黄土丘陵区退耕草地群落优势种叶片光合生理对氮磷添加的响应

以黄土丘陵区退耕草地群落典型优势种白羊草、长芒草和达乌里胡枝子为研究对象,测定不同氮磷（主区N：0,50,100 kg N hm^-2 a^-1;副区P：0,40,80 kg P₂O₅ hm^-2 a^-1）添加处理下叶片光合气体交换参数、叶绿素荧光参数、叶片氮磷含量、光合氮利用效率（PNUE）和光合磷利用效率（PPUE）,探究不同优势种光合生理特征对外源氮磷添加的响应特征与物种差异。结果表明：单独氮添加下,3个优势种的净光合速率（P_n）、蒸腾速率、水分利用效率（WUE_i）、气孔限制值（Ls）和光系统Ⅱ（PSⅡ）活性高于或显著高于未施肥处理,白羊草和达乌里胡枝子的PNUE和PPUE以及长芒草的PPUE显著增加。单独磷添加下,仅达乌里胡枝子的P_n、WUE_i、Ls和PNUE相比未施肥处理显著增加。氮磷共同添加下,50 kg N hm^-2 a^-1处理下,施磷后达乌里胡枝子的P_n以及三个优势种的PNUE显著增加,白羊草的PPUE显著降低;100 kg N hm^-2 a^-1处理下,施磷后达乌里胡枝子的P_n和PNUE均显著增加,白羊草PPUE和长芒草的PNUE的仅在40 kg P₂O₅ hm^-2 a^-1处理下显著增加。3个优势种的P_n均与PSⅡ最大光化学效率和叶片氮含量呈显著正相关关系,长芒草和达乌里胡枝子的P_n与PSⅡ潜在活性和叶片磷含量呈显著正相关关系。总体表明,氮磷对黄土丘陵区退耕草地优势种光合生理具有一定的交互作用,物种属性和施肥水平影响其光合生理特征响应程度。     

镍离子对尖叶莴苣氮素吸收和生长生理的影响

以‘全年油麦菜’尖叶莴苣为试验材料,采用水培方式,研究3个浓度(0 mg·L^-1、0.1 mg·L^-1、1 mg·L^-1)Ni²⁺在22.4 mg·L^-1 N处理下对尖叶莴苣氮素吸收的生长及生理影响。结果显示：(1)尖叶莴苣根系和地上部生物量随处理时间的增加呈上升趋势。与对照T₁(0 mg·L^-1 Ni²⁺、112 mg·L^-1 N)相比,T₂处理(0 mg·L^-1 Ni²⁺、22.4 mg·L^-1 N)对尖叶莴苣根系及叶片生长具有一定抑制作用,植株鲜重、干重、根冠比、根系长度、平均直径、表面积、体积、根尖数、分根数、叶片表面积和体积在T3处理(0.1 mg·L^-1 Ni²⁺、22.4 mg·L^-1 N)下显著高于对照,T₄处理(1 mg·L^-1 Ni²⁺、22.4 mg·L^-1 N)对尖叶莴苣根系及其叶片生长具有一定促进作用,但对其根尖数和分根数表现出一定抑制性。(2)随着Ni²⁺浓度的增加,尖叶莴苣叶片叶绿素a、叶绿素b和总叶绿素含量呈先升后降的变化规律,且均在T₃处理下显著提高。(3)随着处理时间的增加,尖叶莴苣叶片的净光合速率(P_n)、气孔导度(G_s)和蒸腾速率(T_r)逐渐上升,胞间CO₂浓度(C_i)逐渐下降,且T₃处理叶片的G_s显著高于对照,其C_i最低,P_n最大。(4)施加Ni²⁺对尖叶莴苣有机酸、可溶性蛋白和可溶性糖含量以及SOD和POD活性有显著影响,在T₃处理下有机酸含量降低,可溶性糖和可溶性蛋白含量显著增加,SOD和POD活性显著提高。(5)T₃处理尖叶莴苣根系中N及叶片中B和Ca含量较高;根系中Ni含量高于叶片,T₃处理叶片中的Ni含量较低,Mg含量较高;植株体内Cu含量随Ni²⁺浓度增加而下降。研究表明,外源Ni²⁺处理能影响低氮条件下(22.4 mg·L^-1 N)尖叶莴苣幼苗生长及生理状况,适宜浓度(0.1 mg·L^-1)Ni²⁺可有效提高尖叶莴苣根系对氮素的吸收利用效率,减少氮素施用量,促进尖叶莴苣根系和地上部叶片生长,增加光合色素含量,并提高净光合速率,进而改善植株的产量和营养品质。     

