磷酸甘油转移酶基因在水稻白叶枯病菌致病力中的作用研究

稻黄单胞菌稻致病变种(Xanthomonas oryzae pv.oryzae,Xoo)引起的水稻白叶枯病,是水稻上的重要致灾病害之一。前期基因组学分析发现,Xoo菌株PXO99^A的PXO＿04062基因可能编码一个磷酸甘油转移酶,推测与病原菌的致病力有关。本研究采用自杀质粒pK18mobsacB介导的方法,构建了该基因的缺失突变体,并对突变体进行了反式互补。水稻上致病性测定发现,与野生型菌株相比,PXO＿04062基因的缺失突变体DM04062在寄主水稻日本晴上的致病力显著下降,并且其胞外多糖产量、运动性、生物膜形成等均明显降低。进一步采用RNA-seq方法比较突变体转录组的变化,发现突变体中的差异表达基因共有533个,其中包括多个致病相关基因,如胞外多糖合成和细胞运动性等相关基因。与野生型相比,这些基因在突变体中的表达均显著下调。这些结果说明,白叶枯病菌的磷酸甘油转移酶基因是通过影响多种致病力相关因子的合成,来影响病菌在水稻上的致病力。     

水稻白叶枯病菌鞭毛基体基因fliExoo缺失突变分析

 为了阐明水稻白叶枯病菌(Xanthomonas oryzae pv. oryzae,简称Xoo)鞭毛基体组分蛋白基因fliExoo的生物学功能,本研究用Gm^R抗性基因标记交换法成功构建了基因缺失突变体△fliExoo。与野生型菌株PXO99^A相比,△fliExoo鞭毛缺失,菌体易沉降,在0.3%半固体培养基上运动能力明显降低;尽管生长速率和胞外纤维素酶活性无明显变化,但胞外多糖产生和生物膜形成能力明显下降;对水稻品种日本晴的致病性和对非寄主烟草的致敏性反应明显减弱。基因互补可以使上述突变表型恢复。因此,鞭毛基因fliExoo突变不仅影响了细菌鞭毛依赖的运动性,而且对病原菌毒性相关的表型也产生了影响。     

不同培养基继代培养蜡蚧轮枝菌对产孢量和烟粉虱毒力的影响

为了探究不同培养基继代培养的蜡蚧轮枝菌对烟粉虱的作用效果,通过继代培养方法测定JMC-01、JMC-02菌株的产孢量、对烟粉虱的毒力及其几丁质酶活性。结果表明,JMC-01、JMC-02菌株在添加蝉蜕和烟粉虱的PDA培养基中连续培养5代的产孢量、对烟粉虱的毒力、几丁质酶活性均高于无添加的PDA培养基,且JMC-01菌株的各项参数均高于JMC-02菌株。JMC-01菌株在添加蝉蜕的PDA培养基中T_1和T_3代的产孢量最大,均为1.279×10~7个/mL,T_3代对烟粉虱的毒力最大,为88.33%,T_5代的几丁质酶活最高,为0.268 3 U/mL;在添加烟粉虱的PDA培养基中,JMC-01菌株的最大产孢量、对烟粉虱的最高毒力和最高几丁质酶活分别出现在S_2、S_4和S_5代,分别为1.270×10~7个/mL、88.33%和0.325 4 U/mL。回归分析结果显示,JMC-01、JMC-02菌株几丁质酶活性与烟粉虱校正死亡率呈显著的正相关,且拟合优度较好。本试验表明JMC-01、JMC-02菌株均对烟粉虱具有较强的毒力,添加蝉蜕和烟粉虱的培养基可以维持菌株生长特性、增加产孢量、延缓菌株退化、增强菌株的几丁质酶活性,为烟粉虱的生物防治提供了新的菌种资源。     

50%醚菌酯干悬浮剂对香蕉尾孢菌叶斑病的毒力测定与防治试验

香蕉尾孢菌叶斑病是香蕉上的重要病害,每年夏秋季节在华南蕉区普遍发生,危害严重.2003～2004年进行新型杀菌剂50%醚菌酯干悬浮剂对香蕉尾孢菌的毒力测定和防治叶斑病药效试验,结果表明,50%醚菌酯干悬浮剂对香蕉尾孢菌的抑菌效果明显,对香蕉叶斑病有良好的防治效果,对香蕉安全;第3次施药后15 d,50%醚菌酯干悬浮剂1 500倍的防治效果达到82.67%,显著优于对照药剂25%敌力脱乳油1 500倍的防效,建议在生产上推广使用.     

