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1.
表达的hrmA蛋白质具有诱导水稻细胞产生防卫反应的活性   总被引:2,自引:0,他引:2  
 将丁香假单胞菌hrmA基因构建到原核表达载体pET 29中,得到重组载体pET hrmA。将重组载体转化大肠杆菌BL21(DE3),经IPTG诱导,获得了以包含体形式存在的融合蛋白。复性的融合蛋白能诱导水稻悬浮细胞的活性氧迸发,处理20min后达到峰值。Northern杂交结果表明hrmA蛋白能显著地诱导PBZ1基因在水稻悬浮细胞中表达,在所观察的时间内PBZ1基因表达丰度随处理时间延长而增加,用hrmA处理水稻植株,诱导了PAL基因的表达。实验结果表明hrmA融合蛋白具有激活水稻细胞防卫反应的活性。  相似文献   

2.
 通过沸水浴、硫酸铵沉淀、非变性电泳、割胶电洗脱等方法,从一种疫霉的培养液中分离得到分子量约31kDa的蛋白类激发子。该激发子在低浓度下即可诱发烟草叶片发生过敏反应,处理烟草悬浮细胞12h能导致24.6%的细胞死亡,处理24h后细胞的死亡率达67.8%。用该激发子处理烟草悬浮细胞24h后细胞基因组受到明显降解,但无DNAlad-dering现象。用该激发子处理1h即可显著降低烟草悬浮细胞中的ATP水平,而用氧化磷酸化的解偶联剂CCCP处理在降低胞内ATP水平的同时也能显著诱导烟草悬浮细胞死亡,因此抑制胞内ATP合成可能是该激发子诱导烟草细胞死亡的重要机制之一。  相似文献   

3.
中生菌素对水稻悬浮细胞过氧化物酶基因转录表达的影响   总被引:1,自引:0,他引:1  
用白叶枯菌(HB84—17)、10μg/ml中生菌素和10μg/ml中生菌素+白叶枯菌处理水稻抗、感白叶枯病近等位基因系CBB4和沈农1033悬浮细胞,采用斑点杂交的方法研究了中生菌素对水稻悬浮细胞过氧化物酶基因转录表达的影响。结果表明,白叶枯菌、中生菌素和中生菌素+白叶枯菌处理都能诱导悬浮细胞中过氧化物酶基因的转录表达。对于CBB4,白叶枯菌处理3h时过氧化物酶基因开始诱导转录表达,6h达到高峰,诱导表达时间持续至24h;中生菌素处理3h时开始大量表达,6h达到,高峰,大量表达持续至72h。对于沈农1033,白叶枯菌处理48h时过氧化物酶基因才开始诱导转录表达;中生菌素处理,3h就开始大量转录表达,并持续至48h。中生菌素+白叶枯菌处理的感、抗病品种过氧化物酶基因的诱导转录表达强度和模式与单用中生菌素处理相同。  相似文献   

4.
 转录因子基因OsBTF3在水稻品种日本晴悬浮细胞中受白叶枯病菌(Xanthomonas oryzae pv.oryzae,Xoo)诱导表达。为了阐明OsBTF3在水稻叶组织中的表达特征,本研究利用RT-Q-PCR技术,对经3种亲和性病原菌[水稻白叶枯病菌(Xoo)、水稻条斑病菌(Xooc)和稻瘟病菌(Mg)]接种和4种信号分子[脱落酸(ABA)、水杨酸(SA)、茉莉酸甲酯(MeJA)、乙烯(ETH)]诱导处理的水稻叶片中OsBTF3的转录本进行了定量分析。结果表明,OsBTF3对Xoo、Xooc和Mg侵染的基因表达反应均显著地受到诱导,但反应速度和强度略有差异。而4种信号分子对OsBTF3表达的诱导作用差异较大,ABA诱导活性最强,MeJA和ETH次之,SA诱导作用不显著。因此,OsBTF3基因表达不仅具有病原菌Xoo、Xooc和Mg的诱导性,而且也具有信号分子MeJA、ETH和ABA的应答性。  相似文献   

