垂丝海棠应对盐碱复合胁迫的生理响应

盐碱复合胁迫是影响植物生长的主要环境因子。为了探究垂丝海棠（Malus halliana Koehen）响应盐碱复合胁迫的生理特性,利用主成分分析法明确耐盐碱阈值,以2年生垂丝海棠苗为试材,将NaCl与NaHCO₃按1：1的比例混合,通过盆栽浇灌Hogland营养液的方法,模拟5种不同浓度（0、50、100、150、200 mmol/L）盐碱复合胁迫对其光合色素、光合参数、叶绿素荧光参数及渗透调节物质的影响,并对其相关指标进行相关性和主成分分析。结果表明,随着胁迫强度的增大,叶绿素（Chl T）合成受阻,净光合速率（P_n）、气孔导度（G_s）、蒸腾速率（T_r）、胞间CO₂浓度（C_i）、叶片含水量（WC）下降,最大光化学效率（F_v/F_m）、表观光合电子传递速率（ETR）、PSⅡ实际化学效率（φPSⅡ）、光化学猝灭系数（qP）被抑制;类胡萝卜素（Car）含量、水分利用效率（WUE）、电解质外渗率、非调节性能量耗散（Y（NO））升高;胁迫40 d时,初始荧光（F₀）显著升高,而调节性能量耗散Y（NPQ）、非光化学荧光淬灭系数（qN）急剧降低;脯氨酸、可溶性糖、有机酸显著积累,并在100 mmol/L浓度下达到峰值。植株的P_n与G_s、C_i、ETR、φPSⅡ呈极显著正相关,与T_r、WC、qP、Chl T呈显著正相关,与电导率、脯氨酸、Y（NO）呈显著负相关。因此,盐碱复合胁迫下,垂丝海棠叶片主要通过降低G_s、C_i、T_r、WC,提高WUE、大量积累渗透调节物质、启动热耗散机制来保持光合系统伤害与修复的动态平衡。根据主成分分析法,浓度100 mmol/L为耐盐碱阈值,同时G_s、C_i、T_r、Chl T、ETR、φPSⅡ、Y（NO）、WC、qP、电解质渗透率、脯氨酸可作为评价垂丝海棠耐盐碱能力的有效指标。     

干旱胁迫下喷施黄腐酸对燕麦光合及其抗氧化酶活性的影响

为了揭示黄腐酸对干旱胁迫下燕麦光合及其抗氧化酶活性的影响机理,该研究选用燕麦品种‘燕科2号’为材料,采用盆栽试验,以正常供水(田间持水量的75%)为对照(CK),设干旱胁迫处理(田间持水量的45%,D0)、D0 + 喷施不同浓度黄腐酸(0、200、400、600、800、1 000 mg/L)处理(D1~D5),测定各处理燕麦干鲜重、光合性能及其抗氧化酶活性。结果表明:(1)与CK相比,干旱胁迫下燕麦幼苗地上部鲜重和干重、叶片光合色素含量、净光合速率(P_n)、蒸腾速率(T_r)、气孔导度(G_s)均显著降低,并导致叶片F_v/F_m、qP、ETR和Φ_PSⅡ显著下降,使叶片抗氧化酶 SOD、POD、CAT活性分别显著提高25.68%、19.98%和7.29%。(2)与D0相比,D0 +喷施600 mg/L黄腐酸后,燕麦幼苗地上部鲜重和干重分别显著提高了28.59%和39.13%,叶片叶绿素a、叶绿素a+b、类胡萝卜素和P_n、G_s、T_r及F_v/F_m、Φ_PSⅡ、ETR分别显著增加了25.17%、21.03%、47.37%和74.38%、26.47%、43.34%及6.49%、69.57%、70.71%,C_i和F_o、NPQ分别显著降低了19.52%和13.32%、43.75%。(3)干旱胁迫下喷施不同浓度的黄腐酸均使幼苗叶片中SOD、POD、CAT活性较D0处理显著增加,其中喷施600 mg/L黄腐酸的叶片SOD、POD、CAT活性最高,分别较D0处理显著增加了12.19%、76.57%和55.26%。研究认为,叶面喷施适宜浓度黄腐酸能够显著提高干旱胁迫下燕麦幼苗的光合作用及其抗氧化能力,缓解干旱对燕麦幼苗的伤害,进而促进燕麦幼苗的生长,且以叶面喷施600 mg/L黄腐酸效果最佳。     

