高温处理防治黄瓜霜霉病的生理生化机制

以黄瓜感病品种‘长春密刺’为试材,通过室内盆栽试验研究高温对已感染霜霉病菌的黄瓜幼苗生理生化特征的影响.结果显示:(1)在黄瓜幼苗接种霜霉菌后8h,用45℃高温处理90min防治效果最明显.(2)与只接种霜霉病菌的黄瓜幼苗相比,接种霜霉菌后进行高温处理可显著提高叶片叶绿素含量,降低丙二醛含量;与对照相比,接种霜霉菌后进行高温处理可显著提高叶片几丁质酶活性,Western blotting验证了几丁质酶的活性变化.(3)SDS-PAGE结果表明,霜霉菌侵染可诱导一种28kD蛋白表达,高温处理后28kD蛋白的表达量降低.研究结果表明高温处理可能在杀死病原菌的同时,还诱导黄瓜幼苗对霜霉病产生了部分抗性.     

高温调控对黄瓜霜霉病菌侵染的影响

采取温湿度相结合的方法来研究高温处理对黄瓜霜霉病菌（Pseudoperonospora cubensis Rostov)侵染的影响,比较研究了35－50℃4个温度梯度、50％－90％5个湿度组合处理对病菌致病性的影响。同一相对湿度条件下,随着温度的上升病菌致病性降低;40－50℃的高温范围内,在同一温度下随着湿度的不断升高,受处理病菌的致病能力逐渐下降。在RH80％以上、温度40℃以上时,病菌的致病力随着处理时间的延长而变弱;45℃以下的高温高湿处理病菌超过1h,病菌基本上没有致病性。通过高温控制苗期黄瓜霜霉病的研究,确定高温高湿防治苗期黄瓜霜霉病的最佳温湿度为45℃1h(RH80%)。通过高温致死菌诱导植株抗性的研究,初步明确高温处理致死的病菌可以短期诱导植株的抗性。     

拟康氏木霉胞壁多糖对黄瓜抗病性的诱导作用

为了更全面地认识拟康氏木霉菌（Trichoderma pseudokoningii SDTP1）的抗病机理,为该菌的开发和科学的施用提供依据,通过灌根处理和人工接病的方式研究了拟康氏木霉菌胞壁多糖（TPWS）对黄瓜幼苗抗病性的诱导作用。结果表明用浓度为200mg／L的TPWS处理后对人工接种尖镰孢分生孢子悬液的黄瓜幼苗的保护作用达到53．8％,浓度进一步提高保护作用升高不明显;用200mg／L的TPWS处理后,黄瓜幼苗下胚轴内苯丙氨酸解氨酶（PAL）和多酚氧化酶（POD）的活性及木质素的含量分别于6、30、30h后开始迅速升高,最高值时分别是对照的2．2、3．1、4．4倍;用200mg／L的TPWS处理后需要4d黄瓜幼苗才能表现出抗病性,不仅降低了伤根接种尖孢镰刀菌分生孢子悬液幼苗的枯萎病发病率,也能降低在茎部和叶面穿刺接种灰葡萄孢分生孢子悬液幼苗的灰霉病发病率。而TPWS在体外对两种病原菌无抑制作用,我们认为经处理的幼苗枯萎病和灰霉病发病率的降低与TPWS诱导了幼苗的抗病性有关,且这种抗性是系统的、非特异性的。抗病性可能与木质素含量的升高有关。     

草酸诱导黄瓜幼苗对霜霉病的抗性与H2O2的关系

以长春密刺黄瓜幼苗为材料,对经草酸处理或霜霉菌接种后黄瓜叶片的过氧化物酶(POD)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性及H2O2含量的变化进行了研究.结果表明:草酸处理或霜霉菌接种均可诱导黄瓜幼苗叶片H2O2含量显著增加,且草酸预处理后接种的叶片比相应对照叶片能更快地积累H2O2;草酸处理后叶片SOD和POD活性均升高,而CAT活性却受到一定程度的抑制.研究发现,H2O2参与了幼苗对霜霉病的抗性诱导;叶片H2O2含量的增加与其SOD、POD活性升高、CAT活性下降有关;通过调节黄瓜叶片H2O2的含量来调控有关黄瓜霜霉病抗性的防御基因表达是草酸诱导抗性的机制之一.     

