Strawberry Genes and Genomics

Despite its value as a crop and potential utility as an experimental system, relatively little is known about the molecular-genetic aspects of inheritance or physiology in the cultivated strawberry (Fragaria xananassa). This lack of information exists at a time when biotechnology may offer important remedies to address traditional and contemporary challenges that growers face. An improved understanding of genome structure will hasten the development of molecular markers and unveil clues to the composition of this unique, octoploid genome. Definition of gene function will guide the generation of transgenic resources for research use and possibly toward cultivar development. This review seeks to compile and present the current knowledge state of the molecular-genetic basis of cultivated strawberry genomic form and function. Ongoing studies promise to expand the use of genomic tools and appropriate model systems to rapidly discern the structural and functional basis for traits of interest to agriculture, such as those associated with disease, ripening, and volatile production. Together these studies bring new molecular tools to dissect complex traits, implement marker-assisted selection and address important physiological questions in the cultivated strawberry, the Fragaria genus, and the Rosaceae family.     

冻结速率对血小板冷冻干燥保存的影响

冷冻干燥法是使血小板能够长期保存的一种理想方法。冻结过程对血小板的冻干保存至关重要。采用梯度降温、搁板预冷、液氮冻结等三种冻结方式,研究了冻结速率对血小板冷冻干燥保存恢复率的影响。实验结果表明用搁板预冷的方式冻结并干燥的血小板复水后的恢复率最高,达到(93.0.2)%,此时的冻结速度约为10℃/min。扫描电镜照片显示冻干复水后的血小板保持了完整的细胞结构,但与新鲜血小板相比略呈球形。冻干复水后的血小板对1U/ml凝血酶的最大聚集率接近于新鲜血小板,但聚集速度比新鲜血小板慢。     

"两步法"体外培养山羊卵母细胞的超微结构观察

有假说认为,卵母细胞在体外培养过程中,如果延长GV期,可促进卵母细胞进一步成熟,因而提高发育潜能。采用山羊半卵泡和卵母细胞共培养,抑制卵母细胞GVBD发生,从而延长GV期。比较了共培养前后和恢复成熟培养后卵母细胞的超微结构变化,其目的从亚细胞水平寻找卵母细胞进一步成熟的证据。研究发现,常规成熟培养:有卵周隙存在,但不贯通,局部区域卵膜与透明带结合紧密;部分皮质区尚有细胞器存在;微绒毛大部分从透明带中撤出,倒伏于质膜表面,数量较多,形态较为粗大;皮层颗粒质膜下部分单层分布,部分散布于皮质区;线粒体均匀散布于卵质中央区。共培养前:卵母细胞的卵周隙尚未形成,微绒毛没有从透明带中撤出;线粒体等细胞器分布于皮质区,皮层颗粒成簇状分布,皮质区富含细胞器。共培养后:局部形成卵周隙,微绒毛已自透明带中撤出,数量较多,垂直或倒伏于卵膜表面;线粒体以簇状分批开始内移,皮层颗粒已部分单层分布于质膜下,部分皮质区缺乏细胞器。恢复成熟培养后:卵周隙进一步扩大并且贯通,微绒毛数量减少并且绝大多数垂直于卵膜;线粒体在卵质中央区均匀分布,皮层颗粒卵膜下单层分布,大部分皮质区无细胞器存在。利用“两步法”培养得到的卵母细胞与体外常规成熟培养的卵母细胞相比,更有利于皮层颗粒的质膜下单层分布,卵母细胞卵周隙的形成与贯通,微绒毛数量减少和垂直于卵膜表面,无细胞器皮层区的进一步形成。因此,更有利于卵母细胞胞质的进一步成熟。     

A new approach for freezing of aqueous solutions under active control of the nucleation temperature