球孢白僵菌MZ041016菌株对平菇厉眼蕈蚊的毒力

利用球孢白僵菌MZ041016菌株5种孢子悬浮液浓度(3.6×104～3.6×108个/mL)对平菇厉眼蕈蚊成虫和幼虫进行室内毒力测定。在3.6×108个/mL浓度下成虫死亡率最高达86.72%,幼虫死亡率最高达87.69%,成虫的LC50为2.319×105个/mL,幼虫的LC50为3.082×105个/mL。在3.6×104～3.6×108个/mL浓度处理下成虫的LT50依次为8.57、6.14、5.18d和4.28d,幼虫的LT50分别为8.09、6.03、4.96d和4.53d。球孢白僵菌MZ050724菌株在室内对平菇厉眼蕈蚊的成虫和幼虫有较强的毒力。     

The Influence of Co-Suppressing Tomato 1-Aminocyclopropane-1-Carboxylic Acid Oxidase Ⅰ on the Expression of Fruit Ripening-Related and Pathogenesis-Related Protein Genes

The purpose of this study is to explore the influence of co-suppressing tomato ACC oxidase Ⅰ on the expression of fruit ripening-related and pathogenesis-related protein genes, and on the biosynthesis of endogenous ethylene and storage ability of fruits. Specific fragments of several fruit ripening-related and pathogenesis-related protein genes from tomato （Lycopersicon esculentum） were cloned, such as the l-aminocyclopropane-1-carboxylic acid oxidase 1 gene （LeAC01）, 1- aminocyclopropane-l-carboxylic acid oxidase 3 gene （LeAC03）, EIN3-binding F-box 1 gene （LeEBF1）, pathogenesis-related protein 1 gene （LePR1）, pathogenesis-related protein 5 gene （LePR5）, and pathogenesis-related protein osmotin precursor gene （LeNP24） by PCR or RT-PCR. Then these specific DNA fragments were used as probes to hybridize with the total RNAs extracted from the wild type tomato Ailsa Craig （AC＋＋） and the LeAC01 co-suppression tomatoes （V1187 and T4B）, respectively. At the same time, ethylene production measurement and storage experiment of tomato fruits were carded out. The hybridization results indicated that the expression of fruit ripening-related genes such as LeACO3 and LeEBF1, and pathogenesis-related protein genes such as LePR1, LePR5, and LeNP24, were reduced sharply, and the ethylene production in the fruits, wounded leaves decreased and the storage time of ripening fruits was prolonged, when the expression of LeACO1 gene in the transgenic tomato was suppressed. In the co-suppression tomatoes, the expression of fruit ripening-related and pathogenesis-related protein genes were restrained at different degrees, the biosynthesis of endogenous ethylene decreased and the storage ability of tomato fruits increased.     

致仔猪水肿病大肠杆菌减毒株的毒力测定及其在仔猪肠道内增殖的分析

在已构建致仔猪水肿病大肠杆菌S451521菌株的2个毒素敲除菌株（F1、F2）基础上,进一步测定了其毒力及在仔猪肠道内的繁殖能力。选择30日龄的ICR小鼠并随机分成4组,A、B、C试验组分别灌胃接种不同浓度的野生菌株S451521及其毒素敲除菌株F1、F2,D组以灭菌生理盐水作阴性对照;结果表明减毒菌株F1和F2毒力均比野生菌株弱,其中F1株减弱了60.73%、F2株减弱了84.38%。选择24日龄未经仔猪水肿病疫苗免疫过的仔猪共14头,并随机分为2组;第1组口服接种减毒菌株F2,第2组以灭菌生理盐水作阴性对照,定期采集仔猪粪样并进行细菌监测;结果显示,仔猪排出减毒菌株F2的数量随时间不断增多,在接种后28d可达到每克粪样含目的菌约70 000cfu。这说明减毒菌株F2能够在仔猪体内定植并大量繁殖。     

新城疫病毒作为疫苗载体的研究进展

在综述新城疫病毒(NDV) 的分子生物学特征、致病本质、复制机理以及利用反向遗传操作技术获得NDV的感染性克隆的基础上,着重介绍了以NDV作载体,将报告基因、功能基因包括鸡传染性法氏囊病毒VP2基因及流感病毒HA基因等插入NDV基因组的不同位置构建重组新城疫病毒,并对外源基因进行成功表达的研究进展。同时对DNV作为新的疫苗载体表达外源基因的可行性及未来应用前景进行了初探。     

Parent‐of‐origin effects on carcass traits in Japanese Black cattle

Variances caused by the differential expression of paternally and maternally imprinted genes controlling carcass traits in Japanese Black cattle were estimated in this study. Data on marbling score (BMS), carcass weight, rib thickness, rib‐eye area (REA) and subcutaneous fat thickness (SFT) were collected from a total of 13,115 feedlot steers and heifers in a commercial population. A sire–maternal grandsire model was used to analyse the data, and then, imprinting parameters were derived by replacing the genetic effect of the dam with the effect of the maternal grandsire in the imprinting model to calculate the genetic parameter estimates. The proportions of the total genetic variance attributable to imprinted genes ranged from 8.7% (SFT) to 35.2% (BMS). The remarkably large imprinting variance of BMS was mainly contributed by maternally expressed inheritance because the maternal contribution of the trait was much larger than that of the paternal trait. The parent‐of‐origin effect originating from maternal gene expression was also observed for REA. The results suggested the existence of genomic imprinting effects on the traits of the Japanese Black cattle. Hence, the parent‐of‐origin effect should be considered for the genetic evaluation of Japanese Black cattle in breeding programmes.