生防细菌对水稻的促生性及诱导抗性研究

研究了4株生防菌对水稻白叶枯病的抑制作用和菌悬液浸种、浸芽、浇苗和包衣4种处理对水稻生长的促进作用,及对水稻体内过氧化物酶POD、多酚氧化酶PPO、苯丙氨酸解氨酶PAL 3种保护酶的诱导表达作用。结果表明,4个菌株均对水稻幼苗有促生及诱导抗病性的作用。其中,WY2菌株诱导水稻抗病性和对水稻的促生性都要优于其他3株菌株。水稻幼苗接种生防细菌24 h后再接种病原菌,生防细菌能促进植物体内保护酶PAL、POD、PPO活性的提高,进而诱导植物抗病性的提高。     

亚精胺诱导黄瓜幼苗对白粉病抗性的研究

以黄瓜品种‘长春密刺’幼苗为材料,研究了亚精氨(Spd)诱导黄瓜幼苗对白粉病的抗性,并测定Spd处理和白粉菌接种对黄瓜叶片4种防御酶活性及3种防卫基因表达的影响。结果显示:(1)0.2～1.0mmol.L-1 Spd对黄瓜幼苗白粉病抗性均有不同程度的诱抗效果,并以0.8mmol.L-1 Spd处理效果最明显,诱导效率可达55.3%。(2)喷施Spd或接种白粉菌均可提高黄瓜叶片过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)、几丁质酶和β-1,3-葡聚糖酶的活性,且诱导并接种处理的植株叶片上述酶活性均比只诱导不接种处理的上升速度更快;同时,Spd处理和接种白粉菌可以提高植株叶片中POX、PAL、PR-1a基因的表达量。研究表明,Spd处理可以诱导防卫基因表达的增强,提高防御酶活性,显著降低病情指数,增强黄瓜幼苗对白粉病的抗性。     

纯氧对采后杨梅果实腐烂的抑制与抗病相关酶的诱导

为研究高氧对抑制果实腐烂的作用及其与抗病相关酶活性诱导的关系,将杨梅果实采后在5℃用纯氧或空气(对照)处理12 d.结果表明,纯氧处理可显著抑制果实腐烂发生,贮藏12 d后对照果实的腐烂指数达到54%,而处理果实仅为17%.纯氧处理在贮藏前期可诱导杨梅果实几丁质酶和β-1,3-葡聚糖酶活性的升高,并在第6天时达到高峰.另外,纯氧处理增加了苯丙氨酸解氨酶和过氧化物酶的活性及总酚含量,并在整个贮藏期间一直高于对照水平.这些结果表明,高氧抑制杨梅果实腐烂的作用与诱导与抗病相关的酶的活性升高密切相关,抗病性诱导是高氧抑制杨梅果实腐烂的重要原因.     

水杨酸和霜霉病诱导黄瓜PCD过程中线粒体相关基因的表达分析

为研究水杨酸(SA)和霜霉病两种处理诱导黄瓜PCD过程中线粒体相关基因的表达差异及其引发的黄瓜叶片PCD发生方式。选择长出4片真叶的黄瓜植株为试材,用10 mmol/L SA处理叶片,用5×105-10×105个孢子囊/m L的霜霉菌菌液接种叶片,分别取SA处理0、3、12和24 h及接种菌液0、36、72和96 h的叶片提取RNA,用q RT-PCR技术分析16个线粒体相关基因的表达动态。实验结果表明:16个基因在SA处理和霜霉病接种后都表达,呈现相似或相悖的表达趋势,说明SA诱导的PCD相关基因与霜霉病诱导的PCD信号通路有明显的相互联系,其中11个基因在霜霉病处理中结果与SA处理相似皆为上调,5个基因在SA处理中为上调表达,在霜霉病处理过程中却是下调表达,这也表明SA和霜霉病引发抗病信号途径存在差异,从而使黄瓜叶片PCD发生方式不同。通过查找文献分析16个基因的功能及研究现状,这些基因属于4个功能类群,主要包括参与线粒体膜转运、细胞呼吸作用、蛋白质合成、折叠和转运、线粒体信号转导以及其他或未知功能。     

黄瓜霜霉病菌侵染若干因子的研究

研究了温、湿度条件对黄瓜霜霉病菌致病性的影响,结果表明,25～35 ℃最适宜黄瓜霜霉病的发生,15／35 ℃的交替温度变化最有利于霜霉病菌的侵染,但35 ℃以上的高温对霜霉病菌具有杀伤作用;2 h的湿度条件就足以引起侵染,一旦侵入寄主,环境湿度条件对病害的发展影响不大.-20 ℃低温冷冻保存10个月和干燥放置7 d的霜霉菌种仍具致病力.发病的黄瓜叶片可以连续产生孢子囊,但随着发病时间的延长产生孢子囊量逐渐减少.活体叶片单位面积上产生的孢子囊量比离体叶片大,且显症天数与叶片单位面积产生孢囊量呈抛物线型关系.     

