Bioactivity-Guided Fractionation of an Antidiarrheal Chinese Herb Rhodiola kirilowii (Regel) Maxim Reveals (-)–Epicatechin-3-Gallate and (-)–Epigallocatechin-3-Gallate as Inhibitors of Cystic Fibrosis Transmembrane Conductance Regulator

Cystic fibrosis transmembrane conductance regulator (CFTR) is the principal apical route for transepithelial fluid transport induced by enterotoxin. Inhibition of CFTR has been confirmed as a pharmaceutical approach for the treatment of secretory diarrhea. Many traditional Chinese herbal medicines, like Rhodiola kirilowii (Regel) Maxim, have long been used for the treatment of secretory diarrhea. However, the active ingredients responsible for their therapeutic effectiveness remain unknown. The purpose of this study is to identify CFTR inhibitors from Rhodiola kirilowii (Regel) Maxim via bioactivity-directed isolation strategy. We first identified fractions of Rhodiola kirilowii (Regel) Maxim that inhibited CFTR Cl^- channel activity. Further bioactivity-directed fractionation led to the identification of (-)–epigallocatechin-3-gallate (EGCG) as CFTR Cl^- channel inhibitor. Analysis of 5 commercially available EGCG analogs including (+)–catechins (C), (-)–epicatechin (EC), (-)–epigallocatechin (EGC), (-)–epicatechin-3-gallate (ECG) and EGCG revealed that ECG also had CFTR inhibitory activity. EGCG dose-dependently and reversibly inhibited CFTR Cl^- channel activity in transfected FRT cells with an IC₅₀ value around 100 μM. In ex vivo studies, EGCG and ECG inhibited CFTR-mediated short-circuit currents in isolated rat colonic mucosa in a dose-dependent manner. In an intestinal closed-loop model in mice, intraluminal application of EGCG (10 μg) and ECG (10 μg) significantly reduced cholera toxin-induced intestinal fluid secretion. CFTR Cl^- channel is a molecular target of natural compounds EGCG and ECG. CFTR inhibition may account, at least in part, for the antidiarrheal activity of Rhodiola kirilowii (Regel) Maxim. EGCG and ECG could be new lead compounds for development of CFTR-related diseases such as secretory diarrhea.     

Phylogeography of Rhodiola kirilowii (Crassulaceae): A Story of Miocene Divergence and Quaternary Expansion

The evolution and current distribution of the Sino-Tibetan flora have been greatly affected by historical geological events, such as the uplift of the Qinghai-Tibetan Plateau (QTP), and Quaternary climatic oscillations. Rhodiola kirilowii, a perennial herb with its distribution ranging from the southeastern QTP and the Hengduan Mountains (HM) to adjacent northern China and central Asia, provides an excellent model to examine and disentangle the effect of both geological orogeny and climatic oscillation on the evolutionary history of species with such distribution patterns. We here conducted a phylogeographic study using sequences of two chloroplast fragments (trnL-F and trnS-G) and internal transcribed spacers in 29 populations of R. kirilowii. A total of 25 plastid haplotypes and 12 ITS ribotypes were found. Molecular clock estimation revealed deep divergence between the central Asian populations and other populations from the HM and northern China; this split occurred ca. 2.84 million year ago. The majority of populations from the mountains of northern China were dominated by a single haplotype or ribotype, while populations of the HM harbored both high genetic diversity and high haplotype diversity. This distribution pattern indicates that HM was either a diversification center or a refugium for R. kirilowii during the Quaternary climatic oscillations. The present distribution of this species on mountains in northern China may have resulted from a rapid glacial population expansion from the HM. This expansion was confirmed by the mismatch distribution analysis and negative Tajima''s D and Fu''s F _S values, and was dated to ca. 168 thousand years ago. High genetic diversity and population differentiation in both plastid and ITS sequences were revealed; these imply restricted gene flow between populations. A distinct isolation-by-distance pattern was suggested by the Mantel test. Our results show that in old lineages, populations may harbour divergent genetic forms that are sufficient to maintain or even increase overall genetic diversity despite fragmentation and low within-population variation.     

