Sip18 hydrophilin prevents yeast cell death during desiccation stress

Aims: For this study, we performed a genetic screen of S. cerevisiae’s deletion library for mutants sensitive to dehydration stress, with which we aimed to discover cell dehydration–tolerant genes. Methods and Results: We used a yeast gene deletion set (YGDS) of 4850 viable mutant haploid strains to perform a genome‐wide screen for the identification of desiccation stress modifiers. SIP18 is among the genes identified as essential for cells surviving to drying/rehydration process. Deletion of SIP18 promotes the accumulation of reactive oxygen species and enhances apoptotic and necrotic cell death in response to dehydration/rehydration process. Conclusions: SIP18p acts as an inhibitor of apoptosis in yeast under dehydration stress, as suggested by its antioxidative capacity through the ROS accumulation reduction after an H₂O₂ attack. Significance and Impact of the Study: To our knowledge, this is the first systematic screen for the identification of putative genes essential to overcoming cell dehydration process. A broad range of identified genes could help to understand why some strains of high biotechnological interest cannot cope with the drying and rehydration treatments. Dehydration sensitivity makes these strains not suitable to be commercialized by yeast manufactures.     

New national and regional bryophyte records, 8

Abstract

Monoclea forsteri Hook. is a thalloid liverwort species that is found in damp habitats and can, therefore, be expected to be sensitive to dehydration. It does, however, have some unique chemical constituents and anatomical features that could play a role in dealing with the adverse effects of water deficits. Corresponding to the habitat, M. forsteri lost its turgor at high relative water content (RWC≈0.90) and did not survive drying below 20% RWC. Moreover, the gametophytes showed an increase in malondialdehyde content and a depletion of the ascorbate pool during dehydration, indicating oxidative damage. Cellular constituents did not affect turgor pressure during drying and electrolyte leakage from the cells was greatly increased at RWC<0.20. Photosynthetic processes seemed not to be affected by the loss of turgor, but a decline appeared to correlate with an increase in electrolyte leakage. A speedy and fully sustained recovery from dehydration was realized from water contents above 30% and seemed only to be possible if membrane integrity could be preserved. Anatomical characteristics within M. forsteri gametophytes deserve further investigation to better understand their physiological functions.     

Effects of dehydration rate on physiological responses and survival after rehydration in larvae of the anhydrobiotic chironomid

Strategies to combat desiccation are critical for organisms living in arid and semi-arid areas. Larvae of the Australian chironomid Paraborniella tonnoiri resist desiccation by reducing water loss. In contrast, larvae of the African species Polypedilum vanderplanki can withstand almost complete dehydration, referred to as anhydrobiosis. For successful anhydrobiosis, the dehydration rate of P. vanderplanki larvae has to be controlled. Here, we desiccated larvae by exposing them to different drying regimes, each progressing from high to low relative humidity, and examined survival after rehydration. In larvae of P. vanderplanki, reactions following desiccation can be categorized as follows: (I) no recovery at all (direct death), (II) dying by unrepairable damages after rehydration (delayed death), and (III) full recovery (successful anhydrobiosis). Initial conditions of desiccation severely affected survival following rehydration, i.e. P. vanderplanki preferred 100% relative humidity where body water content decreased slightly. In subsequent conditions, unfavorable dehydration rate, such as more than 0.7 mg water lost per day, resulted in markedly decreased survival rate of rehydrated larvae. Slow dehydration may be required for the synthesis and distribution of essential molecules for anhydrobiosis. Larvae desiccated at or above maximum tolerable rates sometimes showed temporary recovery but died soon after.     

Differential mechanisms to induce dehydration tolerance by abscisic acid and sucrose in Spathoglottis plicata (Orchidaceae) protocorms

Abscisic acid (ABA) and sucrose are known to induce dehydration tolerance of in vitro plant cells and tissues. The present study reports the presence of different mechanisms by which sucrose and ABA improve dehydration tolerance of Spathoglottis plicata (orchid) protocorms. Orchid protocorms were generated aseptically from seeds on Murashig and Skoog medium, and then treated for 7 d in medium containing 10 mg L^?1 ABA and/or 10% (w/v) sucrose. Dehydration tolerance of protocorms was determined at ～25 °C under various drying conditions at relative humidity from 7 to 93%. The actual rate of water loss (i.e. drying rate) was determined using the rate constant of tissue water loss during drying according to the first‐order kinetics. Drying rate affected dehydration tolerance. ABA treatment reduced drying rate and increased dehydration tolerance of protocorms at all relative humidity values tested. However, when compared on the basis of actual drying rates, there was no difference in dehydration tolerance between control and ABA‐treated protocorms, suggesting that ABA‐induced tolerance was correlated with the drying rate reduction. Sucrose treatment was more effective than ABA treatment for the induction of dehydration tolerance. Interestingly, sucrose only slightly affected drying rate. ABA treatment significantly enhanced the synthesis of dehydrin, whereas sucrose treatment primarily resulted in sucrose accumulation. Sucrose treatment also affected protein turnover during drying, causing a significant decrease in protein content in protocorms. Slow drying promoted the degradation of high molecular weight proteins and enhanced the synthesis of low molecular weight dehydrin. The data suggest that different physiological mechanisms are probably involved in the induction of dehydration tolerance by ABA and sucrose treatment.     