NO和钙信使系统在草酸诱导黄瓜叶片抗霜霉病中的作用

通过草酸及其与不同抑制剂亚甲基蓝、EGTA、氯丙嗪和Li+组合处理黄瓜叶片,研究了草酸与抑制剂不同处理组合方式对黄瓜叶片POD活性和叶片病情指数的影响,探讨NO、钙信使系统在草酸诱导叶片抗霜霉病中的作用.结果显示,10～70mmol/L草酸均能不同程度诱导黄瓜叶片POD活性的升高,提高叶片对黄瓜霜霉病的抗病性,降低叶片病情指数,并以30mmol/L效果最好.4种抑制剂分别与30mmol/L草酸同时或先于草酸处理,或草酸处理后一定时间再用抑制剂处理,均明显抑制黄瓜叶片POD活性的升高及病情指数的降低.研究表明,NO、Ca2+、钙调素(CaM)和磷酸肌醇均可能参与了草酸诱导黄瓜霜霉病抗性的信号转导过程.     

Development of formulations of biological agents for management of root rot of lettuce and cucumber

The effect of various carrier formulations of Bacillus subtilis and Pseudomonas putida were tested on germination, growth, and yield of lettuce and cucumber crops in the presence of Pythium aphanidermatum and Fusarium oxysporum f.sp. cucurbitacearum, respectively. Survival of B. subtilis and P. putida in various carriers under refrigeration (about 0 degree C) and at room temperature (about 22 degrees C) was also studied. In all carrier formulations, B. subtilis strain BACT-0 survived up to 45 days. After 45 days of storage at room temperature (about 22 degrees C), populations B. subtilis strain BACT-0 were significantly higher in vermiculite, kaolin, and bacterial broth carriers compared with other carriers. Populations of P. putida were significantly higher in vermiculite, peat moss, wheat bran, and bacterial broth than in other carriers when stored either under refrigeration (about 0 degree C) or at room temperature (about 22 degrees C) for 15 or 45 days. Germination of lettuce seed was not affected in vermiculite, talc, kaolin, and peat moss carriers, but germination was significantly reduced in alginate and bacterial broth carriers of B. subtilis compared to the non-treated control. Germination of cucumber seed was not affected by any of the carriers. Significantly higher fresh lettuce and root weights were observed in vermiculite and kaolin carriers of B. subtilis compared with P. aphanidermatum-inoculated control plants. Lettuce treated with vermiculite, and kaolin carriers of B. subtilis, or non-inoculated control lettuce plants had significantly lower root rot ratings than talc, peat moss, bacterial broth, and P. aphanidermatum-inoculated control plants. Growth and yield of cucumber plants were significantly higher in vermiculite-based carrier of P. putida than the other carriers and Fusarium oxysporum f.sp. cucurbitacearum-inoculated plants.     

Effect of temperature on the efficiency of the thermo- and mesophilic aerobic batch biodegradation of high-strength distillery wastewater (potato stillage)

The objective of the study was to assess the effect of temperature on the extent of aerobic batch biodegradation of potato stillage with a mixed culture of bacteria of the genus Bacillus. The experiments were performed in a 5-l stirred-tank reactor at 20, 30, 35, 40, 45, 50, 55, 60, 63 and 65 degrees C with the pH of 7. Only at 65 degrees C, no reduction in chemical oxygen demand (COD) was found to occur. Over the temperature range of 20-63 degrees C, the removal efficiency was very high (with an extent of COD reduction following solids separation that varied between 77.57% and 89.14% after 125 h). The process ran at the fastest rate when the temperature ranged from 30 to 45 degrees C; after 43 h at the latest, COD removal amounted to 90% of the final removal efficiency value obtained for the process. At 20, 55, 60 and 63 degrees C, a 90% removal was attained after 80 h. Two criteria were proposed for the identification of the point in time when the process is to terminate. One of these consists in maximising the product of the extent of COD reduction and the extent of N-NH4 content reduction. The other criterion is a simplified one and involves the search for the minimal value of N-NH4 concentration.     

