Plant cryobanks play a significant role in modern science and breeding. They contribute to the recovery of lost species, the emergence of new plant varieties, and help preserve and explore the diversity of the plant world. The IPPRAS Cryobank collection is constantly supplemented with new samples, while, at the same time, the stored samples are being monitored. In order to test seed germination, seeds of Allium and Veratrum species were thawed. Rare Allium species seeds, such as A. nutans, A. schoenoprasum, and A. victorialis were stored in liquid nitrogen for 17, 19, and 30 years, respectively. Long-term cryopreservation decreased germination rates for A. nutans from 96.55 to 50.00%, for A. schoenoprasum from 72.00 to 62.75%, and for A. victorialis from 90.00 to 83.05%. Seeds of a rare medicinal species, Veratrum lobelianum, were stored in liquid nitrogen for 18 years; the seed germination rate during this storage period has been significantly decreased from 75.00 to 14.81%. V. nigrum seeds were also collected and frozen in liquid nitrogen for 3 days. Short-term cryopreservation did not result in a statistically significant change in germination rates (from 79.71 to 82.69%). The seeds of an endangered ornamental species, Cypripedium calceolus, were collected and kept frozen for 3 days. After cryopreservation, the seeds were planted on three different media, as follows: ½ MS, MS with 10% coconut milk, and BM1. On ½ MS medium, 24.98% seeds formed protocorms, while on MS medium with 10% coconut milk, this number was 10.02%, and on BM1 medium, it was 15.02%, respectively; however, after 2.5 months, all of the protocorms died. Thus, it appears that the existing protocol for seed cryopreservation of C. calceolus needs further improvement. The size, weight, and free water content (WC) of six previously cryopreserved Stipa species and three Allium species were measured. For all the Allium and Stipa species studied, we found no correlation between seed size, WC, and cryotolerance. We also found no correlation between the life form, which reflects the water requirement of the species, and cryotolerance.

Diverse secondary metabolites in plants, with their rich biological activities, have long been important sources for human medicine, food additives, pesticides, etc. However, the large-scale cultivation of host plants consumes land resources and is susceptible to pest and disease problems. Additionally, the multi-step and demanding nature of chemical synthesis adds to production costs, limiting their widespread application. In vitro cultivation and the metabolic engineering of plants have significantly enhanced the synthesis of secondary metabolites with successful industrial production cases. As synthetic biology advances, more research is focusing on heterologous synthesis using microorganisms. This review provides a comprehensive comparison between these two chassis, evaluating their performance in the synthesis of various types of secondary metabolites from the perspectives of yield and strategies. It also discusses the challenges they face and offers insights into future efforts and directions.

The cultivation of meat using in vitro grown animal stem cells offers a promising solution to pressing global concerns around climate change, ethical considerations, and public health. However, cultivated meat introduces an unprecedented necessity: the generation of mass scales of cellular biomaterial, achieved by fostering cell proliferation within bioreactors. Existing methods for in vitro cell proliferation encounter substantial challenges in terms of both scalability and economic viability. Within this perspective, we discuss the current landscape of cell proliferation optimization, focusing on approaches pertinent to cellular agriculture. We examine the mechanisms governing proliferation rates, while also addressing intrinsic and conditional rate limitations. Furthermore, we expound upon prospective strategies that could lead to a significant enhancement of the overall scalability and cost-efficiency of the cell proliferation phase within the cultivated meat production process. By exploring knowledge from basic cell cycle studies, pathological contexts and tissue engineering, we may identify innovative solutions toward optimizing cell expansion.

The systematical characterization and understanding of the metabolic behaviors are the basis of the efficient plant metabolic engineering and synthetic biology. Genome-scale metabolic networks (GSMNs) are indispensable tools for the comprehensive characterization of overall metabolic profile. Here we first constructed a GSMN of tobacco, which is one of the most widely used plant chassis, and then combined the tobacco GSMN and multiomics analysis to systematically elucidate the impact of in-vitro cultivation on the tobacco metabolic network. In-vitro cultivation is a widely used technique for plant cultivation, not only in the field of basic research but also for the rapid propagation of valuable horticultural and pharmaceutical plants. However, the systemic effects of in-vitro cultivation on overall plant metabolism could easily be overlooked and are still poorly understood. We found that in-vitro tobacco showed slower growth, less biomass and suppressed photosynthesis than soil-grown tobacco. Many changes of metabolites and metabolic pathways between in-vitro and soil-grown tobacco plants were identified, which notably revealed a significant increase of the amino acids content under in-vitro condition. The in silico investigation showed that in-vitro tobacco downregulated photosynthesis and primary carbon metabolism, while significantly upregulated the GS/GOGAT cycle, as well as producing more energy and less NADH/NADPH to acclimate in-vitro growth demands. Altogether, the combination of experimental and in silico analyses offers an unprecedented view of tobacco metabolism, with valuable insights into the impact of in-vitro cultivation, enabling more efficient utilization of in-vitro techniques for plant propagation and metabolic engineering.