5.
 本研究对水稻白叶枯病菌与水稻悬浮细胞非亲和互作中蛋白类激发子进行了分离纯化和鉴定.白叶枯病菌JXOV与水稻IRBB4和IR24悬浮细胞互作36 h后的上清液,经Q-Sepharose阴离子交换层析柱分离,对分离的各组分进行抗病性诱导测定,结果表明JXOV与IRBB4非亲和互作的上清液中存在蛋白类激发子.有活性的蛋白组分经阴离子交换层析柱Mono-Q进一步纯化后,SDS-PAGE分析鉴定出2个具激发活性的蛋白,其分子量分别为17.2 kD和49.2 kD,等电点分别为5.8和6.2.利用上述激发子处理水稻能减少病斑长度并诱导水稻防卫酶活性的增加.  相似文献   

6.
激活蛋白PeaT1在烟草细胞膜上的结合位点及其特性   总被引:1,自引:0,他引:1  
 激活蛋白PeaT1是从极细链格孢菌(Alternaria tenuissima)中分离的,能诱导植物产生系统获得抗性的蛋白激发子。为了阐明PeaT1诱导植物提高抗性的分子机制,本文采用RT-PCR技术研究了PeaT1诱导烟草悬浮细胞后不同时间PR-1a基因的转录活性,在烟草悬浮细胞中加入20μg/mL的PeaT18h后,PR-1a基因转录活性达到最高;相同浓度的PeaT1处理烟草植株,烟草叶片中酸性PR蛋白表达量升高。激光共聚焦显微镜观察到FITC荧光素标记的PeaT1可与烟草悬浮细胞表面结合;免疫荧光法进一步证明PeaT1可与烟草细胞质膜结合;使用共价交联剂BS3证明125I-PeaT1可与烟草细胞质膜上的2个分子结合,而与加热或蛋白酶处理的质膜不能发生交联,说明与PeaT1结合的分子具有蛋白特性,分子量分别为20kDa和30kDa。上述实验结果证明在烟草细胞质膜上存在着激发子PeaT1受体样的结合位点。  相似文献   

7.
HarpinXoo诱发烟草过敏反应早期H2O2变化及有关基因表达   总被引:2,自引:0,他引:2  
 HarpinXoo是水稻黄单胞细菌(Xanthomonas oryzae pv.oryzae)产生的蛋白类激发子,用harpinXoo注射烟草最早在处理后20h出现肉眼可见的细胞死亡(macro-HR),分别经埃文斯蓝(Evans blue)和台盼兰(Trypan blue)染色观察,发现诱导后4-8h即开始有少量细胞死亡,表明在macro-HR发生前烟草细胞已陆续发生微细胞死亡(micro-HR);对二氨基联苯胺(diamino benzidine,DAB)染色显示,在烟草细胞死亡早期,伴有H2O2的明显积累;定量RT-PCR法对H2O2代谢相关基因的表达进行了分析,发现harpinXoo诱导后1h编码NADPH氧化酶的基因rboh即开始表达,3~7h表达较强,10h开始减弱,表达趋势与H2O2积累趋势一致;编码交替氧化酶的基因aoxl表达较迟,sodA基因则在诱导后表达减弱。这些基因的不同表达模式说明,它们在harpinXoo诱导的H2O2代谢过程中可能具有不同的作用。  相似文献   

8.
 采用改进的差异显示方法由稻瘟菌激发子处理的水稻悬浮培养细胞中得到7个诱导表达的蛋白激酶cDNA片段。为获得这些蛋白激酶的编码全长序列,构建了稻瘟菌激发子处理的水稻细胞cDNA文库。以上述一个cDNA片段作为探针筛选文库,得到一个含有完整阅读框的cDNA克隆OsEPK1分析OsEPK1的推定氨基酸序列的结果表明:它由N端信号肽、胞外区域、跨膜结构和胞内蛋白激酶结构域等几部分组成,可能为一受体类似蛋白激酶。对比OsEPK1与其它植物受体类似蛋白激酶在胞外区域的相似性,结果显示OsEPK1与小麦叶锈激酶家族同源性较高,推测它们可能属于同一类植物受体类似蛋白激酶。此外,northern杂交显示,OsEPK1可由稻瘟菌激发子迅速瞬时诱导,因而它的诱导可能也是水稻对稻瘟菌侵染做出的一种早期防卫反应。  相似文献   

