Identification of Pathogenicity Defective Mutants from a T-DNA Insertion Mutant Library of Verticillium dahliae     

Cui Weiye  Zhou Lei  Shan Liuying  Zhang Jun  Dai Xiaofeng  Guo Wei 《棉花学报》2018,30(1):29-37

[Objective] Our research aimed to identify pathogenicity defective mutants from a T-DNA insertion mutant library of Verticillium dahliae strain Vd991 and to analyze pathogenicity-related genes. [Method] In total, 294 T-DNA insertion mutants of V. dahliae were tested for their virulence using a cotton infection assay. Southern blot assays were performed to identify the T-DNA insertion copy number of each pathogenicity defective mutant. DNA sequences flanking the T-DNA insertional sites of each mutant were analyzed by high-efficiency thermal asymmetric interlaced PCR. [Result] Based on the virulence assay, the disease indices of cotton plants inoculated with each mutant decreased very significantly in comparison with the index of those inoculated with Vd991. The Southern blot assay revealed that only one mutant contained two T-DNA insertions, while the remaining eight mutants harbored a single T-DNA insert. An analysis of biological characteristics found that the growth and conidial production of these mutants were impaired by the T-DNA insertions compared with the wild type Vd991. The T-DNAs' insertion position and distribution in each mutant were identified by comparison with genome sequences of strain VdLs.17. Furthermore, the pathogenicity-related genes were cloned from strain Vd991. [Conclusion] The screening and identification of T-DNA insertion mutants is an effective method to identify the pathogenicity-related genes of V. dahliae on a genome-wide scale. This laid a foundation for the further breeding of disease-resistant cotton varieties and will promote the study of the pathogenic molecular mechanisms of V. dahliae.  相似文献   

Identification of genes involved in gonadal sex differentiation and the dimorphic expression pattern in <Emphasis Type="Italic">Takifugu rubripes</Emphasis> gonad at the early stage of sex differentiation     

Hongwei Yan  Xufang Shen  Xin Cui  Yumeng Wu  Lianshun Wang  Lei Zhang  Qi Liu  Yusheng Jiang 《Fish physiology and biochemistry》2018,44(5):1275-1290

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四种锌源对珍珠龙胆幼鱼生长性能、抗氧化能力以及矿物元素沉积的影响     

殷彬  林仪彤  迟淑艳  谭北平  姚红梅  姚亚军  董晓慧  杨奇慧  刘泓宇  章双 《水产学报》2018,42(7):1111-1123

为研究硫酸锌(ZnSO_4·7H_2O)、碱式硫酸锌[ZnSO_4·3Zn(OH)2·5H_2O]、甘氨酸锌(C_4H_8N_2O_4Zn)和羟基蛋氨酸锌(C_5H_(11)NO_6S_2Zn)对珍珠龙胆生长性能、肝脏抗氧化能力、肠道发育以及脊椎骨矿物元素沉积的影响,实验选取体质量为(11.00±0.12)g珍珠龙胆360尾,随机分为4组(每组3个重复,每个重复30尾),分别添加ZnSO_4·7H_2O、ZnSO_4·3Zn(OH)2·5H_2O、C4H8N2O4Zn和C5H11NO6S2Zn,记作ZnSO_4、Zn_4(OH)_6SO_4、Zn-Gly和Zn-MHA,使饲料锌水平分别为118.85、116.92、120.66、119.23 mg/kg,制成等氮等脂的实验饲料,饲养8周。结果显示,Zn_4(OH)_6SO_4、Zn-Gly和Zn-MHA组增重率和特定生长率均显著高于ZnSO_4组。Zn_4(OH)_6SO_4、Zn-Gly、Zn-MHA组饲料系数均显著低于ZnSO_4组。Zn-Gly、ZnMHA组肝脏丙二醛含量显著低于ZnSO_4和Zn_4(OH)_6SO_4组,Zn-MHA组Cu Zn-SOD和TSOD活性均显著高于其余各组。与ZnSO_4相比,Zn_4(OH)_6SO_4、Zn-Gly和Zn-MHA均显著提高了前肠和中肠的皱襞厚度;Zn-Gly和Zn-MHA组后肠肌层显著厚于ZnSO_4和Zn_4(OH)_6SO_4组。Zn-Gly组珍珠龙胆脊椎骨Fe和Mn含量显著高于ZnSO_4组。相对于ZnSO_4,以增重率和肝脏CuZn-SOD含量为评价指标时,Zn_4(OH)_6SO_4、Zn-Gly和ZnMHA的相对生物学效价(RBV)分别为119.15%、110.90%,118.43%、113.58%和122.45%、130.84%。研究表明,与ZnSO_4相比,Zn_4(OH)_6SO_4能够显著提高珍珠龙胆幼鱼的生长性能,提高肝脏SOD活性,增强脊椎骨中Fe的沉积;Zn-Gly和Zn-MHA能够显著提高珍珠龙胆幼鱼的生长性能,促进肠道发育,调节脊椎骨矿物元素沉积。  相似文献   

