长江经济带湖南区域外来植物入侵现状及防控对策

[目的]了解长江经济带湖南区域外来入侵植物的种类构成、区系组成和区域分布,推进长江经济带湖南区域的生物多样性保护。[方法]采用实地踏查法开展长江经济带湖南区域外来入侵植物调查,以文献访查法对调查区域的入侵植物物种进行补充分析。[结果]长江经济带湖南区域有外来入侵植物41科102属146种,其中,菊科植物最多(35种),占入侵植物总种数的23.97%;外来入侵植物原产于美洲的最多(99种),占入侵植物总种数的67.81%;从生活型组成看,草本植物是长江经济带湖南区域外来入侵植物的主要组成部分,占比82.88%;从科的分布区系特征看,世界广布类型是入侵植物的主要区系类型(19科),占总科数的46.34%;从属的分布区系特征看,泛热带分布类型最多(32属),占总属数的31.38%。[结论]长江经济带湖南区域外来植物入侵呈现种类多、增速快、危害程度加重等特点,应加强对菊科入侵植物和来源于美洲入侵植物的监管。     

The oxygen delivery response to acute hypoxia during incremental knee extension exercise differs in active and trained males

Background

It is well known that hypoxic exercise in healthy individuals increases limb blood flow, leg oxygen extraction and limb vascular conductance during knee extension exercise. However, the effect of hypoxia on cardiac output, and total vascular conductance is less clear. Furthermore, the oxygen delivery response to hypoxic exercise in well trained individuals is not well known. Therefore our aim was to determine the cardiac output (Doppler echocardiography), vascular conductance, limb blood flow (Doppler echocardiography) and muscle oxygenation response during hypoxic knee extension in normally active and endurance-trained males.

Methods

Ten normally active and nine endurance-trained males (VO_2max = 46.1 and 65.5 mL/kg/min, respectively) performed 2 leg knee extension at 25, 50, 75 and 100% of their maximum intensity in both normoxic and hypoxic conditions (FIO₂ = 15%; randomized order). Results were analyzed with a 2-way mixed model ANOVA (group × intensity).

Results

The main finding was that in normally active individuals hypoxic sub-maximal exercise (25 – 75% of maximum intensity) brought about a 3 fold increase in limb blood flow but decreased stroke volume compared to normoxia. In the trained group there were no significant changes in stroke volume, cardiac output and limb blood flow at sub-maximal intensities (compared to normoxia). During maximal intensity hypoxic exercise limb blood flow increased approximately 300 mL/min compared to maximal intensity normoxic exercise.

Conclusion

Cardiorespiratory fitness likely influences the oxygen delivery response to hypoxic exercise both at a systemic and limb level. The increase in limb blood flow during maximal exercise in hypoxia (both active and trained individuals) suggests a hypoxic stimulus that is not present in normoxic conditions.

     

Compatible solutes mitigate damaging effects of salt stress by reducing the impact of stress-induced reactive oxygen species

