An unusual localization of developmental stages of Cryptosporidium parvum Tyzzer, 1912 in the cells of small intestine of a gnotobiotic piglet

In a gnotobiotic piglet infected with 50.10(6) oocysts of Cryptosporidium parvum, developmental stages of the parasite were found in the duodenum, anterior jejunum and middle jejunum on the first day post infection (1 DPI). During the studies of the ultrastructure, trophozoites and meronts of C. parvum were encountered in the microvillous zone of enterocytes, and the outer membrane of parasites was a continuation of the membrane of microvilli. Unusual was the attachment of C. parvum trophozoite on the opening of a goblet cell, as well as a meront localized in the cytoplasm in the enterocyte of anterior jejunum. These findings show that C. parvum is very adaptive and capable of developing in a still undescribed location.     

利用酵母双杂交筛选与小麦黄花叶病毒P1互作的寄主蛋白

正小麦黄花叶病是我国冬小麦上重要的病毒病害之一,自20世纪90年代以来,每年发生面积都在66万hm~2以上,一般减产10%~30%,严重的达70%以上。该病害在我国河南、江苏、山东、安徽、陕西、湖北、四川等省均有分布,并且呈逐年蔓延趋     

Temporal Variation in Setosphaeria turcica Between 1974 and 1994 and Origin of Races 1, 23, and 23N in the United States

ABSTRACT Setosphaeria turcica causes northern leaf blight, an economically important disease of maize throughout the world. Survey collections of S. turcica isolates from 1974 to 1994 provided a unique opportunity to examine temporal diversity in the eastern United States. Two hundred forty-two isolates of S. turcica from maize were studied with random amplified polymorphic DNA (RAPD) markers, mating type, and virulence on maize differential inbred lines with known Ht resistance genes to examine changes over time. One hundred forty-nine RAPD haplotypes were identified. Nearly 20% of haplotypes recurred in more than one year. Race 0 isolates declined in frequency from 83% in 1974 to near 50% in the 1990s, most likely in response to the widespread deployment of Ht1 in commercial maize hybrids. Races 23 and 23N were present in the collection at low levels throughout the study period and were also found among isolates from Virginia in 1957. The frequency of MAT1-2 isolates increased sharply after 1979 and was associated with the emergence of race 1 during the same period. RAPD markers were used to investigate the genetic diversity among a subset of isolates collected in the United States from 1976 to 1982, the period in which this dramatic shift in race frequency occurred. Multilocus haplotypes were not exclusively associated with known races of S. turcica. Based on shared haplotypes and cluster analysis, race 1 isolates share greater similarity with race 0 than with 23 or 23N isolates, indicating race 1 probably evolved from multiple lineages of race 0. Sorghum spp.-infecting isolates share greater similarity with one another than with maize-infecting isolates and represent a distinct subgroup.     

利用酵母双杂交系统筛选与草莓镶脉病毒移动蛋白P1互作的寄主因子

 构建感染草莓镶脉病毒(SVBV)森林草莓的酵母cDNA文库,利用酵母双杂交系统,筛选出与SVBV P1蛋白互作的15种寄主因子。生物信息学分析发现,这15种寄主因子参与茉莉酸途径、泛素化、光合作用、抗病抗逆、蛋白修饰、蛋白运输和氧化还原等多种生物过程。另外,这些寄主因子还具有其他分子功能,包括氧化还原酶活性、蛋白二硫化物异构酶活性和金属离子结合活性等。本研究初步探讨了P1与寄主因子的互作机理,为揭示SVBV侵染森林草莓以及SVBV在寄主中扩展的分子机制提供理论依据。     