温度和水分变化对冻土区泥炭地土壤氮循环功能基因丰度的影响

以大兴安岭多年冻土区泥炭地为研究对象,通过室内模拟增温实验,研究温度升高对不同深度（0-150 cm）土壤氮循环功能基因丰度的影响。同时针对0-20 cm和20-40 cm土壤设置两个水分处理,分别为土壤原始含水量和淹水状态,研究水分变化对表层土壤氮循环功能基因丰度的影响。结果表明温度升高显著提高了活动层（0-60 cm）、过渡层（60-80 cm）、永冻层（80-100 cm）中nifH、nirK基因丰度,温度升高显著提高了活动层（0-40 cm）和过渡层（60-80 cm）中nirS基因丰度。温度升高显著提高了过渡层（60-80 cm）NH₄⁺-N和较深永冻层（140-150 cm）NO₃^--N的含量,但降低了过渡层（60-80 cm）NO₃^--N和较深永冻层（120-150 cm）NH₄⁺-N的含量,相关性分析表明,NH₄⁺-N含量与nifH和nirS基因丰度呈显著正相关,NO₃^--N含量与nirK基因丰度呈显著正相关,说明温度升高能够通过改变微生物丰度促进过渡层固氮作用和反硝化作用。在增温条件下,淹水处理使表层土壤nirS和nirK基因丰度及NH₄⁺-N含量降低,但提高了NO₃^--N含量,说明淹水造成了过度还原的条件使反硝化底物浓度降低,降低反硝化微生物活性进而抑制了土壤反硝化作用。该结果对于明确未来气候变化影响下冻土区泥炭地土壤氮循环过程具有重要意义。     

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。     

生物炭对褐土旱地玉米季氮转化功能基因、丛枝菌根真菌及N2O释放的影响

以豫西旱地玉米农田为研究对象,设置不同生物炭施用量处理（T0：不施用生物炭;T1：施用生物炭20 t/hm²;T2：施用生物炭40 t/hm²）,采用密闭式静态箱法测定N₂O排放通量和荧光定量PCR法分析丛枝菌根（arbuscular mycorrhizal,AM）真菌、氨单加氧酶（amoA）、亚硝酸盐还原酶（nirS、nirK）以及氧化亚氮还原酶（nosZ）的基因丰度,同时测定土壤理化性状的变化。研究结果表明,随着生物炭施用量的增加,土壤pH和含水量呈增加趋势,土壤有机碳、全氮和铵态氮含量显著提高,土壤容重和硝态氮含量显著降低。T1和T2处理土壤有机碳含量分别较T0显著提高38.44%和71.01%;T1和T2处理土壤铵态氮含量分别较T0显著增加15.89%和30.46%;T2处理土壤全氮含量较T0处理显著提高14.87%;T1和T2处理土壤硝态氮含量分别较T0减少10.57%和21.40%。随着生物炭施用量的增加,AM真菌侵染率显著增加,T1和T2处理分别较T0处理提高71.88%和115.88%;AOA、AOB、nirK和nirS基因丰度显著降低;nosZ基因丰度增加。施加生物炭处理的N₂O排放通量和累积排放量均低于不施生物炭处理,具体表现为：T0 > T1 > T2。相关分析表明,生物炭施用量与AM真菌基因丰度呈显著正相关;与nosZ基因丰度呈正相关;与AOA、AOB、nirK、nirS基因丰度呈极显著负相关。N₂O排放通量与AOA、nirK、nirS基因丰度呈极显著正相关;与土壤含水量和土壤硝态氮含量呈显著正相关;与AM真菌、nosZ基因丰度、易提取球囊霉素含量、铵态氮含量呈极显著负相关。集成推进树（ABT）分析表明,AOA对N₂O排放的影响最大,其次是AM真菌和nirS。总之,生物炭处理改善土壤理化性质、提高土壤AM真菌侵染率、调节硝化、反硝化相关功能基因的丰度,减少N₂O气体排放,为旱地农田合理施用生物炭减少N₂O气体排放提供理论依据。     

