外源绿原酸对苹果抗病相关酶的影响

采用离体培养方法对富士、红星和八棱海棠进行绿原酸处理,研究苹果在外源绿原酸处理下POD、PPO、PAL活性的动态变化以及绿原酸含量的变化。结果表明,富士以0.0001%绿原酸、红星以0.0008%绿原酸、八棱海棠以0.0004%绿原酸处理后,其PAL活性均显著提高,苹果体内也因此而提高了绿原酸水平。     

井冈霉素A对水稻纹枯病菌的毒力和作用机理研究

根据水稻纹枯病菌 Rhizoctonia solani AG- 1IA在含系列浓度井冈霉素 A马铃薯葡萄糖琼脂培养基 (PDA)上的菌丝生长速率 ,计算出药剂的毒力回归方程为:Y=3.2 30 9+0 .872 0 X,r=0 .9910 ,药剂的 EC50 =10 6 .9μg/ m L,EC75=6 34.5 μg/ m L。在含 5 0 0 μg/ m L井冈霉素 A的 PDA上水稻纹枯病菌的菌丝生长抑制率为 74 .1% ,而田间水稻植株上人工接菌的药效试验结果表明 ,5 0 μg/ m L药液喷洒后 7d和 14 d的防效即可分别达到 77.4 %和 76 .7% ,即药剂在 PDA平板上的理论抑制作用仅是田间活体植株上对病菌实际作用效果的 1/ 10。比较室内毒力和田间药效试验的结果可以看出 ,井冈霉素 A具有对病原菌和寄主植物双重作用的特性。研究首次发现 ,在室内培养基上不能有效抑制病菌生长的 1μg/ m L的井冈霉素 A可以在水稻植株未喷药部位产生防御水稻纹枯病的作用 ,且能够持续诱导植物防御反应相关酶——过氧化物酶 (PO)和苯丙氨酸解氨酶 (PAL)的活性增高,表明该药剂可以激发水稻抗性防卫反应的表达。     

锥栗对栗疫病的抗性与苯丙氨酸解氨酶的关系

健康的锥栗(Castanea henryi)皮层中,苯丙氨酸解氨酶(PAL)比活力与抗感病性之间不相关.不同抗病性的锥栗品种接种栗疫菌(Cryphonectria parastttca)后,PAL比活力均上升,接种144 h后,PAL比活力抗病性强的品种比抗病性弱的品种高.接种前以及接种后的各个时间内,木质素含量都是抗病品种比感病品种高.绿原酸的含量与品种的抗病性之间不呈规律性的相关,从绿原酸对栗疫菌菌丝生长影响的测定结果可知,锥栗体内的绿原酸浓度不足以影响栗疫菌.可见,接种后的皮层PAL比活力以及接种前、后的皮层木质素含量可作为衡量锥栗抗病性强弱的指标.     

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。     

Expression of phenylalanine ammonia-lyase cDNA from rice in E. coli BL21DE3

AIM: To study the expression and its kinetics of rice phenylalanine ammonia-lyase gene encoding into E. coli as the basis of treatment for phenylketouria. METHODS: The phenylalanine ammonia-lyase-1-cDNA(rPAL-1-cDNA) from rice was recombined into E. coli high expression vector pET-28c and transformed into E. coli host strain BL21DE3. Engineering bacteria was then inducted by isopropyl-β-D-thiogalactoside (IPTG) for 1, 3, 5, 7 hours, in order to obtain high level expression. RESULTS: After induction, the expression level of fusion protein was 21.40%, 30.60%, 35.40%, 35.43% respectively. The fusion protein exhibited a band of 78.6 kD on SDS-PAGE analysis, but was not found in controls.The target protein was mainly existed in the form of inclusion body. CONCLUSION:Rice PAL gene expressing E. coli was established by gentic engineering technique.     