苹果树腐烂病菌拮抗放线菌JPD-1的筛选及鉴定

为筛选对苹果树腐烂病菌Valsa mali具有拮抗作用的放线菌,从苹果树根际土壤中分离获得放线菌,利用平板对峙法和生长速率法筛选拮抗活性较高的菌株,通过菌株培养特征、形态特征、生理生化特性及16S rDNA序列分析确定其分类地位,并采用离体枝条烫伤接种法测定所筛选获得的拮抗菌株对苹果树腐烂病的防效。结果表明,从苹果树根际土壤中共分离获得28株放线菌,其中菌株JPD-1对苹果树腐烂病菌的拮抗作用最强,抑制率为72.50%,该菌在高氏合成1号培养基上生长最好,培养7 d时菌落生长旺盛,气生菌丝白色,基生菌丝深粉红色,无可溶性色素产生;经形态特征观察、生理生化试验及16S rDNA序列分析,将菌株JPD-1鉴定为公牛链霉菌Streptomyces tau‐ricus。菌株JPD-1发酵滤液对苹果树腐烂病菌的抑制率及对离体枝条上苹果树腐烂病的防效均随稀释倍数的增大而减弱,发酵滤液原液使苹果树腐烂病菌菌丝膨大、末端畸形,抑制率为78.14%,对离体枝条上苹果树腐烂病的防效为76.37%。表明分离到的放线菌菌株JPD-1可以用作苹果树腐烂病的生防材料,具有很好的开发及应用前景。     

高寒草地青藏苔草拮抗内生细菌筛选、鉴定及其促生作用测定

为从青藏苔草Carex moorcroftii内生细菌中开发优势拮抗菌种资源,采用对峙培养法测定其叶片内生细菌对黄瓜枯萎病菌Fusarium oxysporum、马铃薯炭疽病菌Colletotrichum coccodes、孜然根腐病菌F. solani等9种病原真菌的拮抗作用,对所筛选的优良拮抗内生细菌进行形态学及16S rDNA和gyr-B基因序列分析鉴定,并将其经固体发酵制成菌剂,采用生防菌剂接种方法测定其对黄瓜和辣椒的促生作用。结果显示,从青藏苔草叶片中共分离得到14株内生细菌;以黄瓜枯萎病菌作为指示菌初筛得到8株具有拮抗作用的菌株,其中菌株1Y4、1Y5、和1Y7的抑菌率均为57.53%,菌株1Y11的抑菌率为58.44%,均高于平均抑菌率56.73%;菌株1Y4具有广谱抑菌活性,对孜然根腐病和马铃薯炭疽病菌的抑菌率分别达73.33%和71.43%,且具有固氮作用;结合形态学和分子生物学鉴定,确定菌株1Y4为西姆芽胞杆菌Bacillus siamensis;以0.5%1Y4发酵菌剂处理黄瓜和辣椒,株高分别增加153.06%和117.33%,促生效果显著。表明青藏苔草内生细菌1Y4的防病促生作用良好,有开发为微生物菌剂的潜力。     

大豆对灰斑病菌15号小种的抗病基因定位及标记检测

为明确大豆对灰斑病菌15号小种的抗性位点,以大豆抗病品种垦丰16、感病品种绥农10及其杂交F₂、F₃代群体为试验材料,在接种鉴定的基础上,运用SSR标记技术及分离群体组群分析法(BSA法)对垦丰16抗病基因进行了定位,并应用108份大豆新品系对标记进行了符合性检测。结果表明,垦丰16对15号小种的抗性受1对显性基因控制,抗病基因位于大豆染色体组的J连锁群上,将该基因定名为Rcs15。用Mapmaker/Exp 3.0 b进行连锁分析,获得了5个与抗病基因紧密连锁的SSR标记:Satt 529、Satt 431、Sat_151、Satt 547和Sat_224,标记与抗病基因间的排列顺序和遗传距离为Sat_151-10.7 cM-Satt 529-18.5 cM- Rcs15-6.7 cM-Satt 547-7.8 cM-Sat_224-10.7 cM-Satt 431。标记符合性检测结果显示,Satt 547和Sat_224的检测准确率达到85%以上,可用于分子标记辅助选择育种和抗源筛选。     

烟粉虱细胞色素P450基因CYP6JM1的克隆及其在噻虫嗪抗性中的作用

烟粉虱Bemisia tabaci是全球性农业害虫之一, 目前采用的防治手段多为化学防治。噻虫嗪作为一种新烟碱类杀虫剂, 现被广泛应用于田间烟粉虱的防治, 但烟粉虱已对其产生了抗性。本文通过RT-qPCR对烟粉虱噻虫嗪抗性与敏感种群的细胞色素P450基因CYP6JM1表达水平进行了比较, 结果显示, 烟粉虱噻虫嗪抗性种群CYP6JM1基因表达水平明显升高。基因克隆和序列分析表明, CYP6JM1基因具备昆虫P450基因的典型特征。RT-qPCR检测结果表明, 该基因在烟粉虱1~2龄时期及腹部高水平表达。RNAi结果表明, 降低烟粉虱体内CYP6JM1基因的表达量能够显著提高其对噻虫嗪的敏感性, 表明CYP6JM1基因可能与烟粉虱对噻虫嗪的抗性有关。     

