A comparison of heat shock protein genes from cultured cells of the cabbage armyworm, Mamestra brassicae, in response to heavy metals

Heat shock protein (HSP) genes, hsp90, hsp70, hsc70, hsp20.7, and hsp19.7, were cloned and sequenced from cultured cells of the cabbage armyworm, Mamestra brassicae. Analyses of the cDNA sequences revealed open reading frames of 2,151, 1,914, 1,962, 540, and 465 bp in lengths, which encode proteins with calculated molecular weights of 82.5, 69.9, 71.6, 20.7, and 19.7 kDa, respectively. An increased expression was observed in all five genes after exposure to a high temperature. The induction of gene expression was not observed during a low temperature exposure, but was observed when the cells were recovered at ambient temperature. Expression of hsp90, hsp70, and hsp20.7 was induced after exposure to 2 microM of cadmium, while the minimum cadmium concentration for induction of hsp19.7 was 5 microM. The induction of hsp90 expression was somewhat masked by basal levels of expression. Only hsp20.7 expression was induced by exposure to copper. Lead did not induce expression of any of the HSP genes tested. Cadmium-induced up-regulation of hsp70 expression was lasted longer than heat-induced one. These results suggest that hsp70 could be useful to assess the cellular distress or injury induced by cadmium.     

Function of phytochelatin synthase in catabolism of glutathione-conjugates

Detoxification of xenobiotic compounds and heavy metals is a pivotal capacity of organisms, in which glutathione (GSH) plays an important role. In plants, electrophilic herbicides are conjugated to the thiol group of GSH, and heavy metal ions form complexes as thiolates with GSH-derived phytochelatins (PCs). In both detoxification processes of plants, phytochelatin synthase (PCS) emerges as a key player. The enzyme is activated by heavy metal ions and catalyzes PC formation from GSH by transferring glutamylcysteinyl residues (gamma-EC) onto GSH. In this study with Arabidopsis, we show that PCS plays a role in the plant-specific catabolism of glutathione conjugates (GS-conjugates). In contrast to animals, breakdown of GS-conjugates in plants can be initiated by cleavage of the carboxyterminal glycine residue that leads to the generation of the corresponding gamma-EC-conjugate. We used the xenobiotic bimane in order to follow GS-conjugate turnover. Functional knockout of the two PCS of Arabidopsis, AtPCS1 and AtPCS2, revealed that AtPCS1 provides a major activity responsible for conversion of the fluorescent bimane-GS-conjugate (GS-bimane) into gamma-EC-bimane. AtPCS1 deficiency resulted in a gamma-EC-bimane deficiency. Transfection of PCS-deficient cells with AtPCS1 recovered gamma-EC-bimane levels. The level of the gamma-EC-bimane conjugate was enhanced several-fold in the presence of Cd2+ ions in the wild type, but not in the PCS-deficient double mutant, consistent with a PCS-catalyzed GS-conjugate turnover. Thus AtPCS1 has two cellular functions: mediating both heavy metal tolerance and GS-conjugate degradation.     

萝卜乙烯合成途径基因的鉴定及对胁迫的响应

该研究通过生物信息学方法,对萝卜(Raphanus sativus L.)乙烯合成途径关键结构基因(MAT、ACS和ACO)家族成员进行鉴定,并利用转录组数据和荧光定量方法探究其组织表达和对生物及非生物胁迫的响应。结果表明：(1)萝卜基因组中包含8个MAT、16个ACS和7个ACO基因,均可分为3个亚家族。(2)这些基因启动子中至少含有1个光响应顺式作用元件;除ACO1.1和ACO3外,其余基因启动子中均至少含有一种响应植物激素的顺式作用元件;多个基因启动子中含有响应生物及非生物胁迫的顺式作用元件。(3)转录组数据分析发现,萝卜所有MAT、ACS6.1和ACO2/3/4在叶片中均具较高的表达量;根癌农杆菌侵染诱导抗病萝卜的MAT2 4、ACS2/7.1/7.2和ACO1.1/1.2/4显著上调表达,而感病材料多个基因下调表达;铅、镉和铬胁迫均显著促进MAT4.2和ACO1.1/4的表达,但抑制MAT1和ACO5.1的表达;4 ℃低温显著抑制MAT2.1/2.2/4.1和ACO1.1/5.2的表达。(4)qRT PCR分析表明,NaCl和PEG 6000均显著促进ACO5.1/5.2的表达,但抑制MAT4.1和ACO4的表达;果糖和蔗糖可能参与了萝卜对PEG 6000胁迫的响应。该结果为研究萝卜乙烯合成途径基因家族成员的功能奠定了基础。     