调控东亚飞蝗体温的主要环境因子分析     

刘银民  阳积文  范春斌  尚素琴  Bryony Taylor  李红梅 《植物保护学报》2018,45(6):1296-1301

为明确东亚飞蝗Locusta migratoria manilensis体温在常年发生区与环境因子之间的关系,于2015—2016年采用直接抓取和网捕方法在天津市北大港蝗区捕捉东亚飞蝗的秋蝗和夏蝗,测量其瞬时体温以及周围4类8个环境指标,应用因子分析法评价东亚飞蝗体温与环境因子的关系。结果表明,东亚飞蝗白天体温比夜间高,平均体温高于26.5℃;且体温在白天出现2次高峰,分别是上午11点左右的49.0℃和下午14点左右的47.2℃。无论是夏蝗还是秋蝗,其体温均随着龄期增加呈现上升趋势,2～5龄蝗蝻、成虫的平均体温依次为36.9、37.0、37.6、37.2、38.0℃。因子分析结果表明,植被冠层温度是影响飞蝗体温的关键因子,飞蝗所处地面温度对其体温影响最大;风速和植株高度是影响飞蝗体温的重要分因子;植株高度与体温成负相关。表明植被冠层温度与东亚飞蝗的体温变化最为密切。  相似文献   

Baseline sensitivity of <Emphasis Type="Italic">Phytophthora capsici</Emphasis> to the strobilurin fungicide benzothiostrobin and the efficacy of this fungicide     

Dicheng Ma  Jiamei Zhu  Leiming He  Kaidi Cui  Wei Mu  Feng Liu 《European journal of plant pathology / European Foundation for Plant Pathology》2018,151(3):723-734

Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) has seriously threatened global banana production. Recently, identification of Foc TR4 in the Caribbean and Mozambique has aroused widespread concern because it may be directly related to food security for millions of people in Africa and Latin America. To identify a resistant or highly resistant germplasm, 129 accessions were evaluated for Foc TR4 resistance in greenhouses and 100 of them were also assessed in the field. In total, 10 accessions were highly resistant (HR) to Foc TR4; these include four from the AA group, two from the BB group, one of the East African highland bananas (EAHBs), two plantains, and one from wild relatives of banana. Of these, Pahang (AA), Calcutta 4 (AA), and Musa itinerans exhibited the highest degree of resistance with an index of disease (ID2) less than 10, and Foc TR4 could not infect any of them in the field. Furthermore, 31 accessions from AA, AB, AAB, AAAB, and ABB groups and wild relatives were identified as a resistant (R) germplasms. All tested EAHBs and plantains exhibited resistance or high resistance to Foc TR4 infection. Our results provide valuable genetic resources for banana breeding and for studying the mechanisms underlying Fusarium wilt resistance. Furthermore, evaluation of EAHBs and plantains provided a rational for local banana producers and researchers to grow EAHBS and plantains.  相似文献   