水稻稻瘟病拮抗细菌WH1G的筛选鉴定及其抑菌活性

采用平板对峙法,从安徽水稻根际土壤样品中筛选对稻瘟病菌具有较强抑菌活性的拮抗细菌,通过菌落形态观察、生理生化特征、gyrB基因序列分析对该菌株进行鉴定,测定抑菌谱,对其抑菌机制进行初步研究。结果表明,拮抗菌株WH1G对稻瘟病菌具有较强的抑制作用,抑制率达91.79%,经鉴定为解淀粉芽胞杆菌(Bacillus amyloliquefaciens)。该菌可产生蛋白酶和纤维素酶,不产生几丁质酶和β-1,3-葡聚糖酶;用代谢产物无菌滤液处理稻瘟病菌,可造成孢子萌发率降低,菌丝扭曲异常、分支明显减少并产生大量泡囊。该菌对常见的20种植物病原真菌及细菌均具有不同程度的拮抗作用,表现出广谱抗菌活性。     

Modulation of Mouse P450 Isoforms CYP1A2, CYP2B10, CYP2E1, and CYP3A by the Environmental Chemicals Mirex, 2,2-Bis(p-chlorophenyl)-1,1-dichloroethylene, Vinclozolin, and Flutamide

Several environmental chemicals are disruptive to the reproductive and endocrine systems of many species, including humans. Mechanisms for endocrine disruption are presently under scrutiny. Xenobiotic inducible mammalian cytochrome P450 (CYP) enzymes metabolize a variety of substrates including environmental chemicals, pesticides, and drugs. The metabolism, and thus the effect, of endogenous chemicals including steroid hormones, vitamins, etc. that are transformed by CYP enzymes can be influenced by environmental exposure to CYP-inducing chemicals. This study demonstrated that structurally diverse environmental chemicals including mirex, 2,2-Bis(p-chlorophenyl)-1,1-dichloroethylene (DDE), vinclozolin, and flutamide are capable of inducing several mouse liver CYP isozymes. As demonstrated by Western blotting, mirex induced CYP1A2, 2B10, 2E1, and 3A and vinclozolin induced 1A2 and 2B10. The only isoforms significantly induced by DDE and flutamide were 3A and 1A2, respectively. Since some of these isoforms are known to be involved in metabolism of endogenous hormones, we also studied the effects of these CYP inducers on testosterone metabolism and seminal vesicle weights. Mirex and DDE treatments had profound effects on the metabolism of testosterone, resulting in 2.5- to 3-fold more hydroxylated products than controls. Lesser, but significant, increases in specific metabolites of testosterone were also observed following treatment with vinclozolin and flutamide. Seminal vesicle weights were lower for all treatment groups except DDE. Results of this study demonstrate that, due to their CYP-inducing potential, these chemicals may significantly impact testosterone metabolism and this may be a contributing factor in their antiandrogenic effects.     

Two cytochrome P450 genes are involved in imidacloprid resistance in field populations of the whitefly, Bemisia tabaci, in China

The sweet potato whitefly, Bemisia tabaci (Gennadius) (Hemiptera:Aleyrodidae), is an invasive and damaging pest of field crops worldwide. The neonicotinoid insecticide imidacloprid has been widely used to control this pest. We assessed the species composition (B vs. Q), imidacloprid resistance, and association between imidacloprid resistance and the expression of five P450 genes for 14–17 B. tabaci populations in 12 provinces in China. Fifteen of 17 populations contained only B. tabaci Q, and two populations contained both B and Q. Seven of 17 populations exhibited moderate to high resistance to imidacloprid, and eight populations exhibited low resistance to imidacloprid, compared with the most susceptible field WHHB population. In a study of 14 of the populations, resistance level was correlated with the expression of the P450 genes CYP6CM1 and CYP4C64 but not with the expression of CYP6CX1, CYP6CX4, or CYP6DZ7. This study indicates that B. tabaci Q has a wider distribution in China than previously reported. Resistance to imidacloprid in field populations of B. tabaci is associated with the increased expression of two cytochrome P450 genes (CYP6CM1 and CYP4C64).     