早春首次降雨时间及降雨量对古尔班通古特沙漠两种短命植物形态特征与叶绿素荧光的影响

早春降雨事件是影响荒漠短命植物生长的重要因子, 为了解古尔班通古特沙漠积雪完全融化后早春首次降雨时间及降雨量对短命植物光合生长的影响。该研究选取两种优势短命植物尖喙牻牛儿苗(Erodium oxyrhinchum)和琉苞菊(Centaurea pulchella)为研究对象, 设置3个首次降雨时间(积雪完全融化后第10、20和30天)和3个降雨水平(5、10和15 mm), 探究两物种的形态、生物量及光化学效率的响应差异。结果显示: 积雪完全融化后第10天进行首次降雨处理时, 随着首次降雨量的增加, 尖喙牻牛儿苗的叶面积和总生物量呈显著增加趋势, 最大光化学效率呈下降趋势。在第20和30天进行首次降雨处理时, 尖喙牻牛儿苗和琉苞菊的叶面积均呈显著增加趋势。在积雪完全融化后5 mm降雨处理时, 随着首次降雨时间的延迟, 尖喙牻牛儿苗和琉苞菊的最大光化学效率呈先下降后上升趋势; 10 mm降雨处理时, 尖喙牻牛儿苗的冠根比降低, 根长和总生物量呈先增加后下降趋势, 琉苞菊的根长呈显著增加趋势; 15 mm降雨处理时, 尖喙牻牛儿苗的叶面积、冠根比和最大光化学效率呈显著增加趋势; 琉苞菊的冠根比和最大光化学效率呈下降趋势, 总生物量呈显著增加趋势。尖喙牻牛儿苗和琉苞菊的根长分别与0-5 cm和5-10 cm土壤含水量呈显著正相关关系。这说明早春首次降雨量比首次降雨时间更能影响植株生长, 但不同物种对降雨格局变化的响应不一致, 这可能通过影响早春短命植物的形态特征, 进而影响早春荒漠生态系统的稳定性。     

微藻-细菌共生体系在废水处理中的应用

在微藻-细菌协同共生的过程中,藻类光合作用释放的氧气被异养微生物利用来矿化水体中的污染物,细菌呼吸为藻类提供二氧化碳作为碳源。近年来,藻类-细菌协同共生体系在污水处理中的应用得到了广泛的研究。本文重点综述了菌藻协同共生体系中微藻与细菌之间的三种相互作用,以及菌藻协同共生体系在废水处理中的应用。菌藻协同共生体系中的微藻与细菌通过营养交换、信号转导及基因转移等相互作用实现共赢。该体系广泛用于处理富营养化、重金属、药物、多环芳烃(polycyclicaromatic hydrocarbons,PAHs)、石油烃化合物等难降解的有机污染的水体。对于氮、磷等营养物质的去除,其主要机理涉及同化作用、厌氧氨氧化作用、硝化与反硝化作用、磷酸化作用等。对重金属、药物、石油烃化合物及其他有机化合物的去除机制主要是生物吸附、生物富集及细胞内外的生物降解。     

Physiological and molecular mechanisms of heavy metal accumulation in nonmycorrhizal versus mycorrhizal plants