基于体细胞胚发生的细叶百合和兰州百合多倍体诱导及鉴定     

孙红梅  付麟岚  王志平  盖美竹  王春夏 《园艺学报》2018,45(6):1136-1146

以细叶百合（Lilium pumilum DC. Fisch.）和兰州百合（Lilium davidii var. unicolor）的鳞片和胚性愈伤组织作为诱导材料,用不同浓度秋水仙素以浸泡和混培两种处理方式进行多倍体诱导。以形态学观测、根尖染色体计数、气孔观测对多倍体植株进行倍性鉴定,分别获得了19株细叶百合四倍体植株和6株兰州百合四倍体植株,最佳处理方式为0.1%秋水仙素浸泡胚性愈伤组织24 h。使用ISSR对经体细胞胚发生的二倍体和多倍体再生植株进行遗传分析,发现两种百合二倍体植株遗传较稳定,而多倍体植株均发生遗传变异,其中细叶百合和兰州百合遗传变异率分别为15.48%和9.75%。  相似文献   

菊花抗白色锈病相关基因CmDREBa-2的克隆及表达分析     

熊超明  赵兴华  贾红梅  延昕  毛洪玉 《园艺学报》2018,45(5):977-987

以菊花（Chrysanthemum morifolium）免疫白色锈病品种‘C029’为试验材料,基于白色锈病病原菌诱导的菊花转录组数据库,采用RACE（rapid-amplification of cDNA ends）方法克隆得到菊花DREB（Dehydration responsive element binding protein）基因,命名为CmDREBa-2。对其进行生物信息学分析和表达分析,结果表明：CmDREBa-2开放阅读框（ORF）全长291 bp,编码96个氨基酸;预测CmDREBa-2蛋白相对分子质量为11 159.04,pI为11.34,是一个定位于细胞核、亲水的不稳定蛋白;氨基酸序列和结构分析显示该蛋白含有1个由49个氨基酸残基组成的AP2保守结构域,所以该蛋白属于AP2/EREBP家族;进化树分析表明CmDREBa-2属于DREB家族的A-1组,CmDREBa-2蛋白与菊花的其他两个DREB类蛋白CmDREBa和CmDREBb的同源性较高;菊花‘C029’接种白色锈病病原菌6 h后,CmDREBa-2表达量达到最高,约为未接菌对照的34倍;用水杨酸（SA）、茉莉酸甲酯（MeJA）和乙烯利（ETH）处理‘C029’菊花叶片,结果表明CmDREBa-2受3种激素的诱导表达。  相似文献   

Highly specific assays to detect isolates of Pseudomonas syringae pv. actinidiae biovar 3 and Pseudomonas syringae pv. actinidifoliorum directly from plant material          下载免费PDF全文

M. T. Andersen  M. D. Templeton  J. Rees‐George  J. L. Vanneste  D. A. Cornish  J. Yu  W. Cui  T. J. Braggins  K. Babu  J. F. Mackay  E. H. A. Rikkerink 《Plant pathology》2018,67(5):1220-1230

Pseudomonas syringae pv. actinidiae (Psa) is responsible for bacterial canker of kiwifruit. Biovar 3 of Psa (Psa3) has been causing widespread damage to yellow‐ and green‐fleshed kiwifruit (Actinidia spp.) cultivars in all the major kiwifruit‐producing countries in the world. In some areas, including New Zealand, P. syringae pv. actinidifoliorum (Pfm), another bacterial pathogen of kiwifruit, was initially classified as a low virulence biovar of Psa. Ability to rapidly distinguish between these pathovars is vital to the management of bacterial canker. Whole genome sequencing (WGS) data were used to develop PCR assays to specifically detect Psa3 and Pfm from field‐collected material without the need to culture bacteria. Genomic data from 36 strains of Psa, Pfm or related isolates enabled identification of areas of genomic variation suitable for primer design. The developed assays were tested on 147 non‐target bacterial species including strains likely to be found in kiwifruit orchards. A number of assays did not proceed because although they were able to discriminate between the different Psa biovars and Pfm, they also produced amplicons from other unrelated bacteria. This could have resulted in false positives from environmental samples, and demonstrates the care that is required when applying assays devised for pure cultures to field‐collected samples. The strategy described here for developing assays for distinguishing strains of closely related pathogens could be applied to other diseases with characteristics similar to Psa.  相似文献   