黄瓜叶片胞间隙几丁质酶与抗霜霉菌侵染关系的研究

采用高抗霜霉病的‘津春4号’和易感霜霉病的‘长春密刺’黄瓜的温室穴盘幼苗为材料,研究了接种霜霉菌对黄瓜叶片胞间隙几丁质酶累积变化及幼苗生长的影响.结果显示:(1)与易感品种相比,抗病品种叶片胞间隙液蛋白浓度升高快、含量高;同时几丁质酶活性上升的速度快、幅度大,于接种后第2天达到峰值,且高活性维持至第7天;(2) SDS-PAGE电泳分析发现,抗病品种有一种27 kD的酸性几丁质酶在接种后第2天迅速积累,并在接种后第4、7天呈现出较高的表达量;Western blotting的结果也证实了上述胞间隙几丁质酶积累的变化.(3)与未接种对照相比,接种处理后第4、7天,易感黄瓜幼苗鲜重、干物质积累量均出现显著降低,但在接种后第7天抗病品种的上述生长指标要显著高于易感病品种.研究表明,接种霜霉菌后,抗病黄瓜叶片胞间隙几丁质酶迅速累积且活性快速升高,以减轻霜霉病对幼苗生长的侵害,这可能是黄瓜抗霜霉菌侵染的一种防御机制.     

Thermal adaptation in CHO cells at 40 degrees C: the influence of growth conditions and the role of heat shock proteins

The kinetics of thermal adaptation at the nonlethal temperature of 40 degrees C was studied in CHO (Chinese hamster ovary) cells in vitro. Thermal resistance, demonstrated as an increase in mean 45 degrees C killing time or as an increase in the shoulder of the 45 degrees C survival curve, was fully developed by 2 h. Control cells in early logarithmic phase were more heat sensitive than those in stationary phase. Corresponding 45 degrees C killing time frequency distributions were unimodal with an increase in mean killing time from early logarithmic to stationary phase. Cells which were thermally adapted at 40 degrees C for 6 h had biphasic 45 degrees C killing time frequency distributions, and as cells progressed from early logarithmic to stationary phase the heat-sensitive subpopulation progressively declined. Exposure to 40 degrees C produced a 30% increase in total protein synthesis. Proteins with molecular weights 72, 89, and 109 kDa which correspond to those induced by lethal heat shock were synthesized at 40 degrees C, but there was no close temporal correlation between the development of heat resistance at 40 degrees C and synthesis of the heat shock proteins. Cycloheximide (100 micrograms/ml) reduced the mean 45 degrees C killing time but did not totally prevent the development of heat resistance at 40 degrees C.     

黄瓜对霜霉病抗性的显微和超微结构的研究

利用显微及透射和扫描电镜方法,研究了黄瓜对霜霉菌[Pseudoperonosporacubensis(Berk.et Curt.)Rostow.]的抗性机理,结果如下:抗病与感病品种叶表面的气孔密度和大小无明显区别,而病菌分生孢子在叶表面萌发情况有差异。抗病品种被病菌侵染后,细胞迅速颗粒化,与病菌一起死亡,两个侵染点之间的叶肉细胞大量繁殖,细胞中叶绿体减少,膜系统受到破坏,壁增厚,叶表面出现少量很小的病斑;中抗品种表现为少量菌丝蔓延,并产生分生孢子囊梗和孢子,受侵细胞的线粒体和叶绿体膜系统被破坏,随后内细胞和菌丝死亡,但比抗病品种慢,在叶表面形成很多病斑;感病品种表现为叶肉细胞内有大量菌丝蔓延,并产生分生孢子囊梗和孢子,受侵细胞被破坏、解体成碎片,最后与菌丝一起死亡,在叶表面联成大量的病斑。     

Thermoregulatory physiology of the Crested Pigeon<Emphasis Type="Italic"> Ocyphaps lophotes</Emphasis> and the Brush Bronzewing<Emphasis Type="Italic"> Phaps elegans</Emphasis>

