The Driving Forces of Anoplophora glabripennis Have Spatial Spillover Effects

Anoplophora glabripennis Motschulsky, 1854 (Asian longhorned beetle) does serious damage to forests. It has a long history and wide distribution area in China and is spreading there and elsewhere. Extreme climate events, such as cold surges and droughts, have had a promotive impact on Anoplophora glabripennis occurrence, but the spatial spillover effect of extreme climate events and other environmental factors on the occurrence of this pest has not yet been clarified. Two indices, namely, Standardized Precipitation Evapotranspiration Index (SPEI) and Low Temperature Index (LTI), were used to quantify the effects of drought and low-temperature freezing damage. Based on spatial panel data modeling, this study calculated the spatial spillover effect of environmental factors on the incidence of Anoplophora glabripennis in 666 counties in China’s central plains from 2002 to 2009. The factors examined included LTI, SPEI, average wind speed, hours of sunlight, Gross Domestic Product (GDP) of regional primary industry, population density, Normalized Difference Vegetation Index (NDVI), and pest control rate. Study results indicated that the impacts of environmental factors on the incidence rate of Anoplophora glabripennis are different. Low-temperature freezing damage, drought, wind speed, and pest control rate had a driving impact on pest incidence rates. Overall, the direct effect accounts for about 85% of the total effect, while the indirect effect accounts for about 15% of the total effect.

A method for obtaining field wheat freezing injury phenotype based on RGB camera and software control

Background Low temperature freezing stress has adverse effects on wheat seedling growth and final yield. The traditional method to evaluate the wheat injury caused by the freezing stress is by visual observations, which is time-consuming and laborious. Therefore, a more efficient and accurate method for freezing damage identification is urgently needed. Results A high-throughput phenotyping system was developed in this paper, namely, RGB freezing injury system, to effectively and efficiently quantify the wheat freezing injury in the field environments. The system is able to automatically collect, processing, and analyze the wheat images collected using a mobile phenotype cabin in the field conditions. A data management system was also developed to store and manage the original images and the calculated phenotypic data in the system. In this experiment, a total of 128 wheat varieties were planted, three nitrogen concentrations were applied and two biological and technical replicates were performed. And wheat canopy images were collected at the seedling pulling stage and three image features were extracted for each wheat samples, including ExG, ExR and ExV. We compared different test parameters and found that the coverage had a greater impact on freezing injury. Therefore, we preliminarily divided four grades of freezing injury according to the test results to evaluate the freezing injury of different varieties of wheat at the seedling stage. Conclusions The automatic phenotypic analysis method of freezing injury provides an alternative solution for high-throughput freezing damage analysis of field crops and it can be used to quantify freezing stress and has guiding significance for accelerating the selection of wheat excellent frost resistance genotypes.

Application of Supercooling for the Enhanced Shelf Life of Asparagus (Asparagus officinalis L.)

Freezing extends the shelf-life of food by slowing down the physical and biochemical reactions; however, ice crystal formation can result in irreversible damage to the cell’s structure and texture. Supercooling technology has the potential to preserve the original freshness of food without freezing damage. In this study, fresh asparagus was preserved in a supercooled state and its quality changes such as color, weight loss, texture, chlorophyll and anthocyanin content, and enzymatic activities (superoxide dismutase and catalase) were evaluated. The asparagus samples were successfully supercooled at −3 °C with the combination treatment of pulsed electric field (PEF) and oscillating magnetic field (OMF), and the supercooled state was maintained for up to 14 days. Asparagus spears preserved in the supercooled state exhibited lower weight loss and higher levels of quality factors in comparison to the frozen and refrigerated control samples.

