3种荒漠植物光合及叶绿素荧光对干旱胁迫的响应及抗旱性评价

利用盆栽试验结合人工浇水后自然耗水的方法测定干旱胁迫对梭梭、白刺、沙蒿3种荒漠植物叶片水分、光合及叶绿素荧光参数的影响,探讨各指标在干旱胁迫过程中的变化特征、响应机制及其与土壤水分的定量关系,并用隶属函数法对其进行抗旱性排序。结果表明：(1)3种植物叶片相对含水量(RWC)随干旱胁迫天数增加持续降低,最大水分亏缺(RWD)呈波动式上升趋势。(2)3种植物总叶绿素含量(Chl)和叶绿素a(Chla)、叶绿素b(Chlb)含量,以及梭梭、白刺类胡萝卜素含量均随胁迫天数增加而降低;沙蒿类胡萝卜素随土壤含水率降低逐渐升高。(3)梭梭、白刺、沙蒿叶片净光合速率(P_n)、蒸腾速率(T_r)、水分利用效率(WUE)等主要光合气体交换参数对土壤水分表现出明显的阈值响应,适宜的土壤含水率分别为8.04%～19.33%、4.17%～19.10%、6.48%～17.51%。(4)3种植物 PSⅡ最大光化学效率(Fv/F_m)、实际光化学效率(F_v′/F_m′)及光化学淬灭系数(qP)均随干旱胁迫天数增加和光照强度增大而降低,非光化学淬灭系数(NPQ)则呈逐渐上升趋势;干旱胁迫中后期,梭梭、沙蒿的F_v/F_m及F_v′/F_m′均下降,光合机构光合活性遭到破坏,电子传递受阻,PSⅡ反应中心受损,表现出光抑制,而白刺调节自身PSⅡ反应中心免受伤害的能力较强。(5)隶属函数法综合分析表明,3种植物耐旱能力大小依次为白刺>梭梭>沙蒿。研究发现,3种荒漠植物均可通过调节 PSⅡ反应中心开放程度与活性,对干旱胁迫表现出较强的耐性,胁迫后期植物PSⅡ反应中心关闭或不可逆失活,表现出光抑制。     

干旱胁迫对燕麦生长及叶片光系统Ⅱ活性的影响

以‘燕科2号’燕麦品种为试验材料,采用盆栽控水的方式分别设置正常供水(75%田间持水量)、中度干旱胁迫(60%田间持水量)、重度干旱胁迫(45%田间持水量)3个水分处理,利用叶绿素荧光技术研究了不同水分梯度下燕麦生长和叶片光反应系统Ⅱ(PSⅡ)功能的变化,探讨干旱胁迫对燕麦叶片光合性能的影响。结果表明：(1)干旱胁迫导致燕麦株高变矮,叶片数、主茎数、穗数减少,叶片失绿发黄及籽粒产量显著下降。(2)与正常供水相比,重度干旱胁迫下燕麦叶片最大光化学效率(F_v/F_m)和光合性能指数(PI_ABS)显著降低。(3)重度干旱胁迫导致燕麦叶片单位反应中心吸收的光能(ABS/RC)和单位反应中心耗散掉的能量(DI₀/RC)明显下降,单位反应中心用于电子传递的能量(ET₀/RC)和单位反应中心捕获的光能(TR₀/RC)明显升高;有活性反应中心的开放程度(Ψ₀)和电子传递链的量子产额(φ_E0)明显下降、非光化学淬灭最大量子产额(φ_D0)明显升高,V_J、V_K、V_L 3个位点的相对荧光强度明显升高,OJIP曲线初始斜率M_o明显升高。研究发现,燕麦叶片PSⅡ对中度干旱胁迫具有较强的适应能力,而重度干旱胁迫严重伤害其叶片PSⅡ反应中心,导致其反应中心能流分配失衡,电子传递受阻和PSⅡ稳固性减弱,进而影响燕麦光合作用,最终导致燕麦生长受到抑制。     

