Storage Protocols Research Articles

Strawberry Desiccation and Gloss in Refrigerated Storage Depend on the Interaction between Cultivar and Time after Harvest

The objectives of this study were to characterize strawberry cultivars grown under two production systems for gloss at harvest as a proxy for freshness and assess cultivar differences regarding the loss of gloss and changes in desiccation scores during refrigerated postharvest storage. Nine strawberry cultivars were grown in an annual plasticulture system for 3 years, harvested twice weekly, and packaged for refrigerated storage. Four of these cultivars also were grown in a low tunnel production system and harvested, packaged, and evaluated similarly. In both cases, berry gloss was measured at harvest and again after 1 and 2 weeks of refrigerated storage. Subjective desiccation scores were assigned to the berries during refrigerated storage. For both traits, effects were primarily determined by the interaction between the cultivar and time of measurement relative to harvest. For either gloss or desiccation, there was no evidence of differences in cultivar rankings based on the production system or packaging event. Cordial and Sweet Charlie were among cultivars that did not show loss of gloss or changes in desiccation scores during storage; therefore, they would not be good choices for studies that test alternative postharvest storage protocols. Keepsake and USDA Lumina were among the glossiest cultivars and those with the highest gloss levels and desiccation scores after both 1 week and 2 weeks of refrigerated storage. The variability of these traits among cultivars indicate that improvement can be made when developing new cultivars.

In Vitro conservation and genetic diversity analysis of rare species Ribes janczewskii

Ribes janczewskii is a rare and valuable plant known for its resistance to spring frosts, pests, and diseases. It is used in hybridization to develop resistant currant varieties but is on the verge of extinction, listed in Kazakhstan Red Book. This study developed a micropropagation and slow-growth storage protocol for conservation. Genotypes were identified through DNA barcode analysis (rbcL, ITS, and matK) and sequences uploaded to the National Center for Biotechnology Information database. Genetic diversity was assessed using iPBS primers, generating 98 fragments with 88–94% polymorphic bands. Biochemical analysis of fruits showed vitamin C content from 4.64 to 5.61 mg/100 g, vitamin E from 2.26 to 3.16 mg/100 g, vitamin B5 from 3.18 to 4.93 mg/100 g, and quercetin up to 12.5 mg/100 g. Micropropagation stages were optimized with 12% hydrogen peroxide for surface sterilization, achieving up to 73.3% explant viability. Effective hormonal combinations for in vitro culture included WPM with BAP 0.2 mg L−1 and GA 0.5 mg L−1, and for propagation, BAP 0.25 mg L−1, GA 0.5 mg L−1, and IBA 0.5 mg L−1. Mannitol (20 g L−1) was used for slow-growth storage, keeping explants viable for 4 months without re-cultivation.

Optimising the PVS2 treatment duration for cryopreserving seeds of the endangered rheophytic orchid Epipactis flava Seidenf

ABSTRACT Epipactis flava Seidenf., an endangered Thai rheophytic orchid, urgently requires protection to prevent its extinction. This study aimed to develop an effective vitrification cryopreservation protocol for the long-term storage of E. flava seeds. Mature seeds were pretreated with a loading solution and dehydrated with plant vitrification solution 2 (PVS2) for 0, 30, 60, 90, and 120 min at 25 ± 2°C before immersion in liquid nitrogen for 24 hours. Post-cryopreservation, the seeds were germinated on BM-1 medium for 8 weeks, and then the protocorms were transferred to semi-solid MS medium for another 8 weeks. Seeds treated with PVS2 for 90 min and exposed to liquid nitrogen showed the highest viability (33.3%), while 120 min of dehydration yielded the highest germination rates (17.8%) and 5.8% of the protocorms reached advanced developmental stages (stage 4 and 5). The highest percentage of protocorms at stage 5 (3.3%) was also observed at 120 min of PVS2. No germination occurred in the cryopreserved seeds that had not been exposed to PVS2 prior to the liquid nitrogen treatment. Thus, our results showed that the optimal PVS2 exposure time for cryopreservation of E. flava seeds using the vitrification method was 120 min.

Cold storage of Ganaspis kimorum (Hymenoptera: Figitidae) immature stages for optimized classical biological control of Drosophila suzukii (Diptera: Drosophilidae).

