

Cannabis Sativa ◽

Pollen Viability ◽

Growth Stages ◽

Long Term Storage ◽

Whole Wheat ◽

Germination Assay ◽

And Storage ◽

Cryopreservation Method ◽

Term Storage

Pollen viability and storage is of great interest to cannabis breeders and researchers to maintain desirable germplasm for future use in breeding or for biotechnological and gene editing applications. Here, we report a simple and efficient cryopreservation method for long-term storage of Cannabis sativa pollen. Additionally, the bicellular nature of cannabis pollen was identified using DAPI (4′,6-diamidino-2-phenylindole) staining. A pollen germination assay was developed to assess cannabis pollen viability and used to demonstrate that pollen collected from different principal growth stages exhibited differential longevity. Finally, a simple and efficient method that employs pollen combined with baked whole wheat flour and subsequent desiccation under vacuum was developed for the long-term cryopreservation of C. sativa pollen. Using this method, pollen viability was maintained in liquid nitrogen after four months, suggesting long-term preservation of cannabis pollen.

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Modelling of Moisture Movement in Wood during Outdoor Storage

Nonlinear Analysis Modelling and Control ◽

10.15388/na.2001.6.1.15210 ◽

2001 ◽

Vol 6 (2) ◽

pp. 3-14 ◽

Cited By ~ 23

Author(s):

R. Baronas ◽

F. Ivanauskas ◽

I. Juodeikienė ◽

A. Kajalavičius

Keyword(s):

Experimental Data ◽

Finite Difference ◽

Climatic Conditions ◽

Two Dimensional ◽

Moisture Movement ◽

Finite Difference Technique ◽

Long Term Storage ◽

Space Formulation ◽

Term Storage

A model of moisture movement in wood is presented in this paper in a two-dimensional-in-space formulation. The finite-difference technique has been used in order to obtain the solution of the problem. The model was applied to predict the moisture content in sawn boards from pine during long term storage under outdoor climatic conditions. The satisfactory agreement between the numerical solution and experimental data was obtained.

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