9.
棉疫病菌90kD胞外蛋白激发子诱导烟草过敏性反应的研究   总被引:4,自引:2,他引:4  
 就棉疫病菌90 kD胞外蛋白激发子诱导烟草过敏反应(HR)过程中细胞死亡和防卫反应酶系活性变化及病程相关蛋白PR5的诱导进行研究。结果是,以10 nmol/L激发子溶液注射处理W38烟草叶片,HR枯斑周围5 mm宽组织在UV光下呈现蓝色荧光,对处理部位进行Evans blue染色测定结果是至20 h处理部位细胞全部死亡;激发子可诱导烟草防卫反应中苯丙氨酸解氨酶(PAL)的活性提高;可快速诱导PR5基因的转录。上述结果表明90 kD蛋白激发子可诱发烟草的细胞死亡、苯丙烷代谢和PR基因的表达等多条信号途径。  相似文献   

10.
为探究水稻磷酸核酮糖激酶基因OsPRK在水稻诱导抗虫反应中的功能,以水稻秀水110为材料克隆OsPRK基因的全长,通过生物信息学软件分析其序列特征,并应用实时荧光定量PCR技术分析OsPRK基因在水稻不同组织中的分布情况以及在虫害诱导、激素和机械损伤处理水稻中的表达特征。结果显示,水稻OsPRK基因序列全长为1 212 bp,编码403个氨基酸,分子量为44.86 kD,具有1个磷酸核酮糖激酶保守结构域。OsPRK蛋白亚细胞定位结果显示其定位于叶绿体。OsPRK基因在水稻中的表达具有组织特异性,其在内叶、外叶、内叶鞘、外叶鞘和根系这5个组织中相对于内参基因ACTIN的表达量分别为35.83、20.53、6.25、3.21和0.03。与对照相比,二化螟Chilo suppressalis为害能够强烈抑制水稻茎秆中OsPRK基因的表达;褐飞虱Nilaparvata lugens怀卵雌成虫为害1.5、24 h、白背飞虱Sogatella furcifera怀卵雌成虫为害1、8、24 h以及机械损伤处理3、6、24 h均能显著诱导水稻茎秆中OsPRK基因的表达;而OsPRK基因的表达量在茉莉酸处理6、12 h时以及水杨酸处理0.5、1.5 h时被显著抑制,在茉莉酸处理48 h和水杨酸处理24 h时被显著诱导。表明OsPRK基因可能参与了水稻对害虫的诱导防御反应。  相似文献   

11.
 采用从福建省稻田分离纯化的纹枯病菌(Rhizoctonia solani)菌株FJ-15接种籼稻9311,分析了水稻在纹枯病菌侵染致病过程中,编码病程相关蛋白的基因表达动态,并观察了症状的变化。Northern blot分析表明:PR1在接种12 h后开始表达,在之后的4个时间段其表达量逐步增强;而PBZ1也在12 h开始表达,在48 h表达量激剧增强几乎与72 h表达量相当。组织和症状观察表明,接种12 h后叶鞘表面菌丝纵横分枝,接种36 h后出现零星病斑,接种48 h后表现典型的受害症状,接种72 h后病斑继续扩大,并可蔓延到非接种叶鞘。结果表明,PR1PBZ1的表达与水稻和纹枯病菌亲和互作的过程存在对应关系。  相似文献   

12.
Melon plants locally infected with Colletotrichum lagenarium display a marked increase in chitinase activities (exo- and endo-activities) throughout the whole plant. This increase begins 3 days after inoculation in the inoculated cotyledon, and then occurs sequentially in the non-infected tissues.Both fungal elicitors and plant endogenous elicitors induce a rapid increase in chitinase activity in the treated cotyledon. In other organs, chitinase activity is stimulated, to a lesser extent and after a lag period, only by fungal elicitors.The earlier, more rapid, systemic induction of chitinase activity, produced by treatment with the fungal elicitor is correlated by the increased resistance of the tissues to infection by the pathogen.  相似文献   