Increased activities of superoxide dismutase and catalase are not the mechanism of desiccation tolerance induced by hardening in the moss Atrichum androgynum

Abstract

The effects of treatments that increase desiccation tolerance were tested on the activity of the enzymes superoxide dismutase (SOD) and catalase (CAT) in the moss Atrichum androgynum subjected to a drying/wetting cycle. Hardening by both abscisic acid (ABA) pretreatment and partial dehydration significantly increased the rate of recovery of photosynthesis during rehydration following desiccation. Hardening treatments had little effect on SOD activity. In non-hardened plants, SOD activity increased three-fold during desiccation for 32 h at 52% rh, but hardened material tended to display smaller increases in activity. During rehydration, SOD activities rapidly declined to their initial values in all treatments. Hardening by partial dehydration, but not ABA, reduced CAT activity. After desiccation for 32 h, material from all treatments displayed about half the initial CAT activity, and activity did not change during subsequent rehydration. Results show that, while the induction of SOD appears to play a role in desiccation tolerance, a similar induction occurred in both hardened and non-hardened mosses. Induction of greater activities of enzymes that scavenge reactive oxygen species is not responsible for the added tolerance induced by hardening treatments.     

Nuclear and Wolbachia-based multimarker approach for the rapid and accurate identification of tsetse species

Background

Tsetse flies (Diptera: Glossinidae) are solely responsible for the transmission of African trypanosomes, causative agents of sleeping sickness in humans and nagana in livestock. Due to the lack of efficient vaccines and the emergence of drug resistance, vector control approaches such as the sterile insect technique (SIT), remain the most effective way to control disease. SIT is a species-specific approach and therefore requires accurate identification of natural pest populations at the species level. However, the presence of morphologically similar species (species complexes and sub-species) in tsetse flies challenges the successful implementation of SIT-based population control.

Results

In this study, we evaluate different molecular tools that can be applied for the delimitation of different Glossina species using tsetse samples derived from laboratory colonies, natural populations and museum specimens. The use of mitochondrial markers, nuclear markers (including internal transcribed spacer 1 (ITS1) and different microsatellites), and bacterial symbiotic markers (Wolbachia infection status) in combination with relatively inexpensive techniques such as PCR, agarose gel electrophoresis, and to some extent sequencing provided a rapid, cost effective, and accurate identification of several tsetse species.

Conclusions

The effectiveness of SIT benefits from the fine resolution of species limits in nature. The present study supports the quick identification of large samples using simple and cost effective universalized protocols, which can be easily applied by countries/laboratories with limited resources and expertise.

     

Dehydration rate and time of desiccation affect recovery of the lichenic algae <Emphasis Type="Italic">Trebouxia erici</Emphasis>: alternative and classical protective mechanisms

The mechanisms involved in desiccation tolerance of lichens and their photobionts are still poorly understood. To better understand these mechanisms we have studied dehydration rate and desiccation time in Trebouxia, the most abundant chlorophytic photobiont in lichen. Our findings indicate that the drying rate has a profound effect on the recovery of photosynthetic activity of algae after rehydration, greater than the effects of desiccation duration. The basal fluorescence (F′_o) values in desiccated algae were significantly higher after rapid dehydration, than after slow dehydration, suggesting higher levels of light energy dissipation in slow-dried algae. Higher values of PSII electron transport were recovered after rehydration of slow-dried Trebouxia erici compared to rapid-dried algae. The main component of non-photochemical quenching after slow dehydration was energy dependent (q _E), whereas after fast dehydration it was photoinhibition (q _I). Although q _E seems to play a role during desiccation recovery, no significant variations were detected in the xanthophyll cycle components. Desiccation did not affect PSI functionality. Classical antioxidant activities like superoxide dismutase or peroxidase decreased during desiccation and early recovery. Dehydrins were detected in the lichen-forming algae T. erici and were constitutively expressed. There is probably a minimal period required to develop strategies which will facilitate transition to the desiccated state in this algae. In this process, the xanthophyll cycle and classical antioxidant mechanisms play a very limited role, if any. However, our results indicate that there is an alternative mechanism of light energy dissipation during desiccation, where activation is dependent on a sufficiently slow dehydration rate.     