Under abiotic stress conditions, rapid increases in reactive oxygen species (ROS) levels occurs within plant cells. Although their role as a major signalling agent in plants is now acknowledged, elevated ROS levels can result in an impairment of membrane integrity. Similar to our previous findings on imposition of salt stress, application of the hydroxyl radical (OH^•) to Arabidopsis roots results in a massive efflux of K⁺ from epidermal cells. This is likely to cause significant damage to cell metabolism. Since K⁺ loss also occurs after salt application and salt stress leads to increased cellular ROS levels, we suggest that at least some of the detrimental effects of salinity is due to damage by its resulting ROS on K⁺ homeostasis. We also observed a comparative reduction in K⁺ efflux by compatible solutes after both oxidative and salt stress. Thus, we propose that under saline conditions, compatible solutes mitigate the oxidative stress damage to membrane transporters. Whether this amelioration is due to free-radical scavenging or by direct protection of transporter systems, warrants further investigation.Key words: compatible solutes, hydroxyl radical, potassium efflux, reactive oxygen species, salt stressReactive oxygen species (ROS) are continuously produced as by-products of various metabolic pathways.¹ Under unstressed steady-state conditions, cellular ROS levels are kept in check by the sophisticated antioxidant defence system.² However, under adverse environmental conditions, the balance between ROS production and its subsequent scavenging may be perturbed, leading to a rapid increase in ROS levels.³ Although significant progress has been made in defining ROS as a major signalling agent in plants,³ ROS can react with a large variety of biomolecules, causing lipid peroxidation and impairing membrane integrity.^4,5 One such abiotic stress is salt stress,⁶ with ROS generation occurring within minutes of salt application.⁷ Alleviation of oxidative damage may be, therefore, an important strategy of plant salt tolerance.⁸One of the earliest measurable responses to salt stress is a massive K⁺ efflux from plant roots.^9,10 Such K⁺ efflux is initiated within seconds of acute salt stress and may last for several hours^11,12 reducing the intracellular K⁺ pool^13,14 and significantly impairing cell metabolism. Consistent with the key role of K⁺ homeostasis in salt tolerance mechanisms¹⁵ a reduction of K⁺ efflux correlates with increased salt tolerance.^11,12We have previously reported that hydroxyl radical (OH^•) application to Arabidopsis roots also results in a rapid efflux of K⁺ from the epidermis.¹⁶ In this report, we find a similar K⁺ efflux response.¹⁷ As is the case for salt stress,⁹ we found that membrane depolarisation could be responsible for a substantial part of this efflux. However, an observed discrepancy between the membrane depolarisation and the pattern of K⁺ efflux indicates that voltage-dependence is not the only factor influencing K⁺ loss from the root cells after oxidative stress. Demidchik et al.¹⁶ demonstrated that stress-induced K⁺ efflux could be mediated by activation of K⁺ outward rectifying channels directly by OH^•. This direct effect on K⁺ transporters could also account for our observed delay before the peak efflux of K⁺ is measured, indicating that a certain amount of time is required before maximal direct damage by OH^• to transporters occurs. Because both K⁺ channel blockers and non-selective cation channel blockers reduce this efflux, it indicates non-specificity in OH^• attack. Furthermore, combinations of these channel blockers were effective in reducing K⁺ efflux implying that, at least in the short term, the damaging effects of OH^• is due to compromising the transporter systems as opposed to lipid peroxidation. Certainly, K⁺ channels harbour reactive groups, thus are expected to be sensitive to ROS.¹⁸We have previously shown that the exogenous application of low concentrations of a variety of compatible solutes reduces the salt-induced K⁺ efflux.^19,20 Plants, when confronted with a saline environment, respond with a significant elevation in their compatible solute levels. This ameliorates the detrimental effects of salinity.²¹ However, their original proposed role in cellular osmoregulation is under question: their concentration in transgenic plants overexpressing osmolyte biosynthetic genes is not significant for osmotic adjustment, despite showing improved salt tolerance.⁸ Furthermore, one hallmark of the detoxification effect is its lack of specificity, that is, transgenic plants have increased tolerance not only to high salt, but also to drought, cold and heat shock,^22,23 stresses that also result in ROS production.³ Certainly, ecotopic expression studies suggest that compatible solutes increase stress tolerance by protection of membranes and proteins against ROS.⁶We show that in this work that exogenous application of low concentrations of a range of compatible solutes significantly reduces OH^•-induced K⁺ efflux,¹⁷ a similar effect to that we reported after salt application to barley roots¹⁹ and also observed in Arabidopsis (Fig. 1). Interestingly, we found that not only known free-radical scavenging osmolytes,²⁴ but also glycine betaine, previously found to be non-effective in ROS scavenging,²⁴ were effective in reducing OH^•-induced K⁺ efflux. Indeed, glycine betaine showed a greater mitigation of OH^•-induced K⁺ efflux compared to that induced by 50 mM NaCl (Fig. 1). However, it is open to speculation as to whether this mitigation is via direct channel blocking, a direct protection of ion channel proteins or by some other protective mechanism.Open in a separate windowFigure 1Effects of exogenous supply of compatible solutes on net peak K⁺ efflux after application of either 1 mM Cu/a or 50 mM NaCl. Roots were preincubated for 1 h in 5 mM concentration of a number of compatible solutes prior to treatment, Mean ± SE (n = 6-8).In our further investigations we have found that salt-tolerant barley show a reduced ROS-induced K⁺ efflux compared to sensitive varieties.²⁵ This superior ability of salt-tolerant barley cultivars of preventing K⁺ loss further indicates a possible causal link between salt and oxidative stress tolerance. We propose that upon the imposition of salt stress, the instantaneously resulting membrane depolarisation9 results in activation of depolarisation activated K⁺ outward-rectifying channels, leading to the initial massive K⁺ efflux. Over the longer term, ROS levels within the plant cell increase,⁷ resulting in direct damage to K⁺ transporters and the longer-term sustained loss of K⁺ from the cell. Due to mitigation of both NaCl- and OH^•-induced K⁺ efflux by compatible solutes, we propose that one of their primary amelioratory effects is through reducing the damaging effects of salt-produced ROS on K⁺ transporter, and by this means, reducing the effects of stress damage. Whether this amelioration is achieved through free-radical scavenging or due to a direct protection of membrane transports warrants further investigation.     

