Tansley review
Sodium transport in plants:a critical review
reactive oxygen species是什么意思Author for correspondence: Herbert J.Kronzucker
Tel:+14162877436
Email:herbertk@utsc.utoronto.ca Received:6August2010 Accepted:27September2010Herbert J.Kronzucker and Dev T.Britto
Department of Biological Sciences,University of Toronto,1265Military Trail,Toronto,ON M1C 1A4,Canada
New Phytologist(2011)189:54–81 doi:10.1111/j.1469-8137.2010.03540.x Key words:channels,influx,potassium, sodium toxicity,sodium transport.Summary
Sodium(Na)toxicity is one of the most formidable challenges for crop production world-wide.Nevertheless,despite decades of intensive research,the pathways of Na+entry into the roots of plants under high salinity are still not definitively known. Here,we review critically the current paradigms in thisfield.In particular,we explore the evidence supporting the role of nonselective cation channels,pota
s-sium transporters,and transporters from the HKT family in primary sodium influx into plant roots,and their possible roles elsewhere.We furthermore discuss the evi-dence for the roles of transporters from the NHX and SOS families in intracellular Na+partitioning and removal from the cytosol of root cells.We also review the literature on the physiology of Na+fluxes and cytosolic Na+concentrations in roots and invite critical interpretation of seminal published data in these areas.The main focus of the review is Na+transport in glycophytes,but reference is made to literature on halophytes where it is essential to the analysis.
Contents
Summary54 I.Introduction55 II.The role of nonselective cation channels in primary sodium influx–a solid consensus.How solid is the evidence?
55 III.Low-affinity cation transporter1–a forgotten link?61 IV.Are potassium transporters implicated in sodium influx?62 V.HKT:a saga of twists and turns–where do we stand?64 VI.SOS:an ambiguous tale67VII.Vacuolar storage via NHX:some lingering questions68 VIII.Other pathways–the apoplast and possibilities of symport with chloride
69
IX.‘Toxic’Na+fluxes,Na+‘homeostasis’,and the question of cytosolic Na+
71 X.Concluding remarks72 Acknowledgements73 References
73
New
Phytologist
54New Phytologist(2011)189:54–wphytologist
Ó2011The Authors New PhytologistÓ2011New Phytologist Trust
Abbreviations:AAG,amino-acid-gated;AKT,Arabidopsis K+transporter;AVP, Arabidopsis vacuolar pyrophosphatase;CCC,cation-chloride cotransporters;CNGC, cyclic-nucleotide-gatedchannel;DEPC,diethylpyrocarbonate;DA,depolarization-activated; esb,enhanced suberin;GLR,glutamate receptor;HA,hyperpolarization-activated; HAK,high-affinity K+transporter;HKT,high-affinity K+transporter;KcsA,Strepto-myces K+channel;K i,inhibition constant;K m,Michaelis constant;Kna,K+⁄Na+ discrimination locus;KT,K+transporter;KUP,K+uptake permease;LCT,low-affinity cation transporter;Nax,Na+exclusion;NHX,Na+⁄H+exchanger;NSCC,nonselective cation channel;PTS,8-hydroxy-1,3,6-pyrenetrisulphonic acid;QTL,quantitative trait loci;ROS,reactive oxygen species;SBFI,sodium-binding benzofuran isophthalate; SKC,shoot K+concentration;SOS,salt overly sensitive;TEA,tetraethyl ammonium; Trk,K+transporter;VI,voltage-insensitive.