云南省元阳县籼/粳型稻瘟病菌无毒基因Avr-Pia、Avr-Pita1和Avr-Pii多样性分析

为明确云南省元阳县籼/粳型稻瘟病菌中无毒基因的多样性及差异,采用接种鉴别品种方法和PCR技术对9株粳型菌株和21株籼型菌株中无毒基因Avr-Pia、Avr-Pita1和Avr-Pii的多样性进行分析。结果显示,30株稻瘟病菌菌株中无毒基因Avr-Pia、Avr-Pita1和Avr-Pii的平均出现频率分别为33.3%、76.7%和0,Avr-Pia、Avr-Pita1在籼型菌株和粳型菌株中的出现频率分别为4.8%和100.0%、66.7%和100.0%。Avr-Pia主要为存在/缺失的多样性,籼/粳型菌株对水稻品种Achiasahi(Pia)的致病率分别为100.0%和11.1%;Avr-Pita1可划分为3个类群,除菌株CH1195属于PO类群外,籼型菌株均属于J1类群,粳型菌株则属于J2/J3类群;籼/粳型菌株对水稻品种K1(Pita)的致病率分别为100.0%和0,且籼/粳型菌株Avr-Pita1蛋白序列在第83位和第192位氨基酸差异明显。Avr-Pii在30株稻瘟病菌菌株中均未检出,籼/粳型菌株对水稻品种Fujisaka No. 5(Pii)的致病率分别为100.0%和0。表明Avr-Pia和Avr-Pita1在粳型菌株中的出现频率高于籼型菌株且变异程度低,粳型菌株中可能存在其它无毒基因使其不能侵染Fujisaka No. 5。     

Distribution of major clonal lineages EU_13_A2, EU_2_A1, and EU_23_A1 of Phytophthora infestans associated with potato late blight across crop seasons and regions in Algeria

Potato is one of the most important agricultural crops in Algeria and worldwide. Each year, potato late blight, caused by Phytophthora infestans, is responsible for significant damage that leads to large production losses, and is thus a direct threat to food security in Algeria. In this study, 131 isolates of P. infestans and 92 DNA fingerprints captured on FTA cards were sampled from commercial and seed production fields in three major potato production regions (western, eastern, and central) during the main-season and late-season in Algeria over six cropping seasons (2010–2016). Genotypes of P. infestans and population genetic diversity were analysed using a 17-plex simple-sequence repeat (SSR) marker assay, and the mating type of all isolates was characterized. Both mating types (A1 and A2) were found, and often occurred in the same field. Differences in mating type proportion were observed between regions and between sampling periods. Analysis with SSR markers showed the prevalence of the EU_13_A2 lineage (70%) over EU_2_A1 (16%), EU_23_A1 (10%), and 4% of unknown multilocus lineage (MLL). The EU_13_A2 showed differentiation within the group. EU_23_A1 was found mainly in late-season crops. However, the cropping region did not influence the distribution of lineages due to the dispersal of the pathogen in Algeria by seeds. Genetic structure did not reveal a clear variation in distribution of the three lineages throughout the sampling regions. These data provide important new information on the composition and change over time of P. infestans populations in Algeria and open the way for a better understanding of the local epidemiology of this important pathogen.     

环塔里木盆地枣树氮磷钾合理配施技术研究

以新疆环塔里木盆地阿克苏红旗坡农场5年生红枣树为研究对象,在全年施肥量（纯N 675 kg·hm^-2、P₂O₅ 540 kg·hm^-2、K₂O 135 kg·hm^-2）一定条件下,通过田间施肥试验,研究氮磷钾不同配施比例对红枣树生理特性及果实品质的影响。结果表明：萌芽前期施纯N、P₂O₅、K₂O肥270、180、67.5 kg·hm^-2,开花期施纯N、P₂O₅肥202.5、180 kg·hm^-2,果实膨大期施纯N、P₂O₅、K₂O肥202.5、180、67.5 kg·hm^-2的综合效果佳,有利于枣树茁壮生长和果实品质的提高。与常规施肥相比,红枣树开花期、幼果期、果实膨大期和成熟期的叶绿素SPAD值增幅分别为6%、4%、5%和11%;各测量时段09∶00、11∶00、13∶00、15∶00、17∶00、19∶00的净光合速率分别提高11%、14%、16%、82%、15%、44%;气孔导度分别提高34%、25%、30%、132%、129%、96%;胞间CO₂浓度分别降低13%、22%、14%、18%、24%、15%;蒸腾速率分别降低50%、45%、28%、24%、33%、52%;果实维生素C、还原糖、蛋白质含量最高,分别为6.41 mg/100g、19.33%和2.53%,差异显著。     