Programs to mass rear and release Ganaspis kimorum Buffington (Hymenoptera: Figitidae) are ongoing in many countries to manage its host Drosophila suzukii (Matsumura) (Diptera: Drosophilidae). To optimize parasitoid production and synchronize field releases with pest outbreaks, we investigated a cold storage protocol by exposing different immature G. kimorum stages to 2 storage temperatures (10 or 15°C) for 2, 4, 6, or 8wk. We further studied those temperature/exposure time combinations that allowed survival but prevented parasitoid emergence before the end of each cold exposure, or the cold storage suitability (CSS). The impact of cold storage on G. kimorum emergence time, total development time, and emergence rate was then evaluated for temperature/exposure time combinations with CSS > 60%. We also assessed fitness costs in terms of longevity and parasitism rates of emerging parasitoids. Results showed that only G. kimorum larvae and pupae were suitable for cold storage under this methodology. Cold exposure significantly delayed the emergence time and total development time of larvae and pupae, whereas the emergence rate was significantly reduced only for larvae at 10°C for 6wk. The longevity of emerging male and female parasitoids was significantly reduced when stored cold as larvae, while no effects were reported for the parasitism rate by emerged females. These findings provide new insights into G. kimorum cold sensitivity and offer valuable options to better schedule mass rearing and wasp releases for biological control of D. suzukii.

Assessing the impact of different donor milk treatments on infant health and growth: a systematic review protocol

IntroductionDonor human milk (DHM) is the first alternative if mother’s own milk is unavailable or contraindicated. Much DHM research has focused on its nutritional, immunological and biochemical composition in response...

A Case Study on Integrating an Eco-Design Tool into the Construction Decision-Making Process

To enhance the sustainability of construction and meet the sector’s environmental agenda, it is essential to comprehensively scrutinize the environmental, social, and economic impacts of construction projects from the project’s design stage. Such assessment is of utmost importance to minimize the impacts of both new construction and rehabilitation projects and is particularly critical during the selection of building materials and construction solutions. This work reports improvements in functionality and user-friendliness of an eco-design tool (UAveiroGreenBuilding) targeting the construction/rehabilitation sector and previously developed within our research group. The optimized version of the eco-design tool underwent validation through the assessment of competitive window frame materials (e.g., wood, PVC, and aluminum) for potential implementation in a rehabilitation project. Windows with PVC frames were identified as the preferred window configuration due to their superior environmental performance and favorable economic profile. Additionally, a digital communication interface was developed to connect the eco-design tool with building information modeling (BIM) projects, achieved through a routine integrated using a Dynamo application. Such successful integration not only streamlined and expedited the data transfer process by obviating the need for manual input but it also enabled the storage of environmental data throughout the life cycle of the project using a simple and reliable data storage protocol.

Continuously tunable single-photon level nonlinearity with Rydberg state wave-function engineering

Extending optical nonlinearity into the extremely weak light regime is at the heart of quantum optics, since it enables the efficient generation of photonic entanglement and implementation of photonic quantum logic gate. Here, we demonstrate the capability for continuously tunable single-photon level nonlinearity, enabled by precise control of Rydberg interaction over two orders of magnitude, through the use of microwave-assisted wave-function engineering. To characterize this nonlinearity, light storage and retrieval protocol utilizing Rydberg electromagnetically induced transparency is employed, and the quantum statistics of the retrieved photons are analyzed. As a first application, we demonstrate our protocol can speed up the preparation of single photons in low-lying Rydberg states by a factor of up to∼40. Our work holds the potential to accelerate quantum operations and to improve the circuit depth and connectivity in Rydberg systems, representing a crucial step towards scalable quantum information processing with Rydberg atoms.

Optimizing adipose-derived stromal vascular fraction storage: Temperature and time impact on cell viability in regenerative medicine.