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。     

盐胁迫下赤霉素对黄瓜种子萌发及幼苗生长的影响

研究外源GA₃对盐胁迫下黄瓜种子萌发和幼苗生长的影响。结果表明,添加不同质量浓度GA₃的各处理,其发芽率、发芽势和发芽指数均显著高于NaCl胁迫处理,其中以100 mg/L GA₃处理的发芽势、发芽率和发芽指数最高,幼苗的叶面积、根长、根冠比也最大,同时叶片中叶绿素含量最高,幼苗叶片的光合速率(P_n)、气孔导度(G_s)、胞间CO₂摩尔分数(C_i)及蒸腾速率(T_r)等均达到最大;而当赤霉素的质量浓度为50 mg/L时,叶片中的POD活性为2 005 U/(g·min）,达最大值。     

外源H2O2对盐碱胁迫下苹果矮化砧木幼苗生理特性的影响

以2年生苹果矮化砧木M9 T337为试材,采用盆栽试验法,设置浇灌清水(CK)和盐碱胁迫(0.1 mol/L NaCl+NaHCO₃溶液)+ 喷施5种浓度的H₂O₂ ［0(T₁)、0.2 mmol/L(T₂)、0.4 mmol/L(T₃)、0.6 mmol/L(T₄)、0.8 mmol/L(T₅)］ 处理,测定各处理叶片叶绿素含量、光合气体交换参数、渗透调节物质含量、抗氧化酶活性和细胞膜透性,并利用相关性与主成分分析进行综合评价,以探讨外源过氧化氢(H₂O₂)增强其盐碱耐性的生理机制。结果表明：(1)随着盐碱胁迫(T₁)的时间延长,M9 T337幼苗叶片叶绿素a(Chl a)含量、叶绿素b (Chl b)含量、叶绿素总量(Chl t)、净光合速率(P_n)、气孔导度(G_s)、蒸腾速率(T_r)、可溶性蛋白(SP)含量均呈逐渐下降趋势;胞间CO₂浓度(C_i)、可溶性总糖(TSS)含量、脯氨酸(Pro)含量、过氧化氢酶(CAT)活性、抗坏血酸过氧化物酶(APX)活性、相对电导率(REC)、丙二醛(MDA)含量均呈上升趋势;超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性均呈先升后降趋势。(2)与CK相比,盐碱胁迫+外源H₂O₂(T₂- T₅)处理后M9 T337幼苗叶片各指标均呈现不同幅度变化,且存在明显浓度效应,并以T₃(0.4 mmol/L H₂O₂)处理叶片的Chl a、Chl b、Chl t、SP和G_s降幅最小,C_i、REC、MDA升幅最小,TSS、Pro、APX升幅最大。(3)M9 T337幼苗叶片P_n与T_r、G_s、Chl a、Chl b、Chl t、SP、SOD、POD呈显著正相关,与C_i、MDA、CAT、APX、REC呈显著负相关。(4)综合评价表明,各处理对M9 T337幼苗叶片生理特性的效应依次为：CK>T₃>T₄>T₂>T₅>T₁。研究发现,叶面喷施适宜浓度H₂O₂可有效改善盐碱胁迫下M9 T337幼苗光合能力,显著提高抗氧化酶活性和渗透调节物质的含量,降低细胞膜透性,从而达到缓解盐碱胁迫的作用,并以0.4 mmol/L H₂O₂处理效果最佳。     