An experimental setup for controlled freezing of aqueous solutions is introduced. The special feature is a mechanism to actively control the nucleation temperature via electrofreezing: an ice nucleus generated at a platinum electrode by the application of an electric high voltage pulse initiates the crystallization of the sample. Using electrofreezing, the nucleation temperature in pure water can be precisely adjusted to a desired value over the whole temperature range between a maximum temperature Tn(max) close to the melting point and the temperature of spontaneous nucleation. However, the presence of additives can inhibit the nucleus formation. The influence of hydroxyethylstarch (HES), glucose, glycerol, additives commonly used in cryobiology, and NaCl on Tn(max) were investigated. While the decrease showed to be moderate for the non-ionic additives, the hindrance of nucleation by ionic NaCl makes the direct application of electrofreezing in solutions with physiological salt concentrations impossible. Therefore, in the multi-sample freezing device presented in this paper, the ice nucleus is produced in a separate volume of pure water inside an electrode cap. This way, the nucleus formation becomes independent of the sample composition. Using electrofreezing rather than conventional seeding methods allows automated freezing of many samples under equal conditions. Experiments performed with model solutions show the reliability and repeatability of this method to start crystallization in the test samples at different specified temperatures. The setup was designed to freeze samples of small volume for basic investigations in the field of cryopreservation and freeze-drying, but the mode of operation might be interesting for many other applications where a controlled nucleation of aqueous solutions is of importance.     

Differential responses of Arabidopsis ecotypes to cold, chilling and freezing temperatures

Arabidopsis plants show an increase in freezing tolerance in response to exposure to low nonfreezing temperatures, a phenomenon known as cold acclimation. In the present study, we evaluated the physiological and morphological responses of various Arabidopsis ecotypes to continuous growth under chilling (14°C) and cold (6°C) temperatures and evaluated their basal freezing tolerance levels. Seedlings of Arabidopsis plants were extremely sensitive to low growth temperatures: the hypocotyls and petioles were much longer and the angles of the second pair of true leaves were much greater in plants grown at 14°C than in those grown at 22°C, whereas just intermediate responses were observed under the cold temperature of 6°C. Flowering time was also markedly delayed at low growth temperatures and, interestingly, lower growth temperatures were accompanied by longer inflorescences. Other marked responses to low temperatures were changes in pigmentation, which appeared to be both ecotype specific and temperature dependent and resulted in various visual phenotypes such as chlorosis, necrosis or enhanced accumulation of anthocyanins. The observed decreases in chlorophyll contents and accumulation of anthocyanins were much more prominent in plants grown at 6°C than in those grown at 14°C. Among the various ecotypes tested, Mt‐0 plants markedly accumulated the highest levels of anthocyanins upon growth at 6°C. Freezing tolerance examination revealed that among 10 ecotypes tested, only C24 plants were significantly more sensitive to subzero temperatures. In conclusion, Arabidopsis ecotypes responded differentially to cold (6°C), chilling (14°C) and freezing temperatures, with specific ecotypes being more sensitive in particular traits to each low temperature.     

张家界山区角倍蚜两虫态过冷却点比较及人工植藓保蚜越冬技术

为了解张家界地区所产角倍蚜Schlechtendalia chinensis(Bell)秋迁蚜及越冬若蚜的耐寒性及两虫态间在耐寒性方面的差异,应用过冷却点测定仪,抽样测定了上述两虫态个体的过冷却点和体液结冰点,结果表明:秋迁角倍蚜过冷却点和体液结冰点的平均值、最低值和最高值分别依次为-13.32±0.77,-15.69,-11.12℃和-13.00±0.72,-14.54,-10.80℃;而无翅若蚜过冷却点和体液结冰点的平均值、最低值和最高值分别依次为-18.95±0.82,-20.24,-17.02℃和-18.70±0.82,-20.07,-16.95℃。方差分析表明,越冬若蚜的过冷却点和体液结冰点都明显低于秋迁蚜的过冷却点和体液结冰点,进而推测,越冬若蚜的耐寒能力要显著强于秋迁蚜的耐寒能力。试验还表明,张家界地区角倍蚜的次生寄主苔藓(侧枝匐灯藓Plagiomnium maximoiczii)通过人工植藓可在小型聚酯瓶(polyester bottle)内正常生长,接种后的秋迁蚜可顺利生产若蚜,若蚜也能顺利产织蜡球进而度过寒冷的冬天直至下一年春迁蚜产生。     

Preservative fluid and storage conditions alter body mass estimation in a terrestrial insect