The adipose-derived stromal vascular fraction (SVF) plays a crucial role in regenerative medicine owing to its regenerative and immunomodulatory properties. However, the effective utilization of SVF in therapeutic applications requires careful consideration of storage conditions to maintain cell viability. We conducted a research on 43 patients of different ages and sexes who were older than 18 years. This study explored the impact of different temperatures (-80, -20, and 4 °C) on SVF storage in platelet-poor plasma for 1 and 6 months. SVF extracted using a semi-UNISTATION™ system was subjected to rigorous analysis of cell count and viability using a LUNA-STEM™ Dual Fluorescence Cell Counter. The results indicated a significant correlation between the storage conditions and SVF viability. Notably, storing SVF at 4 °C demonstrated the highest cell viability and count, while -80 °C storage exhibited the least favorable outcomes. This study emphasizes the importance of minimizing storage time to preserve SVF viability, as evidenced by a decline in both cell count and viability over a 6-month period. Comparisons with the existing literature underscore the need for precise protocols for SVF storage, with considerations for temperature and cryoprotective agents. These findings provide valuable insights for developing optimal SVF storage protocols to enhance therapeutic outcomes and reduce the need for repeated adipose tissue harvesting. Despite the limitations of the study, such as the use of a cell counter instead of flow cytometry, the results establish the foundation for further research on refining SVF storage methods. The ideal storage temperature is from 4 °C, while the length of storage time inversely affects the viability of SVF; the longer the storage time, the lower the number and the viability of SVF cells, regardless of the temperature at which they are preserved.

Roadmap to success: Blueprint for enterprise-wide deployment of a point-of-care ultrasound platform, inclusive of governance, policy, education, credentialing, and quality assurance.

Point-of-care ultrasound (POCUS) has emerged as a cost-effective diagnostic tool that significantly augments physical examinations. Positioned as an extension of traditional examination methods, particularly appealing to the upcoming generation of clinicians, it holds promise in potentially replacing the stethoscope in various medical assessments and procedures. The University of Rochester is deploying 2500 POCUS machines, enhancing compliance for image storage and documentation. Halfway through our 4-year deployment plan, we will discuss our governance structure, educational initiatives, and credentialing strategies. While POCUS adoption has seen organic growth over the past decade, our aim is to implement a comprehensive strategy ensuring adherence to established protocols for image storage and documentation. At present, we have successfully deployed 789 probes, with integration across 64 departments or divisions into our IT platform. Notably, this implementation has resulted in a remarkable 116% increase in hospital charges, underscoring the tangible impact of POCUS integration. However, achieving compliance and education among established providers has proven to be challenging.

IN VITRO SLOW-GROWTH STORAGE OF THE ENDANGERED SPECIES MALUS NIEDZWETZKYANA FOR BIODIVERSITY CONSERVATION

The conservation of biodiversity requires efficient ex situ methods for storing plant genetic material outside its natural habitat. This approach will maintain valuable, endemic, and endangered plant species for extended periods. One widely used method is in vitro slow-growth storage. Our study describes the results of slow-growth storage of micropropagated shoots of the endangered and endemic Malus niedzwetzkyana. In vitro shoots of annual shoots of the Malus niedzwetzkyana were used as the object of the study. The shoots were obtained using a previously developed protocol of micropropagation. To develop slow-growth storage in in vitro culture of shoots, we studied the influence of osmotic agents, light, and temperature regimes. We investigated cultivation conditions such as temperature (4°C and 24°C) and photoperiod (16/8 and growth in the dark), as well as the composition of nutrient media to slow growth (sucrose, mannitol, abscisic acid). The research results showed that the optimal condition for medium-term storage of shoots is the QL nutrient medium with the addition of 3% or 6% sucrose under standard conditions. The optimized protocol allowed us to preserve the shoots of Malus niedzwetzkyana for 5 months without re-cultivation. This result confirms the effectiveness of the selected protocol for slow-growth storage of Malus niedzwetzkyana shoots. The explants stopped growing but remained viable, and the recovery period took 35 days (1 passage). The developed protocol enabled us to preserve plant material for several months without intermediate cultivation. As a result of this work, an in vitro collection was created to conserve the rare and endangered species of Malus niedzwetzkyana. The research is conducted in the framework of the program BR18574125 with financial support from the Ministry of Science and Higher Education of the Republic of Kazakhstan (duration: 2023-2024).

Quality Assessment of Long-Term Cryopreserved Human Bone-Derived Marrow Mesenchymal Stromal Cell Samples: Experience from the Texas Heart Institute Biorepository and Biospecimen Profiling Core.