不同退化红砂荒漠草地的水分分配格局

研究了内蒙古阿拉善盟不同过牧退化红砂草地的土壤 植物 大气系统的水分分配格局、不同退化草地和主要植物种的水分利用效率 .2 0 0 1年降雨量 12 4 .3mm ,其中试验期 119.4mm .1m深土壤水分结果表明 ,10～ 4 0cm土层受蒸散影响最大 ;由于主要共存种红砂和无芒隐子草根系分布和蒸腾强度不同等 ,含水量在 10～ 2 0cm土层以中度退化区显著低于其它样区 (P <0 .0 5 ) ,而 2 0～ 4 0cm土层以轻度退化区较低 .样地年均蒸发量为 30 .6mm ,红砂种群的年均蒸腾量为 11.9mm .随着草地退化加剧 ,裸地的蒸发量和退化指示种匍根骆驼蓬种群的蒸腾量增加 ,而红砂种群的蒸腾量降低 .与较轻度退化区比 ,中度和重度退化区的水分利用率分别下降了 14 .6 %和 4 6 .1% ,红砂水分利用率分别下降了 37.8%和 73.8% .     

Ghrelin在小鼠体内早期胚胎发育中的表达

为了探讨Ghrelin是否在昆明系小白鼠(Mus musculus)体内早期胚胎发育中发挥作用。运用激素超排卵技术及动物解剖学方法获取小鼠体内不同发育阶段的早期胚胎,应用实时定量PCR技术检测了Ghrelin mRNA的相对表达量。结果表明,Ghrelin mRNA在小鼠各期胚胎中均有表达;同时,GhrelinmRNA的表达量呈现一定规律性变化,2-细胞期表达量最低,8-细胞期表达量达到最高值,总体趋势是先降低后升高之后又降低。研究结果揭示,Ghrelin可能在小鼠早期胚胎发育中发挥一定的作用。     

我国巴拉巴按蚊形态描述

白踝按蚊种团是由很多形态近似的蚊种组成,其中巴拉巴按蚊和白踝按蚊均为东南亚一带的重要传疟媒介。我国过去资料均记载为白踝按蚊,但未从形态上详细鉴定。为了确定该种的分类地位,1963年作者等在海南岛万宁县兴隆附近山林区采集“白踝按蚊”全套标本10批共150多套,1971年后从云南省采集少数标本进行形态鉴定,发现其与Colless(1956,1957)所描述的白踝按蚊Anopheles leucosphyrus leucosphyrus Donitz(1901)有很明显的差别,而与巴拉巴按蚊A.balabacensis balabacensis Balsas(1936)除某些特征略有差异外,余均完全一致。按Colless的分类法,过去我国记载的“白踝按蚊”,应鉴定为巴拉巴按蚊A.balabacensis balabacensis Baisas。     

