稻瘟菌细胞壁热解物诱导水稻抗稻瘟病的研究

以水稻近等基因系和广东省稻瘟病菌优势小种ZC13的细胞壁热解物(HDS)为材料进行诱导抗病性的研究。HDS通过细胞壁121°C 20 m in热解获得。HDS中葡萄糖质量浓度为176.83 m g/mL。HDS在体外对稻瘟病菌的孢子萌发和菌丝生长均无抑制作用,但处理水稻后可以显著提高水稻对稻瘟病菌的抗病性。在亲和性品系中,诱导抗病效果最高可达82.47%,且浓度与活性间存在正相关趋势;在非亲和品系中,热解物处理引起HR反应。HDS处理后,水稻过氧化物酶(POD)和苯丙氨酸解氨酶(PAL)活力明显升高。     

无机铜试剂对棉花苯丙氨酸解氨酶活性的影响

利用无机铜试剂（CIE）在新陆早17和新陆中26两棉花品种的3叶期和蕾期进行叶面喷施,检测棉株叶片内的苯丙氨酸解氨酶（PAL）的活性变化。结果表明,两品种在3叶期和蕾期均表现为PAL活性显著增高,第2天后即开始产生对PAL活性诱导作用,第10天后PAL的活性开始回落至对照组水平;两品种蕾期PAL的诱导活性均明显高于3叶期,且PAL高活性水平持续天数较3叶期长;棉花不同生育期使用CIE,均可增强棉叶PAL的活性;两品种在不同生育期的PAL活性出现峰值的时间、强度不一,显示出品种抗性的差异性。试验表明,CIE可诱导增强棉叶中PAL的活性。     

Response dynamics of three defense related enzymes in cotton leaves to the interactive stress of Helicoverpa armigera (Hübner) herbivory and omethoate application

In order to explore the response dynamics of the activities of defense related enzymes in cotton leaves towards the interactive stress of Helicoverpa armigera herbivory and omethoate application, the activities of phenylalanine ammonia-lyase(PAL), lipoxygenase(LOX), and polyphenol oxidase(PPO) were examined from 6 to 126 h after cotton leaves were treated 12 h of H. armigera herbivory, and then sprayed with 800 mg L–1 omethoate. The results showed that the changes in the activities of PAL, LOX and PPO that occured under the interactive stress of H. armigera herbivory and omethoate application reflected the interactive effects of the two stresses on cotton defense. The similarity between the response dynamics of PAL, LOX, and PPO activities in cotton leaves under the interactive stress and that under H. armigera herbivory treatment alone showed that the induction of H. armigera herbivory on the activities of PAL, LOX and PPO in cotton leaves played a leading role in the interactive effects, and the effect of omethoate application played only a minor role. A joint factor analysis was performed according to a method which has been used to analyze the joint toxicity of pesticides; this analysis sought to clarify if there was a synergistic, antagonistic, or additive effect on PAL, LOX, and PPO activity in cotton leaves resulting from the interactive H. armigera herbivory and omethoate treatment. In the interactive effect on the response of PAL activity in cotton leaves, antagonistic effects of the omethoate application towards H. armigera herbivory were observed at 6 and 12 h. Synergistic effects were then observed at 18 and 30 h. Antagonistic effects were observed from 54 to 78 h and synergistic effects were finally observed at 126 h. The correlation between H. armigera herbivory and omethoate application in the interactive effect on cotton defense responses of LOX activity also fluctuated from synergism to antagonism during the time course. In the interactive effect on PPO activity, only antagonism was observed between H. armigera herbivory and omethoate application. In the interactive stress of H. armigera herbivory and omethoate application on cotton defense responses, omethoate affected the defense responses of cotton to H. armigera herbivory by producing antagonistic and synergistic effects. These results will be useful to understand the relationship between host plant and herbivorous pest.     

薇甘菊萎蔫病毒感染对薇甘菊光合特性和4种酶活性的影响

为明确薇甘菊萎蔫病毒(Mikania micrantha wilt virus,MMWV)对薇甘菊(Mikania micrantha)叶片生理生化的影响,探讨了MMWV侵染对薇甘菊叶片超氧化物岐化酶(superoxide dismutase,SOD)、过氧化物酶(peroxidase,POD)、多酚氧化酶(polyphenol oxidase,PPO)和苯丙氨酸解氨酶(phenylalanine ammonia-lyase,PAL)活性及叶绿素总含量和光合特性的影响.结果表明:MMWV感染薇甘菊16d内叶片的SOD活性均比对照组高,其中第16天达最大值,但接种后第24和32天SOD活性分别比对照低了13.28％和25.37％;POD活性在接种后16-32d均显著高于对照.PPO和PAL变化趋势相似,在接种后第8天这两个酶的活性分别比对照高了77.75％和23.58％,而在第32天分别比对照减少了14.27％和20.53％.随着侵染时间的增加,感病薇甘菊叶片中叶绿素含量减少,叶绿素a/b值逐步降低;同时最大净光合速率(Pmax)和光饱和点(LSP)分别比健康对照减少了32.34％和12.52％,而对暗呼吸速率(Rd),光补偿点(LCP)和表观量子效率(AQY)无显著影响.表明MMWV侵染可减低薇甘菊叶片光合作用的效率.     