柴胡幼苗越冬抗寒性及其相关生理指标筛选

以北柴胡、地产柴胡、三岛柴胡的根为试验材料,以早春植株成活率为越冬抗寒性指标,测定越冬期不同柴胡品种根系生理生化特征,进行柴胡抗寒性综合评价,并对主要抗寒生理指标进行通径分析和相关性分析,以探讨冬季自然低温条件下柴胡生理特性与抗寒性的关系,筛选适合柴胡越冬抗寒性鉴定的生理指标。结果表明:(1)北柴胡、地产柴胡、三岛柴胡返青期的返青率依次为80%、50%、10%,且各品种间差异显著。(2)3个品种柴胡越冬期根系可溶性糖含量、游离脯氨酸含量、可溶性蛋白含量、根系活力与返青率大小顺序一致,其中北柴胡根系游离脯氨酸平均含量分别为地产柴胡和三岛柴胡的2.44倍和4.49倍,且可溶性蛋白含量显著高于地产柴胡、三岛柴胡,但3个品种柴胡的相对电导率、丙二醛含量水平与返青率大小顺序相反。(3)自然越冬过程中,各柴胡品种的根系活力均呈下降趋势,且以三岛柴胡根系活力下降幅度最大,达80%,而北柴胡根系活力下降幅度最小为52.04%。(4)越冬期3个柴胡品种的越冬抗寒性综合排序为:北柴胡地产柴胡三岛柴胡;柴胡根中脯氨酸含量、可溶性蛋白含量与综合评价值呈极显著正相关关系,相对电导率与综合评价值呈显著负相关关系;生理指标决策系数的大小顺序依次为脯氨酸含量可溶性蛋白含量电导率。研究表明,越冬期柴胡根系游离脯氨酸含量是影响其抗寒性的主要影响因素,其含量可作为评价柴胡抗寒性的指标。     

低温胁迫时间对4种幼苗生理生化及光合特性的影响

以盐肤木(Rhus chinensis)、假连翘(Duranta repens)、老鸭嘴(Thunbergia erecta)和葛藤(Pueraria lobata)4种幼苗为试验材料,研究了人工模拟下的低温胁迫环境(6℃)对幼苗叶片生理生化及光合特性的影响.结果表明:(1)随低温胁迫的延长,假连翘幼苗的叶绿素含量先升后降,其余3种幼苗持续下降,低温解除后均显著回升;4种幼苗的脯氨酸含量持续上升或波动;葛藤幼苗的可溶性蛋白质含量先升后降,其余幼苗显著上升,且低温解除后有所增加;4种幼苗叶片可溶性糖含量以及丙二醛含量呈增加趋势,SOD活性持续上升或先升后降.低温解除后,盐肤木幼苗的SOD活性略有上升,其余幼苗显著下降.(2)随低温胁迫的延长,4种幼苗的净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)均持续下降,假连翘的胞间CO2浓度(Ci)持续上升,其余幼苗先降后升.低温解除后4种幼苗叶片Pn、G、Tr均有不同程度的回升,Ci有不同程度的下降.低温胁迫和恢复48 h期间,4种幼苗的脯氨酸、可溶性糖、可溶性蛋白质含量的增加和SOD活性的稳定或增加,减轻了幼苗叶片细胞的膜脂过氧化程度,维持了细胞膜的完整性,是幼苗对低温胁迫适应性反应的重要调节机制.恢复48 h时的Pn、Gs和Tr均有不同程度的回升,Ci有不同程度的下降,说明4种幼苗有一定的抗寒能力.(3)用主成分分析法分别对生理生化指标和光合生理指标进行分析,均得出4种幼苗的抗寒性顺序为葛藤＞盐肤木＞老鸭嘴＞假连翘,这一结果可为采矿石废弃地生态恢复的植物筛选提供科学依据.     

Loss of Trx-2 enhances oxidative stress-dependent phenotypes in Drosophila

Overexpression of thioredoxin (TRX) confers oxidative stress resistance and extends lifespan in mammals and insects. However, less is known about phenotypes associated with loss of TRX. We investigated loss-of-function phenotypes of Trx-2 in Drosophila, and found that the mutant flies are hyper-susceptible to paraquat, a free radical generator, but not to hydrogen peroxide. They contain a high amount of protein carbonyl, which dramatically increases with age. Trx-2 mutants express high levels of anti-oxidant genes, such as superoxide dismutase, catalase, and glutathione synthetase. This is the first demonstration of biochemical and physiological consequences caused by loss of Trx-2 in Drosophila.     