Artificial diet based investigation on the impact of purified Cry1Ac,Cry1Fa and Cry2Ab on the survival and reproductive performance of adult green lacewing, <Emphasis Type="Italic">Chrysopa pallens</Emphasis> (Rambur) (Neuroptera: Chrysopidae)     

Intazar Ali  Zhang Shuai  Asif Sajjad  Hafiz Mahmood ur Rehman  Cui Jin-Jie 《Phytoparasitica》2018,46(1):127-135

The green lacewing Chrysopa pallens (Rambur) (Neuroptera: Chrysopidae) is a common and abundant predator in many cropping systems in palearctic realm and it’s conservation is helpful in sustainable pest management in agro-ecosystem. Prior to commercialization of Bt crops in any agro- ecosystem, it is necessary to evaluate the impact of Cry proteins upon non-target organisms especially biological control agents (BCA). In present study an artificial diet consisting of shrimp, beef, beef liver and egg yolk was developed to mass-rear C. pallens for its use as biological control agents in sustainable pest management. Moreover, an artificial diet based risk assessment protocol was developed to investigate the impact of Cry1Ac, Cry1Fa and Cry2Ab on the survival and reproductive performance of C. pallens adults. C. pallens was fed on diets incorporated with Cry proteins and without addition of Cry proteins (control). The same diet containing boric acid was served as a positive control. Temporal stability, bioactivity and intake of Cry proteins by C. pallens were confirmed using double-antibody sandwich, enzyme-linked immunosorbent assay and bioactivity verification bioassays. Survival and reproductive performance of C. pallens, e.g., pre-oviposition period, daily fecundity, total fecundity and 30-day old adults dry weights, exhibited non-significant differences (p?>?0.05) for the diets containing Cry1Ac, Cry1Fa and Cry2Ab (50 μg/g) against Control. However, significant reduction in survival and reproductive performance (p?<?0.05) was observed in positive control. Our findings reveal that artificial diet is a good source of nutritional requirement with enhanced survival and reproductive performance of C. pallens and can be used for mass rearing of predator in case of natural diet scarcity and Cry proteins are safe for adult C. pallens and Bt crops cultivation help in predators conservation in sustainable agriculture.  相似文献   

金龟子绿僵菌在草原羊草和克氏针茅根际的种群动态和根内宿存鉴定     

蔡霓  王峰  农向群  李红梅  王广君  黄训兵  崔伯阳  王露  涂雄兵  张泽华 《植物保护》2018,44(6):32-37

绿僵菌防治草原蝗虫效果显著,其在草原田间的存活能力影响其持续控害效果。除了侵染昆虫,绿僵菌还具有在植物根际宿存和根内生的生活方式,但相关宿存规律和内生性的研究报道很少。本文研究了绿僵菌在内蒙古草原两种优势种植物羊草和克氏针茅根际的种群数量变化并对其在这两种草的根内宿存进行了鉴定。结果表明,在干旱的内蒙古草原,绿僵菌施用后种群数量在30 d内快速下降,但能够以低密度在羊草、克氏针茅根际土壤中至少延续宿存75 d,羊草根际环境比较利于绿僵菌生存。对菌株egfp基因标记的特异PCR检测证明了绿僵菌在羊草和克氏针茅根内宿存。试验数据为指导植保生物防治中充分利用绿僵菌的昆虫病原性和植物内生特性提供理论基础,也将成为绿僵菌物种生活方式多样性、与植物互作及共进化研究的重要基础。  相似文献