昆仑山北坡不同海拔塔里木沙拐枣的光合生理生态特性

在2008年7月25日-8月6日的连续晴天中,选择昆仑山北坡塔里木沙拐枣自然分布区3个海拔高度(2100,2300,2500m),利用便携式光合测定仪LI-6400测定塔里木沙拐枣的光合生理生态特性。结果表明:2100m处塔里木沙拐枣的光补偿点(LCP)和光饱和点(LSP)与2300m,2500m处差异分别达到显著,而3者间的最大净光合速率(P_max)差异均达显著。表观量子效率(AQY)在3个海拔之间差异均不显著,但在2100m处的羧化效率(CE)分别与2300m和2500m处的差异显著。相同海拔下塔里木沙拐枣的暗呼吸速率值(R_day)要高于光呼吸速率值(R_p),且2500m处的暗呼吸速率分别与2100m和2300m处的有显著的差异,2100m处的光呼吸速率分别与2300m和2500m差异显著。3个海拔塔里木沙拐枣的净光合速率(P_n)、蒸腾速率(T_r)和气孔导度(G_s)的日变化均为单峰曲线。随着海拔的升高,塔里木沙拐枣P_n,T_r,G_s和L_s的日均值降低,但光能利用率(LUE)和水分利用效率(WUE)却显著增加。塔里木沙拐枣的P_n与叶温(T_l)、大气温度(T_a)和光照强度(PPFD)具有极显著的正相关关系,与海拔呈显著负相关,与空气相对湿度(RH)和大气CO₂浓度(C_a)之间均不具有显著相关性。T_r与T_a,T_l和PPFD具有极显著的正相关性,与RH之间存在显著的负相关。G_s只与C_a之间呈极显著的负相关。P_n与T_r,L_s,G_s,WUE和V_pdl分别具有极显著的正相关性,与C_i呈极显著的负相关。通过对不同海拔高度塔里木沙拐枣光合生理参数与光、温等生态因子关系的对比分析表明:塔里木沙拐枣对山地荒漠草地自然环境变化的温度和光照有很好的生态适应性。     

遮阴和施氮对黄檗幼苗叶片光合特性和碳氮计量特征的影响

选取大田环境下3年生黄檗幼苗,采用人工控制双因素随机区组试验,在不同光照［全光照(S₀)、轻度遮光21.4%(S₁)和重度遮光8.7%(S₂)］和不同氮添加［无添加对照(F₀)、轻度添加(F₁)和重度添加(F₂)］条件下,测定黄檗幼苗叶片的相对叶绿素含量(SPAD值)、气体交换参数及碳氮化学计量特征,探讨黄檗幼苗对遮阴和施氮的响应机制。结果表明：(1)随着遮光程度增强,黄檗幼苗叶片的SPAD值、蒸腾速率(T_r)、气孔导度(G_s)、碳氮比(C∶N)和瞬时光合氮利用率(PNUE)均呈现先增后降的趋势,两种遮光条件下SPAD均显著高于全光照环境;黄檗幼苗叶片的净光合速率(P_n)、水分利用率(WUE)和气孔限制值(L_s)逐渐降低;而叶片氮(N)和碳(C)含量均呈先降后升的趋势。(2)随氮添加量增加,黄檗幼苗叶片的P_n、WUE、L_s、N和C含量均呈先增后降趋势,T_r、G_s和PNUE则逐渐下降,而C∶N逐渐增加。(3)黄檗幼苗叶片的P_n在各光氮组合处理间均无显著差异;SPAD含量以S₁F₀、S₂F₀和S₁F₂处理组合显著较高,而以全光照(S₀)处理组合显著最低;T_r和G_s以轻度遮光(S₁)处理组合明显较高,而以S₂F₁、S₀F₂、S₂F₂明显较低;WUE和L_s均以S₀F₂显著最高,S₂F₂处理组合显著最低。黄檗幼苗叶片N和C含量在重度遮光/轻度氮添加(S₂F₁)时具有较大值,而其C∶N和PNUE在轻度遮光/无氮添加(S₁F₀)时具有较大值。(4)隶属函数综合评价认为,黄檗幼苗对光氮复合作用总体属中等耐受型,轻度遮光时不添加氮肥(S₁F₀)和轻度氮添加(S₁F₁)及全光照时轻度氮添加(S₀F₁)为适于幼苗生长的光氮组合。研究发现,光环境是影响黄檗幼苗光合作用和更新的主导因子,但黄檗苗期能耐受一定的遮阴胁迫;光照不受限制时,适当增加氮肥有利于黄檗幼苗生长;光照受限(重度遮光)时,施氮则抑制叶片叶绿素合成,降低了幼苗光能利用率,不利于其生长。     

