Involvement of a vascular hypersensitive response in quantitative resistance to Ralstonia solanacearum on tomato rootstock cultivar LS‐89          下载免费PDF全文

K. Nakaho  S. Seo  K. Ookawa  Y. Inoue  S. Ando  Y. Kanayama  S. Miyashita  H. Takahashi 《Plant pathology》2017,66(1):150-158

Ralstonia solanacearum causes bacterial wilt disease in Solanaceae spp. Expression of the Phytophthora inhibitor protease 1 (PIP1) gene, which encodes a papain‐like extracellular cysteine protease, is induced in R. solanacearum‐inoculated stem tissues of quantitatively resistant tomato cultivar LS‐89, but not in susceptible cultivar Ponderosa. Phytophthora inhibitor protease 1 is closely related to Rcr3, which is required for the Cf‐2‐mediated hypersensitive response (HR) to the leaf mould fungus Cladosporium fulvum and manifestation of HR cell death. However, up‐regulation of PIP1 in R. solanacearum‐inoculated LS‐89 stems was not accompanied by visible HR cell death. Nevertheless, upon electron microscopic examination of inoculated stem tissues of resistant cultivar LS‐89, several aggregated materials associated with HR cell death were observed in xylem parenchyma and pith cells surrounding xylem vessels. In addition, the accumulation of electron‐dense substances was observed within the xylem vessel lumen of inoculated stems. Moreover, when the leaves of LS‐89 or Ponderosa were infiltrated with 10⁶ cells mL^?1 R. solanacearum, cell death appeared in LS‐89 at 18 and 24 h after infiltration. The proliferation of bacteria in the infiltrated leaf tissues of LS‐89 was suppressed to approximately 10–30% of that in Ponderosa, and expression of the defence‐related gene PR‐2 and HR marker gene hsr203J was induced in the infiltrated tissues. These results indicated that the response of LS‐89 is a true HR, and induction of vascular HR in xylem parenchyma and pith cells surrounding xylem vessels seems to be associated with quantitative resistance of LS‐89 to R. solanacearum.  相似文献   

Optimization of 0-8 Aspergillus oryzae Solid State Fermentation Conditions and the Effect of Fermented Product on Nutrient Digestion of Broiler     

MA Jian-qing  GUO Yan-li  ZHANG Tie-ying 《中国畜牧兽医》2017,44(11):3179-3186

The high-yield protease was obtained by screening high-yield protease Aspergillus oryzae and optimizing its solid-state fermentation conditions, and then the effect of fermented product on nutrient digestion in broiler was studied. After the screening and re-screening of 8 different Aspergillus oryzae spores, a new Aspergillus oryzae strain 0-8 with high acid protease was obtained. The fermentation conditions of acid protease were optimized by single factor and orthogonal test experiment. The results showed that the optimum temperature was 30℃, the most suitable fermentation time was 66 h, the initial water content was 47.40%, the inoculation quantity was 1.5×10⁷ spores/mL, the content of bran was 80%, C/N was 1/3, the optimized vigor of acid protease reached 14 416.64 U/g, which was 157.02% higher than that before optimization. 500 mg/kg Aspergillus oryzae 0-8 fermented product was added in the broiler diet, and the mutrient digestibility of broiler were measured after 10 d of adaptation and 5 d of test period. The results of the nutrient digestion of broilers showed that feeding Aspergillus oryzae 0-8 fermentation increased the CP and energy digestibility of broiler in a certain degree compared to control group, but there's no significant difference (P>0.05). Dietary supplementation of Aspergillus oryzae fermentation was beneficial to the production of broilers, but its additive dose or feeding pattern should be further studied.  相似文献   

Screening and Identification of Protease-producing Bacillus amyloliquefaciens from Pig's Intestines and the Preliminary Study of Its Enzymatic Properties     

NING Yu-chang  ZHANG Xiao-jing  QIAO Hong-xing 《中国畜牧兽医》2017,44(8):2398-2407

A Bacillus strains,MY-6 strain which secreted high amount of protease was isolated from healthy pig's intestines,and it was identified as Bacillus amyloliquefaciens by the methods of morphological observation, biochemical tests,16S rDNA sequence amplification and comparison, biolog identification. In order to get an initial understanding of the enzymatic properties of its protease, the dynamic relationship between protease activity and bacterial life cycle, effects of pH,temperature,metal ions and inhibitors on protease activity,and the protease hydrolysis of different substrates were investigated. The results showed that,the protease synthesis of MY-6 strain was synchronized with its spore formation which began in the early stage of the exponential growth phase. The optimum conditions for the protease was 40℃ and pH 7.0,Ca²⁺ and Mn²⁺ had a intensive effects on the protease activity, PMSF could completely inhibit the activity of protease, indicating that the protease was a serine protease.The content of TCA soluble peptides for protein hydrolyzate fish protein, soybean protein, corn protein and casein was 0.83,0.67,0.58 and 0.38 μmol/g,respectively. In conclusion,Bacillus amyloliquefaciens MY-6 strain could be used as an initial selection of probiotic Bacillus to improve protein digestion.  相似文献   