Cold-Triggered Induction of ROS- and Raffinose Metabolism in Freezing-Sensitive Taproot Tissue of Sugar Beet

Sugar beet (Beta vulgaris subsp. vulgaris) is the exclusive source of sugar in the form of sucrose in temperate climate zones. Sugar beet is grown there as an annual crop from spring to autumn because of the damaging effect of freezing temperatures to taproot tissue. A collection of hybrid and non-hybrid sugar beet cultivars was tested for winter survival rates and freezing tolerance. Three genotypes with either low or high winter survival rates were selected for detailed study of their response to frost. These genotypes differed in the severity of frost injury in a defined inner region in the upper part of the taproot, the so-called pith. We aimed to elucidate genotype- and tissue-dependent molecular processes during freezing and combined analyses of sugar beet anatomy and physiology with transcriptomic and metabolite profiles of leaf and taproot tissues at low temperatures. Freezing temperatures induced strong downregulation of photosynthesis in leaves, generation of reactive oxygen species (ROS), and ROS-related gene expression in taproots. Simultaneously, expression of genes involved in raffinose metabolism, as well as concentrations of raffinose and its intermediates, increased markedly in both leaf and taproot tissue at low temperatures. The accumulation of raffinose in the pith tissue correlated with freezing tolerance of the three genotypes. We discuss a protective role for raffinose and its precursors against freezing damage of sugar beet taproot tissue.

Experimental Study on Critical Membrane Water Content of Proton Exchange Membrane Fuel Cells for Cold Storage at −50 °C

Membrane water content is of vital importance to the freezing durability of proton exchange membrane fuel cells (PEMFCs). Excessive water freezing could cause irreversible degradation to the cell components and deteriorate the cell performance and lifetime. However, there are few studies on the critical membrane water content, a threshold beyond which freezing damage occurs, for cold storage of PEMFCs. In this work, we first proposed a method for measuring membrane water content using membrane resistance extracted from measured high frequency resistance (HFR) based on the finding that the non-membrane resistance part of the measured HFR is constant within the range of membrane water content of 2.98 to 14.0. Then, freeze/thaw cycles were performed from −50 °C to 30 °C with well controlled membrane water content. After 30 cycles, cells with a membrane water content of 8.2 and 7.7 exhibited no performance degradation, while those higher than 8.2 showed significant performance decay. Electrochemical tests revealed that electrochemical surface area (ECSA) reduction and charge transfer resistance increase are the main reasons for the degradation. These results indicate that the critical membrane water content for successful cold storage at −50 °C is 8.2.

P–054 Apoptosis related-microRNAs in Oligoasthenoteratozoospermia and Azoospermia men may reveal novel study of freezing damage