基于叶绿体基因组数据的芸香科属间系统发育初步分析（专题约稿）

该研究在人工控制水分条件下,设置3个干旱胁迫处理,选用3个主栽油菜品种‘陇油10号’、‘陇油2号’、‘青杂5号’幼苗进行盆栽试验,测定干旱胁迫条件下叶片相对含水量、叶绿素含量、光合气体交换参数及叶绿素荧光参数等指标,考察各指标在干旱胁迫过程中的变化特征,并通过主成分分析(PCA)和隶属函数法评价品种的抗旱性及其主要响应因子,以揭示西北地区油菜幼苗响应干旱胁迫的光合调控机制。结果表明：(1)各品种油菜幼苗的叶片相对含水量(RWC)均随干旱胁迫程度的递增而逐渐降低,最大水分亏缺(WSD)却逐渐上升。(2)各品种油菜幼苗叶片的叶绿素a含量、叶绿素总含量随着干旱胁迫程度的递增而先增加后递减,且同一种幼苗在不同处理间差异显著。(3)各品种油菜幼苗叶片的净光合速率(P_n)、水分利用效率(WUE)、单株生物量、气孔导度(G_s)、胞间CO₂浓度(C_i)均在受到干旱胁迫时迅速降低,且同一品种幼苗在不同处理间差异显著,而其叶片蒸腾速率(T_r)在干旱胁迫下无显著变化。(4)各品种油菜幼苗叶片光化学猝灭系数(qP)和非光化学猝灭系数(NPQ)随着干旱胁迫程度的递增先增加后递减,最大光化学效率(F_v/F_m)和电子传递速率(ETR)在受到干旱胁迫时迅速降低,且同一品种幼苗在不同处理间差异显著。(5)主成分分析结果表明,在油菜幼苗受到干旱胁迫时RWC、C_i、G_s、P_n、WUE、叶绿素总含量、叶绿素a含量和NPQ起主要调控作用;隶属函数法综合评价表明,3个品种油菜幼苗耐旱能力由高到低依次为‘陇油10号’＞‘陇油2号’＞‘青杂5号’。     

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。     

模拟酸雨胁迫下硅对髯毛箬竹光合特性的影响

以髯毛箬竹为试验材料,研究了模拟酸雨胁迫(pH 3.0)下硅对髯毛箬竹叶片叶绿素相对含量、光合作用日变化以及叶绿素荧光特性等的影响.结果表明:20和100 mg·L^-1 Na₂SiO₃预处理可以不同程度地抑制酸雨胁迫下髯毛箬竹叶片叶绿素含量的显著下降,且以100 mg·L^-1浓度处理效果最佳,叶绿素含量可提高22.7%,而高浓度(500 mg·L^-1)预处理则无缓解作用-酸雨胁迫下,髯毛箬竹光合“午休”现象加重,日平均净光合速率(P_n)、气孔导度(G_s)和气孔限制值(L_s)明显降低,而胞间CO₂浓度(C_i)增大,经过20～100 mg·L^-1硅预处理后,P_n、G_s、L_s不同程度增加,而C_i有所降低,且以100 mg·L^-1硅预处理效果最佳,日平均P_n增加39.2%.酸雨胁迫下,箬竹PSⅡ最大荧光(F_m)、最大光化学效率(F_v/F_m)、潜在活性(F_v/F_o)、有效光化学效率(F_v′/F_m′)、最大荧光产额(F_m′)、光化学淬灭系数(q_P)、非光化学淬灭系数(q_N)、PSII实际光化学效率(Φ_PSⅡ)降低,暗适应和光适应下的最小荧光产额F_o、F_o′ 则升高;而100 mg·L^-1硅预处理明显抑制了胁迫下各荧光参数的变化,F_v/F_m、F_v/F_o、F_v′/F_m′和Φ_PSⅡ分别增加35.2%、146.2%、55.0%和24.3%.说明外源适宜浓度硅预处理能有效地缓解酸雨胁迫导致的髯毛箬竹光合活性下降和光合系统损伤,从而提高胁迫下植物的光合能力.     

Comparison of parameters estimated from A/Ci and A/Cc curve analysis

The parameters estimated from traditional A/C _i curve analysis are dependent upon some underlying assumptions that substomatal CO₂ concentration (C _i) equals the chloroplast CO₂ concentration (C _c) and the C _i value at which the A/C _i curve switches between Rubisco- and electron transport-limited portions of the curve (C _i-t) is set to a constant. However, the assumptions reduced the accuracy of parameter estimation significantly without taking the influence of C _i-t value and mesophyll conductance (g _m) on parameters into account. Based on the analysis of Larix gmelinii’s A/C _i curves, it showed the C _i-t value varied significantly, ranging from 24 Pa to 72 Pa and averaging 38 Pa. t-test demonstrated there were significant differences in parameters respectively estimated from A/C _i and A/C _c curve analysis (p<0.01). Compared with the maximum ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) carboxylation rate (V_cmax), the maximum electron transport rate (J_max) and J_max/V_cmax estimated from A/C _c curve analysis which considers the effects of g _m limit and simultaneously fits parameters with the whole A/C _c curve, mean V_cmax estimated from A/C _i curve analysis (V_cmax-C _i) was underestimated by 37.49%; mean Jmax estimated from A/C _i curve analysis (J_max-C _i) was overestimated by 17.8% and (J_max-C _i)/(V_cmax-C _i) was overestimated by 24.2%. However, there was a significant linear relationship between V_cmax estimated from A/C _i curve analysis and V_cmax estimated from A/C _c curve analysis, so was it J_max (p<0.05).     

