Molecular Mechanisms Underlying Freezing Tolerance in Plants: Implications for Cryopreservation.

Cryopreservation is a crucial technique for the long-term ex situ conservation of plant genetic resources, particularly in the context of global biodiversity decline. This process entails freezing biological material at ultra-low temperatures using liquid nitrogen, which effectively halts metabolic activities and preserves plant tissues over extended periods. Over the past seven decades, a plethora of techniques for cryopreserving plant materials have been developed. These include slow freezing, vitrification, encapsulation dehydration, encapsulation-vitrification, droplet vitrification, cryo-plates, and cryo-mesh techniques. A key challenge in the advancement of cryopreservation lies in our ability to understand the molecular processes underlying plant freezing tolerance. These mechanisms include cold acclimatization, the activation of cold-responsive genes through pathways such as the ICE-CBF-COR cascade, and the protective roles of transcription factors, non-coding RNAs, and epigenetic modifications. Furthermore, specialized proteins, such as antifreeze proteins (AFPs) and late embryogenesis abundant (LEA) proteins, play crucial roles in protecting plant cells during freezing and thawing. Despite its potential, cryopreservation faces significant challenges, particularly in standardizing protocols for a wide range of plant species, especially those from tropical and subtropical regions. This review highlights the importance of ongoing research and the integration of omics technologies to improve cryopreservation techniques, ensuring their effectiveness across diverse plant species and contributing to global efforts regarding biodiversity conservation.

Characteristic of the gene candidate SecARS encoding alkylresorcinol synthase in Secale.

BACKGROUND: Alkylresorcinols (ARs) are compounds belonging to the class of phenolic lipids. A rich source of ARs are cereal grains such as rye, wheat, triticale or barley. ARs found in plants are characterized by a variety of biological properties such as antimicrobial, antifungal and cytotoxic activity. Moreover, they are proven to have a positive influence on human health. Here, we aimed to find and characterize the gene with ARs synthase activity in the species Secale cereale. METHODS AND RESULTS: Using BAC library screening, two BAC clones containing the gene candidate were isolated and sequenced. Bioinformatic analyses of the resulting contigs were used to examine the structure and other features of the gene, including promoter, intron, 3'UTR and 5'UTR. Mapping using the FISH procedure located the gene on the 4R chromosome. Comparative analysis showed that the gene is highly similar to sequences coding for type III polyketide synthase. The level of gene expression in various parts of the plant was investigated, and the biochemical function of the gene was confirmed by heterologous expression in yeast. CONCLUSIONS: The conducted analyses contributed to a better understanding of the processes related to ARs synthesis. Although the research concerned the rye model, the knowledge gained may help in understanding the genetic basis of ARs biosynthesis in other species of the Poaceae family as well.

Software simulation of tumour motion dose effects during flattened and unflattened ITV-based VMAT lung SBRT.

PURPOSE: Restricted studies comparing different dose rate parameters are available while ITV-based VMAT lung SBRT planning leads to perform the analysis of the most suitable parameters of the external beams used. The special emphasis was placed on the impact of dose rate on dose distribution variations in target volumes due to interplay effects. METHODS: Four VMAT plans were calculated for 15 lung tumours using 6 MV photon beam quality (flattening filter FF vs. flattening filter free FFF beams) and maximum dose rate of 600 MU/min, 1000 MU/min and 1400 MU/min. Three kinds of motion simulations were performed finally giving 180 plans with perturbed dose distributions. RESULTS: 6FFF-1400 MUs/min plans were characterized by the shortest beam on time (1.8 ±â€¯0.2 min). Analysing the performed motion simulation results, the mean dose (Dmean) is not a sensitive parameter to related interplay effects. Looking for local maximum and local minimum doses, some discrepancies were found, but their significance was presented for individual patients, not for the whole cohort. The same was observed for other verified dose metrics. CONCLUSIONS: Generally, the evaluation of VMAT robustness between FF and FFF concepts against interplay effect showed a negligible effect of simulated motion influence on tumour coverage among different photon beam quality parameters. Due to the lack of FFF beams, smaller radiotherapy centres are able to perform ITV-based VMAT lung SBRT treatment in a safe way. Radiotherapy department having FFF beams could perform safe, fast and efficient ITV-based VMAT lung SBRT without a concern about significance of interplay effects.

Large Prostate Volume Does Not Negatively Impact Health-Related Quality of Life in Patients with Prostate Cancer Treated with Ultrahypofractionated Stereotactic Body Radiotherapy.

BACKGROUND: Survival outcomes after primary radiotherapy for localized prostate cancer (PCa) are excellent, regardless of the specific treatment modality. For this reason, health-related quality of life (HRQOL) has come to play an ever more important role in treatment selection. Stereotactic body radiation therapy (SBRT) is increasingly used to treat patients with PCa. However, the impact of prostate volume on HRQOL is not clear. In this study, we aimed to determine whether a large prostate volume negatively influences HRQOL outcomes in patients undergoing ultrahypofractionated SBRT. MATERIAL AND METHODS: We conducted a prospective study of 530 men with low- and intermediate-risk localized PCa. All patients were treated from 2013 to 2017 with SBRT (Cyberknife system). HRQOL data were collected at baseline (pre-treatment), immediately after treatment, and at 12 and 24 months. QOL variables were assessed with the European Organization for Research and Treatment of Cancer QLQ-C30 and PR-25 module. Differences in the QLQ-C30 scales were considered clinically relevant when the change was >10 points. For the analysis, patients were classified into two groups according to prostate volume (≤60 vs. >60 cm3). RESULTS: The prostate volume was ≤60 cm3 in 415 patients (78.3%) and >60 cm3 in 115 (21.7%). No between-group differences were observed at baseline for any of the following variables: clinical stage; hormonal therapy; marital status; educational level; or employment status. No clinically-significant deterioration (functional and symptom scales) was observed in either group between the baseline and 24-month assessment. There were no clinically-relevant differences between the groups on any of the HRQOL variables, regardless of the prostate volume. CONCLUSIONS: This study shows that a large prostate volume (>60 cm3) does not appear to negatively impact HRQOL outcomes at two years in patients with localized prostate cancer treated with ultrahypofractionated SBRT administered with the CyberKnife system.