Study question Could choosing of non-apoptotic spermatozoa by biological biomarkers such as microRNAs promote post-thaw fertilization ability? Summary answer Biological alterations in correlation with apoptosis and oxidative stress markers such as microRNAs may preserve the function and fertility of spermatozoa during cryopreservation. What is known already Biological changes of cryopreserved spermatozoa such as microRNAs against cryo-injury were investigated. It was presented that several sperm parameters such as motility and abstinence period can impact the percentage of post-thaw sperm survival. Recent study, reported that microRNAs related to process of motility, sperm structure and apoptosis were associated with different expression after cryopreservation. More comprehensive study needed to fully mention the effect of microRNAs and their correlations with other biomarkers in cryopreservation. Study design, size, duration Our study was performed on 58 men who were 24–40 years old. Their ejaculated samples were classified as sever (concentrations less than 5 million sperm/mL) Oligoasthenoteratozoospermia (SOAT), mild (concentrations 5 million – 10 million sperm/ mL) Oligoasthenoteratozoospermia (MOAT), obstructive azoospermai (OA), Non obstructive azoospermai (NOA) (absence of spermatozoa in the semen) and normal group (concentrations more than 15 million sperm/ mL). Then each sample was grouped into fresh and cryopreserved one. Participants/materials, setting, methods Density Gradient centrifugation (DGC) was performed to obtain high quality sperm without round cells after freeze-thawing. Biopsy of testicular tissue was prepared after Testicular Sperm Extraction (TESE) surgery. Then biological biomarkers were examined before and after cryopreservation including microRNA–122 (miR–122), miR–383, miR–15b, miR–184, miR–34c and target genes such as P53, Caspase9 and CYCLIN D1, using Quantitative real-time polymerase chain reaction (RT-PCR). Glutathione peroxidase (GPx), Superoxide dismutase (SOD) and malondialdehyde (MDA) using imaging multi-mode reader. Main results and the role of chance There was a significant reduction in sperm total motility and morphology in Cryopreserved-infertile groups (MOAT and SOAT) compared with the Fresh-infertile groups. Decreased level of GPx activity was associated with increased concentration of MDA during freeze-thawing procedure in oligoasthenoteratozoospermia. Also increasing levels of SOD, and DNA fragmentation were showed. Our data demonstrated that reduction of CYCLIN D1 in MOAT-Cryopreserved (P = 0.0174) and NOA-Cryopreserved (P = 0.0001) groups were considerable compared with their fresh ones. We observed high level of Capase9 and in cryopreserved groups (P = 0.01).The expression of miR–34c was increased significantly in NOA-Cryo (P = 0.0064), and OA-Cryo (P = 0.0441) in comparison with their fresh groups. The expression of miR–184 (P = 0.0275) was enhanced in NOA-Cryo as compared to NOA-Fresh. Quantitative RT-PCR demonstrated meaningful decrease level of miR–383 expression in SOAT-Cryopreserved as compared with SOAT-Fresh (P = 0.0223). On the other hand, expression level of miR–383 was increased in NOA group significantly (P = 0.0437) and in OA group non-significantly during freezing. There was non-significant decrease of miR–122 and miR–15b in MOAT and SOAT-Cryopreserved groups in comparison to their Fresh groups. We observed reduced expression of miR–122 (P = 0.0109) and miR–15b (P = 0.0322) in OA group after freezing. Also, there was meaningful increased level of miR–15b (P = 0.0234) in NOA-Cryo compared with Fresh. Limitations, reasons for caution Because of the ethical principle, we can not obtain testicular samples from normal groups. So, we analyzed NO and OA groups with each other. Wider implications of the findings: Our study documented that total motility can be interfered by microRNAs. This phenomenon effects on the total motility of post-thaw spermatozoa. Also the increase level of MDA may disturb microRNAs regulation in the infertile cases. These non-coding RNAs may be known as fertility biomarker to development of freeze-thawing strategies. Trial registration number 60961

P-054 Pre-selected for an award: Apoptosis related-microRNAs in Oligoasthenoteratozoospermia and Azoospermia men may reveal novel study of freezing damage