Characterization of cytochrome P-450SCC-containing liposomes

Purified cytochrome $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ from bovine adrenocortical mitochondria was incorporated into liposomes by the cholate-dilution method utilizing either dialysis or Sephadex gel filtration. Among synthetic phospholipids tested, dioleoylglycerophosphocholine showed the best stability during the incorporation of $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ into liposomes. A maximum amount of heme was incorporated into liposomes at a molar ratio of phospholipid to the cytochrome of approx. 200. When $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ was incorporated into the dioleoylglycerophosphocholine liposomes by the cholate-filtration method, the $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}\text{-}\text{containing liposomes}$ showed two major populations on the elution pattern of the Sepharose 4B gel filtration, and were seen at a diameter of 200–600 Å and its aggregated forms. When the cytochrome was incorporated into dioleoylglycerophosphocholine liposomes or cholesterol-free adrenocortical mitochondrial liposomes, $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ was less stable than $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ in aqueous solution. Cholesterol or adrenodoxin markedly stabilized the liposomal $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$. Liposomal $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ required cholesterol for its optimum reduction with adrenodoxin, adrenodoxin reductase, and NADPH in the presence of CO. About 70% of the total heme in the dioleoylglycerophosphocholine liposomes was reduced by the enzymatic reduction in the presence of cholesterol, indicating that 70% of the total molecules are exposed to the surface of the outer monolayer. In order to see the location of the heme in membrane, the dioleoylglycerophosphocholine-liposomal $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ was subjected to $\text{p-}\text{chloromercuriphenyl sulfonic acid}$ treatment. This reagent destroyed the liposomal $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$. These results suggest that the heme is located in the proximity of the $\text{p-}\text{chloromercuriphenyl sulfonic acid}$ reacting sites which are exposed to the surface, or located on the vincinity of polar heads of the membrane.     

Occurrence of N-malonyl-D-alanine in pea seedlings

One of the ninhydrin-negative alanine conjugates isolated from pea seedlings was identified as $\text{N-}\text{malonyl}\text{-}\text{D}\text{-}\text{alanine}$.The identification of this conjugate was carried out by a comparison of its gas-liquid chromatographic and mass spectrometric properties, and its nuclear magnetic resonance and infrared spectra with those of synthetic $\text{N-}\text{malonyl}\text{-}\text{D}\text{-}\text{alanine}$. The alanine in the conjugate was shown to be present as the D-isomer by enzymatic and chromatographic analyses.     

The effects of multiple features of alternatively spliced exons on the KA/KSratio test

Background  

The evolution of alternatively spliced exons (ASEs) is of primary interest because these exons are suggested to be a major source of functional diversity of proteins. Many exon features have been suggested to affect the evolution of ASEs. However, previous studies have relied on the K _A /K _S ratio test without taking into consideration information sufficiency (i.e., exon length > 75 bp, cross-species divergence > 5%) of the studied exons, leading to potentially biased interpretations. Furthermore, which exon feature dominates the results of the K _A /K _S ratio test and whether multiple exon features have additive effects have remained unexplored.     

ATP/ADP exchange activity of gastric (H+ + K+)-ATPase

The ATP/ADP exchange is shown to be a partial reaction of the $\text{(}\text{H}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ by the absence of measurable nucleoside diphosphokinase activity and the insensitivity of the reaction to $\text{P}{}^1$, $\text{P}{}^5$ -di(adenosine-5′) pentaphosphate, a myokinase inhibitor. The exchange demonstrates an absolute requirement for Mg²⁺ and is optimal at an ADP/ATP ratio of 2. The high ATP concentration ($\text{K}{}_{0.5}\text{= 116 μ}\text{M}$) required for maximal exchange is interpreted as evidence for the involvement of a low affinity form of nucleotide site. The ATP/ADP exchange is regarded as evidence for an ADP-sensitive form of the phosphoenzyme. In native enzyme, pre-steady state kinetics show that the formation of the phosphoenzyme is partially sensitive to ADP while modification of the enzyme by pretreatment with 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) in the absence of Mg²⁺ results in a steady-state phosphoenzyme population, a component of which is ADP sensitive. The ATP/ADP exchange reaction can be either stimulated or inhibited by the presence of K⁺ as a function of pH and Mg²⁺.     