In biomedical research, biorepositories are pivotal resources that safeguard and supply clinical samples for scientific investigators. Proper long-term cryopreservation conditions are essential to maintain biospecimen quality. In this study, we analyzed the efficacy of sample cryopreservation at the Texas Heart Institute Biorepository and Biospecimen Profiling Core (THI-BRC). Our assessments included a thorough review of internal processes, quality reports, and both internal and external audit outcomes. We examined the integrity of human bone marrow-derived multipotent mesenchymal stromal cells (BM-MSCs) that were cryopreserved for over 5 years. These samples originated from randomly selected clinical trial participants or commercially sourced cell lines. Parameters such as cell viability, DNA and RNA integrity, population doubling time, sterility, and BM-MSC-specific attributes such as surface antigen expression and differentiation potential were studied. BM-MSC samples cryopreserved for ∼6 months served as our control. Our results demonstrated that the 5-year cryopreserved samples maintained their integrity compared with the shorter-term stored control samples. Moreover, THI-BRC has met accreditation agency standards and has not received any repeated deficiencies over 7 years. Collectively, our findings affirm that THI-BRC's biospecimen storage protocols align with accepted standards as confirmed by the quality assessment of long-term stored clinical samples.

Why should a bioengineering lab have engineers and health professionals?

Through the description of the methodology of the development of a bite force measurement device it will be shown how interdisciplinary work of Engineers and Health Professionals bring enhance of life quality to general population. Bite force measurement is a reliable exam to check stomatognathic system (SS) conditions. In order to provide a reliable, low cost and do-it-yourself gnathodynamometer a Dentist joined Bioengineering Laboratory (LabBio) at UFMG. The development of a 3D printed resin structure was made using CAD/CAM and tested by means of FEM. A Carbon Nano Tube (CNT) extensometer developed at CTNANO at UFMG to capture the structure deformation were fixed in two geometries that showed good results in FEM simulation and will be tested in an EMIC universal mechanical testing machine (DL1000) equipped with a 1 kN load cell. The electrical response of the extensometers was monitored using a Keithley 2000 digital multimeter (Tektronix), connected to a computer and remotely controlled by a LabView application (View Point Systems), a data storage protocol in a SD card and in the clouds using IoT and an data acquisition system will be tested. In bench tests both geometries showed good results with deformation capturable from 40 N. Data was codified using an Arduino Nano and a program was developed for data acquisition and storage. The interdisciplinary work generates prototypes with promising results in bench tests. The fruit of the team work may generate a toll that improves life quality of population by allowing more people to be tested and lowering health costs.

Sub-zero non-freezing of vascularized composite allografts in a rodent partial hindlimb model

Ischemia is a major limiting factor in Vascularized Composite Allotransplantation (VCA) as irreversible muscular injury can occur after as early as 4–6 h of static cold storage (SCS). Organ preservation technologies have led to the development of storage protocols extending rat liver ex vivo preservation up to 4 days. Development of such a protocol for VCAs has the added challenge of inherent ice nucleating factors of the graft, therefore, this study focused on developing a robust protocol for VCA supercooling. Rodent partial hindlimbs underwent subnormothermic machine perfusion (SNMP) with several loading solutions, followed by a storage solution with cryoprotective agents (CPA) developed for VCAs. Storage occurred in suspended animation for 24h and VCAs were recovered using SNMP with modified Steen. This study shows a robust VCA supercooling preservation protocol in a rodent model. Further optimization is expected to allow for its application in a transplantation model, which would be a breakthrough in the field of VCA preservation.

Analysis of the morphology of Sus scrofa domesticus oocyte-cumulus complexes exposed to low and ultra-low temperatures

Relevance. The priority task of reproductive technologies in animal husbandry is the development of effective storage protocols for female gametes. One way to preserve gametes is vitrification, which reduces damage to intracellular organelles by increasing viscosity upon cooling and minimizing crystallization.Methods. Various additives to cryoprotectants are used for optimization. In our work, titanium tetrapolyethylene glycolate was used in a 10-fold molar excess of polyethylene glycol (TTPEG*10PEG), the nature of the effect of which on cells was studied during short-term storage at low (5 °С) and ultra-low temperatures (-196 °С).The purpose of the study is a comprehensive analysis of the morphology of gametes and somatic cells (cumulus) of ovarian follicles of pigs after exposure to low and ultra-low temperatures (vitrification) when included in the TTPEG*10PEG low-temperature storage and vitrification protocol.Results. Exposure to TTPEG*10PEG at 5 °С caused an increase in the level of oocytes with subsequent morphological degenerations in points with control (from 13% to 5%, р = 0.005). After vitrification with TTPEG*10PEG: the level of gametes with low cumulus cells expansion increased to 35% in terms of the proportion of control gametes (23%); the proportion of denudated gametes was reduced to 50% compared to the control 65% (р < 0.05); the level of morphologically degenerated gametes corresponded to that of native oocytes (8% each) and was lower than in the control (17%), р < 0.005. The obtained data indicate the protective and cryoprotective effects of TTPEG*10PEG at a concentration of 2%, which suggests the possibility of its use in the technology of intraovarian vitrification of female reproductive cells.