外源硅对自毒作用下黄瓜幼苗生长及光合特性的影响

以‘绿博6号’黄瓜幼苗为试材,采用营养液培法研究不同浓度外源硅(0、0.5、1.0、1.5和2.0 mmol/L)对肉桂酸(cinnamic acid,CA)模拟自毒胁迫(3.0 mmol/L CA)下黄瓜幼苗生长、光合参数和叶绿素荧光参数的影响。结果表明：(1)自毒胁迫显著抑制了黄瓜幼苗的生长、根系形态建成和生物量的积累,显著降低了净光合速率(P_n)、蒸腾速率(T_r)、气孔导度(G_s)和叶绿素(Chl a、Chl b和Chl t)含量,并显著降低了叶片中PSⅡ的电子传递速率(ETR)、PSⅡ实际光化学效率［Y_(Ⅱ)］和光化学淬灭系数(qP)。(2)添加适宜浓度外源硅可有效缓解自毒胁迫对黄瓜幼苗生长的影响,并提高其P_n、G_s、T_r和叶绿素含量,在一定程度上维持叶片光合系统的稳定。(3)添加适宜浓度外源硅能使自毒胁迫下黄瓜叶片的F_v/F_m、ETR、Y_(Ⅱ)和qP显著升高,而非光化学淬灭系数(NPQ)则显著下降。研究发现,添加适宜浓度外源硅能提高自毒胁迫下黄瓜幼苗的叶绿素含量、F_v/F_m、ETR、Y_(Ⅱ)和qP,使光合机构趋于稳定,抑制P_n的下降,缓解自毒胁迫对光合系统的损伤,从而增强黄瓜幼苗对自毒胁迫的抗性,并以1.0 mmol/L外源硅处理的效果最佳。     

逆境胁迫对菜豆黄化苗转绿过程中交替呼吸途径与叶绿素荧光特征的影响

以菜豆黄化幼苗作为试验材料,探讨了铅(Pb)或PEG(聚乙二醇)胁迫下交替呼吸途径在植物转绿过程中对叶绿素含量以及叶绿素荧光特性的影响,以阐明逆境胁迫下植物交替呼吸途径的生理学作用。结果显示：(1)与菜豆黄化幼苗正常转绿过程(对照)相比,Pb或PEG胁迫导致菜豆黄化幼苗的叶绿素含量积累延迟,使叶片PSⅡ潜在最大光化学量子效率(F_v/F_m)、光适应下最大光化学效率(F_v′/F_m′)、PSⅡ光适应下实际光化学效率(Y(Ⅱ))和光化学荧光猝灭系数(qP)显著下降,而非光化学猝灭系数(NPQ)则显著增加。(2)在菜豆黄化幼苗转绿过程中,Pb或PEG胁迫导致其交替呼吸途径容量较对照均显著上升。(3)Pb或PEG胁迫下,交替呼吸途径抑制剂［水杨基氧肟酸(SHAM,1 mmol/L)］使菜豆黄化幼苗转绿过程中叶绿素含量、F_v/F_m、F_v′/F_m′、Y(Ⅱ)和qP进一步下降, NPQ却进一步增加,说明抑制交替呼吸途径会加剧Pb或PEG胁迫对PSⅡ反应中心活性的进一步抑制,使还原力积累加剧,造成热耗散进一步增加。研究表明,Pb或PEG胁迫均显著降低了菜豆黄化幼苗PSⅡ对光能的利用率,进而阻碍了菜豆黄化幼苗转绿进程;交替呼吸途径有助于在胁迫条件下缓解PSⅡ的过度还原,可能在一定程度上缓解了Pb或PEG胁迫对其转绿进程的阻碍作用。     

水分胁迫对极小种群东兴金花茶幼苗光合特性的影响

为了解东兴金花茶幼苗对水分胁迫的适应能力和响应机制,该文以东兴金花茶1年生实生苗为材料,采用盆栽控水试验,研究不同控水时间处理对东兴金花茶幼苗的生理生态特性的影响。结果表明:随着控水时间的延长,水分胁迫的程度不断加剧,东兴金花茶叶片净光合速率(P_n)、气孔导度(G_s)、蒸腾速率(T_r)呈现显著下降趋势; 胞间CO₂浓度(C_i)呈现先低后高的变化趋势,水分利用效率(WUE)呈现先高后低的变化趋势。土壤含水率和叶片相对含水量均呈现不断下降的趋势,丙二醛呈现先降后升的变化趋势; 东兴金花茶幼苗的荧光参数F_v/F_m和F_v/F_o呈现先增加后不断下降趋势,分别从0.806下降至0.754和4.17下降至3.08,表明水分胁迫降低了PS Ⅱ原初光能转化效率,光合作用原初反应过程受到抑制。基于水分胁迫的生理生态指标和叶片生物性状的变化表明,控水时间在4 d情况下东兴金花茶可以提高自身水分利用效率来抵抗干旱,说明东兴金花茶幼苗对水分胁迫具有一定的适应性和响应机制。控水8～12 d,东兴金花茶的光合指标下降显著,土壤含水率下降至14.157%～15.065%,其叶片萎蔫、打卷,低于此水平东兴金花茶幼苗会因过度干旱而死亡,表明东兴金花茶幼苗对水分胁迫的耐受极限土壤含水率为14.157%～15.065%。研究结果有助于营建适宜的环境以保证东兴金花茶的正常生长和繁殖,对东兴金花茶迁地保护、引种培育和回归自然具有重要的科学指导意义。     