Metabolism of the herbicides chlorotoluron, diuron, linuron, simazine, and atrazine by CYP1A9 and CYP1C1 from Japanese eel (Anguilla japonica)

Through the use of a number of bioconversion experiments we demonstrated that P450 proteins (CYP1A9 and CYP1C1) from Japanese eel (Anguilla japonica) metabolized a number of herbicides and the drug phenacetin. We performed bioconversion experiments in which substrates were added directly to incubation medium. The resulting metabolites were extracted and analyzed by high-performance liquid chromatography. Proteins CYP1A9 and CYP1C1 metabolized 50 nmol of the drug phenacetin to yield 12.1 and 1.1 nmol of product (acetaminophen), respectively. Further incubation of CYP1A9 with 50 nmol of the herbicides chlorotoluron, diuron, linuron, simazine, or atrazine yielded 16.5, 18.5, 7.3, 1.6, or 0.8 nmol of product, respectively. CYP1C1 also metabolized linuron, diuron, and simazine yield 5.4, 4.6, or 0.7 nmol of product, respectively. Next, polyclonal antibody was isolated by immunizing with two conjugated-peptides (amino acid residues 272–290 and 294–310) of CYP1A9. This antibody did not recognize human CYP1A2 or CYP1C1. Western blotting using the antibody revealed one band in the livers of Japanese eel and tilapia (Oreochromis niloticus). Theses results suggest that CYP1A9 and CYP1C1 metabolize herbicides, and that CYP1A9 is an useful biomarker of contamination when detected with this antibody.     

烟粉虱细胞色素CYP6EM1基因的克隆及对吡虫啉抗性的作用

烟粉虱是一种世界性农业害虫,其防治手段以化学防治为主,新烟碱类杀虫剂吡虫啉常年用于防治烟粉虱,田间烟粉虱已经形成严重的抗药性。本研究通过分析烟粉虱吡虫啉抗性和敏感种群,发现细胞色素CYP6EM1基因在吡虫啉抗性品系中上调了4.7倍,进而克隆了其全长基因,进行了荧光定量PCR分析,发现该基因在吡虫啉抗性烟粉虱3龄若虫期和雄虫成虫期过量表达,并且在抗性成虫胸部和腹部过量表达。最后通过RNA干扰的方法使成虫的CYP6EM1基因表达量下降了54.8%,之后发现当烟粉虱暴露于吡虫啉时死亡率显著升高了39.65%,这表明CYP6EM1与烟粉虱对吡虫啉抗性的形成相关。研究结果对于揭示烟粉虱对吡虫啉产生抗性的机制有帮助,也为烟粉虱抗性水平田间监测及烟粉虱综合治理提供理论依据。     

转cry1Ab/1Ac基因水稻对野生褐家鼠Rattus norvegicus取食、体重及器官重量的影响

为评价转Bt基因水稻对野生啮齿类动物的影响,以野生褐家鼠Rattus norvegicus为试验对象,分别饲喂转cry1Ab/1Ac基因水稻华恢1号(HH1)及其亲本非转基因水稻明恢63(MH63),通过取食选择试验及110 d连续饲喂试验,测定转基因水稻对褐家鼠取食量、体重及器官的影响。结果显示,野生褐家鼠雌鼠日平均取食MH63和HH1分别为5.52 g和5.37 g,雄鼠日平均取食分别为7.00 g和6.52 g,在选择取食MH63和HH1稻谷上无显著差异;在连续饲喂试验的不同时期,褐家鼠体重及每日取食稻谷量在对照组与试验组间均无显著差异;饲喂试验结束后解剖观察,褐家鼠脾脏、肾脏、肾上腺,雌性卵巢和子宫以及雄性睾丸、附睾和储精囊的绝对重量及相对重量在对照组与试验组间均无显著差异。研究表明,与MH63相比,转cry1Ab/1Ac基因水稻对野生褐家鼠的取食、体重以及主要器官均无显著性影响。