纤细齿梗孢丝氨酸蛋白酶cDNA克隆及其序列分析和表达     

刘连盟  张亚波  李安娜  李多川 《中国生物防治》2010,26(3):327-334

纤细齿梗孢Olpitrichum tenellum是一种寄生在轮枝镰孢菌Fusarium verticillioides上的活体营养菌寄生真菌。丝氨酸蛋白酶在菌寄生过程可能起到重要作用。根据丝状真菌丝氨酸蛋白酶的同源保守序列设计简并引物,通过RT-PCR及RACE的方法,克隆得到1845bp的全长cDNA片段。其可读框为1554bp（GenBank登录号为EU368754）,编码517个氨基酸的蛋白质。比对分析发现该蛋白是一种分泌型的丝氨酸蛋白酶,属于丝氨酸蛋白酶S8家族的枯草杆菌蛋白酶subtilisin,具有典型的信号肽-前肽-成熟肽的结构。将该基因可读框除去信号肽插入酵母表达载体pPIC9K,转化毕赤酵母Pichia pastoris GS115,在甲醇诱导下成功分泌出具有生物活性的重组蛋白酶,诱导120h后酶活性可达153U/ml。重组蛋白酶经DEAE-Sepharose层析进行纯化,SDS-PAGE分析其分子量为39kD。其反应的最适温度为60℃,最适pH为8。  相似文献   

蚕蛹蛋白高效分解菌的分离筛选及部分酶学性质的研究(英文)     

孔军  涂国全  邝哲师  赵祥杰  叶明强  罗国庆 《农业科学与技术》2010,11(2)

[目的]获得蚕蛹蛋白高效分解菌,探索其有关酶学性质。[方法]从腐败蚕蛹中采用稀释平板分离蛋白高效分解菌。先进行菌株的初筛:取采集的样品2g,分别加入装有LB培养基的摇瓶中,置摇床上180r/min,37℃培养4～5d。将发酵液稀释一定倍数,涂布法进行分离,根据酪素培养基中单菌落透明圈和菌落直径比值(H/C)大小,选取H/C比值较大的菌株进行复筛。将初筛得到的待测菌株接种于发酵培养基中,32℃摇床培养60h。发酵液离心(4000r/min,5min)后取上清,测定酶活力,并比较不同菌株发酵液蛋白酶pH、温度作用范围。对筛选出的菌株所产中性蛋白酶再进行pH值条件下酶活力稳定性测验,酶活力热稳定性试验,及不同金属离子对中性蛋白酶活力的影响试验。蚕蛹粉采用筛选出的菌株固体发酵后,于60℃烘箱烘干,测定其中小分子蛋白含量。[结果]经筛选得到1株产中性蛋白酶的KB细菌,在摇瓶发酵条件下其酶活达到589.08U/ml,发酵蚕蛹粉后中小分子蛋白含量达到15.09%,比未发酵的对照和实验室现用菌株发酵分别提高了95.72%和15.67%,这表明KB菌株在蚕蛹粉发酵中优于实验室现用菌株。KB菌株所产中性蛋白酶在pH值6.5～9.0条件较稳定,pH值8.0为其最适pH值,在pH值8.0、温度55℃条件下处理0～30min时的酶活较稳定,残余酶活力在90.0%以上。[结论]初步获得蚕蛹蛋白高效分解菌。  相似文献   

温和气单胞菌TL97528株丝氨酸蛋白酶基因片段克隆与序列分析   总被引：1，自引：0，他引：1  

沈锦玉  李新华  潘晓艺  尹文林  曹铮 《上海海洋大学学报》2010,(2)

中华鳖(Trionyx sinensis)细菌性疾病主要由气单胞菌感染引起的,通过脱脂奶平板检测及酶活性试验得出温和气单胞菌TL97528株分泌的胞外蛋白酶为丝氨酸蛋白酶,而丝氨酸蛋白酶是气单胞菌的主要毒力因子。根据已发表的嗜水气单胞菌的丝氨酸蛋白酶基因序列保守区域设计引物,PCR扩增出一长度为809 bp大小的特异片段,该序列在Genbank上的登录号为FJ357446。对该序列进行分析发现,已发表的温和气单胞菌(Aeromonas sobria)丝氨酸蛋白酶基因(GenBank登录号为AF253471)同源性为98%、与其他几株已发表的嗜水气单胞菌(Aeromonas hydrophila)丝氨酸蛋白酶基因(GenBank登录号分别为AF126213、AY841795、DQ127822、CP000462、CP000644、AF159142)同源性分别为98%、82%、82%、82%、81%、81%,与杀鲑气单胞菌(Aeromonas salmonicida)丝氨酸蛋白酶基因(GenBank登录号为X67043)同源性为80%。用DNAstar软件分析,与鳖源温和气单胞菌TK961010株的同源性为98%,...  相似文献   