短枝木麻黄家系对青枯病的抗性评价与选择

为探讨短枝木麻黄(Casuarina equisetifolia)对青枯病(Ralstonia solanacearum)侵染后的生理生化响应,利用其生理生化响应指标与抗病性特征的相关性开展短枝木麻黄家系的抗病评价和选择。结果表明,26个短枝木麻黄家系褐梗小枝接种青枯病后,病情指数(DI)和相对病害强度(RDI)均存在极显著差异(P<0.01)。在接种青枯病后,家系间的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)、总酚(TP)和类黄酮(Flav)差异均达显著或极显著水平,说明参试家系对青枯病抗性的变异丰富,具有较大的选择潜力。短枝木麻黄家系的病情症状与6个生理生化指标间均呈显著或极显著的负相关关系,说明青枯病侵染后防御酶活性或次生代谢物含量越高,木麻黄的病害程度越低。利用2个病情特征性状和6个生理生化指标对短枝木麻黄不同家系进行综合评价和选择,按照30%的入选率初步选出201、206、198、207、202、213、208和195号等8个抗病家系。这些抗病家系的RDI和DI比总体均值分别低25.94%和28.93%,SOD、CAT...     

Biogeographic position and body size jointly set lower thermal limits of wandering spiders

Most species encounter large variations in abiotic conditions along their distribution range. The physiological responses of most terrestrial ectotherms (such as insects and spiders) to clinal gradients of climate, and in particular gradients of temperature, can be the product of both phenotypic plasticity and local adaptation. This study aimed to determine how the biogeographic position of populations and the body size of individuals set the limits of cold (freezing) resistance of Dolomedes fimbriatus. We compared D. fimbriatus to its sister species Dolomedes plantarius under harsher climatic conditions in their distribution range. Using an ad hoc design, we sampled individuals from four populations of Dolomedes fimbriatus originating from contrasting climatic areas (temperate and continental climate) and one population of the sister species D. plantarius from continental climate, and compared their supercooling ability as an indicator of cold resistance. Results for D. fimbriatus indicated that spiders from northern (continental) populations had higher cold resistance than spiders from southern (temperate) populations. Larger spiders had a lower supercooling ability in northern populations. The red‐listed and rarest D. plantarius was slightly less cold tolerant than the more common D. fimbriatus, and this might be of importance in a context of climate change that could imply colder overwintering habitats in the north due to reduced snow cover protection. The lowest cold resistance might put D. plantarius at risk of extinction in the future, and this should be considered in conservation plan.     

Functional Characterization of Hevea brasiliensis CRT/DRE Binding Factor 1 Gene Revealed Regulation Potential in the CBF Pathway of Tropical Perennial Tree

Rubber trees (Hevea brasiliensis) are susceptible to low temperature and therefore are only planted in the tropical regions. In the past few decades, although rubber trees have been successfully planted in the northern margin of tropical area in China, they suffered from cold injury during the winter. To understand the physiological response under cold stress, we isolated a C-repeat binding factor 1 (CBF1) gene from the rubber tree. This gene (HbCBF1) was found to respond to cold stress but not drought or ABA stress. The corresponding HbCBF1 protein showed CRT/DRE binding activity in gel shift experiment. To further characterize its molecular function, the HbCBF1 gene was overexpressed in Arabidopsis. The HbCBF1 over expression (OE) line showed enhanced cold resistance and relatively slow dehydration, and the expression of Arabidopsis CBF pathway downstream target genes, e.g. AtCOR15a and AtRD29a, were significantly activated under non-acclimation condition. These data suggest HbCBF1 gene is a functional member of the CBF gene family, and may play important regulation function in rubber tree.     

Could Behaviour and Not Physiological Thermal Tolerance Determine Winter Survival of Aphids in Cereal Fields?