玉米叶片中次生代谢物丁布含量和苯丙氨酸解氨酶活性的关系研究

[目的]分析比较不同玉米品种在不同生育时期丁布含量和苯丙氨酸解氨酶活性变化。[方法]采用高效液相色谱法对各个生育时期倒1叶、倒3叶丁布含量进行检测及PAL含量的测定。[结果]结果表明:不同玉米品种倒1叶、倒3叶丁布含量随着生育期的增加而减少,并且倒1叶减少的速度大于倒3叶;而苯丙氨酸解氨酶活性随着生育时期的增加而升高。[结论]玉米叶片随着生育时期增加苯丙氨酸解氨酶活性与丁布含量之间存在负相关关系。     

拮抗菌与病原菌处理对采后桃果实多酚氧化酶、过氧化物酶及苯丙氨酸解氨酶的诱导

 Pichiamembranefaciens能有效抑制桃果实采后软腐病。笔者观测和比较接种拮抗酵母菌P .mem branefaciens和软腐病菌Rhizopusstolonifer对采后桃果实多酚氧化酶 (PPO)、过氧化物酶 (POD)和苯丙氨酸解氨酶(PAL)活性的诱导情况。试验结果表明 ,桃果实接种P .membranefaciens +R .stolonifer 2 4h后 ,PPO和PAL活性开始升高 ,并在整个试验过程中一直保持较高的水平。只接种R .stolonifer也能诱导桃果实PPO和PAL活性的增加 ,但效果不如接种拮抗菌 +病原菌的好。然而 ,接种拮抗菌 +病原菌和只接种病原菌对诱导POD活性都没有明显的作用。可以认为 ,拮抗菌诱导抗性相关酶活性的提高是发挥抑病作用的一种重要方式     

大豆品种的抗病性与叶片内苯丙氨酸解氨酶活性关系的研究

在抗病品种东农9674，东农1572和感病品种黑农39，东农95-165上接种大豆灰斑病菌的1号，2号，4号，6号，7号等5个生理小种，测定从接种到显症前与抗性有关的苯丙氨酸解氨酶活性的变化。研究结果表明，抗病的大豆品种接种不同生理小种的灰斑病菌后，PAL的活性分别从第6-7天起吴上升趋势且明显高于对照；相反感病品种从8-9天起PAL活性明显低于对照，因此说明PAL的活性与大豆抗灰斑病性呈正相关关系。     

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。     

Differential structure and physiological response to canker challenge between ‘Meiwa’ kumquat and ‘Newhall’ navel orange with contrasting resistance

Asiatic citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is one of the most detrimental diseases affecting citrus production. ‘Meiwa’ kumquat (Fortunella crassifolia) and ‘Newhall’ navel orange (Citrus sinensis) are two genotypes with contrasting Xcc resistance. In the current study, efforts were made to perform pairwise comparison between them in terms of structural difference and biochemical response to Xcc attack. Pinprick inoculation demonstrated that ‘Meiwa’ had lower disease incidence rate and smaller lesion size, concurrent with significantly less bacterial number at the inoculation sites relative to ‘Newhall’. Microscopic observation demonstrated that stomatal density and size of ‘Meiwa’ were significantly smaller than ‘Newhall’. Stomatal apertures of ‘Meiwa’ were smaller than ‘Newhall’ measured at 2 h after their leaves were immersed in Xcc suspension, consistent with the bacteria number in the internal tissues counted at 4 d. Epicuticular wax content of ‘Meiwa’ was significantly higher than that of ‘Newhall’, while no difference in palisade and spongy tissue was observed between them. Enzymatic activity assay showed that after Xcc inoculation, ‘Meiwa’ exhibited significantly higher activities of catalase (CAT), peroxidase (POD) and phenylalanine ammonia-lyase (PAL) in comparison with ‘Newhall’ at most of the sampling time points. Our data showed that the two genotypes with contrasting Xcc resistance exhibited clear difference in the structure and physiological response, which may provide information to facilitate our understanding of Xcc resistance.