I.Introduction
Soil salinity is a global environmental challenge,affecting crop production on over800million hectares,or a quarter to a third of all agricultural land on earth(Szabolcs,1989; Rengasamy,2010).The problem is particularly severe in irrigated areas(Flowers,1999;Zhu,2001),where as much as one-third of global food production takes place(Munns, 2002;Munns&Tester,2008;Zhang et al.,2010)and where i
nfiltration of highly saline sea water(Flowers,2004) is common.However,salinity is also increasing in dryland agriculture in many parts of the world(Wang et al.,1993; Rengasamy,2006).While saline soils contain numerous salts at elevated concentrations,NaCl typically dominates (Zhang et al.,2010),and it is believed that the harmful effects of saline conditions on most species are principally brought about by a combination of osmotic stress and ionic stress exerted by the sodium component of NaCl (Blumwald,2000;Hasegawa et al.,2000;Munns&Tester, 2008).Only in the cases of some woody species,such as in the genera Citrus and Vitis(grapevine),does chloride appear to be the more toxic ion(White&Broadley,2001).It is for this reason that decades of research activity have been dedi-cated to the characterization of Na+transport and distribu-tion in plants,and in particular itsfirst entry into plant roots.In recent years,this endeavour has been augmented by the search for molecular candidates for Na+transport, with some remarkable successes,but not without significant controversies.In this review,we will take a critical look at the main classes of transporters that have been identified, chiefly by means of electrophysiological and molecular tech-niques,and will discuss these achievements in the context of the whole plant and of plant cultivation in thefield,to which significant discoveries must ultimately relate.We par-ticularly focus on aspects where conclusions may have been drawn prematurely,and point out discrepancies that require further discussion or experimentation to achieve progress. We shall show that the link between electrophysiological evidenc
e of Na+transport via nonselective cation channels (NSCCs)in protoplasts and artificial bilayer systems on the one hand,and in planta‘toxic’Na+fluxes on the other,may have been accepted prematurely;that many published Na+flux values under saline conditions in plant roots are ener-getically difficult to explain,and may require a new inter-pretation;that participation in Na+uptake by transporters such as low-affinity cation transporter1(LCT1)and K+ transporters from the KUP⁄HAK⁄KT and AKT families, and as yet poorly characterized‘back-up’systems of K+ acquisition,cannot be discounted at this point;that evi-dence for the role of HKT2transporters in primary Na+ uptake under K+deprivation conditions is strong,as is evidence for the role of HKT1transporters in controlling internal Na+distribution between the root and the shoot, while evidence for their roles in primary Na+uptake under saline conditions is limited;that evidence for the role of Salt Overly Sensitive1(SOS1)in Na+efflux back into the exter-nal medium is not as clear as frequently indicated,and its role in root–shoot Na+transfer is obscure;that evidence for the role of NHX in vacuolar Na+sequestration and subse-quent rescue from Na+toxicity is strong,but important questions remain;and that a proper evaluation of the role of cytosolic Na+,and,in particular,the cytosolic Na+:K+ ratio,is hampered by a scarcity of direct measurements (these are summarized here)and its utility,as well as that of total-tissue Na+accumulation,as a predictor of sodium stress may not be as great as is often stated.
II.The role of nonselective cation channels in primary sodium influx–a solid consensus.How solid is the evidence?
1.The functional subclasses of NSCCs
Even though no definitive molecular candidates have thus far emerged,a strong consensus has developed in recent years,largely based on electrophysiological studies,that var-ious classes of NSCCs catalyse primary influx of Na+under saline conditions.NSCCs are thoroughly characterized
in
New
Phytologist Tansley review
Ó2011The Authors
New PhytologistÓ2011New Phytologist Trust New Phytologist(2011)189:54–wphytologist
animals,and their functions are well understood in their cellular signaling,vascular endothelial function,Ca 2+influx in response to store depletion,and renal ion homeostasis (Kaupp &Seifert,2002;Clapham,2003;Firth et al.,2007;Venkatachalam &Montell,2007;Kauer &Gibson,2009).In plants,several categories of NSCCs have also been iden-tified,and these have been subdivided (Demidchik &Tester,2002;Demidchik &Maathuis,2007),according to their response to changes in membrane electrical potential,into the following major classes:(1)depolarization-activated NSCCs (DA-NSCCs),(2)hyperpolarization-activated NSCCS (HA-NSCCs),and (3)voltage-insensitive NSCCs (VI-NSCCs).Additional classification systems distinguish NSCCs by their reponsiveness to certain ligands and physi-cal stimuli and include cyclic-nucleotide-gated NSCCs (CNGCs),amino-acid-gated NSCCs (AAG-NSCCs),and reactive-oxygen-species-activated NSCCs (ROS-NSCCs).These may well constitute representatives of subclasses (1)through (3),as may other minor types of NSCCs not discussed here (see Demidchik &Maathuis,2007).