盐分胁迫下杠柳叶片光合特性的光响应研究

利用CID型便携式光合作用仪测定不同NaCl浓度下杠柳叶片净光合速率(P_n)、蒸腾速率(T_r)、气孔导度(G_s)、水分利用效率（WUE）及光能利用率(LUE)生理参数的光响应过程,阐明盐分胁迫下其对光照响应的规律,探讨有利于杠柳正常生长的盐分浓度和光照条件。结果表明:（1）各盐分浓度下杠柳叶片光补偿点（LCP）在21.89～65.05 μmol·m^-2·s^-1之间变动,介于阴性植物与阳性植物之间;杠柳随土壤盐分的不同,其光合作用参数对光照强度表现出一定的适应性和可塑性;50 mmol·L^-1盐分浓度下杠柳光合同化能力最强,最有利于其干物质的积累,表现出一定的耐盐性。（2）轻度的盐分胁迫（小于50 mmol·L^-1）可以提高杠柳叶片的P_n、G_s、WUE和LUE,而盐分胁迫对杠柳的T_r有抑制作用,并随着盐分浓度的增加其抑制作用愈强烈。（3）维持杠柳正常生长的土壤盐分浓度小于50 mmol·L^-1,最佳PAR为1 000～2 000 μmol·m^-2·s^-1;而保持杠柳最大WUE和LUE的光照强度分别为800和100 μmol·m^-2·s^-1。     

武夷山国家公园不同林地土壤呼吸动态变化及其影响因素

探明亚热带山岳型国家公园不同林地利用方式下土壤呼吸（R_s）的动态变化规律以及影响因素,对准确评价和预测该区域以国家公园为主体的自然保护地体系的碳收支具有重要的现实意义。以武夷山国家公园为研究对象,利用Li-8100开路式土壤碳通量测定系统对茶园、锥栗（Castanea henryi（Skam） Rehd.et Wils.）林、马尾松（Pinus massoniana Lamb.）林和裸地的土壤呼吸及近地面气温、土壤温度、土壤湿度、土壤养分和土壤微生物碳（MBC）、氮（MBN）进行测定。结果显示：（1）与近地面气温、土壤温度和土壤湿度相同,不同林地的R_s均呈现夏 > 春 > 秋 > 冬的季节动态,R_s的季节均值按大小排序为茶园（3.10 μmol m^-2 s^-1） > 马尾松（2.96 μmol m^-2 s^-1） > 锥栗（2.32 μmol m^-2 s^-1） > 裸地（1.43 μmol m^-2 s^-1）,锥栗和裸地之间、锥栗与马尾松之间均差异显著（P<0.01）。除马尾松林外,其他林地水热因子（近地面气温、土壤温度和土壤湿度）的单因子二次多项式模型对R_s的拟合度最高。水热因子共同建立的复合模型中,土壤温度、湿度的幂-指数模型对茶园R_s的拟合度较高,土壤温度和土壤湿度能够解释R_s变化的80%,马尾松林的R_s较适用于土壤温度、湿度建立的对数函数模型,而三因子线性模型（进入回归法）对锥栗林和裸地的R_s的拟合度最优,R²分别为0.565和0.281。（2）茶园和锥栗林的碳、氮、磷含量均高于马尾松林和裸地,MBN含量茶园 > 马尾松 > 锥栗 > 裸地。茶园的R_s与全磷（TP）、有效磷（AP）、全钾（TK）、速效钾（AK）含量呈极显著（P<0.01）正相关,马尾松林的R_s受TP、TK、AK含量的影响极显著（P<0.01）,锥栗林的R_s与TK、AK、MBN含量呈现显著（P<0.05）正相关,裸地的R_s受MBN含量影响较为显著（P<0.05）,4种林地土壤呼吸与养分的多元逐步回归方程R²均接近1。综上,茶园和马尾松林土壤呼吸速率较高,且所有林地的土壤呼吸均呈现夏 > 春 > 秋 > 冬的季节动态。温度和湿度与土壤呼吸的相关性强,是水热条件丰富的亚热带山岳地区土壤呼吸季节变化的主导因素,其中武夷山茶园土壤呼吸对水热因子的响应在4种林地中最为敏感。除温度和湿度外,各林地土壤呼吸受P、K元素的影响较大,其中茶园主要受P元素影响,马尾松林地受K元素影响较多。     