Body mass is a frequently used trait in ecological and evolutionary research. In the present study, I demonstrate that sampling and storage conditions affect wet and dry weights in an insect predator, Anchomenus dorsalis (Pontoppidan) (Coleoptera: Carabidae). Live beetles were placed in one of five preservative fluids for 1 month to simulate sampling by pitfall traps. Sodium chloride solution, ethylene glycol, ethylene glycol + detergent, and propylene glycol caused significant increases in both wet and dry weights compared with control (short‐term frozen) specimens, whereas formaldehyde did not. In a separate experiment, four methods of long‐term (6 months) sample storage (freezing, ethanol, propylene glycol, and ethyl acetate vapour) all caused significant changes in wet weight compared with the control treatment. The dry weight of the specimens preserved in ethanol decreased significantly in contrast to the long‐term frozen specimens and those in propylene glycol and ethyl acetate vapour, whose dry weight did not differ significantly from the control specimens. The combination of formaldehyde as the preservative fluid and freezing as the storage method thus appears to be an optimal combination for studies in which the body mass of dead insects is considered.     

Improved cryopreservation of in vitro-produced bovine embryos using a chemically defined freezing medium

This study evaluates a new synthetic substitute (CRYO3, Ref. 5617, Stem Alpha, France) for animal-based products in bovine embryo cryopreservation solutions. During the experiment, fetal calf serum (FCS) and bovine serum albumin (BSA) were used as references. A combination of a thermodynamic approach using differential scanning calorimetry and a biological approach using in vitro-produced bovine embryo slow-freezing was used to characterize cryopreservation solutions containing CRYO3, FCS and BSA. The CRYO3 and fetal calf serum (FCS) slow-freezing solutions were made from Dulbecco's phosphate-buffered saline containing 1.5 m ethylene glycol, 0.1 m sucrose and 20% (v.v⁻¹) of CRYO3 or FCS. The bovine serum albumin (BSA) solution was made by adding 0.1 m sucrose to a commercial solution containing 1.5 m ethylene glycol and 4 g L⁻¹ BSA. These solutions were evaluated using three characteristics: the end of melting temperature, the enthalpy of crystallization (thermodynamic approach) and the embryo survival and hatching rates after in vitro culture (biological approach). The CRYO3 and FCS solutions had similar thermodynamic properties. In contrast, the thermodynamic characteristics of the BSA solution were different from those of the FCS and CRYO3 solutions. Nevertheless, the embryo survival and hatching rates obtained with the BSA and FCS solutions were not different. Similar biological properties can thus be obtained with slow freezing solutions that have different physical properties within a defined range. The embryo survival rate after 48 h of in vitro culture obtained with the CRYO3 solution (81.5%) was higher than that obtained with the BSA (42.2%, P = 0.000 12) and FCS solutions (58%, P = 0.016). Similarly, the embryo hatching rate after 72 h of in vitro culture was higher with the CRYO3 solution (61.1%) than with the BSA (31.1%, P = 0.0055) and FCS solutions (36%, P = 0.018). We conclude that CRYO3 can be used as a chemically defined substitute for animal-based products in in vitro-produced bovine embryo cryopreservation solutions.     

Winter cold tolerance and the geographic range separation of Bromus tectorum and Bromus rubens,two severe invasive species in North America

The invasive grasses Bromus rubens and Bromus tectorum are responsible for widespread damage to semiarid biomes of western North America. Bromus. tectorum dominates higher and more northern landscapes than its sister species B. rubens, which is a severe invader in the Mojave desert region of the American Southwest. To assess climate thresholds controlling their distinct geographic ranges, we evaluated the winter cold tolerance of B. tectorum and B. rubens. Freezing tolerance thresholds were determined using electrolyte leakage and whole‐plant mortality. The responses of the two species to winter cold and artificial freezing treatments were similar in 2007–2008 and 2009–2010. When grown at minimum temperatures of 10 °C, plants of both species had cold tolerance thresholds near ?10 °C, while plants acclimated to a daily minimum of ?10 to ?30 °C survived temperatures down to ?31 °C. In the winter of 2010–2011, a sudden severe cold event on December 9, 2010 killed all B. rubens populations, while B. tectorum was not harmed; all tested plants were 7–8 weeks old. Controlled acclimation experiments demonstrated that 8‐week‐old plants of B. rubens had a slower acclimation rate to subzero temperatures than B. tectorum and could not survive a rapid temperature drop from 1 to ?14 °C. Four‐month‐old B. rubens populations were as cold tolerant as B. tectorum. Our results show that severe and sudden freeze events in late autumn can kill young plants of B. rubens but not B. tectorum. Such events could exclude B. rubens from the relatively cold, Intermountain steppe biome of western North America where B. tectorum predominates.