Traits of physiological thermotolerance are commonly measured in the laboratory as predictors of the field success of ectotherms at unfavourable temperatures (e.g. during harsh winters, heatwaves, or under conditions of predicted global warming). Due to being more complicated to measure, behavioural thermoregulation is less commonly studied, although both physiology and behaviour interact to explain the survival of ectotherms. The aphids Metopolophium dirhodum, Rhopalosiphum padi and Sitobion avenae are commercially important pests of temperate cereal crops. Although coexisting, these species markedly differ in winter success, with R. padi being the most abundant species during cold winters, followed by S. avenae and lastly M. dirhodum. To better understand the thermal physiology and behavioural factors contributing to differential winter success, the lethal temperature (physiological thermotolerance) and the behaviour of aphids in a declining temperature regime (behavioural thermotolerance) of these three species were investigated. Physiological thermotolerance significantly differed between the three species, with R. padi consistently the least cold tolerant and S. avenae the most cold tolerant. However, although the least cold tolerant of the study species, significantly more R. padi remained attached to the host plant at extreme sub-zero temperatures than S. avenae and M. dirhodum. Given the success of anholocyclic R. padi in harsh winters compared to its anholocyclic counterparts, this study illustrates that behavioural differences could be more important than physiological thermotolerance in explaining resistance to extreme temperatures. Furthermore it highlights that there is a danger to studying physiological thermotolerance in isolation when ascertaining risks of ectotherm invasions, the establishment potential of exotic species in glasshouses, or predicting species impacts under climate change scenarios.     

Traditional Chinese medicinal herbs as potential AChE inhibitors for anti-Alzheimer’s disease: A review

Alzheimer’s disease (AD) is a progressive neurodegenerative disease affecting 25 million people worldwide, and cholinergic hypothesis is considered as an important hypotheses in the processes of improving cognitive function and recognition skills in recent years. For the long-term treatment of AD, traditional Chinese medicine are particularly suitable for drug discovery. In this review, we sum up six traditional Chinese medicinal herbs concerned with development of AChEIs, including Herba Epimedii, Coptis Chinensis Franch, Rhizoma Curcumae Longae, Green tea, Ganoderma, Panax Ginseng. The listed compounds based on these herbs are belonging to six classes Flavonoids, Alkaloids, Ketones, Polyphenols, Terpenoid and Saponins, respectively. These compounds could be very promising agents in the search for potent anti-Alzheimer’s drugs.     

干旱胁迫对青藏高原6种植物生理指标的影响

以青藏高原6种植物:红花岩黄芪(Hedysarum multijugum Maxim.)、西北沼委陵菜(Comarum salesovianum(Steph.)Asch.et Graebn.)、鲜黄小檗(Berberis diaphana Maxim.)、鲜卑花(Sibiraea laevigata(Linn.)Maxim.)、树锦鸡儿(Caragana arborescens(Amm.)Lam.)、砂生槐(Sophora moorcroftiana(Benth.)Baker)为试材进行干旱胁迫模拟盆栽试验,通过对比6种植物叶片中丙二醛(MDA)、脯氨酸、可溶性蛋白质、叶绿素、可溶性糖、淀粉含量的变化,对供选植物进行抗旱性能分析与评价,为在青藏高原地区筛选优良耐旱植物提供参考依据。设置土壤含水量占田间持水量80%以上为正常水平,50%—70%为轻度干旱,30%—50%为中度干旱,低于30%为重度干旱。研究表明,6种供试植物叶片MDA含量逐渐升高,当土壤含水量占田间持水量12%左右时达到最大值,树锦鸡儿的增幅较另5种苗木最小,而峰值最大。脯氨酸含量都呈增加趋势,其中砂生槐最高,为9981.67mg/g,峰值与对照值相比,砂生槐增加最多为37.43倍,鲜卑花最小为0.49倍。苗木叶片中可溶性蛋白质含量总体表现出现增大后减小的规律,且所有测定值都高于正常水分条件下的含量。6种苗木的叶绿素含量在干旱胁迫下总体呈现先降低再升高最后再降低的趋势,24 d时均低于对照值,下降最多的为西北沼委陵菜(15.43 mg/g)。由于干旱下苗木的光合产物转化受抑,以淀粉形式的累积量减少,所以6种苗木的可溶性糖含量呈上升趋势,而淀粉含量呈下降趋势,通过积累可溶性糖以提高细胞的渗透调节能力。砂生槐、树锦鸡儿的可溶性糖含量增幅与淀粉含量的减少明显高于其余苗木,使其在较低叶水势下仍保持较高的细胞膨压,以减缓水分胁迫的不利影响。主成分分析表明,6种苗木抗旱性由强到弱依次为:树锦鸡儿、红花岩黄芪、砂生槐、鲜黄小檗、鲜卑花、西北沼委陵菜。     