超氧化物歧化酶模拟化合物的生物活性研究

根据天然超氧化物歧化酶(SOD)活性部位结构合成了含苯并咪唑的5种配体及其32种分别含Cu（Ⅱ）、Fe（Ⅲ）、Mn（Ⅱ）、Co（Ⅱ）的模拟化合物.经光谱、电化学测试证明这些化合物具有拟S3D活性,其50％抑制率浓度（IC50）为10-6～10-8mol·L-1,催化超氧离子自由基（O-·2）歧化的反应速率常数kq为106～108mol-1·L·s-1.同时观察到几种模拟化合物有抗肿瘤活性或增强水稻抗寒性.     

广东省豆科植物结瘤固氮及根瘤菌资源的初步研究

于2000～2001年在广东省境内54个县（市、区）进行了豆科植物结瘤固氮资源的调查及根瘤菌的采集工作。共采集到豆科植物根瘤样品484份,隶属于37属78种;根瘤的形状以圆形、椭圆形、珊瑚状和姜状居多,大小一般在1～10 mm之间,颜色多为淡红色或黄色。用乙炔还原法对24种93份根瘤样品进行了固氮酶活性测定,结果表明,大多数样品的固氮酶活性在1～10 μmol C₂H₄·g^-1 fresh nodule·h^-1之间。从采集到的根瘤中分离纯化出410株根瘤菌,对其中312株进行了回接试验,回接成功率为93.3 %。调查发现广东省现有栽培豆科作物种类与《广州植物志》的记载相似,所以本研究在豆科栽培作物方面有一定的代表性。在调查采集到根瘤的豆科植物中,三尖叶猪屎豆（Crotalaria micans）、毛排钱草（Phyllodium elegans）、细长柄山蚂蝗（Podorcarpium leptopus）等在本研究之前未见到有结瘤固氮的报道。     

鄱阳湖沉水植物区不同深度土壤甲烷排放对温度水分变化的响应

采集鄱阳湖沉水植物区0-10 cm和10-30 cm土壤样品,通过设置2个温度（18℃和28℃）和2个水分（淹水2 cm和土柱取出水面后的实际土壤水分含量）处理组合,进行持续2年的甲烷（CH₄）排放室内培养实验,以探讨不同深度土壤CH₄排放对温度、水分变化的响应差异,以及温度、水分和土层对湿地土壤CH₄排放的交互影响。结果表明：0-10 cm和10-30 cm土壤CH₄排放速率变化范围分别为0.01-3.63 μgCH₄-C kg^-1d^-1、0.02-1.99 μgCH₄-C kg^-1d^-1;均值分别为0.72和0.15 μgCH₄-C kg^-1d^-1。温度、水分和土层3因素及其交互作用均对土壤CH₄排放有显著影响（P<0.01）,且土层的影响最大。两水分处理下的CH₄排放对温度变化的敏感性均表现为0-10 cm（Q₁₀为1.78、3.26）高于10-30 cm土层（Q₁₀为1.04、1.08）。CH₄平均排放速率及累计排放量均表现为0-10 cm显著高于10-30 cm土层,且培养前期高于培养后期,显示基质有效性对土壤CH₄排放的重要影响。     