适度紫外辐射增强对白鲜光合特性和药用活性成分的影响

次生代谢成分环境调控是药用植物优质栽培的理论和实践基础。然而,迄今为止,短期紫外光诱导对药用活性成分积累效应方面的研究仍然比较薄弱。该文以光环境敏感植物白鲜(Dictamnus dasycarpus)为研究对象,探索短期不同强度(低剂量和中剂量)紫外(UV)辐射增强对其根、茎和叶中黄柏酮、梣酮、白鲜碱和柠檬苦素4种有效成分的诱导效应。结果表明:(1)无论是低剂量还是中剂量UV-A和UV-B辐射条件下,白鲜叶片光系统Ⅱ(PS Ⅱ)最大光化学量子产量(F_v/F_m)均大于0.76; PS Ⅱ实际光合量子产量Y(Ⅱ)、调节性能量耗散的量子产量Y(NPQ)、基于“湖泊模型”光化学淬灭系数(qL)和非光化学淬灭系数(NPQ)与对照(未经紫外辐射处理)相比均没有显著差异; 低剂量与中剂量UV-B辐射均显著增加白鲜PS Ⅱ的非调节性能量耗散的量子产量Y(NO)。(2)适度短期紫外辐射增强能够诱导白鲜药用活性成分快速积累,根中4种有效成分最高可提升51%,主要在白鲜根中积累。其中,中剂量UV-A辐射和低剂量UV-B辐射效果最明显,不仅根中黄柏酮、梣酮、白鲜碱和柠檬苦素4种活性成分明显提升,还促进茎中白鲜碱和叶中梣酮的积累。综上所述,该研究结果表明短期紫外辐射增强能有效诱导白鲜药用活性成分的积累,并通过提升白鲜PS Ⅱ非光化学效率来提升白鲜光强耐受性。     

干旱胁迫对黄柳雌雄扦插苗生长和生理特性的影响

以黄柳雌雄扦插苗为试验材料,通过称重控水法研究干旱胁迫下其生长和生理特性,以揭示黄柳雌雄植株对干旱胁迫的生理适应性及抗逆能力差异。结果显示：(1)在对照处理下,黄柳雌雄扦插苗在株高、基径、总叶片数、单叶面积、茎干重、根干重、光合色素含量(除叶绿素a)、气体交换、叶绿素荧光、叶片相对含水量(RWC)、丙二醛(MDA)含量、脯氨酸含量和可溶性糖含量等方面均没有显著的性别差异。(2)干旱胁迫显著降低了黄柳雌雄扦插苗的生长和光合作用,扰乱了其渗透调节功能和抗氧化酶系统;与雄株相比,雌株具有较高的株高、总叶片数、单叶面积、茎干重、根干重、总叶绿素含量、净光合速率(P_n)、PSⅡ最大化学效率(F_v/F_m)、PSⅡ有效光量子产量(Φ_PSⅡ)和叶片RWC,以及较低的非光化学淬灭系数(qN)、MDA含量、脯氨酸含量和可溶性糖含量;且雌株叶干重、叶绿素a含量、超氧化物歧化酶(SOD)活性和过氧化物酶(POD)活性始终大于雄株。研究表明,干旱胁迫对黄柳雌雄扦插苗的生长和生理特性均具有严重的抑制作用,且雌雄株之间的响应存在显著的性别差异;雌株通过维持较高的光合作用以保证植株的生长,从而表现出比雄株更强的抗旱性,这也是其天然种群性比偏雌的重要原因之一。     

盐胁迫对朴树和速生白榆幼苗光合特性及叶绿素荧光参数的影响

以1年生的朴树和速生白榆幼苗为试材,采用山东省林业科学研究院东营分院34.6‰的地下天然盐水配制所需盐水浓度,定期定量浇灌,使土壤分别达0（CK）、2‰、2.5‰、3.5‰和5‰的含盐量,各处理胁迫45 d后测定其生长量（地径、苗高）、叶绿素含量、光合参数以及叶绿素荧光特性的变化,研究不同盐胁迫处理对朴树和速生白榆生长及光合作用的影响,探究其耐盐能力。结果显示: （1）整个盐胁迫过程中速生白榆的生长量较朴树高,但随着盐胁迫强度的增加朴树幼苗叶绿素含量呈先升高后降低的变化趋势,速生白榆幼苗则低于CK呈下降趋势。（2）朴树和速生白榆初始荧光（F_o）、最大荧光（F_m）和PSⅡ最大光能转化效率(F_v/F_m)均表现出下降趋势,与朴树相比速生白榆的F_o、F_m和F_v/F_m下降不明显,但在＞2.5‰盐胁迫下速生白榆的非光化学猝灭系数（NPQ）较朴树增加更明显。研究表明,朴树叶绿素含量在每个盐胁迫浓度下均显著高于速生白榆,但在大于2.5‰浓度胁迫下速生白榆的荧光参数变化较朴树占优势;在非环境胁迫及较低盐胁迫条件下（≤2.5‰）,朴树较速生白榆具有更好的光化学性能,但在较高盐胁迫条件下（＞2.5‰）,速生白榆光合参数及荧光参数较朴树变化稳定,能更好的适应盐胁迫环境。     