Study question Could choosing of non-apoptotic spermatozoa by biological biomarkers such as microRNAs promote post-thaw fertilization ability? Summary answer Biological alterations in correlation with apoptosis and oxidative stress markers such as microRNAs may preserve the function and fertility of spermatozoa during cryopreservation. What is known already Biological changes of cryopreserved spermatozoa such as microRNAs against cryo-injury were investigated. It was presented that several sperm parameters such as motility and abstinence period can impact the percentage of post-thaw sperm survival. Recent study, reported that microRNAs related to process of motility, sperm structure and apoptosis were associated with different expression after cryopreservation. More comprehensive study needed to fully mention the effect of microRNAs and their correlations with other biomarkers in cryopreservation. Study design, size, duration Our study was performed on 58 men who were 24-40 years old. Their ejaculated samples were classified as sever (concentrations less than 5 million sperm/mL) Oligoasthenoteratozoospermia (SOAT), mild (concentrations 5 million – 10 million sperm/ mL) Oligoasthenoteratozoospermia (MOAT), obstructive azoospermai (OA), Non obstructive azoospermai (NOA) (absence of spermatozoa in the semen) and normal group (concentrations more than 15 million sperm/ mL). Then each sample was grouped into fresh and cryopreserved one. Participants/materials, setting, methods Density Gradient centrifugation (DGC) was performed to obtain high quality sperm without round cells after freeze-thawing. Biopsy of testicular tissue was prepared after Testicular Sperm Extraction (TESE) surgery. Then biological biomarkers were examined before and after cryopreservation including microRNA-122 (miR-122), miR-383, miR-15b, miR-184, miR-34c and target genes such as P53, Caspase9 and CYCLIN D1, using Quantitative real-time polymerase chain reaction (RT-PCR). Glutathione peroxidase (GPx), Superoxide dismutase (SOD) and malondialdehyde (MDA) using imaging multi-mode reader. Main results and the role of chance There was a significant reduction in sperm total motility and morphology in Cryopreserved-infertile groups (MOAT and SOAT) compared with the Fresh-infertile groups. Decreased level of GPx activity was associated with increased concentration of MDA during freeze-thawing procedure in oligoasthenoteratozoospermia. Also increasing levels of SOD, and DNA fragmentation were showed. Our data demonstrated that reduction of CYCLIN D1 in MOAT-Cryopreserved (P = 0.0174) and NOA-Cryopreserved (P = 0.0001) groups were considerable compared with their fresh ones. We observed high level of Capase9 and in cryopreserved groups (P = 0.01).The expression of miR-34c was increased significantly in NOA-Cryo (P = 0.0064), and OA-Cryo (P = 0.0441) in comparison with their fresh groups. The expression of miR-184 (P = 0.0275) was enhanced in NOA-Cryo as compared to NOA-Fresh. Quantitative RT-PCR demonstrated meaningful decrease level of miR-383 expression in SOAT-Cryopreserved as compared with SOAT-Fresh (P = 0.0223). On the other hand, expression level of miR-383 was increased in NOA group significantly (P = 0.0437) and in OA group non-significantly during freezing. There was non-significant decrease of miR-122 and miR-15b in MOAT and SOAT-Cryopreserved groups in comparison to their Fresh groups. We observed reduced expression of miR-122 (P = 0.0109) and miR-15b (P = 0.0322) in OA group after freezing. Also, there was meaningful increased level of miR-15b (P = 0.0234) in NOA-Cryo compared with Fresh. Limitations, reasons for caution Because of the ethical principle, we can not obtain testicular samples from normal groups. So, we analyzed NO and OA groups with each other. Wider implications of the findings Our study documented that total motility can be interfered by microRNAs. This phenomenon effects on the total motility of post-thaw spermatozoa. Also the increase level of MDA may disturb microRNAs regulation in the infertile cases. These non-coding RNAs may be known as fertility biomarker to development of freeze-thawing strategies. Trial registration number 60961

Standardization of electrolyte leakage data and a novel liquid nitrogen control improve measurements of cold hardiness in woody tissue

Background A variety of basic and applied research programs in plant biology require the accurate and reliable determination of plant tissue cold hardiness. Over the past 50 years, the electrolyte leakage method has emerged as a popular and practical method for quantifying the amount of damage inflicted on plant tissue by exposure to freezing temperatures. Numerous approaches for carrying out this method and analyzing the resultant data have emerged. These include multiple systems for standardizing and modeling raw electrolyte leakage data and multiple protocols for boiling or autoclaving samples in order to maximize leakage as a positive control. We compare four different routines for standardization of leakage data and assess a novel control method—immersion in liquid nitrogen in lieu of traditional autoclaving—and apply them to woody twigs collected from 12 maple (Acer) species in early spring. We compare leakage data from these samples using each of four previously published forms of data analysis and autoclaving vs. liquid nitrogen controls and validate each of these approaches against visual estimates of freezing damage and differential thermal analysis. Results Through presentation of our own data and re-analysis of previously published findings, we show that standardization of raw data against estimates of both minimum and maximum attainable freezing damage allows for reliable estimation of cold hardiness at the species level and across studies in diverse systems. Furthermore, use of our novel liquid nitrogen control produces data commensurate across studies and enhances the consistency and realism of the electrolyte leakage method, especially for very cold hardy samples. Conclusion Future leakage studies that relativize data against minimum and maximum leakage and that employ our updated liquid nitrogen control will contribute generalizable, repeatable, and realistic data to the existing body of cold hardiness research in woody plants. Data from studies conducted using a liquid nitrogen (and not an autoclaving) control can still be compared to previously published data, especially when raw data are standardized using the best-performing approach among those we assessed. Electrolyte leakage of woody twigs emerges as a useful technique for quickly assessing the probability of tissue death in response to freezing in dormant plants. Differential thermal analysis may provide different and complementary information on cold hardiness.