The first definitive demonstration,using patch-clamp approaches,of NSCC-type conductances in plants dates to 1989,when Stoeckel and Takeda reported constitutive cation fluxes across the plasma membranes of triploid endosperm cells in species from the genera Haemanthus and Clivia that displayed minimal selectivity for various alkali,and some earth alkali,ions,and could be activated follow-
ing depolarizations of the membrane potential (Stoeckel &Takeda,1989).Despite some constitutive activity,these types of NSCCs have thus been classified in category 1above.DA-NSCC operation has since been confirmed in a large number of experimental systems,including leaf and root cell preparations from Arabidopsis thaliana ,Thlaspi arvense and T.caerulescens ,Hordeum vulgare and Phaseolus vulgaris (Cerana &Colombo,1992;Spalding et al.,1992;
de Boer &Wegner,1997;Pei et al.,1998;Pin
˜eros &Kochian,2003;Zhang et al.,2004).Their main function appears to be in conducting Ca 2+(White &Ridout,1999;White et al.,2000),although a role in catalyzing K +release from root cells under sudden imposition of saline condi-tions has also been proposed (Shabala et al.,2006).By con-trast,the role of DA-NSCCs in catalyzing primary Na +fluxes under salt stress conditions has been much less con-clusively demonstrated.Nevertheless,in a major review on the topic (Demidchik &Maathuis,2007),it was suggested that members of this depolarization-activated class of NSCCs may well be involved in this function.The proposal was based upon reference to a series of comparative electro-physiological studies conducted in Arabidopsis thaliana and its natural halophyte relative Thellungiella halophila (Volkov et al.,2004;Volkov &Amtmann,2006;Wang et al.,2006);studies that,however,concluded that the predominant Na +conductances observed were voltage-insensitive ,not depolarization-activated.A role
for the
subclass of depolarization-activated NSCCs in catalyzing significant Na +fluxes under saline conditions therefore remains purely speculative at this point.
NSCC category 2(HA-NSCCs)can be excluded from further in-depth discussion in the context of primary Na +fluxes under salinity,as hyperpolarization of the plasma membrane,inherent to the gating properties of these chan-nels (Gelli &Blumwald,1997;Hamilton et al.,
2000;Ve
´ry &Davies,2000;Demidchik et al.,2007),does not typically accompany the imposition of salinity,neither in short-term nor in long-term applications of Na +(Laurie et al.,2002;Carden et al.,2003;Shabala et al.,2006;Volkov &Amtmann,2006;Malagoli et al.,2008).2.VI-NSCCs:the current consensus
In contrast to the above categories,a substantial number of studies support a role for VI-NSCCs (category 3)in cataly-zing Na +fluxes across the plasma membrane,in particular in roots (some reports have also focused on shoots:
Elzenga &van Volkenburgh,1994;Ve
´ry et al.,1998),and it is here where more extensive discussion is warranted.CNGCs,AAG-NSCCs and ROS-NSCCs may well repre-sent subclasses of this type of NSCC (Demidchik &Maathuis,2007).The earliest demonstration of VI-NSCCs was in wheat (Triticum aestivum ;Moran et al.,1984;see also:Tyerman et al.,1997;Buschmann et al.,2000;Davenport &Tester,2000),followed by extensive work in rye (Secale cereale ;White &Tester,1992;White &Lemtiri-Chlieh,1995;White &Ridout,1995;White,1996),maize (Zea mays ;Roberts &Tester,1997),barley (Hordeum vulgare ;Amtmann et al.,1997),A.thaliana (Maathuis &Sanders,2001;Demidchik &Tester,2002;Shabala et al.,2006;Volkov &Amtmann,2006),Thellungiella halophila (Volkov et al.,2004;Volkov &Amtmann,2006;Wang et al.,2006),and Capsicum annuum (Murthy &Tester,2006).Common features unite the observations in this large body of studies:VI-NSCCs are so named because their open probability is not significantly,or at best weakly,modulated by membrane