Endomembrane Ca2+-ATPases play a significant role in virus-induced adaptation to oxidative stress

Although the role of Ca²⁺ influx channels in oxidative stress signaling and cross-tolerance in plants is well established, little is known about the role of active Ca²⁺ efflux systems in this process. In our recent paper,¹⁷ we reported Potato Virus X (PVX)-induced acquired resistance to oxidative stress in Nicotiana benthamiana and showed the critical role of plasma membrane Ca²⁺/H⁺ exchangers in this process. The current study continues this research. Using biochemical and electrophysiological approaches, we reveal that both endomembrane P_2A and P_2B Ca²⁺-ATPases play significant roles in adaptive responses to oxidative stress by removing excessive Ca²⁺ from the cytosol, and that their functional expression is significantly altered in PVX-inoculated plants. These findings highlight the crucial role of Ca²⁺ efflux systems in acquired tolerance to oxidative stress and open up prospects for practical applications in agriculture, after in-depth comprehension of the fundamental mechanisms involved in common responses to environmental factors at the genomic, cellular and organismal levels.Key words: cytosolic calcium, reactive oxygen species, cross-tolerance, calcium pumpThe phenomenon of cross-tolerance to a variety of biotic and abiotic stresses is well-known.^1,2 Some of the demonstrated examples include the correlation between oxidative stress tolerance and pathogen resistance.^3–5 At the mechanistic level, changes in cytosolic Ca²⁺ levels [Ca²⁺]_cyt, have long been implicated as a quintessential component of this process.⁶ The rise in [Ca²⁺]_cyt is proven to be essential for the development of the oxidative burst required for triggering the activation of several plant defense reactions.^7,8 The observed elevation in H₂O₂ level is believed to result from Ca²⁺-dependent activation of the NADPH oxidase,⁸ which then causes a further increase in [Ca²⁺]_cyt via a positive feedback mechanism. This process is further accomplished by defense gene activation, phytoalexin synthesis and eventual cell death.⁹ Downstream from the stimulus-induced [Ca²⁺]_cyt elevation, cells possess an array of proteins that can respond to a message. Such proteins include calmodulin (CaM),¹⁰ Ca²⁺-dependent protein kinases¹¹ and CaM binding proteins.¹² Of note is that when Ca²⁺ channels are blocked, biosynthesis of ROS is prevented.¹³While the role of Ca²⁺ influx channels in oxidative stress signaling and cross-tolerance in plants is well established, little is known about the involvement of active Ca²⁺ efflux systems in this process. In contrast, in animal systems the essential role of re-establishing [Ca²⁺]_cyt to resting levels is widely reported. A sustained increase in [Ca²⁺]_cyt in the alveolar macrophage is thought to be the consequence of membrane Ca²⁺-ATPase dysfunction.¹⁴ In endothelial cells, inhibition of the Ca²⁺/Na⁺ electroneutral exchanger of the mitochondria was named as one of the reasons for [Ca²⁺]_cyt increases.¹⁵ A significant loss of the plasma membrane Ca²⁺-ATPase (PMCA) activity was reported in brain synapses in response to oxidative stress,¹⁶ suggesting that PMCA may be a downstream target of oxidative stress.In our recently published paper¹⁷ we reported the phenomenon of Potato Virus X (PVX)-induced acquired resistance to oxidative stress in Nicotiana benthamiana plants and showed the critical role of plasma membrane Ca²⁺/H⁺ exchangers in this process. Nonetheless, questions remain, is this transporter the only active Ca²⁺ efflux system involved in this process?In addition to Ca²⁺/H⁺ exchangers, active Ca²⁺ extrusion could also be achieved by Ca²⁺-ATPases. Two major types of Ca²⁺-ATPases that differ substantially in their pharmacology and sensitivity to CaM are known.¹⁸ Type P_2A pumps (also called ER-type or ECA^19,20) are predominantly ER-localized,¹⁹ although they are also present at other endomembranes (e.g., tonoplast and Golgi). Four members of this group have been identified in the Arabidopsis genome (named AtECAs 1 to 4).^18,21 These pumps lack an N-terminal autoregulatory domain, are insensitive to CaM and suppressed by cyclopropiazonic acid (CPA).¹⁹ P_2B (or ACA) pumps contain an autoinhibitory N-terminal domain that possesses a binding site for Ca²⁺-CaM.¹⁸ Ten members are known in Arabidopsis (termed AtACA1, 2, 4 and 7 to 13).²¹ Plant P_2B pumps are located at the plasma membrane²⁰ as well as in inner membranes such as tonoplast (e.g., ACA4), ER (e.g., ACA2) and plastids.