The metabolic physiology of the Crested Pigeon (Ocyphaps lophotes) and the Brush Bronzewing (Phaps elegans) is generally similar to that expected for birds of their size, but the Crested Pigeon has a number of characteristics which would aid survival in hot and dry regions. Body temperature increased similarly for the Crested Pigeon (from 38.8 degrees C to 41.5 degrees C) and the Brush Bronzewing (39.3 degrees C to 41.4 degrees C) over ambient temperatures (T(a)s) from 10 degrees C to 35 degrees C. Both species became hyperthermic (body temperature, T(b)>42 degrees C) at T(a)=45 degrees C. Basal metabolic rate of the Crested Pigeon (0.65 ml O(2) g(-1) h(-1) at 40 degrees C) was approximately 71% of that predicted for a columbid bird, while BMR of the Brush Bronzewing (0.87 ml O(2) g(-1) h(-1) at 20 degrees C to 40 degrees C) was approximately 102% of predicted. Total evaporative water loss increased exponentially with T(a) for both species, from <1 mg H(2)O g(-1) h(-1) at 10 degrees C to >12 mg H(2)O g(-1) h(-1) at 45 degrees C. It was similar and low for both species at T(a)<30 degrees C, but was higher for the Brush Bronzewing than the Crested Pigeon at T(a)>30 degrees C. Ventilatory minute volume matched oxygen consumption, such that oxygen extraction efficiency did not change with T(a) and was similar for both species (approximately 20%). Expired air temperature was considerably lower than T(b) for both species at T(a)<35 degrees C, potentially reducing respiratory water loss by approximately 65% at T(a)=10 degrees C to approximately 30% at T(a)=35 degrees C. Cutaneous evaporative cooling was significant for both species, with skin resistance decreasing as T(a) increased. The Crested Pigeon had a lower skin resistance than the Brush Bronzewing at T(a)=45 degrees C. The Brush Bronzewing had apparently reached its maximum cutaneous water loss at 30 degrees C and relied on panting to cool at higher T(a).     

Acute heat stress protects rats against endotoxin shock.

The purpose of this study was to determine 1) whether prior (24-h) heat stress could render rats cross-resistant to the lethal activity of bacterial lipopolysaccharide (LPS) and 2) whether this acquired state of resistance is associated with endotoxemia during the heat stress event. Four groups (n = 7/group) of rats were examined: 1) saline treated, 2) LPS treated, 3) heat stressed and saline treated, and 4) heat stressed and LPS treated. Saline or LPS (Escherichia coli, serotype 0111:B4, 20 mg/kg body wt) was given intravenously 24 h after exposure to heat (ambient temperature 47-50 degrees C, relative humidity 30%) for heat-stressed rats and at the same time of day for nonheated rats; survival was monitored for 48 h. Thermal responses were similar (P > 0.05); values for maximum core temperature (Tc) and time above Tc of 40 degrees C were 42.7 +/- 0.1 and 42.6 +/- 0.1 degrees C (SE) and 44.0 +/- 2.1 and 47.9 +/- 3.7 (SE) min for the heat-stressed saline-treated and heat-stressed LPS-treated rats, respectively. Administration of LPS to nonheated rats resulted in 71.4% (5 of 7 rats) lethality. In contrast, all (7 of 7) rats subjected to a single nonlethal heat stress event 24 h before LPS treatment survived (P < 0.05). Endotoxin was not detected in arterial plasma immediately after heat stress in rats (n = 6) exposed to a Tc of 42.9 +/- 0.1 degrees C. These findings demonstrate that acute heat stress can protect rats from the lethal activity of LPS.     

Storage effects on genomic DNA in rolled and mature coca leaves

Rolled and mature leaf tissue was harvested from Erythroxylum coca var. coca Lam. (coca) to determine a method for storage that would maintain DNA with high quality and content up to 50 days. Harvesting coca leaf tissue under Andean field conditions often requires storage from 3 to 10 days before extraction where tissue integrity is lost. All samples of rolled and mature coca leaf tissue were harvested and separately stored fresh in RNAlater for 50 days at 4 degrees, -20 degrees, and 23 degrees C, while similar samples were air-dried for 72 h at 23 degrees C or oven-dried for 72 h at 40 degrees C after storage, before extraction. Triplicate samples of each tissue type were extracted for DNA at 10-day intervals and showed that DNA integrity and content were preserved in leaf tissue stored at 4 degrees and -20 degrees C for 50 days. Rolled and mature leaf tissue stored at 4 degrees, -20 degrees, and 23 degrees C showed insignificant degradation of DNA after 10 days, and by day 50, only leaf tissue stored at 4 degrees and -20 degrees C had not significantly degraded. All air- and oven-dried leaf tissue extracts showed degradation upon drying (day 0) and continuous degradation up to day 50, despite storage conditions. Amplified fragment length polymorphism analysis of DNA from rolled and mature leaf tissue of coca stored at 4 degrees and -20 degrees C for 0, 10, and 50 days showed that DNA integrity and content were preserved. We recommend that freshly harvested rolled or mature coca leaf tissue be stored at 4 degrees, -20 degrees, and 23 degrees C for 10 days after harvest, and if a longer storage is required, then store at 4 degrees or -20 degrees C.