Substrate specificities of a bacterial sialidase and rat liver ganglioside GM3 sialyltransferase

Sialidases cleave off sialic acid residues from the oligosaccharide chain of gangliosides in their catabolic pathway while sialyltransferases transfer sialic acid to the growing oligosaccharide moiety in ganglioside biosynthesis. Ganglioside G_M3 is a common substrate for both types of enzymes, for sialidase acting on ganglioside G_M3 as well as for ganglioside G_D3 synthase. Therefore, it is possible that both enzymes recognize similar structural features of the sialic acid moiety of their common substrate, ganglioside G_M3. Based on this idea we used a variety of G_M3 derivatives as glycolipid substrates for a bacterial sialidase (Clostridium perfringens) and for G_D3 synthase (of rat liver Golgi vesicles). This study revealed that those G_M3 derivatives that were poorly degraded by sialidase also were hardly recognized by sialyltransferase (G_D3 synthase). This may indicate similarities in the substrate binding sites of these enzymes.     

Inhibition of oxygen release by anoxia in a C3-plant (Nicotiana tabacum cv. Wisconsin 38). Comparison with C4-plants

Tobacco leaves (Nicotiana tabacum var. Wisconsin 38) submitted to anaerobic conditions behave in a manner similar to that of maize, sugarcane, or sorghum leaves (C₄-plants); more precisely, a lag time in O₂ release is exhibited when the leaves are exposed to light after treatment in the dark under pure nitrogen. Although the conditions for the appearance of this phenomenon in tobacco are somewhat different, the main features are identical to those observed with maize: abolition of the lag time upon immediate exposure to light, release of CO₂ under light (illumination burst of CO₂), photochemical nature of the reactions involved in the abolition of the lag time, activation of oxygen release by far-red light, and the antagonistic effect of red and far-red light on the lag time. The high CO₂ compensation point of tobacco leaves permits the classification of this plant among the C₃ group. A comparison of these experimental results with others from the literature suggests than the distinguishing features between C₃- and C₄-plants are not as sharp as generally thought.     

Regulation of Metabolic and Electron Transport Pathways in the Freshwater Bacterium Beggiatoa leptomitiformis D-402

The biomass yield of freshwater filamentous sulfur bacteria of the genus Beggiatoa, when grown lithoheterotrophically or mixotrophically, has been shown to increase 2 to 2.5 times under microaerobic conditions (0.12 mg/l oxygen) as compared to aerobic conditions (9 mg/l oxygen). The activity of the glyoxylate cycle key enzymes have been found to increase two to three times under microaerobic conditions (at an O₂ concentration of 2 mg/l), and the activities of the sulfur metabolism enzymes increased three to five times (at an O₂ concentration of 0.1–0.5 mg/l). It has also been found that, under microaerobic conditions, thiosulfate was almost completely oxidized to sulfate by the bacteria, without accumulation of intermediate metabolites. At the same time, a 2- to 15-fold decrease in the activities of the tricarboxylic acid cycle enzymes involved in the reduction of NAD and FAD was observed. Reorganization of the respiratory chain after changes in aeration and type of nutrition was also observed. It has been found that, in cells grown heterotrophically, the terminal part of the respiratory chain contained an aa ₃-type oxidase, whereas, during mixotrophic, lithoheterotrophic, and autotrophic growth, aa ₃-type oxidase synthesis was inhibited, and the synthesis of a cbb ₃-type oxidase, which is induced under microaerobic conditions, was activated. The gene of the catalytic subunit CcoN of the cbb ₃-type oxidase was sequenced and proved to be highly homologous to the corresponding genes of other proteobacteria.__________Translated from Mikrobiologiya, Vol. 74, No. 4, 2005, pp. 452–459.Original Russian Text Copyright © 2005 by Muntyan, Grabovich, Patritskaya, Dubinina.     

