Induction of an antioxidant enzyme system and other oxidative stress markers associated with compatible and incompatible interactions between chickpea (Cicer arietinum L.) and Fusarium oxysporum f. sp.ciceris

To ascertain if active oxygen species play a role in fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris, the degree of lipid peroxidation (malondialdehyde formation) and the activity levels of diamine oxidase (DAO), an apoplastic H₂O₂-forming oxidase, and several antioxidant enzymes, namely ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guaiacol-dependent peroxidase (GPX) and superoxide dismutase (SOD), were determined spectrophotometrically in roots and stems of ‘WR315’ (resistant) and ‘JG62’ (susceptible) chickpea cultivars inoculated with the highly virulent race 5 of the pathogen. Moreover, APX, CAT, GPX and SOD were also analysed in roots and stems by gel electrophoresis and activity staining; and the protein levels of APX and SOD in roots were determined by Western blotting. In roots, infection by the pathogen increased lipid peroxidation and CAT and SOD activities, although such responses occurred earlier in the incompatible compared with the compatible interactions. APX, GPX and GR activities were also increased in infected roots, but only in the compatible interaction. In stems, infection by the pathogen increased lipid peroxidation and APX, CAT, SOD and GPX activities only in the compatible interaction, and DAO activity only in the incompatible one. In general, electrophoregrams agreed with the activity levels determined spectrophotometrically and did not reveal any differences in isoenzyme patterns between cultivars or between infected and non-infected plants. Further, Western blots revealed an increase in the root protein levels of APX in the compatible interaction and in those of SOD in both compatible and incompatible interactions. In conclusion, whereas enhanced DAO activity in stems, and earlier increases in lipid peroxidation and CAT and SOD activities in roots, can be associated with resistance to fusarium wilt in chickpea, the induction of the latter three parameters in roots and stems along with that of APX, GR (only in roots) and GPX (only in stems) activities are rather more associated with the establishment of the compatible interaction.     

水稻与稻瘟病菌非亲和性互作中重要防御酶活性变化规律研究

 选以CO39为背景的水稻抗稻瘟病近等基因系,与稻瘟菌生理小种ZC₁₃(菌株97-151a)组成的3类典型非亲和性互作,以亲和性互作为对照,对各互作中过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)、几丁质酶及β-1,3-葡聚糖酶的活性变化规律进行了系统研究。完全非亲和性互作C101A51/97-151a、高度非亲和性互作C101L AC/97-151a及中度非亲和性互作C104 PKT/97-151a,POD比活性接种后即开始明显升高,48h前达到高峰,升高趋势一直持续到7d完全显症时,幅度基本与各互作非亲和程度呈正相关;亲和性互作CO39/97-151a接种后40 h POD比活性才开始升高,4~6 d达到高峰,峰值也较大。3类非亲和性互作PAL比活性在接种后0 h或16 h开始较明显升高,整个互作中形成3~4个较明显的峰;亲和性互作中PAL比活性一直明显下降。3类非亲和性互作外切几丁质酶比活性接种后即开始升高,基本一直保持升高趋势,在40 h前幅度较大,并形成1~3个较高的峰;亲和性互作外切几丁质酶比活性接种后即开始大幅度升高直至完全显症,48h后幅度远高于非亲和性互作。3类非亲和性互作β-1,3-葡聚糖酶比活性在24 h内开始较明显升高,在48h前形成2~3个较明显的峰;亲和性互作在接种后β-1,3-葡聚糖酶比活性即开始升高,在48h后显著高于非亲和性互作。讨论了POD、PAL、几丁质酶及β-1,3-葡聚糖酶参与水稻抗稻瘟病的可能性。     

Identification of ALS inhibitor‐resistant Amaranthus biotypes using polymerase chain reaction amplification of specific alleles

Summary Polymerase chain reaction (PCR) amplification of specific alleles (PASA) was adapted as a molecular marker‐based method for the rapid detection of point mutations in Amaranthus retroflexus and Amaranthus rudis leading to ALS inhibitor resistance. Two pairs of primers were designed for the specific amplification of alleles of the ALS gene of susceptible and resistant biotypes. The allele‐specific primer matched the desired allele, but mismatched the different allele at its 3′ end. Differentiation was carried out by comparison of the amplified DNA fragments in gel electrophoresis after PASA‐PCR. In A. rudis, differentiation was possible with one PCR and genomic DNA as probe. A ‘nested’ PCR was necessary for the differentiation of sensitive and resistant A. retroflexus. PASA is useful for the identification of resistant weed biotypes and also as a monitoring tool to map resistance occurrence and distribution. Advantages include the fast and clear separation of those plants with and without mutations at an early stage of development, its easy and consistent performance and quick results compared with existing resistance detection tests. These advantages, when combined with management strategies, enable further activities to reduce herbicide resistance.     