Uptake, translocation, detoxification, and sequestration of heavy metals (HMs) are key processes in plants to deal with excess amounts of HM. Under natural conditions, plant roots often establish ecto‐ and/or arbuscular‐mycorrhizae with their fungal partners, thereby altering HM accumulation in host plants. This review considers the progress in understanding the physiological and molecular mechanisms involved in HM accumulation in nonmycorrhizal versus mycorrhizal plants. In nonmycorrhizal plants, HM ions in the cells can be detoxified with the aid of several chelators. Furthermore, HMs can be sequestered in cell walls, vacuoles, and the Golgi apparatus of plants. The uptake and translocation of HMs are mediated by members of ZIPs, NRAMPs, and HMAs, and HM detoxification and sequestration are mainly modulated by members of ABCs and MTPs in nonmycorrhizal plants. Mycorrhizal‐induced changes in HM accumulation in plants are mainly due to HM sequestration by fungal partners and improvements in the nutritional and antioxidative status of host plants. Furthermore, mycorrhizal fungi can trigger the differential expression of genes involved in HM accumulation in both partners. Understanding the molecular mechanisms that underlie HM accumulation in mycorrhizal plants is crucial for the utilization of fungi and their host plants to remediate HM‐contaminated soils.     

Weather effects on birds of different size are mediated by long‐term climate and vegetation type in endangered temperate woodlands

Species occurrence is influenced by a range of factors including habitat attributes, climate, weather, and human landscape modification. These drivers are likely to interact, but their effects are frequently quantified independently. Here, we report the results of a 13‐year study of temperate woodland birds in south‐eastern Australia to quantify how different‐sized birds respond to the interacting effects of: (a) short‐term weather (rainfall and temperature in the 12 months preceding our surveys), (b) long‐term climate (average rainfall and maximum and minimum temperatures over the period 1970–2014), and (c) broad structural forms of vegetation (old‐growth woodland, regrowth woodland, and restoration plantings). We uncovered significant interactions between bird body size, vegetation type, climate, and weather. High short‐term rainfall was associated with decreased occurrence of large birds in old‐growth and regrowth woodland, but not in restoration plantings. Conversely, small bird occurrence peaked in wet years, but this effect was most pronounced in locations with a history of high rainfall, and was actually reversed (peak occurrence in dry years) in restoration plantings in dry climates. The occurrence of small birds was depressed—and large birds elevated—in hot years, except in restoration plantings which supported few large birds under these circumstances. Our investigation suggests that different mechanisms may underpin contrasting responses of small and large birds to the interacting effects of climate, weather, and vegetation type. A diversity of vegetation cover is needed across a landscape to promote the occurrence of different‐sized bird species in agriculture‐dominated landscapes, particularly under variable weather conditions. Climate change is predicted to lead to widespread drying of our study region, and restoration plantings—especially currently climatically wet areas—may become critically important for conserving bird species, particularly small‐bodied taxa.     

Autumn bird migration phenology: A potpourri of wind,precipitation and temperature effects

Climate change has caused a clear and univocal trend towards advancement in spring phenology. Changes in autumn phenology are much more diverse, with advancement, delays, and ‘no change' all occurring frequently. For migratory birds, patterns in autumn migration phenology trends have been identified based on ecological and life‐history traits. Explaining interspecific variation has nevertheless been challenging, and the underlying mechanisms have remained elusive. Radar studies on non‐species‐specific autumn migration intensity have repeatedly suggested that there are strong links with weather. In long‐term species‐specific studies, the variance in autumn migration phenology explained by weather has, nevertheless, been rather low, or a relationship was even lacking entirely. We performed a spatially explicit time window analysis of weather effects on mean autumn passage of four trans‐Saharan and six intra‐European passerines to gain insights into this apparent contradiction. We analysed data from standardized daily captures at the Heligoland island constant‐effort site (Germany), in combination with gridded daily temperature, precipitation and wind data over a 55‐year period (1960–2014), across northern Europe. Weather variables at the breeding and stopover grounds explained up to 80% of the species‐specific interannual variability in autumn passage. Overall, wind conditions were most important. For intra‐European migrants, wind was even twice as important as either temperature or precipitation, and the pattern also held in terms of relative contributions of each climate variable to the temporal trends in autumn phenology. For the trans‐Saharan migrants, however, the pattern of relative trend contributions was completely reversed. Temperature and precipitation had strong trend contributions, while wind conditions had only a minor impact because they did not show any strong temporal trends. As such, understanding species‐specific effects of climate on autumn phenology not only provides unique insights into each species' ecology but also how these effects shape the observed interspecific heterogeneity in autumn phenological trends.     