Microhabitat Preference of Egernia napoleonis in Undisturbed Jarrah Forest,and Availability and Introduction of Microhabitats to Encourage Colonization of Restored Forest

An animal's microhabitat requirements can impact its ability to colonize restored areas, particularly species requiring slow developing microhabitats, such as logs and woody debris piles. Introduction of these microhabitats may be required to facilitate colonization by some species. Restored bauxite mine‐pits in the Jarrah (Eucalyptus marginata) forest of south‐western Australia contain introduced log piles at densities of 1 ha^?1. However, these have not facilitated colonization by Napoleon's skink (Egernia napoleonis), which rely on logs for habitat and are largely absent from restored sites. We radio‐tracked 12 skinks in unmined forest to determine their microhabitat preferences and examined differences in vegetation structure, and microhabitat and food availability, between restored and unmined forests to identify reasons for their absence. Restored and unmined forests differed in canopy, mid‐ and understory cover and ground substrates, which were all potential barriers to colonization. Food availability was similar between restored and unmined forest, thus not a barrier to colonization. Skinks primarily utilized long logs, large woody debris piles, and large trees; microhabitats that were scarce or absent in restored sites and, therefore, potential barriers to colonization. Using this information, we introduced small woody debris piles into restored sites in close proximity to unmined areas containing skinks to facilitate skink colonization. This showed early signs of success and suggested that the lack of logs and woody debris were barriers to colonization. However, further monitoring is required to accurately determine the long‐term value of woody debris piles in facilitating skink colonization.     

The use of plant stress biomarkers in assessing the effects of desiccation in zygotic embryos from recalcitrant seeds: challenges and considerations

Zygotic embryos from recalcitrant seeds are sensitive to desiccation. In spite of their sensitivity, rapid partial dehydration is necessary for their successful cryopreservation. However, dehydration to water contents (WCs) that preclude lethal ice crystal formation during cooling and rewarming generally leads to desiccation damage. This study investigated the effects of rapid dehydration on selected stress biomarkers (electrolyte leakage, respiratory competence, rate of protein synthesis, superoxide production, lipid peroxidation, antioxidant activity and degree of cellular vacuolation) in zygotic embryos of four recalcitrant‐seeded species. Most biomarkers indicated differences in the levels of stress/damage incurred by embryos dried to WCs < and >0.4 g·g^?1, within species; however, these changes were often unrelated to viability and percentage water loss when data for the four species were pooled for regression analyses. Dehydration‐induced electrolyte leakage was, however, positively related with percentage water loss, while biomarkers of cellular vacuolation were positively related with both percentage water loss and viability. This suggests that electrolyte leakage and degree of cellular vacuolation can be used to quantify dehydration‐induced stress/damage. Biomarkers such as superoxide production, whilst useful in establishing the nature of the dehydration stress incurred may not be able to distinguish the effects of different WCs/drying times. Irrespective of which biomarker is used, the data suggest that understanding differences in desiccation sensitivity across recalcitrant‐seeded species will remain a challenge unless these biomarkers are related to a generic desiccation stress index that integrates the effects of percentage water loss and drying time.     

马鞍菌属3个中国新记录

【背景】目前国内学者已发表60个马鞍菌属物种,但山西省尚缺乏马鞍菌属真菌分布的系统性报道。【目的】调查我国山西省马鞍菌属的种类组成及地理分布。【方法】采集标本,对其进行详细的形态特征描述,并提取DNA,测定热激蛋白(heat shock protein, hsp)和核糖体大亚基(large subunit, LSU)序列,基于贝叶斯法(Bayesian inference, BI)和最大似然法(maximum likelihood, ML)构建分子系统发育树。【结果】鉴定结果表明这3个马鞍菌属物种分别为类帽状马鞍菌(Helvella capucinoides)、丹麦马鞍菌(H. danica)和柔毛马鞍菌(H. pubescens)。它们在形态上与模式标本的描述基本一致。基于hsp和LSU的系统发育关系分析也支持形态学的鉴定结果。【结论】参考我国截至目前已报道的马鞍菌属物种,确定这3个种均为中国新记录种。     