Characterization of marine bacteria highly resistant to mercury exhibiting multiple resistances to toxic chemicals

Several strains of bacteria unusually highly resistant to mercury were isolated from seawater and marine sediment samples and identified by 16S rDNA sequencing and were also characterized by a battery of biochemical and morphological tests. The bacterial isolates were identified to belong to the genera Pseudomonas, Alcaligenes, Brevibacterium and Bacillus. Many of the chosen isolates were tested for growth in the presence of different heavy metals and a variety of xenobiotics. Growth curves of all six bacteria highly resistant to mercury examined for growth at different concentrations of Hg exhibited prolonged lag phase, during which time necessary physiological adaptations to toxic milieu were undergone. All the strains tested for antibiotic resistance showed little to no effect of antibiotics on their normal growth. Results of this study demonstrate the occurrence of diverse groups of marine prokaryotes capable of high tolerance to mercury with a potential to degrade a variety of toxic heavy metals and xenobiotics.     

Trichosanhemiketal A and B: Two 13,14-seco-13,14-epoxyporiferastanes from the root of Trichosanthes kirilowii Maxim.

Of the 32 Trichosanthes species in China, T. kirilowii Maxim. is the most renowned species used in traditional Chinese medicine and has diverse pharmacological properties. However, most of the phytochemical studies of T. kirilowii have focused on the fruits and seeds. In our investigation of the chemical constituents of T. kirilowii roots, two previously undescribed sterols [trichosanhemiketal A and B (1 and 2)], together with 13 known compounds, were isolated and their structures were elucidated. To the best of our knowledge, this represents the first isolation of compounds with a 13,14-seco-13,14-epoxyporiferastane (1–2) skeleton from the Cucurbitaceae family. The anti-inflammatory activity of the isolated compounds was determined through an analysis of their inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in macrophage RAW264.7 cells. Of the compounds, 4, 5, 6, and 8 showed significant inhibitory activities, with IC₅₀ values of 8.5, 15.1, 25.4, and 28.5 µM, respectively. In addition, compound 4 inhibited inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 expression in a concentration-dependent manner.     

木芙蓉对淹水胁迫的响应及其耐涝性综合评价

为挖掘木芙蓉(Hibiscus mutabilis)耐涝种质资源,对13个木芙蓉品种水淹15 d后的形态和生理生化指标进行研究,并构建耐涝评价体系.结果表明,水淹胁迫后,木芙蓉品种各性状的变化趋势和幅度均不同,部分性状间的相关性显著(P<0.05).14个指标按贡献率大小归因为形态生长指标、不定根形态指标、膜脂过氧化指...     

Life‐history traits and physiological limits of the alpine fly Drosophila nigrosparsa (Diptera: Drosophilidae): A comparative study

Interspecific variation in life‐history traits and physiological limits can be linked to the environmental conditions species experience, including climatic conditions. As alpine environments are particularly vulnerable under climate change, we focus on the montane‐alpine fly Drosophila nigrosparsa. Here, we characterized some of its life‐history traits and physiological limits and compared these with those of other drosophilids, namely Drosophila hydei, Drosophila melanogaster, and Drosophila obscura. We assayed oviposition rate, longevity, productivity, development time, larval competitiveness, starvation resistance, and heat and cold tolerance. Compared with the other species assayed, D. nigrosparsa is less fecund, relatively long‐living, starvation susceptible, cold adapted, and surprisingly well heat adapted. These life‐history characteristics provide insights into invertebrate adaptations to alpine conditions which may evolve under ongoing climate change.     