Role of polymerase β in DNA metabolism and genomic instability

The major role of DNA polymerase β was thought to be limited in its involvement in short patch base excision repair by removing 5’-deoxyribose phosphate and base insertion. However, the recent researches indicate that polymerase β might take part in a wide spectrum of DNA metabolism reactions, including long patch base excision repair, DNA replication, recombination, meiosis and transleisional DNA synthesis. Because of its wide and important cellular function, an inappropriate intracellular polymerase β level might be associated with genomic instability. Down-regulation or mutation of polymerase β is mutagenic due to deficient in DNA repair, while overexpression of this error-prone β polymerase might perturb the normal function of other accurate polymerases and cause genomic instability as well.     

Se-containing spirulina phycocyanin attenuated liver injury induced by carbon tetrachloride in mice

AIM:To investigate the effect of Se-containing spirulina phycocyanin (Se-SPC) on liver injury of mice induced by carbon tetrachloride (CCl₄). METHODS:The mouse model was conducted by intragastric feeding with 2% CCl₄ oil for three times, meanwhile Se-SPC, spirulina phycocyanin (SPC) and Na₂SeO₃ were injected (ip) to various groups for 7 days. Then selenium (Se), glutathione peroxidase (GPx), superoxide dismutase (SOD), alanine aminotransferase (ALT), malondiaoldehyde (MDA) and nitric oxide (NO) levels in blood and liver were measured. RESULTS:The level of Se,GPx and SOD activities were obviously higher(P<0.05)but ALT activity,MDA and NO₂^-/NO₃^- levels were remarkably lower(P<0.05)in Se-SPC treated groups than those in CCl₄ groups,and effects of high dose Se-SPC on Se,GPx,MDA and NO₂^-/NO₃^- were even more significant(P<0.01).Under the same dose of Se or protein,effects of all selected targets in Se-SPC groups were more efficient than those in SPC groups and inorganic-Se groups.Furthermore,Se levels had a positive correlation with GPx activity(r=01705),which had negative correlation with levels of MDA,NO₂^-/NO₃^- and ALT(r=-0.629,r=-0.336,r=-0.457,respectively), and positive correlations between ALT activity and MDA or NO₂^-/NO₃^- level were found (r=0.519,r=0.641). CONCLUSION:These results indicated that Se-SPC may attenuate liver injury of mice induced by CCl₄ through its anti-inflammatory action and enhancing selenoenzyme expression.     

Effects of angiotensin II-induced hypertension chemotherapy on the activity of interleukin-1β converting enzyme in transplanted intracerebral rat gliomas

AIM: To investigate the activity of interleukin-1β converting enzyme in transplanted intracerebral rat gliomas under angiotensin II-induced hypertension chemotherapy. METHODS: The brain tumor model was produced in Wistar rats by stereotaxic inoculation of C6 glioma cells (1×10¹² /L). Tumor-bearing rats were treated with carmustine, teniposide and lisplatin (chemotherapy) during angiotensin II-induced hypertension. Then, the survival time of tumor-bearing rats, tumor blood flow, the concentration of drug, volume of gliomas and the activity of interleukin-1β converting enzyme in glioma were examined.RESULTS: The survival time of tumor-bearing rats was significantly longer in chemotherapy with angiotensin II-induced hypertension group than that of chemotherapy alone. In addition, regional tumor blood flow, the concentration of chemotherapeutic drug and the activity of interleukin -1β converting enzyme in transplanted rat gliomas were increased, while the volume of gliomas was decreased in hypertention chemotherapy group compared with chemotherapy alone. CONCLUSION: Chemotherapy with angiotensin II-induced hypertension has a enhancing effect on chemotherapy for improving the drug delivery to tumor tissue by a increased tumor blood flow and enhancing activity of interleukin -1β converting enzyme.     