Comparative study of Bifidobacteriumanimalis, Escherichiacoli, Lactobacilluscasei and Saccharomycesboulardii probiotic properties

The present work investigates some probiotic properties of four different microorganisms (Bifidobacterium animalis var. lactis BB-12, Escherichia coli EMO, Lactobacillus casei and Saccharomyces boulardii). In vitro and in vivo tests were carried out to compare cell wall hydrophobicity, production of antagonistic substances, survival capacity in the gastrointestinal tract of germ-free mice without pathological consequence, and immune modulation by stimulation of Küpffer cells, intestinal sIgA and IL-10 levels. In vitro antagonism against pathogenic bacteria and yeast was only observed for the probiotic bacteria B. animalis and L. casei. The hydrophobic property of the cell wall was higher for B. animalis and E. coli EMO, and this property could be responsible for a better ability to colonize the gastrointestinal tract of germ-free mice. Higher levels of sIgA were observed mainly for S. boulardii, followed by E. coli EMO and B. animalis, and only S. boulardii induced a significant higher level of IL-10. In conclusion, for a probiotic use, S. boulardii presented better characteristics in terms of immunomodulation, and B. animalis and L. casei for antagonistic substance production. The knowledge of the different probiotic properties could be used to choice the better microorganism depending on the therapeutic or prophylactic application.     

Dermal cysts participate in reparative regeneration of epidermis in Hrhr/Hrhr mice

One of the phenotypic effects of mutation in the Hr gene in mice is disintegration of hair follicles and their degeneration into open funnel-shaped structures (utricles) opened on skin surface and cysts located in the depth of the dermis. The aim of the current study consists in analysis of the process of reparative regeneration of skin in homozygotous mice with one of the mutant alleles of the Hr gene—Hr ^hr . It is shown that epithelial cells that constitute the inner pavement of cysts take part in the process of epithelization of deep skin wounds. This indicates that the competence of ectodermal cells in relation to inductive signals from injured skin remains in Hr ^hr homozygote mice, in spite of the significant anatomic abnormalities of the hair follicles.     

Two-Electron Reactions S2QB → S0QB and S3QB → S1QB are Involved in Deactivation of Higher S States of the Oxygen-Evolving Complex of Photosystem II

The oxygen-evolving complex of Photosystem II cycles through five oxidation states (S₀-S₄), and dark incubation leads to 25% S₀ and 75% S₁. This distribution cannot be reached with charge recombination reactions between the higher S states and the electron acceptor Q_B⁻. We measured flash-induced oxygen evolution to understand how S₃ and S₂ are converted to lower S states when the electron required to reduce the manganese cluster does not come from Q_B⁻. Thylakoid samples preconditioned to make the concentration of the S₁ state 100% and to oxidize tyrosine Y_D were illuminated by one or two laser preflashes, and flash-induced oxygen evolution sequences were recorded at various time intervals after the preflashes. The distribution of the S states was calculated from the flash-induced oxygen evolution pattern using an extended Kok model. The results suggest that S₂ and S₃ are converted to lower S states via recombination from S₂Q_B⁻ and S₃Q_B⁻ and by a slow change of the state of oxygen-evolving complex from S₃ and S₂ to S₁ and S₀ in reactions with unspecified electron donors. The slow pathway appears to contain two-electron routes, S₂Q_B → S₀Q_B, and S₃Q_B → S₁Q_B. The two-electron reactions dominate in intact thylakoid preparations in the absence of chemical additives. The two-electron reaction was replaced by a one-electron-per-step pathway, S₃Q_B → S₂Q_B → S₁Q_B in PS II-enriched membrane fragments and in thylakoids measured in the presence of artificial electron acceptors. A catalase effect suggested that H₂O₂ acts as an electron donor for the reaction S₂Q_B → S₀Q_B but added H₂O₂ did not enhance this reaction.     