Divergence for tolerance to thermal-stress related traits in two Drosophila species of immigrans group

We investigated associations of the species specific divergence of thermal adaptations of Drosophila nasuta and Drosophila immigrans with seasonal as well as geographical changes in the relative abundance. In D. immigrans, cold resistance and egg-to-adult viability after a cold-shock have shown a positive cline with latitude, whereas no such clinal patterns were evident for heat resistance as well as egg-to-adult viability after heat stress. In contrast, basal heat tolerance and egg-to-adult viability after a heat-shock increased towards the equator i.e. from north to south in D. nasuta. Further, we also found species-specific divergence in seasonal climatic selection responses for heat as well as cold resistance i.e. warmer and higher humidity conditions of the rainy season have evidenced ∼2-fold higher heat tolerance of D. nasuta, but lower temperature as well as lower humidity conditions of the autumn significantly increased (∼3-fold) cold tolerance of D. immigrans. Geographical variations were significant for the absolute and relative hardening capacity for cold tolerance in D. immigrans, but for heat resistance of D. nasuta. Results of multiple regression analysis of changes in thermal tolerance level in latitudinal populations as a simultaneous function of climatic variables (T_ave and RH) have shown that average yearly relative humidity levels significantly affect heat tolerance of D. nasuta, but both T_ave and RH contribute to changes in the cold tolerance of D. immigrans. Thus, our results suggest that climatic selection responses for basal and hardened thermo-tolerance levels affect relative abundance of D. nasuta and D. immigrans across a latitudinal transect as well as seasons.     

Five small heat shock protein genes from Chilo suppressalis: characteristics of gene, genomic organization, structural analysis, and transcription profiles

Small heat shock proteins (sHSPs) are the most diverse but also the most poorly known family of molecular chaperones, and they play essential roles in various biological processes. The striped stem borer, Chilo suppressalis (Insecta: Lepidoptera: Pyralidae), is one of the most serious pests of rice, causing extensive damage and yield loss. In this study, we isolated and characterized five members of the sHSPs family—Cshsp19.8, Cshsp21.4, Cshsp21.5, Cshsp21.7a, and Cshsp21.7b—from C. suppressalis. The cDNAs of these genes encoded proteins of 177, 187, 191, 191, and 191 amino acids with isoelectric points of 7.0, 5.6, 6.1, 6.3, and 6.3, respectively. While Cshsp19.8, Cshsp21.5, and Cshsp21.7b had no introns, Cshsp21.4 and Cshsp21.7a contained one and two introns, respectively. Structural analysis indicated that all five Cshsps possessed conserved arginine and a V/IXI/V motif, which is related to hydrophobic characteristics of sHSPs. The five heat shock proteins can be classified into two main groups: an orthologous type (Cshsp21.4 and Cshsp21.7a) and a species-specific type (Cshsp19.8, Cshsp21.5, and Cshsp21.7b). Real-time quantitative PCR analyses revealed that Cshsp19.8, Cshsp21.5, Cshsp21.7a, and Cshsp21.7b all exhibited their highest expression levels within Malpighian tubules or the hindgut, while such levels were found in the head for Cshsp21.4. The expression of Csshsps at different developmental stages revealed that the mRNA levels of Cshsp19.8, Cshsp21.4, Cshsp21.5, and Cshsp21.7b peaked in adults, whereas the highest level of Cshsp21.7a was observed in first instar larvae. Cshsp19.8 and Cshsp21.7b were both upregulated dramatically by heat and cold, and Cshsp21.5 could be induced by cold stress. Neither Cshsp21.4 nor Cshsp21.7a responded to heat or cold. These results demonstrated that different Csshsps play distinctive roles in the regulation of the physiological activities in C. suppressalis.     