Successfully developed in 1976, the continuous in vitro culture of Plasmodium falciparum has many applications in the field of malaria research. It has become an important experimental model that directly uses a human pathogen responsible for a high prevalence of morbidity and mortality in many parts of the world and is a major source of biological material for immunological, biochemical, molecular, and pharmacological studies. Until present, the basic techniques described by Trager and Jensen and Haynes et al. remain unchanged in many malaria research laboratories. Nonetheless, different factors, including culture media, buffers, serum substitutes and supplements, sources of erythrocytes, and conditions of incubation (especially oxygen concentration), have been modified by different investigators to adapt the original technique in their laboratories or enhance the in vitro growth of the parasites. The possible effects and benefits of these modifications for the continuous cultivation of asexual intraerythrocytic stages of P. falciparum, as well as future challenges in developing a serum-free cultivation system and axenic cultures, are discussed.

Former mine sites can provide habitat for many rare specialised bryophyte species that have adapted to metal-rich soil conditions that are toxic to most other plant species. Some of the bryophyte species found in this habitat are facultative metallophytes, and others are regarded as strict metallophytes, the so-called \'copper mosses\'. It is a general assumption in the literature that Cephaloziella nicholsonii and C. massalongoi, both categorised as Endangered in the IUCN Red List for Europe, are also strict metallophytes and obligate copper bryophytes. This in vitro experiment investigated the growth and gemma production of these two species from different sites in Ireland and Britain on treatment plates of 0 ppm, 3 ppm, 6 ppm, 12 ppm, 24 ppm, 48 ppm and 96 ppm copper. Results show that elevated copper is not an obligate requirement for optimum growth. Differences in response to the copper treatment levels among populations evident within both species could possibly be due to ecotypic variation. A case is also made for the taxonomic revision of the Cephaloziella genus. Implications for the species\' conservation are discussed.

In this study, the ecological conditions of the natural habitat of Lemna minuta Kunth in Morocco were investigated, and the impact of five synthetic growth media (Murashige-Skoog (MS), Schenk-Hildebrand (SH), Hoagland medium (HM), 10X Algal Assay Procedure (AAP), and Swedish Standard Institute medium (SIS)) on the morphophysiological and biochemical parameters was analysed. The morphophysiological parameters included root length, frond surface area, and fresh weight, while the biochemical parameters included photosynthetic pigments, carbohydrates, and protein content. The study was conducted in vitro in two phases: an uncontrolled aeration system (Phase I) and a controlled aeration system (Phase II).The results showed that the pH, conductivity, salinity, and ammonium levels in the natural habitat were within the optimal range for duckweed growth. The measured orthophosphate concentrations were higher compared to previous observations, while the recorded chemical oxygen demand values were low. The study also revealed a significant effect of the culture medium composition on the morphophysiological and biochemical parameters of the duckweed. The fresh weight biomass, relative growth rate in fronds, relative growth rate in surface area, root length, protein content, carbohydrates, chlorophyll (a), chlorophyll (b), total chlorophyll, carotenoids, and the chlorophyll (a/b) ratio were all affected by the culture medium.The most accurate regression models described the growth index GI(F) based on time and in vitro culture conditions in both phases. In Phase I, the best models for MS, SIS, AAP, and SH media were linear, weighted quadratic, cubic, and weighted cubic, respectively. In Phase II, the best models for all growth media were linear. The time coefficients (in days) for Phase II were 0.321, 0.547, 1.232, 1.470, and 0.306 for AAP, HM, MS, SH, and SIS, respectively.Comparing the morphophysiological and biochemical parameters of fronds from different media and analysing the regression model results showed that the SH and MS media were the best among the tested media for the in vitro culture of L. minuta in controlled aeration conditions. However, further research is needed to develop new synthetic media that best promote the growth and maintenance of this duckweed in long-term culture.