Chitosan Stimulates Defense Reactions in Grapevine Leaves and Inhibits Development of Botrytis Cinerea

Chitosan (β-1,4-linked glucosamine oligomer) derived from crab shells conferred a high protection of grapevine leaves against grey mould caused by Botrytis cinerea. Under controlled conditions, it was shown to be an efficient elicitor of some defense reactions in grapevine leaves and to inhibit directly the in vitro development of B. cinerea. Treatment of grapevine leaves by chitosan led to marked induction of lipoxygenase (LOX), phenylalanine ammonia-lyase (PAL) and chitinase activities, three markers of plant defense responses. Dose-response curves show that maximum defense reactions (PAL and chitinase activities) and strong reduction of B. cinerea infection were achieved with 75–150 mg l⁻¹ chitosan. However, greater concentrations of chitosan did not protect grapevine leaves with the same efficiency, but inhibited mycelial growth in vitro. Present results underlined the potency of chitosan in inducing some defense responses in grapevine leaves which in turn might improve resistance to grey mould.     

杨盘二孢激发子的分离及稳定性研究

 从杨盘二孢的培养滤液和菌丝中获得2种激发子粗提物,分别测定其糖和蛋白质的含量,发现滤液激发子粗提物(CFE)糖和蛋白质的含量分别为41.07和40mg/mL,菌丝激发子粗提物(CME)糖和蛋白质含量分别为48.07和55mg/mL。滤液激发子粗提物对温度不敏感而对碱性条件敏感;菌丝激发子粗提物对酸碱不敏感而对温度敏感。用Sephadex G-100柱层析的方法初步纯化2种激发子,并且菌丝激发子粗提物过柱后得到2个活性组分J14和J25;滤液激发子粗提物过柱后获得2个活性组分L9和L16。将4个组分进行烟草叶片过敏反应和I-895杨酶活性的诱导,结果表明,菌丝激发子强活性物质集中在J14组分;而滤液激发子强活性物质集中在L9组分。     

不同毒性玉米大斑病菌侵染对感病玉米叶片PAL活性的影响

利用从自然界中分离纯化的156株玉米大斑病菌菌株中筛选得到的强毒菌株YC和弱毒菌株01-23T分别接种感病玉米叶片,测定玉米叶片苯丙氨酸解氨酶(PAL)活性的动态变化。结果表明,与不接种对照相比,接种强毒菌株YC 1~2 d时,玉米叶片PAL活性显著降低;接种3~4 d时,PAL活性略有上升但差异不显著;接种5 d时,PAL活性又显著降低。接种弱毒菌株01-23T 1~2 d时,玉米叶片PAL活性略有降低但差异不显著,接种3~5 d时,PAL活性上升但差异仍不显著。PAL是玉米抵抗玉米大斑病的一种重要防御酶,在侵染初期,只有降低玉米叶片的PAL活性玉米大斑病菌才能达到成功定殖的目的,可为明确玉米大斑病菌致病性的生化机制提供一定理论依据。     

不同供氮水平对荞麦茎叶中黄酮含量的影响

以普通荞麦品种晋荞1号(Fagopyrum esculentumMoench,cv.Jinq iao 1)为材料,通过盆栽试验研究苗期和开花前不同供氮水平(0(CK)、100、200 kg.hm-2)对不同生育期荞麦茎叶的干物质量、芦丁和黄酮含量的影响。结果表明,与对照相比,经100、200 kg.hm-2的氮肥处理,盛花期叶片中芦丁和黄酮含量分别下降了36%、46%和27%、47%,成熟期分别下降了20%、56%和22%、52%;经100、200 kg.hm-2的氮肥处理,盛花期茎中黄酮含量分别下降了13%和19%,成熟期分别下降了31%和28%。高氮处理对荞麦叶片黄酮含量的负效应比对茎的更为明显。100 kg.hm-2的氮肥处理下单株茎叶干物质量及芦丁和黄酮产量皆最高。适量施用氮肥将有利于单株水平上的干物质产量和黄酮产量的提高。     

梨苯丙氨酸解氨酶基因的生物信息学分析

[目的]对梨苯丙氨酸解氨酶(PAL)的基因进行生物信息学分析。[方法]以梨的苯丙氨酸解氨酶基因为研究对象,运用生物信息学方法对其编码的蛋白质结构、理化性质及功能结构域进行分析。[结果]梨pal基因的全长编码区序列(Coding sequences,CDS)长2 160 bp,编码720个氨基酸;序列比对结果显示,梨的pal基因与葡萄、树莓、甜樱桃、烟草、毛果杨、柠檬、水稻pal基因的同源性分别为86%、90%、94%、85%、88%、88%和73%;系统进化分析结果显示,梨pal基因与甜樱桃的进化关系最为密切;梨苯丙氨酸解氨酶定位于细胞基质,二级结构以随机卷曲和α-螺旋为主要构件,三维建模成功。[结论]为今后研究PAL和梨木质素合成的分子机理提供了一定的理论参考。     

山药PAL基因全长cDNA序列的克隆、表达与分析

为了阐明苯丙氨酸解氨酶(PAL,EC4.3.1.5)基因在山药地下块茎酚类物质积累过程中的分子机理,本研究运用普通PCR、RACE和TAIL-PCR技术从大薯(Dioscorea alataL.)’Zixiao’地下块茎中克隆到同一个PAL基因的3个片段,即长度分别为1 145bp、1 151bp和424bp的中间片段、3’端和5’端的cDNA序列;将3段cDNA序列拼接后获得大小为2 376bp的PAL基因全长cDNA序列,该序列含有一个1986bp的最大开放阅读框(ORF),一个28bp的5’端非翻译区,一个362bp含25 nt的Poly(A+)尾的3’端非翻译区,最大ORF可推测编码一个包括PAL酶活性中心的特征序列(GTITASGDLVPLSYIA)在内的661个氨基酸的多肽,分子量为71.974kDa,等电点6.310;大薯地下块茎的PAL基因分别在核苷酸与氨基酸水平上和GenBank中所选已知其他物种的同源性为73%~77%和76%~82%,说明PAL基因在系统进化上相对保守;RT-PCR分析表明该基因仅在地下块茎中表达,而且表达丰度在收获前逐渐降低。