Physiological and Proteomics Analyses Reveal the Mechanism of Eichhornia crassipes Tolerance to High-Concentration Cadmium Stress Compared with Pistia stratiotes

Cadmium (Cd) pollution is an environmental problem worldwide. Phytoremediation is a convenient method of removing Cd from both soil and water, but its efficiency is still low, especially in aquatic environments. Scientists have been trying to improve the ability of plants to absorb and accumulate Cd based on interactions between plants and Cd, especially the mechanism by which plants resist Cd. Eichhornia crassipes and Pistia stratiotes are aquatic plants commonly used in the phytoremediation of heavy metals. In the present study, we conducted physiological and biochemical analyses to compare the resistance of these two species to Cd stress at 100 mg/L. E. crassipes showed stronger resistance and was therefore used for subsequent comparative proteomics to explore the potential mechanism of E. crassipes tolerance to Cd stress at the protein level. The expression patterns of proteins in different functional categories revealed that the physiological activities and metabolic processes of E. crassipes were affected by exposure to Cd stress. However, when some proteins related to these processes were negatively inhibited, some analogous proteins were induced to compensate for the corresponding functions. As a result, E. crassipes could maintain more stable physiological parameters than P. stratiotes. Many stress-resistance substances and proteins, such as proline and heat shock proteins (HSPs) and post translational modifications, were found to be involved in the protection and repair of functional proteins. In addition, antioxidant enzymes played important roles in ROS detoxification. These findings will facilitate further understanding of the potential mechanism of plant response to Cd stress at the protein level.     

A novel calcium-dependent protein kinase gene from Populus euphratica, confers both drought and cold stress tolerance

Populus species are the most important timber trees over the Northern hemisphere. Most of them are cold- and drought-sensitive except the Populus euphratica Oliv. Here, a calcium-dependent protein kinase (CDPK) gene cloned from P. euphratica, designated as PeCPK10, was rapidly induced by salt, cold, and drought stresses. The protein encoded by PeCPK10 was localized within the nucleus and cytosol, which may be important for its specific regulation in cellular functions. To elucidate the physiological functions of PeCPK10, we generated transgenic Arabidopsis plants overexpressing PeCPK10. The results showed that PeCPK10-transgenic lines experienced better growth than vector control plants when treated with drought. Stronger abscisic acid-induced promotion of stomatal closing has been showed in transgenic lines. Particularly, overexpression of PeCPK10 showed enhanced freezing tolerance. Constitutive expression of PeCPK10 enhanced the expression of several abscisic acid-responsive genes and multiple abiotic stress-responsive genes such as RD29B and COR15A. Accordingly, a positive regulator responsive to cold and drought stresses in P. euphratica is proposed.     

Genome-Wide Identification,Characterization, and Stress-Responsive Expression Profiling of Genes Encoding LEA (Late Embryogenesis Abundant) Proteins in Moso Bamboo (Phyllostachys edulis)

Late embryogenesis abundant (LEA) proteins have been identified in a wide range of organisms and are believed to play a role in the adaptation of plants to stress conditions. In this study, we performed genome-wide identification of LEA proteins and their coding genes in Moso bamboo (Phyllostachys edulis) of Poaceae. A total of 23 genes encoding LEA proteins (PeLEAs) were found in P. edulis that could be classified to six groups based on Pfam protein family and homologous analysis. Further in silico analyses of the structures, gene amount, and biochemical characteristics were conducted and compared with those of O. sativa (OsLEAs), B. distachyon (BdLEAs), Z. mays (ZmLEAs), S. bicolor (SbLEAs), Arabidopsis, and Populus trichocarpa. The less number of PeLEAs was found. Evolutionary analysis revealed orthologous relationship and colinearity between P. edulis, O. sativa, B. distachyon, Z. mays, and S. bicolor. Analyses of the non-synonymous (Ka) and synonymous (Ks)substitution rates and their ratios indicated that the duplication of PeLEAs may have occurred around 18.8 million years ago (MYA), and divergence time of LEA family among the P. edulis-O. sativa and P. edulis–B. distachyon, P. edulis-S. bicolor, and P. edulis-Z. mays was approximately 30 MYA, 36 MYA, 48 MYA, and 53 MYA, respectively. Almost all PeLEAs contain ABA- and (or) stress-responsive regulatory elements. Further RNA-seq analysis revealed approximately 78% of PeLEAs could be up-regulated by dehydration and cold stresses. The present study makes insights into the LEA family in P. edulis and provides inventory of stress-responsive genes for further functional validation and transgenic research aiming to plant genetic improvement of abiotic stress tolerance.