potential,in contrast to the categories of NSCCs discussed above.Currents are constitutive and instantaneous (i.e.per-manently present when ensemble averages,not individual channel traces,are examined),and they lack time-dependent activation (Tyerman et al.,1997;Amtmann &Sanders,1999;White,1999b).VI-NSCCs have been shown,in clas-sic current–voltage relationships,to conduct both inward and outward currents,and thus may constitute both influx and efflux pathways in planta (Shabala et al.,2006;Volkov &Amtmann,2006).VI-NSCCs also exhibit several pharmacological characteristics that separate them fr
om other classes of ion channels (see Demidchik et al.,2002;Demidchik &Maathuis,2007):they are not sensitive to the potassium channel inhibitors Cs +and tetra-ethyl-ammonium (TEA +),are not affected by the alkali cations Li +and Na +,
the
Tansley review
New
Phytologist
Ó2011The Authors
New Phytologist Ó2011New Phytologist Trust
New Phytologist (2011)189:54–wphytologist
sodium channel inhibitor tetrodotoxin(cf.Allen et al., 1995)or the calcium channel inhibitors verapamil a
nd nifedipine,but are greatly inhibited by the trivalent cations lanthanum(La3+)and gadolinium(Gd3+;it should be noted, however,that these two cations are very broad-spectrum;Qu et al.,2007).One class of VI-NSCCs can also be partially blocked by divalent cations,including Ba2+and Zn2+,as well as,especially importantly,Ca2+and Mg2+, while another class is not inhibited by these ions,but instead transports them(Demidchik&Maathuis,2007).Some VI-NSCCs are also inhibited by the organic compound quinine (Demidchik&Tester,2002),but this feature is not universal (White&Lemtiri-Chlieh,1995;White&Broadley,2000). Other treatments,including pH changes(stimulation by alkaline pH and inhibition by acidic pH)and application of the histidine modifier diethylpyrocarbonate(DEPC;strong inhibition),have also been shown to be effective in selected experimental systems,such as A.thaliana(Demidchik& Tester,2002)and rye(White,1999a),but have as yet not been tested widely.Within a given experimental
A.thaliana),such responses,in addition to the more univer-sally exhibited ones,provide valuable gauges for a critical comparative evaluation of physiological results obtained by different methods(for further discussion,see Section II.3 below).
In most studies on VI-NSCCs,clear demonstration of Na+conductance was provided.As suggested by their name, VI-NSCCs are,to a high degree,nonselective for cations, that is,similar permeation of a vari
ety of cations can be observed when such tests are conducted.Nevertheless,ion preferences are still encountered,resulting in selectivity series.Many such series have been published,and,while generally similar,they vary in their detail.In a seminal study on A.thaliana(Demidchik&Tester,2002),the series observed(cation permeabilities are listed relative to Na+) was:K+(1.49)>NH4+(1.24)>Rb+(1.15)>Cs+(1.10) >Na+(1.00)>Li+(0.73)>TEA+(0.47).In rye roots (White&Tester,1992),the series was:K+(1.36)=Rb+ (1.36)>Cs+(1.17)>Na+(1.00)>Li+(0.97)>TEA+ (0.41).In wheat,NH4+(2.06)>Rb+(1.38)>K+(1.23) >Cs+(1.18)>Na+(1.00)>Li+(0.83)>TEA+(0.20)was reported(Davenport&Tester,2000).In other words,in these three benchmark studies(see also Tyerman et al., 1997and Volkov&Amtmann,2006),the macronutrient potassium(and,where tested,also ammonium)was trans-ported to a significantly greater extent than sodium,from equimolar concentrations(see also Zhang et al.,2010). Thus,for this category of NSCCs,the cation selectivity ser-ies appear to follow a more consistent pattern than the frequently cited range of K+:Na+selectivity ratios for NSCCs of0.3to3(Demidchik et al.,2002;Demidchik& Maathuis,2007).The published selectivity series should provide an important gauge for determining the contribu-tion of NSCCs to Na+conductance in planta.