^18,19 These pumps probably constitute the basis for precise cytosolic Ca²⁺ regulation; as the Ca²⁺ concentration increases, CaM is activated and binds to the autoinhibitory domain of the Ca²⁺ pump. This results in the activation of the pump.In our recent study,¹⁷ we found no significant difference between the purified plasma membranes fractions isolated from control and UV-treated tobacco plants (with or without PVX inoculation) either in the Ca²⁺-ATPase activity or in the Ca²⁺-ATPase expression level and its ability to bind CaM. This suggests that the plasma membrane P_2B type pumps (the only pump type known to be expressed at the plasma membrane) play no major role in removing excess Ca²⁺ from the cytosol under oxidative stress conditions. This led to an obvious question: what about endomembrane Ca²⁺-ATPases?To address this issue, microsomal membrane fractions were isolated from tobacco leaves in a manner previously described for plasma membrane fractions¹⁷ (Fig. 1A). Western blot and CaM overlay assays were then made to investigate the role of endomembrane P_2B Ca²⁺-ATPases in our reported phenomena of acquired resistance. The results show that the expression of the P_2B Ca²⁺ pumps in PVX-inoculated plants is significantly higher than in control plants (Fig. 1B), correlating well with the CaM overlay assay (Fig. 1C). As no difference was observed for the P_2B Ca²⁺-ATPase expression levels in the plasma membranes,¹⁷ the observed difference in the microsomal fractions of PVX-infected plants must be due to an increased expression of endomembrane P_2B Ca²⁺-ATPases. Given the fact that Ca²⁺ pumps have a high affinity for calcium, the observed increase in endomembrane P_2B-type Ca²⁺-ATPases expression in PVX-inoculated plants may be advantageous for more efficient Ca²⁺ removal from the cytosol into internal organelles.Open in a separate windowFigure 1Expression of P_2B Ca²⁺ in purified microsomal fractions from tobacco leaves. Measurements were undertaken C = mock controls; C-UV = mock controls treated with UV-light; PVX = PVX infected plants; PVX-UV = PVX inoculated plants treated with UV-light. (A) Coomassie Brilliant Blue-stained gel; (B) Protein blot immunostained with a non isoform-specific polyclonal antibody for P_2B Ca²⁺-ATPases; (C) CaM overlay assay.To decipher the possible role of P_2A Ca²⁺-ATPases in acquired resistance, a series of electrophysiological experiments were conducted using inhibitors of P_2A-type Ca²⁺-ATPases, such as thapsigargin (TG)²² and cyclopiazonic acid (CPA).²³ Ion-selective Ca²⁺ microelectrodes were prepared as described elsewhere in reference ²⁴ and ²⁵, and net Ca²⁺ fluxes were measured from tobacco mesophyll tissue following previously described protocols.¹⁷ Leaf pre-treatment for 2 h in either of these inhibitors dramatically suppressed the net Ca²⁺ efflux measured from tobacco mesophyll cells 2 h after UV light exposure (Fig. 2). Given the specificity of TG and CPA inhibitors for P_2A-type Ca²⁺-ATPases, these results strongly support a hypothesis that both endomembrane P_2A and P_2B Ca²⁺-ATPases play significant roles in plant adaptive responses to oxidative stress. This is achieved by removing excess Ca²⁺ from the cytosol.Open in a separate windowFigure 2Effect of known Ca²⁺-ATPase blockers on light-induced Ca²⁺ flux kinetics after 20 min of UV-C treatment. Leaf mesophyll segments were pre-treated in either 5 µM TG (thapsigargin) or 50 µM CPA (cyclopiazonic acid) for 1–1.5 h prior to exposure to UV-C light. Net Ca²⁺ fluxes were measured 2 h after the end of UV treatment. These were compared with two controls: (1) no pre-treatment/no UV exposure (closed circles) and (2) no pre-treatment/20 min UV exposure (open squares). Mean ± SE (n = 4 to 7).Combining these results with our previously reported observations in reference ¹⁷, the following model is proposed (Fig. 3). Oxidative stress (such as UV) causes increased ROS production in leaf chloroplasts, leading to the elevated [Ca²⁺]_cyt. Several Ca²⁺ efflux systems are involved in restoring basal cytosolic Ca²⁺ levels. Two of these, the plasma membrane Ca²⁺/H⁺ exchanger¹⁷ and endomembrane P_2A and P_2B Ca²⁺-ATPases (as reported in this study) are upregulated in PVX inoculated plants and contribute to the improved tolerance to oxidative stress. Overall, these findings highlight the potential role of Ca²⁺ efflux systems in virus-induced tolerance to oxidative stress in plants. This is consistent with our previous reports on the important role of Ca²⁺ efflux systems in biotic stress tolerance²⁶ and brings forth possibilities for genetic engineering of more tolerant plants by targeting expression and regulation of active Ca²⁺ efflux systems at either the plasma or endomembranes.Open in a separate windowFigure 3The proposed model of oxidative stress signaling and the role of Ca²⁺-efflux systems in acquired resistance and plant adaptation to oxidative stress.Overall, a better adaptation of virus-infected plants to a short wave UV irradiation as compared to uninfected controls may suggest that infection triggers common defense mechanisms that could be efficient against secondary unrelated stresses. This observation may lead to the development of novel strategies to protect plants against complex environmental stress conditions.     