A NEW SPINICAUDATAN GENUS (CRUSTACEA: ‘CONCHOSTRACA’) FROM THE LATE CRETACEOUS OF MADAGASCAR

Abstract: A new spinicaudatan genus and species, Ethmosestheria mahajangaensis gen. et sp. nov., is described from the Anembalemba Member (Upper Cretaceous, Maastrichtian) of the Maevarano Formation, Mahajanga Basin, Madagascar. This is the first spinicaudatan reported from the post‐Triassic Mesozoic of Madagascar. The new species is assigned to the family Antronestheriidae based on the cavernous or sievelike ornamentation on the carapace. Of well‐documented Mesozoic spinicaudatan genera, Ethmosestheria mahajangaensis is most closely related to Antronestheria Chen and Hudson from the Great Estuarine Group (Jurassic) of Scotland. However, relatively poor documentation of the ornamentation of most Gondwanan Mesozoic spinicaudatan species precludes detailed comparison among taxa. Ethmosestheria mahajangaensis exhibits ontogenetic trends in carapace growth: a change in carapace outline from subcircular/subelliptical to elliptical, and from very wide juvenile growth bands to narrow adult growth bands. Ornamentation style, however, does not vary with ontogeny. Ethmosestheria mahajangaensis individuals lived in temporary pools in a broad channel‐belt system within a semiarid environment; preserved desiccation structures on carapaces indicate seasonal drying out of pools within the river system. Specimens of Ethmosestheria mahajangaensis are preserved with exquisite detail in debris flow deposits; these are the first spinicaudatans reported from debris flow deposits. These deposits also contain a varied vertebrate fauna, including dinosaurs, crocodyliforms, turtles, and frogs. Rapid entombment of the spinicaudatan carapaces likely promoted early fossil diagenesis leading to highly detailed preservation.     

Identification of Belgian mosquito species (Diptera: Culicidae) by DNA barcoding

Since its introduction in 2003, DNA barcoding has proven to be a promising method for the identification of many taxa, including mosquitoes (Diptera: Culicidae). Many mosquito species are potential vectors of pathogens, and correct identification in all life stages is essential for effective mosquito monitoring and control. To use DNA barcoding for species identification, a reliable and comprehensive reference database of verified DNA sequences is required. Hence, DNA sequence diversity of mosquitoes in Belgium was assessed using a 658 bp fragment of the mitochondrial cytochrome oxidase I (COI) gene, and a reference data set was established. Most species appeared as well‐supported clusters. Intraspecific Kimura 2‐parameter (K2P) distances averaged 0.7%, and the maximum observed K2P distance was 6.2% for Aedes koreicus. A small overlap between intra‐ and interspecific K2P distances for congeneric sequences was observed. Overall, the identification success using best match and the best close match criteria were high, that is above 98%. No clear genetic division was found between the closely related species Aedes annulipes and Aedes cantans, which can be confused using morphological identification only. The members of the Anopheles maculipennis complex, that is Anopheles maculipennis s.s. and An. messeae, were weakly supported as monophyletic taxa. This study showed that DNA barcoding offers a reliable framework for mosquito species identification in Belgium except for some closely related species.     

Taxonomic re-evaluation of the Azolla genus in Portugal

ABSTRACT

The Portuguese and the European Flora refer to the presence of two or three Azolla species in Portugal: A. filiculoides Lam., A. caroliniana Willd. and/or Azolla mexicana Presl., the latter included in the last edition of Flora Europaea. In the present work, the taxonomy of Azolla species is reviewed using the two most important characters that can distinguish between these two/three species: papillae in the dorsal leaf lobe and perine architecture of the megaspore apparatus. Other characteristics, such as the hyaline border cells of the dorsal leaf lobes and the number of glochidia septa in microsporangium massulae, are also used. All the Azolla specimens, collected from several locations in Portugal, were identified as Azolla filiculoides Lam. This identification disagrees with previous published reports on Azolla taxonomy in Portugal as well as with herbarium identification.     

基于单细菌共焦拉曼光谱的细菌快速检测

【背景】目前利用共焦拉曼光谱技术进行成像和成分鉴别方面的研究较多,但如何快速检测与鉴别多种细菌方面的研究较少。【目的】基于共焦拉曼光谱技术,建立一种在单细菌水平上实现病原微生物快速分类鉴定的方法。【方法】以大肠杆菌为研究对象,利用共焦拉曼光谱技术在单细菌水平上进行了激发波长的优化试验,并研究了大肠杆菌存放时间对单细菌拉曼光谱信息的影响。同时,对白色葡萄球菌、大肠杆菌、金黄色葡萄球菌、沙门氏菌和铜绿假单胞菌进行了共焦拉曼光谱测试,并对5种细菌进行单细菌拉曼光谱的归属分析,设计共焦拉曼光谱技术结合支持向量机(support vector machine,SVM)模型学习算法,进行了5种细菌的快速分类鉴别。【结果】对于单细菌拉曼光谱探测,532、633和785 nm这3种常见的拉曼探测波长中,532 nm具有更好的激发效率和光谱信噪比。结合SVM模型对5种细菌的识别分类,SVM模型的灵敏度和特异性达到了96.00%以上,整体准确率为98.25%。不同存放时间下大肠杆菌拉曼光谱的重复性和稳定性都很好,且SVM模型匹配率均在90.00%以上。【结论】单细菌拉曼光谱结合SVM模型可对5种细菌进行快...     