中生菌素对水稻白叶枯病的防治机制

对不同白叶枯病抗性水稻品种用200μg/ml中生菌素55℃温汤药液浸种，自然降温，秧田3—4叶期和移栽前5d各用30μg/ml中生菌素处理后，于成株期剪叶接种白叶枯病菌。结果表明中生菌素前期处理，于成株期接种白叶枯病菌时，高抗、中等抗性和感病品种中过氧化物酶、苯丙氨酸解氨酶和多酚氧化酶3种酶活性都有不同程度的提高，且以中等抗性品种酶活提高最多，接种24和48h，3种酶活性较清水对照分别增加26．92％、26．74％、24．06％和7．09％、1．31％、1．60％。盆栽试验表明，中生菌素对中抗品种的防治效果最好，达58．4％。说明中生菌素对水稻防御酶活性的诱激作用是其防治白叶枯病的机制之一。     

棉花黄萎病菌致病型的AFLP分析

 选用41个棉花黄萎病菌(Verticillium dahliae)代表菌系,在温室条件下,对4个棉花品种鄂荆1号(感)、中棉所12(耐)、文-5(抗)和唐棉2号(抗)进行致病性测定,结果可将供试菌系分为落叶型与非落叶型2类。选取8对AFLP引物PCR扩增的结果中,统计带型稳定、清晰且有多态性的条带,共169条作系统聚类分析,将上述菌系分为2大类,第一类为非落叶型菌系,包括10个非落叶型菌系和1个过渡菌系;第二类为30个落叶型菌系。根据聚类分析建立树状图,发现菌系与地理来源存在一定的相关性,而依据菌系致病力强弱分类则相关关系不大。选用25对EcoRⅠ和MseⅠ引物组合,对供试的41个V.dahliae进行AFLP扩增,筛选到2对引物E64(GACTGCGTACCAATTCGAC)、M53(GATGAGTCCTGAGTAACCG)和E49(GACTGCGTACCAATTCCAG)、M65(GAT-GAGTCCTGAGTAAGAG),能分别扩增出433bp和110bp2条仅为V.dahliae非落叶型菌系独有的特异片段,可将落叶型与非落叶型菌系分开,这2条特异片段被命名为EM₄₃₃和EM₁₁₀。     

豇豆与锈菌互作中的多酚氧化酶和过氧化物酶活性及其与抗性的关系

测定了具有不同抗性水平的5个豇豆Vigna sesquipdalis Wight品种在受锈菌Uromyces vignae Barcl侵染前和侵染后的若干阶段中的多酚氧化酶(PPO)和过氧化物酶(POD)活性,并分析其与抗性的关系.结果表明,在接种后24h内,免疫和抗病品种的PPO比活性及其变化率均高于感病品种,且前者PPO比活性变化率高峰出现早,后者出现迟.在接种后,各品种的POD比活性及其变化率均上升,但中抗和感病品种的高峰出现早,免疫和高抗品种出现晚.此外,中抗和感病品种的POD比活性及其变化率在接种12h左右出现高峰后立即下降,而高抗品种的则持续上升至24h左右出现高峰,免疫品种的POD比活性也在24h左右出现高峰,但其POD比活性变化率则持续到48h左右达到高峰,且免疫和抗病品种的峰值明显大于感病品种.     

Expression of cyclooxygenase-2 in IL-1β-stimulated mesangial cells is medicated by NF-κB/IκB signal pathway

AIM: To investigate the role of NF-κB/IκB signal pathway in the regulation of cyclooxygenase-2 (COX-2) expression in human mesangial cells (HMC). METHODS: The PGE₂ concentration in supernatants of HMC was measured by radioimmunoassay. COX-2 mRNA and protein expression were determined by RT-PCR and Western blot. Electrophoretic mobility shift assay (EMSA) and Western blot were used to detect the activity of NF-κB and degradation of IκB. RESULTS: IL-1β significantly upregulated COX-2 expression and PGE₂ production in HMC. Significant up-regulation of NF-κB activation, nuclear translocation of p65 subunit, and degradation of IκB α and IκB β were observed in IL-1β-induced HMC. CONCLUSION: Expression of COX-2 in IL-1β-induced HMC is mediated by NF-κB/IκB signal pathway.