Additional subclasses of NSCCs that have been the subject of some discussion in the context of Na+fluxes are cyclic nucleotide-gated and amino-acid-(in particular,glu-tamate-)gated NSCCs(CNGCs
and AAG-NSCCs;see also Demidchik&Maathuis,2007).Among these,CNGCs are perhaps the best studied.They are characterized by gating mediation involving the second messengers cAMP and cGMP,and their role in animal physiology is diverse and has been extensively investigated,in particular within the context of transduction of visual and olfactory stimuli and Ca2+signalling(Kaupp&Seifert,2002;Talke et al.,2003; Gobert et al.,2006;Takeuchi&Kurahashi,2008). However,functional expression of plant CNGCs has proved difficult,and thus little functional consolidation has occurred to date,even though some20CNGCs have been found in the A.thaliana genome(Gobert et al.,2006; Demidchik&Maathuis,2007).However,in a few cases, expression in heterologous systems,including Xenopus laevis oocytes and yeast,has been successful(Leng et al.,2002; Balague´et al.,2003;Gobert et al.,2006),and sensitivity to cAMP and cGMP has been observed,as well as sensitivity to Cs+(Balague´et al.,2003)and Mg2+(Leng et al.,1999). Interestingly,in planta Na+fluxes,in glycophytes under toxic conditions,are typically reported to be insensitive to Cs+(see later discussion onfluxes;also see,however,Kader &Lindberg(2005)for work examining the protoplasts of rice;Wang et al.(2007)for work on the halophyte Sueda maritima,and Voigt et al.(2009)for Na+tissue content data in cowpea(Vigna unguiculata)–these studies present evidence of Cs+sensitivity of Na+uptake).Cesium sensitiv-ity,and the voltage sensitivity seen in many CNGCs,reduce the likelihood of their significant involvement in catalyzing Na+fluxes in whole plants for
extended periods of time(the roles of AtCNGC2,4,11and12in response to pathogen attack,and theflow of Ca2+under such conditions,are,by contrast,well documented;Balague´et al.,2003; Demidchik&Maathuis,2007;Guo et al.,2010).Two CNGCs from the A.thaliana genome,AtCNGC3and AtCNGC10,have nevertheless been linked to primary K+ and Na+fluxes in roots.In the case of the former (AtCNGC3),tissue expression analysis has localized the transporter to root epidermal and cortical cells,and a null mutation in the gene has been shown to reduce the net uptake rate of Na+during the initial(although not the later) stages of NaCl exposure,resulting in slightly enhanced growth on intermediate(40–80mM)NaCl concentrations; the Na+content of mutant seedlings,however,was not dif-ferent from that of the wild type following longer term treatments at high(80–120mM)NaCl concentrations (Gobert et al.,2006).The work may indicate a role for AtCNGC3in Na+uptake in the early phases(the initial few hours)of salt stress.In the case of AtCNGC10,tissue expression studies have also localized the transporter to root tissues,and the gene was able to complement the
reduced
New
Phytologist Tansley review
Ó2011The Authors
New PhytologistÓ2011New Phytologist Trust New Phytologist(2011)189:54–wphytologist
K +uptake phenotype of the A.thaliana akt1;1mutant (see Hirsch et al.,1998;Spalding et al.,1999),establishing a possible role for the transporter in alkali ion fluxes in roots (Li et al.,2005).Other more recent studies,however,have shown a greater role of AtCNGC3in the transport of the earth alkali ions Ca 2+and Mg 2+(Guo et al.,2010),although Na +transport may be involved indirectly (Guo et al.,2008),while other CNGCs,such as AtCNGC2,are strongly selective for K +over Na +(Leng et al.,2002).In support of an in planta involvement of CNGCs in Na +transport under toxic conditions,some studies have indeed reported a sensitivity of unidirectional or net fluxes of Na +to cyclic nucleotides (Maathuis &Sanders,2001;Essah et al.,2003;Rubio et al.,2003;Maathuis,2006;see,how-ever,Section II.3).Additionally,the observation that salt-tolerant varieties of rice down-regulate OsCNGC1to a greater extent than salt-sensitive varieties under saline con-ditions (Senadheera et al.,2009)may also be taken as cir-cumstantial evidence for an involvement
of CNGCs in Na +influx.Based on these findings,therefore,the role of CNGCs in primary Na +fluxes cannot be dismissed at this point,and deserves careful further investigation,but the balance of the evidence does not currently favour a signifi-cant involvement (see also Zhang et al.(2010),who review conflicting information regarding whether CNGCs are blocked,or activated,by cyclic nucleotides).