Species-specific Seedling Responses to Hurricane Disturbance in a Puerto Rican Rain Forest1

Seedling dynamics were followed in a Puerto Rican forest for 20 months following a severe hurricane to study the interactive effects of hurricane debris, nutrients, and light on seedling diversity, density, growth, and mortality. Three treatments (debris removal, an unaltered control with hurricane debris, and chemical fertilization added to hurricane debris) altered levels of forest debris and soil nutrients. Canopy openness was measured twice using hemispherical photographs of the canopy. We examined the demographic responses of six common species to treatments over time. Seedling densities increased for all six species but the only significant treatment effects were increased densities of the pioneer tree Cecropia and the shrub Palicourea in the debris removal treatment. Seedling growth declined with declining light levels for four species but not for the pioneer tree Alchornea or the non‐pioneer tree Dacryodes. Only Cecropia and the non‐pioneer tree Chionanthus had treatment effects on growth. Mortality also differed among species and tended to be highest in the fertilized plots for all but Cecropia and Dacryodes. We found only some of the expected differences between pioneer and non‐pioneer plants, as each species had a unique response to the patchy distributions of organic debris, nutrients, and light following the hurricane. High local species diversity was maintained through the individualistic responses of seedlings after a disturbance.     

Molecular identification of mosquitoes (Diptera: Culicidae) in southeastern Australia

DNA barcoding is a modern species identification technique that can be used to distinguish morphologically similar species, and is particularly useful when using small amounts of starting material from partial specimens or from immature stages. In order to use DNA barcoding in a surveillance program, a database containing mosquito barcode sequences is required. This study obtained Cytochrome Oxidase I (COI) sequences for 113 morphologically identified specimens, representing 29 species, six tribes and 12 genera; 17 of these species have not been previously barcoded. Three of the 29 species ─ Culex palpalis, Macleaya macmillani, and an unknown species originally identified as Tripteroides atripes ─ were initially misidentified as they are difficult to separate morphologically, highlighting the utility of DNA barcoding. While most species grouped separately (reciprocally monophyletic), the Cx. pipiens subgroup could not be genetically separated using COI. The average conspecific and congeneric p‐distance was 0.8% and 7.6%, respectively. In our study, we also demonstrate the utility of DNA barcoding in distinguishing exotics from endemic mosquitoes by identifying a single intercepted Stegomyia aegypti egg at an international airport. The use of DNA barcoding dramatically reduced the identification time required compared with rearing specimens through to adults, thereby demonstrating the value of this technique in biosecurity surveillance. The DNA barcodes produced by this study have been uploaded to the ‘Mosquitoes of Australia–Victoria’ project on the Barcode of Life Database (BOLD), which will serve as a resource for the Victorian Arbovirus Disease Control Program and other national and international mosquito surveillance programs.     

Differential proteomics of dehydration and rehydration in bryophytes: evidence towards a common desiccation tolerance mechanism

All bryophytes evolved desiccation tolerance (DT) mechanisms during the invasion of terrestrial habitats by early land plants. Are these DT mechanisms still present in bryophytes that colonize aquatic habitats? The aquatic bryophyte Fontinalis antipyretica Hedw. was subjected to two drying regimes and alterations in protein profiles and sucrose accumulation during dehydration and rehydration were investigated. Results show that during fast dehydration, there is very little variation in protein profiles, and upon rehydration proteins are leaked. On the other hand, slow dehydration induces changes in both dehydration and rehydration protein profiles, being similar to the protein profiles displayed by the terrestrial bryophytes Physcomitrella patens (Hedw.) Bruch and Schimp. and, to what is comparable with Syntrichia ruralis (Hedw.) F. Weber and D. Mohr. During dehydration there was a reduction in proteins associated with photosynthesis and the cytoskeleton, and an associated accumulation of proteins involved in sugar metabolism and plant defence mechanisms. Upon rehydration, protein accumulation patterns return to control values for both photosynthesis and cytoskeleton whereas proteins associated with sugar metabolism and defence proteins remain high. The current results suggest that bryophytes from different ecological adaptations may share common DT mechanisms.