Determination of femtomol quantities of gibberellic acid by radioimmunoassay

A highly sensitive and specific radioimmunoassay which allows the detection of as little as 5 fmol (2 pg) of gibberellic acid (GA₃) in crude plant extracts is described. Antisera of high affinity and titer were obtained by immunizing rabbits with a conjugate of carboxyl-coupled GA₃ and bovine serum albumin. [¹²⁵I]Gibberellic acid-[N-(p-hydroxybenzyl) putrescine]amide of high specific activity, used as the immunotracer, is readily displaced by gibberellic acid methyl ester but not by free gibberellic acid. Thus, methylation of extracts prior to analysis is required. The assay is very specific; besides GA₃, only the closely related GA₇ is highly immunoreactive. Various gibberellins, related compounds, as well as other classes of plant hormones do not interfere with the assay. Levels of immunoreactive gibberellins (GA₃, GA₇) in actively growing tissues, among them cell suspension cultures of 33 different species, were determined.Abbreviations ABA abscisic acid - GC-MS gas chromatography-mass spectroscopy - TLC thin-layer chromatography - GA gibberellin Part 17 in the Series: Use of Immunoassay in Plant Science     

Incorporation of 14CO2 into C4 acids by leaves of C3-C4 intermediate and C3 species of Moricandia and Panicum at the CO2 compensation concentration

Comparative ¹⁴CO₂ pulse-¹²CO₂ chase studies performed at CO₂ compensation ()-versus air-concentrations of CO₂ demonstrated a four-to eightfold increase in assimilation of ¹⁴CO₂ into the C₄ acids malate and aspartate by leaves of the C₃-C₄ intermediate species Panicum milioides Nees ex Trin., P. decipiens Nees ex Trin., Moricandia arvensis (L.) DC., and M. spinosa Pomel at . Specifically, the distribution of ¹⁴C in malate and aspartate following a 10-s pulse with ¹⁴CO₂ increases from 2% to 17% (P. milioides) and 4% to 16% (M. arvensis) when leaves are illuminated at the CO₂ compensation concentration (20 l CO₂/l, 21% O₂) versus air (340 l CO₂/l, 21% O₂). Chasing recently incorporated ¹⁴C for up to 5 min with ¹²CO₂ failed to show any substantial turnover of label in the C₄ acids or in carbon-4 of malate. The C₄-acid labeling patterns of leaves of the closely related C₃ species, P. laxum Sw. and M. moricandioides (Boiss.) Heywood, were found to be relatively unresponsive to changes in pCO₂ from air to . These data demonstrate that the C₃-C₄ intermediate species of Panicum and Moricandia possess an inherently greater capacity for CO₂ assimilation via phosphoenolpyruvate (PEP) carboxylase (EC 4.1.1.31) at the CO₂ compensation concentration than closely related C₃ species. However, even at , CO₂ fixation by PEP carboxylase is minor compared to that via ribulosebisphosphate carboxylase (EC 4.1.1.39) and the C₃ cycle, and it is, therefore, unlikely to contribute in a major way to the mechanism(s) facilitating reduced photorespiration in the C₃-C₄ intermediate species of Panicum and Moricandia.Abbreviations Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase - PEP phosphoenolpyruvate - CO₂ compensation concentration - 3PGA 3-phosphoglycerate - SuP sugar monophosphates - SuP₂ sugar bisphosphates Published as Paper No. 8249, Journal Series, Nebraska Agricultural Research Division     

Co-function of C3-and C4-photosynthetic pathways in C3, C4 and C3-C4 intermediate Flaveria species