Digestive proteolytic and amylolytic activities of Helicoverpa armigera in response to feeding on different soybean cultivars     

Bahram Naseri  Yaghoub Fathipour  Saeid Moharramipour  Vahid Hosseininaveh  Angharad MR Gatehouse 《Pest management science》2010,66(12):1316-1323

  相似文献   

草鱼鱼肉蛋白酶解物抗氧化性及功能特性研究   总被引：12，自引：3，他引：9  

李雪  罗永康  尤娟 《中国农业大学学报》2011,16(1):94-99

为全面了解水解度(DH)、蛋白酶种类对草鱼鱼肉蛋白酶解产物抗氧化性和功能特性的影响,采用木瓜蛋白酶及Alcalase 2.4L在各自最适条件下进行酶解,制备水解度为10%和20%的酶解产物,对其功能特性进行分析。结果显示:随着水解度升高酶解产物的亚铁离子螯合能力增强,但还原力和清除DPPH自由基的能力下降(P<0.05)。相同水解度下与Alcalase 2.4L酶解产物相比,木瓜蛋白酶酶解产物具有较强的清除DPPH自由基能力和还原力(P<0.05)。2种蛋白酶酶解产物的溶解性、乳化性、起泡性均在pH4时达到最低,而后随pH升高而增大。相同pH下随着水解度的升高酶解产物的溶解性增强,乳化性下降。相同pH及水解度下木瓜蛋白酶酶解产物的溶解性和起泡性小于Alcalase 2.4L酶解产物,但乳化性优于Alcalase 2.4L酶解产物。酶解产物的抗氧化性及功能特性受水解度及蛋白酶种类的影响。  相似文献   

饲料中去皮豆粕替代鱼粉比例对草鱼消化酶活力的影响   总被引：3，自引：0，他引：3  

吴莉芳  秦贵信  赵元  王洪鹤  孙泽威  孙玲 《中国畜牧杂志》2010,46(1)

试验旨在研究去皮豆粕替代鱼粉比例对草鱼消化酶活力的影响。选择健康的草鱼为试验对象,以鱼粉为动物蛋白源,去皮豆粕为植物蛋白源,去皮豆粕分别替代0%、15%、30%、45%和60%的鱼粉蛋白,配制成5种等蛋白、等能的半精制饲料,在室内单循环控温养殖系统中进行8周生长试验。试验结束后,取草鱼的前、中、后段肠道和肝胰脏,分别用福林-酚试剂法、碘-淀粉比色法和脂肪酶试剂盒测定了肠道和肝胰脏蛋白酶、淀粉酶和脂肪酶的活力。结果表明:饲料中去皮豆粕替代鱼粉对草鱼肠道和肝胰脏的蛋白酶活力具有显著影响,随着去皮豆粕替代鱼粉蛋白水平的增加,草鱼肝胰脏和肠道蛋白酶活力逐渐降低,60%组与对照组差异显著(P0.05);去皮豆粕替代鱼粉对草鱼肝胰脏、前肠、后肠淀粉酶活力无负面影响,但草鱼中肠淀粉酶活力45%和60%组显著低于对照组(P0.05);去皮豆粕替代鱼粉对草鱼肝胰脏和肠道脂肪酶活力影响不显著(P0.05)。  相似文献   

糙米发芽过程中蛋白酶活力及含氮物质的变化   总被引：5，自引：0，他引：5  

曹晓虹  温焕斌  李翠娟  韩永斌  顾振新 《南京农业大学学报》2010,33(2)

以糙米为原料,研究其发芽过程中蛋白酶活力及蛋白质含量、组成、水解度和肽含量等降解产物的变化。结果表明:糙米经48h发芽,与未发芽前相比,蛋白酶活力增加3.39倍,蛋白质水解度提高5.37倍,肽总量升高38.68%,水解度与肽含量间相关系数达0.9606;清蛋白和球蛋白含量下降74.76%,谷蛋白和醇溶蛋白含量增加95.68%,游离氨基酸含量升高2.17倍,非蛋白氮含量增加。糙米发芽72h时,蛋白酶活力增幅减小,蛋白质水解度及肽含量继续升高,而清蛋白增多,谷蛋白减少。SDS-PAGE测定结果显示,糙米发芽过程中,大分子量的蛋白质组分降解,小分子量的多肽类成分增加,游离氨基酸含量显著升高;发芽48h前蛋白质的转化与降解趋势平缓,之后其变化极显著。因此,糙米发芽48h是其含氮物质变化的重要转折点。  相似文献