A long cold storage protocol for Anisopteromalus calandrae based on promoting prepupal development and arresting early pupal growth under low temperature.

Anisopteromalus calandrae (Howard) shows great promise as an ectoparasitoid for controlling various coleopteran pests in warehouses. However, for a large-scale release, it is crucial to establish an ample supply of A. calandrae while carefully maintaining their quality and effectiveness. Appropriate cold storage techniques are the key to achieving these goals. Previous studies on cold storage have focused on specific developmental stages and explored cold storage conditions that can be applied only to those stages. Herein, we examined the development, survival and reproductive capacity of A. calandrae at different temperatures (13, 16, and 19°C) and storage durations (30, 60, and 90 d) and evaluated the fitness of the offspring. A. calandrae completed its egg-to-larva development and pupated at 16°C, but its development was arrested at an early pupal stage. Even after 90 d of cold storage at 16°C, the survival rate of A. calandrae remained high at 77%, with no significant impact on reproductive capacity. Furthermore, cold storage showed no negative effect on the F1 generation. In contrast, eggs stored at 13°C failed to hatch, whereas those stored at 19°C developed. Adults emerged after > 60 d. This indicates that storage at 19°C is only suitable for short durations. Our findings highlight the developmental pattern of A. calandrae at 16°C, indicating that the parasitic wasp can be stored for a long time at this temperature across all stages of development before pupation, substantially facilitating its mass reproduction and industrial production.

An improved identity-based public audit protocol for cloud storage

With the rapid development of informatization, a vast amount of data is continuously generated and accumulated, leading to the emergence of cloud storage services. However, data stored in the cloud is beyond the control of users, posing various security risks. Cloud data auditing technology enables the inspection of data integrity in the cloud without the necessity of data downloading. Among these, public auditing schemes have experienced rapid development due to their ability to avoid additional user auditing expenses. However, malicious third-party auditors can compromise data privacy. This paper proposes an improved identity-based cloud auditing scheme that can resist malicious auditors. This scheme is also constructed on an identity-based public auditing scheme using blockchain to prevent malicious auditing. We found the scheme is not secure because a malicious cloud server can forge authentication tags for outsourced data blocks, while our scheme has not these security flaws. Through security proofs and performance analysis, we further demonstrate that our scheme is secure and efficient. Additionally, our scheme has typical application scenarios.

Revisiting serum storage protocols: assessing the impact of temperature variations on indirect immunofluorescence testing for pemphigus and bullous pemphigoid.

Indirect immunofluorescence (IIF) plays a crucial role in the diagnosis of pemphigus and bullous pemphigoid (BP) by detecting the presence of circulating autoantibodies in the serum of patients. The standard serum transportation method requires delivery to laboratories at 2-8 °C within a day and storage at -20 to -80 °C. However, this protocol poses logistical challenges. To carry out a study assessing how temperature variations affect the accuracy of IIF tests. This case-control study analysed 203 serum specimens: 102 from patients with pemphigus and 101 from patients with BP. Specimens were stored at -80 °C (control), 24 °C and 40 °C for 7 days before analysis to investigate variations in IIF titres between test and control conditions. In pemphigus serum, 95% of specimens at 24 °C and 76% at 40 °C showed no titre difference compared with controls. Similarly, 89% of BP serum specimens at 24 °C and 82% at 40 °C matched the control titres. While 57 specimens across both groups experienced reduced titres, the decrease was primarily marginal (a 1-step reduction in 54 cases, 2-step in 3), with no transition from positive to negative results. Storing serum at 24-40 °C for up to 7 days before testing only slightly influences IIF outcomes for pemphigus and BP. These findings could prompt a significant revision in the existing strict transport guidelines, ensuring efficient use of resources without sacrificing the accuracy of diagnostic tests.