13.
 Hyphal cell wall crude elicitor(CWE)of rice blast pathogen could induce hypersensitive response in tobacco and induce other nonhost plants to be resistant to other fungal pathogens.When corn was treated with CWE,they inhibited the infection of Exserohilum turcicum and Curvularia lunata,and when plants of capsicum and cucumber were treated with CWE,they inhibited the infection of Colletotrichum gloeospori-oides.CWE solution showed no bioactivity on spore germination and hyphal growth of the experiment fungi in vitro.Nonhost resistance induced by CWE to other fungal pathogens was not complete resistance.The induced resistant effect(IRE) increased as CWE concentration increased,however,IRE had somehow satura-ted concentration of CWE.Induced nonhost resistance by incompatible pathogen was quantitative to other compatible pathogens.The induced resistance was best at 2 or 3 d after CWE treatment,and then decreased.IRE was about 20 percent in 10 d after CWE treatment.  相似文献   

14.
Priming of defense reactions by an elicitor results in an enhanced ability of the plant to respond to subsequent pathogen challenges. We previously showed that application of lipopolysaccharides (LPS) to potato cell suspensions causes apoplastic acidification, but does not stimulate lipoxygenase (LOX) activity. Here, we tested the ability of various elicitors to prime and elicit defense reactions in potato cell suspensions. Adding 20 microg ml(1) LPS, laminarin, harpin N, or a concentrated culture filtrate (CCF) of Phytophthora infestans to cell cultures 18 h before a second elicitation with LPS did not alter the intensity of apoplastic acidification compared with a single LPS application. Conversely, high concentrations (200 or 400 microg ml(1)) of LPS, laminarin, and harpin N activated LOX in cells pretreated with 1 microg ml(1) CCF, but not in cells pretreated with LPS, laminarin, or harpin N. LOX response was maximal in pretreated cells of potato cv. Bintje when the second elicitation occurred 18 to 24 h after CCF application. These results showed that LOX activation is primed in potato cells by CCF, but not by LPS, harpin N, or laminarin. Finally, bioassays showed a slightly greater reduction of rot weight in half tubers treated with CCF followed by LPS before inoculation with Pectobacterium atrosepticum than in half tubers treated with either preparation alone, indicating a priming effect of CCF on both LOX induction and disease suppression.  相似文献   

15.
Elicitin and a new protein 75 kDa elicitor were purified from the culture filtrate of Phytophthora palmivora, a pathogen of Hevea brasiliensis (rubber plant). Elicitin was obtained by using a one step of DEAE cellulose chromatography and the new elicitor was obtained by two steps of chromatography: a DEAE cellulose column followed by a hydrophobic column. Both elicitors were stable to heat and a wide range of pH values, but were sensitive to ProteaseK. Both elicitors induced scopoletin, peroxidase isozymes (with substrate o-dianisidine and scopoletin) and total phenolic compounds in cell suspension of H. brasiliensis with similar kinetics. In addition, both elicitors induced peroxidase enzyme (o-dianisidine), total phenolic compounds and enhanced local resistance against P. palmivora on young rubber tree seedlings. However, the increase of peroxidase enzyme and total phenolic compounds in rubber tree seedlings was different from those in cell suspension. Furthermore, during the expression of local resistance the zoospore of P. palmivora induced the peroxidase enzyme (o-dianisidine) more rapidly and with higher level than the control plants. H. brasiliensis is more responsive to the new elicitor than elicitin in triggering defense responses. That is the new elicitor was active at a concentration lower than those required for elicitin, about a 30-fold decrease for activation defense responses in cell suspension. For induction of peroxidase enzyme (o-dianisidine), phenolic compounds and local resistance of rubber plants against P. palmivora, the 75 kDa protein was active at about a 2-fold lower concentration when compared to elicitin.  相似文献   