Multiplexed SSR and agronomic data used in an investigation of obsolete diversity of rye (Secale cereale L.).

Rye (Secale cereale L.) is one of the most important cereal crops cultivated in the world due to its ability to produce high yields even when grown under environmental stress conditions. About 27,000 Secale accessions have been collected and preserved in 70 gene banks worldwide. Although the germplasm represents a great source of genetic diversity, the molecular characteristics refers only to the part of them. Here, we present data obtained by the Simple Sequence Repeat markers (SSR) analysis of 100 rye accessions preserved in the gene bank of the Polish Academy of Sciences Botanical Garden - Center for Biological Diversity Conservation in Powsin. Additionally, the data presented in this article refers to evaluation of agronomoic traits and weather conditions measured for 14 years. The data was used in the research article "Investigation of obsolete diversity of rye (Secale cereale L.) using multiplexed SSR fingerprinting and evaluation of agronomic traits" [1].

Looking for Hidden Enemies of Metabarcoding: Species Composition, Habitat and Management Can Strongly Influence DNA Extraction while Examining Grassland Communities.

Despite the raising preoccupation, the critical question of how the plant community is composed belowground still remains unresolved, particularly for the conservation priority types of vegetation. The usefulness of metabarcoding analysis of the belowground parts of the plant community is subjected to a considerable bias, that often impedes detection of all species in a sample due to insufficient DNA quality or quantity. In the presented study we have attempted to find environmental factors that determine the amount and quality of DNA extracted from total plant tissue from above- and belowground samples (1000 and 10,000 cm2). We analyzed the influence of land use intensity, soil properties, species composition, and season on DNA extraction. The most important factors for DNA quality were vegetation type, soil conductometry (EC), and soil pH for the belowground samples. The species that significantly decreased the DNA quality were Calamagrostis epigejos, Coronilla varia, and Holcus lanatus. For the aboveground part of the vegetation, the season, management intensity, and certain species-with the most prominent being Centaurea rhenana and Cirsium canum-have the highest influence. Additionally, we found that sample size, soil granulation, MgO, organic C, K2O, and total soil N content are important for DNA extraction effectiveness. Both low EC and pH reduce significantly the yield and quality of DNA. Identifying the potential inhibitors of DNA isolation and predicting difficulties of sampling the vegetation plots for metabarcoding analysis will help to optimize the universal, low-cost multi-stage DNA extraction procedure in molecular ecology studies.

Investigation of obsolete diversity of rye (Secale cereale L.) using multiplexed SSR fingerprinting and evaluation of agronomic traits.

Common rye (Secale cereale L.) is one of the most important cereals in Europe. Nevertheless, its germplasm collections are among the least numerous compared with cereals. There are only about 27,000 Secale accessions in 70 gene banks around the world. Despite extensive research on the molecular characterization of genetic resources, only a fraction of this collection has been described. The main objective of the presented study was to perform genotypic and phenotypic characterization of an obsolete gene pool represented by 100 accessions originated from 28 countries around the world and preserved in the gene bank of the Polish Academy of Sciences Botanical Garden - Center for Biological Diversity Conservation in Powsin. Genetic analysis using simple sequence repeat markers showed that the obsolete gene pool is relatively large. This indicates that different sources of variability were used in breeding programs. However, the genetic variation is in no way related to the place of origin. Despite the great differences in the genetic make-up, the collection showed a broadly common phenotype. This could result in a low level of interest among breeders in the stored germplasm, undervalued as a source of important but not easily observable traits, e.g., high disease resistance, which was found in some accessions.

Factors influencing quality of life in patients during radiotherapy for head and neck cancer.

INTRODUCTION: Radiotherapy (RT) in combination with chemotherapy is a standard of care for patients with head and neck squamous cell carcinoma (HNSCC). The RT is associated with side effects, which impact on quality of life (QoL). Thus, the aim of this prospective longitudinal study was to investigate the impact of RT on the QoL of patients with HNSCC during RT. MATERIAL AND METHODS: From September 2008 to February 2010, 205 patients with locally advanced HNSCC were enrolled. The data pertaining to their QoL were collected using the EORTC QLQ-C30 and the EORTC Head and Neck Module (QLQ-H&N35) and then all items were transformed to a 0-100 scale according to the guidelines of the EORTC. The following clinical factors were chosen to study their potential influence on the QoL; site of primary, clinical stage, and methods of therapy: RT vs. chemoradiotherapy (CRT). Additionally, the sociodemographic factors (age, gender, education, habit of smoking) were studied. RESULTS: Deterioration of almost all scales and items in the QLQ-C30 and QLQ-H&N35 questionnaire were noted at the end of RT. The following factors negatively influenced the QoL: age < 60 years (p < 0.05), female gender (p < 0.05), habit of smoking (p < 0.01), advanced clinical stage (III and IV) (p < 0.05), site of primary (larynx, hypopharynx) (p < 0.01), and CRT (p < 0.01). CONCLUSIONS: Our study showed that RT significantly negatively influenced QoL at the end of the RT course. Additionally, this study demonstrated that age, gender, smoking habit, tumor site, and clinical stage of disease showed a significant effect on the QoL of HNSCC patients during RT.