Black soldier fly defatted meal as a dietary protein source for broiler chickens: effects on carcass traits,breast meat quality and safety

Finding insect meals as alternative sources of poultry feedstuffs is a recent research topic; therefore, the present study aimed to evaluate the effects of defatted black soldier fly (Hermetia illucens L., HI) larvae meal in broiler chicken diets on the carcass characteristics and meat quality parameters, proximate composition, fatty acid profile and the heavy metal content of the breast meat. Four dietary treatments were designed: a control diet (HI0) and three experimental diets (HI5, HI10 and HI15), corresponding to 50, 100 and 150 g/kg HI inclusion levels, respectively. The inclusion of 50, 100 and 150 g HI meal per kg feed supply 16.56%, 33.01% and 49.63% of required crude protein. The broilers were slaughtered at day 35, the carcasses were weighed and the breast muscles were excised from 16 birds per each feeding group (two birds per replicate pens) and used for meat quality evaluation. Linear and quadratic responses were observed, for increasing HI meal levels, in the live and carcass weights (maximum for HI10). As far as the colour of the breast meat is concerned, redness (a*) showed a linear response, while yellowness (b*) linearly decreased with increasing HI meal levels (minimum for HI15). As the HI larvae meal increased in the diets, the moisture content linearly decreased and the protein content increased. The total saturated fatty acid and total monounsaturated fatty acid proportions rose to the detriment of the polyunsaturated fatty acid fraction. The HI larvae meal, used in the current study, represents a valuable protein source for broiler chickens when included by up to 100 g/kg in their diets, as an improved slaughtering performance was observed without any detrimental effects on meat quality parameters or heavy metal residues in the meat.     

Detection of mitochondrial dysfunction in vitro by laser‐induced autofluorescence

Mitochondrion plays a significant role in a variety of biological functions. Because of their diverse character and location in the cellular systems, mitochondria commonly get exposed to various extrinsic and intrinsic cellular stresses. The present study reports a novel approach to detection of mitochondrial dysfunction based on tryptophan autofluorescence of its proteins in mouse liver, using laser‐induced fluorescence (LIF) as a tool. Mitochondria, isolated from the mouse liver, were initially tested for purity and integrity using lactate dehydrogenase and succinate dehydrogenase (SDH) assays. Mitochondrial stress was induced by treating the isolated mitochondria with heavy metals at 10 and 0.01 mM for sodium arsenite and mercuric chloride, respectively. Upon treatment with the heavy metal, tryptophan autofluorescence quenching was recorded at 281 nm excitation. The functional integrity of the mitochondria treated with heavy metals was evaluated by measuring SDH and cytochrome c oxidase activities at various concentrations of mitochondria, which showed impaired activity as compared to control upto a concentration of 6.25 μg. A significant shift was also observed in the autofluorescence of proteins upto the level below 1 μg, suggesting their conformational change and hence altered structural integrity of mitochondria. Circular dichroism spectroscopy data of the mitochondrial proteins treated with heavy metals further validates their conformational change as compared to untreated control. The present study clearly shows that the LIF can be a novel detection tool to detect altered structural integrity of cellular mitochondria upon stress, and it also possesses the potentiality to combine with other interdisciplinary modalities.