16.
In the present study, the effect of red light on the infection behavior of Bipolaris oryzae on rice leaves and the effects of chemical inhibitors and spore germination fluid (SGF) of B. oryzae on red-light-induced resistance against brown spot disease were investigated. Red light irradiation and natural light did not differ significantly with respect to their effect on spore germination and appressorium formation of B. oryzae 24 h after inoculation. However, formation of infection hyphae was significantly inhibited under red light compared to that under natural light. Pretreatment with the photosynthetic inhibitor 3-(3,4-dichorophenyl)-1,1-dimethylurea (DCMU) or with the aromatic amino acid inhibitor N-(phosphonomethyl) glycine (glyphosate) for 24 h inhibited the development of resistance in rice leaves. To elucidate the elicitor(s) produced during the B. oryzae–rice plant interaction, the effect of SGF prepared in the absence (A-SGF) or presence (P-SGF) of rice leaves on red-light-induced resistance was investigated. When rice plants were pretreated with A-SGF or P-SGF for 24 h, P-SGF had elicitor activity under red light, but not under natural light or with A-SGF. These results suggest that germinating spore of B. oryzae produced an elicitor(s) under red light conditions in a host-dependent manner. In conclusion, we hypothesize that an unidentified elicitor(s) produced at an early stage of fungal infection during the B. oryzae–rice interaction contributes to the inhibition of cell death in rice leaves under red light and enhances resistance-related tryptophan and phenylpropanoid pathways.  相似文献   

17.
稻瘟菌激发子CSB I专化性及相关性质研究   总被引:5,自引:1,他引:5  
 以一套水稻抗稻瘟病近等基因系为材料,接种稻瘟菌(Magnaporthe grisea)细胞壁来源的糖蛋白激发子CSB I,其诱导植保素的积累在高度非亲和性互作水稻远高于亲和性互作水稻;研究同时表明,CSB I可专化性诱导完全非亲和性互作和高度非亲和性互作水稻的过敏性坏死反应;表明该激发子具有小种-品种专化性。经热、胰蛋白酶和过碘酸钠处理后的生物活性检测结果表明,CSB I的活性位点为糖基部分。经pH稳定性检测,CSB I在酸性及相对弱碱性条件下较稳定;而在强碱性条件下,激发子活性下降较多,甚至完全丧失。对CSB I诱导活性的有效浓度测定表明,激发子诱导水稻叶片酶活性升高的最低有效浓度为0.07~0.70 nmol/L。  相似文献   

18.
为研究海洋生境棘孢木霉菌Trichoderma asperellum TCS007对植物的促生长及诱导抗逆作用,以黄瓜和水稻为试材,评价菌株TCS007对植物种子萌发、幼苗和根系生长等生长量及其对叶绿素、可溶性糖和蛋白含量的影响,并在低温和高盐胁迫条件下,研究其对植株生长相关生理指标影响。结果表明:1 × 106 CFU/mL的TCS007孢子悬浮液可以显著促进黄瓜种子萌发,且处理后黄瓜幼苗的株鲜重、叶面积、根鲜重及根系活力分别提升13.53%、17.97%、66.67%和27.30%,总叶绿素、可溶性糖和可溶性蛋白含量分别增加22.75%、18.24%和8.60%;在5 ℃、10 ℃和15 ℃低温胁迫下,1 × 106 CFU/mL的TCS007孢子悬浮液显著提高了黄瓜叶片中过氧化物酶 (POD) 和超氧化物歧化酶 (SOD) 的活性,降低了丙二醛 (MDA) 含量和相对电导率 (REC);在0.1 mol/L的氯化钠 (高盐) 胁迫下,不同含量的TCS007孢子悬浮液均可显著缓解氯化钠对水稻幼苗的盐害作用,且随着孢子含量的增高,缓解效果越为明显,其中添加5% 1 × 106 CFU/mL TCS007孢子悬浮液的水稻幼苗POD、SOD和过氧化氢酶(CAT)活性均显著增加并能降低其REC。综上,棘孢木霉菌TCS007对植物具有良好的促生长效果和诱导抗逆作用,研究结果可为其开发成为新型生物农药提供理论依据。  相似文献   

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