The potential for C₄ photosynthesis was investigated in five C₃-C₄ intermediate species, one C₃ species, and one C₄ species in the genus Flaveria, using ¹⁴CO₂ pulse-¹²CO₂ chase techniques and quantum-yield measurements. All five intermediate species were capable of incorporating ¹⁴CO₂ into the C₄ acids malate and aspartate, following an 8-s pulse. The proportion of ¹⁴C label in these C₄ products ranged from 50–55% to 20–26% in the C₃-C₄ intermediates F. floridana Johnston and F. linearis Lag. respectively. All of the intermediate species incorporated as much, or more, ¹⁴CO₂ into aspartate as into malate. Generally, about 5–15% of the initial label in these species appeared as other organic acids. There was variation in the capacity for C₄ photosynthesis among the intermediate species based on the apparent rate of conversion of ¹⁴C label from the C₄ cycle to the C₃ cycle. In intermediate species such as F. pubescens Rydb., F. ramosissima Klatt., and F. floridana we observed a substantial decrease in label of C₄-cycle products and an increase in percentage label in C₃-cycle products during chase periods with ¹²CO₂, although the rate of change was slower than in the C₄ species, F. palmeri. In these C₃-C₄ intermediates both sucrose and fumarate were predominant products after a 20-min chase period. In the C₃-C₄ intermediates, F. anomala Robinson and f. linearis we observed no significant decrease in the label of C₄-cycle products during a 3-min chase period and a slow turnover during a 20-min chase, indicating a lower level of functional integration between the C₄ and C₃ cycles in these species, relative to the other intermediates. Although F. cronquistii Powell was previously identified as a C₃ species, 7–18% of the initial label was in malate+aspartate. However, only 40–50% of this label was in the C-4 position, indicating C₄-acid formation as secondary products of photosynthesis in F. cronquistii. In 21% O₂, the absorbed quantum yields for CO₂ uptake (in mol CO₂·[mol quanta]^-1) averaged 0.053 in F. cronquistii (C₃), 0.051 in F. trinervia (Spreng.) Mohr (C₄), 0.052 in F. ramosissima (C₃-C₄), 0.051 in F. anomala (C₃-C₄), 0.050 in F. linearis (C₃-C₄), 0.046 in F. floridana (C₃-C₄), and 0.044 in F. pubescens (C₃-C₄). In 2% O₂ an enhancement of the quantum yield was observed in all of the C₃-C₄ intermediate species, ranging from 21% in F. ramosissima to 43% in F. pubescens. In all intermediates the quantum yields in 2% O₂ were intermediate in value to the C₃ and C₄ species, indicating a co-function of the C₃ and C₄ cycles in CO₂ assimilation. The low quantum-yield values for F. pubescens and F. floridana in 21% O₂ presumably reflect an ineffcient transfer of carbon from the C₄ to the C₃ cycle. The response of the quantum yield to four increasing O₂ concentrations (2–35%) showed lower levels of O₂ inhibition in the C₃-C₄ intermediate F. ramosissima, relative to the C₃ species. This indicates that the co-function of the C₃ and C₄ cycles in this intermediate species leads to an increased CO₂ concentration at the site of ribulose-1,5-bisphosphate carboxylase/oxygenase and a concomitant decrease in the competitive inhibition by O₂.Abbreviations PEP phosphoenolpyruvate - PGA 3-phosphoglycerate - RuBP ribulose-1,5-bisphosphate     

Genomics of the<Emphasis Type="Italic"> ccoNOQP</Emphasis>-encoded<Emphasis Type="Italic"> cbb</Emphasis><Subscript>3</Subscript> oxidase complex in bacteria