Proteomic profiling of oil bodies isolated from the unicellular green microalga Chlamydomonas reinhardtii: with focus on proteins involved in lipid metabolism

Oil bodies are sites of energy and carbon storage in many organisms including microalgae. As a step toward deciphering oil accumulation mechanisms in algae, we used proteomics to analyze purified oil bodies from the model microalga Chlamydomonas reinhardtii grown under nitrogen deprivation. Among the 248 proteins (≥ 2 peptides) identified by LC-MS/MS, 33 were putatively involved in the metabolism of lipids (mostly acyl-lipids and sterols). Compared with a recently reported Chlamydomonas oil body proteome, 19 new proteins of lipid metabolism were identified, spanning the key steps of the triacylglycerol synthesis pathway and including a glycerol-3-phosphate acyltransferase (GPAT), a lysophosphatidic acid acyltransferase (LPAT) and a putative phospholipid:diacylglycerol acyltransferase (PDAT). In addition, proteins putatively involved in deacylation/reacylation, sterol synthesis, lipid signaling and lipid trafficking were found to be associated with the oil body fraction. This data set thus provides evidence that Chlamydomonas oil bodies are not only storage compartments but also are dynamic structures likely to be involved in processes such as oil synthesis, degradation and lipid homeostasis. The proteins identified here should provide useful targets for genetic studies aiming at increasing our understanding of triacyglycerol synthesis and the role of oil bodies in microalgal cell functions.