BACKGROUND: Vanilla planifolia is the most widely cultivated species of vanilla with high economic importance. However, seed germination under artificial conditions is difficult and yields low germination percentages. The seeds are adapted to endozoochorous dispersal, and we therefore tried to simulate the conditions in the digestive tract by acid scarification of seeds.
RESULTS: Immature seeds lacking dormancy, used as a control, showed the highest germination percentage. Among the treatments tested for mature seeds, the hydrochloric acid treatments were significantly the best in breaking dormancy and inducing germination, irrespective of the acid concentration and the presence of pepsin. Conventional treatment with a hypochlorite solution induced much lower germination percentage. Sulphuric acid at concentration 50% was too strong and caused damage to the seeds. Important factor is also high cultivation temperature 30 °C as there was nearly no germination at 25 °C.
CONCLUSIONS: Our protocol significantly improves the efficiency of generative propagation of vanilla and allows for significantly higher germination percentages than previously described. The strongly positive effect of hydrochloric acid may be related to the adaptation of seeds to endozoochorous dispersal.

BACKGROUND: Due to the lack of tumor suppressor function of the fragile histidine triad (FHIT) gene product, sebaceous gland carcinomas can develop.
OBJECTIVE: The model of the sebocyte cell line SZ95 was used to identify methylated CpG islands at the 5\'-end of the FHIT gene and the decrease of gene expression as well as the increase of double-stranded (ds) DNA breaks were examined.
METHODS: Methylation, immunofluorescence analysis, promotor sequencing and treatment of SZ95 cells with 5‑azacytidine/trichostatin A (TSA).
RESULTS: The cultivation was accompanied by an increasing methylation of the CpG islands, a decrease of the FHIT gene expression and an accumulation of ds-DNA breaks. Treatment with 5‑azacytidine/TSA showed a decrease in DNA methylation and a re-expression of FHIT transcripts.
CONCLUSIONS: Epigenetic changes in the cellular genome are caused by in vitro cell culture. Consequently, a positive selection of sebocytes with an epigenetically inactivated FHIT locus occurs.
UNASSIGNED: HINTERGRUND: Fehlt die Tumorsuppressorfunktion des fragilen Histidin-Trias(FHIT)-Genprodukts, so können Talgdrüsenkarzinome entstehen.
UNASSIGNED: Am Modell der Sebozytenlinie SZ95 sollten methylierte CpG-Inseln am 5′-Ende des FHIT-Gens identifiziert, die Abnahme der Genexpression und die Zunahme von DNA-Doppelstrang(ds)-Brüchen untersucht werden.
METHODS: Methylierungs‑, Immunfluoreszenzanalysen, Promotorsequenzierung sowie Behandlung der SZ95-Zellen mit 5‑Azacytidin/Trichostatin A (TSA).
UNASSIGNED: Die Kultivierung der Sebozyten verlief mit einer zunehmenden Methylierung der CpGs, einer Abnahme der FHIT-Genexpression sowie Anhäufung von DNA-ds-Brüchen. Die Behandlung mit 5‑Azacytidin/TSA zeigte eine Abnahme der DNA-Methylierung sowie Re-Expression von FHIT-Transkripten.
CONCLUSIONS: Durch In-vitro-Zellkultur werden epigenetische Veränderungen im zellulären Genom hervorgerufen. Es folgt eine positive Selektion von Sebozyten mit epigenetischer Inaktivierung des FHIT-Genorts.

Micropropagation of rare Veronica caucasica M. Bieb. was achieved by successful in vitro cultivation of mono-nodal segments on MS medium supplemented with 1.0 mg L-1 6-benzylaminopurine (BA) and then transferring the regenerated plants on hormone free basal MS medium for root development. In vitro multiplicated plants were successively acclimated in a growth chamber and a greenhouse with 92% survival. The number of plastid pigments and the total phenolics content in in vitro cultivated and ex vitro adapted plants were unchanged, and no accumulation of reactive oxygen species (ROS) was detected by staining with 3-3\'-diaminobenzidine (DAB) and 2\',7\'-dichlorofluorescein diacetate (DCF-DA). Nuclear Magnetic Resonance (NMR) fingerprinting allowed for the identification of the major alterations in metabolome of V. caucasica plants during the process of ex situ conservation. Iridoid glucosides such as verproside, aucubin and catalpol were characteristic for in vitro cultivated plants, while in ex vitro acclimated plants phenolic acid-protocatechuic acid and caffeic acid appeared dominant. The successful initiation of in vitro and ex vitro cultures is an alternative biotechnological approach for the preservation of V. caucasica and would allow for further studies of the biosynthetic potential of the species and the selection of lines with a high content of pharmaceutically valuable molecules and nutraceuticals.