不同叶型维西堇菜叶片结构及光合特征比较

为探讨叶片斑纹的光合适应意义,该文选择有斑和无斑两种叶型的维西堇菜(Viola monbeigii)作为研究材料,采用石蜡切片制片和显微镜观察叶片结构及GFS 3000便携式光合测量仪测量光合参数并进行比较。结果表明:(1)两种叶型维西堇菜气孔均为不等型气孔,有斑叶片上、下表皮气孔密度、栅栏组织(PT)厚度、栅栏组织与海绵组织的比值(PT/ST)均显著低于无斑叶片; 而上表皮气孔较大,表现出更适应弱光环境的结构特征。(2)两种叶型维西堇菜暗呼吸速率(R_d)、初始荧光(F_o)、最大荧光产量(F_m)、光化学猝灭系数(q_P)、非光化学猝灭系数(q_N)、PSⅡ最大光化学量子产量(F_v/F_m)、实际光化学反应量子效率(Yield)均无显著差异,有斑植株叶片叶绿素含量、最大净光合速率(P_max)显著低于无斑植株; 有斑叶片表观光合电子传递速率(ETR)在光合有效辐射(PAR)处于400～2 000 μmol·m^-2·s^-1之间时显著低于无斑叶片,无斑植株的光饱和点(LSP)高于有斑植株,而补偿点(LCP)较低。综上认为,无斑植株PAR利用范围较宽,光合适应能力更强,有利于维西堇菜充分利用环境中有限的资源,保障物种生存; 有斑植株具有较适应弱光胁迫的特征,表明叶片斑纹的出现可能是维西堇菜适应林缘弱光环境的一种策略。     

红芽芋驯化苗对盐胁迫的光合及生理响应

为探讨江西铅山红芽芋的耐盐机制,以其组培移栽驯化苗为材料,研究了盐胁迫对其生物量积累、光合特性、荧光特性等抗逆生理特性的影响。结果表明:（1）红芽芋幼苗生物量和根冠比在低盐胁迫下（50 mmol·L^-1）得到显著促进,而在高盐胁迫下（100～250 mmol·L^-1）受到显著抑制。（2）低盐胁迫幼苗的净光合速率（P_n）、气孔导度（G_s）、气孔限制值（L_s）、水分利用效率（WUE）和瞬时羧化效率（CUE）比对照（0 mmol·L^-1）显著增加,细胞间CO₂浓度（C_i）比对照显著下降,蒸腾速率（T_r）与对照无显著差异;高盐胁迫幼苗的P_n、L_s、WUE、CUE和G_s较对照显著下降,C_i比对照显著增加。（3）低盐胁迫幼苗的最大荧光（F_m）、PSⅡ潜在光化学效率（F_v/F₀）和光化学猝灭系数（q_P）比对照显著增加,初始荧光（F₀）较对照显著下降,PSⅡ最大光化学效率（F_v/F_m）、PSⅡ实际光化学效率（Φ_PSⅡ）、开放的PSⅡ反应中心捕获激发能效率（F_v′/F_m′）和非光化学猝灭系数（NPQ）与对照无显著差异;而高盐胁迫幼苗的F₀、F_m、F_v/F_m、F_v/F₀、Φ_PSⅡ、F_v′/F_m′和q_P均较对照显著下降,NPQ比对照显著增加。（4）各盐胁迫幼苗叶片的可溶性蛋白含量以及过氧化物酶、超氧化物歧化酶和过氧化氢酶活性与对照相比先升后降,并以低盐下最高;可溶性总糖和脯氨酸含量均比对照显著增加;丙二醛含量和质膜透性相对值在低盐胁迫下无显著变化,而在高盐下显著增加;叶绿素含量和根系活力在低盐胁迫下无显著变化,而在高盐胁迫后开始显著下降。研究发现,江西铅山红芽芋移栽驯化苗的耐盐阈值为 50 mmol·L^-1,其能够诱导提高叶片可溶性蛋白含量和主要保护酶活性,稳定质膜透性、叶绿素含量和根系活力,增加PSⅡ潜在光化学效率,提高PSⅡ的电子传递活性,维持PSⅡ实际光化学效率,有效启动非辐射热能量耗散机制来保护了光合机构,最终提高净光合速率,增加生物量。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.