Many bacteria adapt to microoxic conditions by synthesizing a particular cytochrome c oxidase (cbb ₃) complex with a high affinity for O₂, encoded by the ccoNOQP operon. A survey of genome databases indicates that ccoNOQP sequences are widespread in all sub-branches of Proteobacteria but otherwise are found only in bacteria of the CFB group (Cytophaga, Flexibacter, Bacteroides). Our analysis of available genome sequences suggests four major strategies of regulating ccoNOQP expression in response to O₂. The most widespread strategy involves direct regulation by the O₂-responsive protein Fnr. The second strategy involves an O₂-insensitive paralogue of Fnr, FixK, whose expression is regulated by the O₂-responding FixLJ two-component system. A third strategy of mixed regulation operates in bacteria carrying both fnr and fixLJ-fixK genes. Another, not yet identified, strategy is likely to operate in the -Proteobacteria Helicobacter pylori and Campylobacter jejuni which lack fnr and fixLJ-fixK genes. The FixLJ strategy appears specific for the -subclass of Proteobacteria but is not restricted to rhizobia in which it was originally discovered.     

Susceptibility of Helicoverpa zea and Heliothis virescens (Lepidoptera: Noctuidae) to Vip3A insecticidal protein expressed in VipCot™ cotton

Susceptibility of laboratory and field colonies of Helicoverpa zea (Boddie) and Heliothis virescens F. to Vip3A insecticidal protein was studied in diet incorporation and diet overlay assays from 2004 to 2008. Responses of field populations were compared to paired responses of University of Arkansas laboratory susceptible H. zea (LabZA) and H. virescens (LabVR) colonies. After 7 d of exposure, observations were made on number of dead larvae (M) and the number of larvae alive but remaining as first instars (L1). Regression estimates using M (LC₅₀) and M plus L1 (MIC₅₀) data were developed for laboratory and field populations. Susceptibility of laboratory and field populations exposed to Vip3A varied among different batches of protein used over the study period. Within the same batch of Vip3A protein, susceptibilities of laboratory colonies of both species (LabZA and LabVR) were similar. Field colonies were significantly more susceptible to Vip3A than the respective reference colonies of both species. Within field populations, susceptibility to Vip3A varied up to 75-fold in H. zea and 132-fold in H. virescens in LC₅₀ estimates. Variabilities in MIC₅₀s were up to 59- and 11-fold for H. zea and H. virescens, respectively.     

Characterization of the oligosaccharide component of alpha3beta1 integrin from human bladder carcinoma cell line T24 and its role in adhesion and migration

Malignant transformation is highly associated with altered expression of cell surface N-linked oligosaccharides. These changes concern integrins, a family of cell surface glycoproteins involved in the attachment and migration of cells on various extracellular matrix proteins. The integrin alpha3beta1 is particularly interesting because of its role in migration and invasion of several types of metastatic tumours. In this study, alpha3beta1 from human bladder T24 carcinoma cells was purified and treated with peptide N-glycosidase F. Then the N-glycans of the alpha3 and beta1 subunits were characterized using matrix-assisted laser desorption ionization mass spectrometry (MALDI MS). In alpha3beta1 integrin the presence of high-mannose, hybrid and predominantly complex type N-oligosaccharides was shown. Unlike to normal epithelium cells, in both subunits of alpha3beta1 integrin from cancer cells, the sialylated tetraantennary complex type glycan Hex7HexNAc6FucSia4 was present. In a direct ligand binding assay, desialylated alpha3beta1 integrin exhibited significantly higher fibronectin-binding capability than untreated integrin, providing evidence that sialic acids play a direct role in ligand-receptor interaction. Moreover, alpha3beta1 integrin was shown to take part in T24 cell migration on fibronectin: anti-alpha3 antibodies induced ca 30% inhibition of wound closure. Treatment of T24 cells with swainsonine reduced the rate of bladder carcinoma cell migration by 16%, indicating the role of beta1,6 branched complex type glycans in this process. Our data show that alpha3beta1 integrin function may be altered by glycosylation, that both subunits contribute to these changes, and that glycosylation may be considered a newly found mechanism in the regulation of integrin function.