Tracking the quality of graphene oxide suspension during long-term storage

Graphene oxide is a popular material because of not only its cost–efficient production but also its unique properties. However, after long-term storage, several changes occur in the structure of graphene oxide obtained using the modified Hummers’ method, affecting its stability and physicochemical properties. Despite the obvious problem, there is only a limited number of works devoted to ageing factors, their impact on the properties and structure of graphene oxide itself, and operational modes of materials and devices based on graphene oxide. Therefore, to facilitate the widespread industrial application of graphene oxide, it is crucial to establish standardized protocols for storage and quality assessment of graphene oxide. In this paper, the prepared samples were extensively characterized using various techniques, including transmission electron microscopy, X-ray photoelectron spectroscopy, UV–Vis and fluorescence spectroscopy techniques. Among the various analytical methods, comprehensive spectroscopic analysis of the absorption and fluorescence properties has proven to be the most sensitive to sample ageing. We investigated the change in optical and physicochemical properties of graphene oxide during its long-term storage under various conditions. It was found that graphene oxide precipitated during storage, and the O/C ratio in graphene oxide decreased slightly from 0.49 to 0.47. Still, the type of oxygen-containing functional groups did not change, which might indicate possible intra-structural changes. Using transmission electron microscopy, molecular dynamic and density functional theory simulation, we showed that the rearrangement of functional groups on the surface leads to the formation of large sp2 domains that interact with each other, leading to aggregation of the graphene oxide.

Effect of short-term storage on cannabinoid content of dried floral hemp (Cannabis sativa L) inflorescence

Floral hemp, a plant rich in cannabinoids, has gained attention for its potential therapeutic properties. It is crucial to have proper storage conditions that will maintain the cannabinoid content. To the best of our knowledge, very little has been done on the storage conditions of dried floral hemp. Hence, this study investigated the effect of storage conditions like moisture content, temperature, storage time, light exposure, and packaging materials, on cannabinoid stability in dried floral hemp. The cultivar ‘Quick Spectrum’ at two moisture levels (8.5 and 4.5 %) was stored over a 6-month storage period. Results indicated that a higher amount of cannabinoids samples was retained at 8.5 ± 1 % moisture compared to samples at 4.5 ± 0.5 %, even though total CBD, total ∆9-THC, and total CBDV content remained statistically the same throughout the storage period, and was not affected by temperature, lighting, and packaging material. In contrast, certain individual cannabinoids, such as CBDVA, CBD, ∆9-THCA, and ∆9-THC were significantly affected by temperature and lighting. Notably, higher storage temperatures (22 °C) and light exposure (F15T8/F5 fluorescent light) increased the degradation and decarboxylation rate of cannabinoids. These findings showed the importance of optimized storage conditions for floral hemp, especially long-term storage. This will help provide standardized storage protocols to aid producers, testing laboratories, distributors, and consumers in making informed decisions about storage practices and advancing quality control in the hemp industry.

Blackberry synthetic seeds storage: effects of temperature, time, and sowing substrate

In vitro propagation, is becoming the predominant method for blackberry propagation due to its advantages compared to agamic traditional propagation methods. Synthetic seed technology represents a promising approach to further enhance the productivity of in vitro propagation facilitating the exchange of plant materials among laboratories and contributing to germplasm conservation efforts. This study aimed to establish an optimal protocol for the storage and sowing of synthetic blackberry seeds obtained through the encapsulation of clump bases. The synthetic seeds were sown without storage (Control) and after storage periods of 30, 60, and 120 days at 4 °C and 25 °C in the dark, in three different substrates (agarised, perlite, and potting). After forty-five days from sowing viability, regeneration rate, shoot and root numbers and lengths, as well as fresh and dry weights of the plantlets, were assessed. Results indicated that agarised substrate consistently exhibited favourable outcomes, with sustained regeneration rates and robust plantlet development even after prolonged storage at 4 °C. Synthetic seeds sown in perlite and potting substrates demonstrated enhanced regeneration rates following storage at 4 °C for 60 and 120 days. On the contrary, storage at 25 °C resulted in a notable decline in regeneration rate, highlighting its inadequacy for blackberry synthetic seed conservation purposes. These findings underscore the importance of sowing substrate selection and storage temperature in optimizing the storage and sowing protocols for synthetic blackberry seeds.