Ligand-dependent reactivity of (Na+ + K+)-ATPase with showdomycin

Showdomycin inhibited pig brain ($\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ with pseudo first-order kinetics. The rate of inhibition by showdomycin was examined in the presence of 16 combinations of four ligands, i.e., Na⁺, K⁺, Mg²⁺ and ATP, and was found to depend on the ligands added. Combinations of ligands were divided into five groups in terms of the magnitude of the rate constant; in the order of decreasing rate constants these were: (1)$\text{Na}{}^{\mathrm{+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{+}\text{ATP}$, (2) $\text{Mg}{}^{\mathrm{2+}}$, $\text{Mg}{}^{\mathrm{2+}}\text{+}\text{K}{}^{\mathrm{+}}$, $\text{K}{}^{\mathrm{+}}$ and none, (3) $\text{Na}{}^{\mathrm{+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{,}\text{Na}{}^{\mathrm{+}}$, $\text{K}{}^{\mathrm{+}}\text{+}\text{Na}{}^{\mathrm{+}}$ and $\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{+}\text{Mg}{}^{\mathrm{2+}}$, (4) $\text{Mg}{}^{\mathrm{2+}}\text{+}\text{K}{}^{\mathrm{+}}\text{+}\text{ATP}\text{,}\text{K}{}^{\mathrm{+}}\text{+}\text{ATP}$ and $\text{Mg}{}^{\mathrm{2+}}\text{+}\text{ATP}\text{, (5)}\text{K}{}^{\mathrm{+}}\text{+}\text{Na}{}^{\mathrm{+}}\text{+}\text{ATP}$, $\text{Na}{}^{\mathrm{+}}\text{+}\text{ATP}$, $\text{Na}{}^{\mathrm{+}}\text{+}\text{ATP}$, $\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{+}\text{ATP}$ and ATP. The highest rate was obtained in the presence of Na⁺, Mg²⁺ and ATP. The apparent concentrations of Na⁺, Mg²⁺ and ATP for half-maximum stimulation of inhibition ($\text{K}{}_{0.5}{}^{\mathrm{<mtext>s</mtext>}}$) were 3 mM, 0.13 mM and 4μM, respectively. The rate was unchanged upon further increase in Na⁺ concentration from 140 to 1000 mM. The rates of inhibition could be explained on the basis of the enzyme forms present, including E₁, E₂, ES, E₁-P and E₂-P, i.e., E₂ has higher reactivity with showdomycin than E₁, while E₂-P has almost the same reactivity as E₁-P. We conclude that the reaction of ($\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ proceeds via at least four kinds of enzyme form (E₁, E₂, E₁ · nucleotide and EP), which all have different conformations.     

Characterization of partially purified (Na+ + K+)-ATPase from porcine lens

The partial purification of $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ from pig lens has been achieved by treatment with deoxycholate followed by density gradient centrifugation. The specific activity of the final preparation, ranging from 300 to 500 nmol/h per mg protein, is increased approx. 100-fold compared to the homogenate. A parallel increase in $\text{p-}\text{nitrophenylphosphatase}$ activity is also observed. Sodium dodecyl sulfate (SDS) gel electrophoresis reveals six major protein bands, one of which is the 93 kDa α subunit of $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ which can be phosphorylated by reaction with [γ-³²P]ATP. A second band contains a glycoprotein which displays an apparent molecular weight of 51 000 and thus appears to be the β subunit of the enzyme. The enzyme is sensitive to ouabain with the $\text{I}{}_{50}$ for $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ and $\text{p-}\text{nitrophenylphosphatase}$ inhibition being 1.2 and 1.3 μM, respectively. Several agents which inhibit $\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ from other tissues such as oligomycin, Ca²⁺, vanadate, $\text{N-}\text{ethylmaleimide}$, $\text{p-}\text{chloromercuribenzenesulfonic acid}$ (PCMBS) and 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB) also inhibit the lens enzyme. Monovalent cations other than K⁺ are partially effective in activating the ($\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}$)-ATPase and $\text{p-}\text{nitrophenylphosphatase}$ activities. The K⁺ congeners were relatively more effective in supporting $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ compared to $\text{p-}\text{nitrophenylphosphatase}$ activity. Other kinetic properties of the lens enzyme are also comparable to those of the enzyme from other tissues. Utilizing the partially purified membrane bound enzyme, discontinuities in Arrhenius plots of $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ activity, $\text{p-}\text{nitrophenylphosphatase}$ activity and fluoresence polarization of the fluidity probe, 1,6-diphenyl-1,3,5-hexatriene (DPH), are observed near the physiological temperature of lens. The possible significance of these observations for the mechanism of cataract formation are discussed.