Another subgrouping of ligand-sensitive NSCCs that may be involved in Na +transport is that of AAG-NSCCs,and,in particular,those gated by glutamate.Precedents for glutamate-activated NSCCs abound in the animal literature (Dingledine et al.,1999;Traynelis et al.,2010),but their role in plant physiology,and under conditions of sodium toxicity,is more obscure (Lam et al.,1998;Davenport,2002;Demidchik &Maathuis,2007).At this time,con-vincing functional analyses of these channels are lacking,despite the fact that,as with CNGCs,some 20AAG-NSCCs have been identified in the A.thaliana genome.The voltage insensitivity and instantaneous activation of currents,along with sensitivity to quinine and lanthanides in one study (Demidchik et al.,2004),suggest that AAG-NSCCs may represent subclasses of VI-NSCCs.While some evidence from Xenopus oocytes indicates the possibil-ity of Na +transport in at least some members of this family (AtGLR1;1,AtGLR 1;4and AtGLR3;7;Roy et al.,2008;Tapken &Hollmann,2008),the preponderance of evi-dence currently supports a role for AAG-NSCCs in Ca 2+transport (Dennison &Spalding,2000;Dub
os et al.,2003;Demidchik et al.,2004)and signalling during development (Kim et al.,2001;Turano et al.,2002;Li et al.,2006;Qi et al.,2006;Walch-Liu et al.,2006),rather than a role in primary Na +fluxes under saline conditions (cf.Essah et al.,2003).Similarly,ROS-NSCCs (perhaps most NSCCs?)appear to be predominantly involved in Ca 2+transport (Demidchik &Maathuis,2007).
3.Linking electrophysiological readings from proto-plasts to fluxes in the whole plant:the challenge It has to be strongly emphasized,and we will return to this critical point later,that essentially all demonstrations of the role of NSCCs,and in particular of VI-NSCCs,in catalyz-ing Na +fluxes have been achieved by patch-clamp analysis with isolated protoplasts or artificial lipid bilayers.By con-trast,the connection between such measurements and Na +fluxes at the level of whole tissues and the whole plant is,in fact,much less secure (Malagoli et al.,2008;Britto &Kronzucker,2009;Zhang et al.,2010),although the oppo-site conclusion is often stated (Davenport,2002;Munns &Tester,2008).Several key studies have attempted to relate Na +currents measured by electrophysiology in protoplasts and artificial lipid bilayer systems to Na +fluxes and accumulation in intact plants and ⁄or plant tissues.Once such set of comparative experiments was carried out in wheat (Davenport &Tester,2000),and another in A.thaliana (Demidchik &Tester,2002;Essah et al.,2003).Both sets of studies employed 22Na +-labelling of excised plants roots alongside electrophysiological exami
na-tions of protoplast and lipid bilayer preparations within a genotype.In the first of these studies,the authors showed that ‘Na +influx through the NSC channel resembled 22Na +influx’(Davenport &Tester,2000),and,indeed,con-cluded,even within the paper’s title,that a ‘nonselective cation channel mediates toxic sodium influx in wheat’.
This attribution was supported in large part by the partial sensitivity of both radiolabelled Na +fluxes and Na +currents to Ca 2+,Mg 2+and Gd 3+,and their insensitivity to other inhibitors,including those specific to potassium channels (TEA +and Cs +;cf.Kader &Lindberg,2005;Wang et al.,2007;Zhang et al.,2010).While Ca 2+sensitivity may indeed link NSCC operation well to the frequently (albeit not universally:see Yeo &Flowers,1985;Schmidt et al.,1993;Malagoli et al.,2008)observed amelioration of Na +toxicity by Ca 2+in whole plants (LaHaye &Epstein,1969;Greenway &Munns,1980;Rengel,1992;Epstein,1998),it should be kept in mind that Ca 2+has a myriad of other effects on plants (Britto et al.,2010;Zhang et al.,2010)and thus can hardly be seen as specific,and that the similarly strong Mg 2+sensitivity documented for NSCC operation (Davenport &Tester,2000;their Fig.4)is not typically reflected in the Na +toxicity rescue of plants (LaHaye &Epstein,1969).In addition,however,other issues deserve discussion.First,Ca 2+sensitivity,while exhibiting similar K i values for electrical currents in bilayer preparations and tracer fluxes in roots (in the range of 610–650l
M;Davenport &Tester,2000;see also White,1999b;cf .Wang et al.,2007;Malagoli et al.,2008),was much more pronounced in sin-gle-channel preparations (>50%)than it was in roots,where,at Ca 2+concentrations above 3mM,c .75%of the influx seen at the lowest [Ca 2+]was still observed,measuring
in
Tansley review
New
Phytologist
Ó2011The Authors
New Phytologist Ó2011New Phytologist Trust
New Phytologist (2011)189:54–wphytologist
版权声明：本站内容均来自互联网，仅供演示用，请勿用于商业和其他非法用途。如果侵犯了您的权益请与我们联系QQ:729038198，我们将在24小时内删除。
发表评论