6种中草药提取物对烟草黑胫病菌的抑制作用

采用生长速率法测定了A(生物碱类)、B(苷类)、C(挥发油类)、D(糖类)、E(黄酮类)、F(香豆素类)6种中草药提取物对烟草黑胫病菌的抑制效果,结果表明:6种中草药提取物对烟草黑胫病菌均有明显的抑制作用,且随使用剂量的增大而增强。A药和B药分别在使用剂量为5 mL/L和9mL/L时抑制率大于50%,与对照化学药剂72%甲霜锰锌3 000倍液和中生菌素500倍液抑制率无明显差异。C药在使用剂量为80 mL/L时,抑制率达到了70.39%,明显高于对照药剂。D药、E药、F药在使用剂量为80 mL/L、8 mL/L、12.5 mL/L时,抑制率分别达到99.22%、86.64%、79.64%,显著高于对照药剂(P<0.05),其中,D药可完全抑制烟草黑胫病菌的菌丝生长。将6种中草药提取物对烟草黑胫病菌的毒力进行比较,E药的有效抑制中浓度(EC50)最小,为2.65mL/L,其后依次为A药、F药、B药、C药、D药,EC50分别为4.56、4.76、5.99、16.55、17.66 mL/L。6种中草药提取物作为防治烟草黑胫病的抑菌活性物值得进一步研究。     

银法利和烯酰吗啉对烟草黑胫病菌的毒力测定及田间防治试验

采用菌丝生长速率法测定了银法利(氟吡菌胺+霜霉威盐酸盐)、烯酰吗啉对7株来自广西不同烟区、对甲霜灵敏感性不同的烟草黑胫病菌的毒力,结果表明银法利、烯酰吗啉对7个烟草黑胫病菌株均具有很好的抑制作用,EC50值范围分别为1.1090～1.9661、0.6218～1.0625 mg/L,两者对甲霜灵敏感型、一般耐药型、耐药型的菌株的毒力无明显差异,说明它们与甲霜灵没有交互抗性.盆栽试验表明银法利和烯酰吗啉对耐甲霜灵菌株引起的黑胫病有良好的防治效果.2008 ～2010年的田间防治试验结果显示,68.75％银法利悬浮剂和50％烯酰吗啉可湿性粉剂对烟草黑胫病具有很好的防效,显著优于常用药剂68％精甲霜灵·代森锰锌水分散粒剂的防效.     

不同抗性板栗品种的防御酶系对栗疫菌Cp-毒素的响应

对照.此外,抗病品种酶活性高于中抗品种和感病品种;防御酶的活性与品种抗病性之间呈正相关,毒素作用后防御酶活性的变化可作为衡量板栗抗病性强弱的指标.     

Chestnut blight fungus in Croatia: diversity of vegetative compatibility types,mating types and genetic variability of associated Cryphonectria hypovirus 1

In order to improve understanding of its diversity, 338 isolates of Cryphonectria parasitica, the causal agent of chestnut blight, were sampled from 10 chestnut populations throughout chestnut‐growing coastal and continental areas of Croatia. Eighteen vegetative compatibility (VC) types were identified. The VC type EU‐1 was the most widespread, comprising 42·9% of the isolates, followed by EU‐2 (21%) and EU‐12 (14·2%). In respect to the occurrence of the main VC types, the C. parasitica populations in Croatia combined features of both northwestern and southeastern European populations. Perithecia and mating‐type ratios of approximately 1 : 1 were found in all populations, suggesting that sexual reproduction of the fungus is common in Croatia. Natural hypovirulence was also evident in all populations, with incidence of hypovirus‐infected isolates ranging from 12·7% in Istria‐Buje to 66·6% in the continental part of the country. A total of 36 hypovirus‐infected isolates sampled throughout Croatia were analysed in ORF‐A and ORF‐B by RT‐PCR/RFLP analysis. All viral isolates belonged to the Italian subtype of Cryphonectria hypovirus 1 (CHV‐1) and were closely related to the isolates found in other European countries. The RFLP patterns found were also identical or similar to the patterns of three isolates collected in Croatia 22 years ago, suggesting a slow evolution of the hypovirus.     

Characterization of mutants of the chestnut blight fungus (<Emphasis Type="Italic">Cryphonectria parasitica</Emphasis>) with unusual hypovirus symptoms

The type virus of the family Hypoviridae, Cryphonectria hypovirus 1 strain EP713 (CHV1-EP713), infects Cryphonectria parasitica, the filamentous causal fungus of chestnut blight, and reduces its virulence. This pathosystem serves as a model to study fungus-mycovirus interactions. We previously developed a genetic screening protocol for host factors associated with symptom induction by CHV1-EP713 and its mutants. In the procedure the standard field fungal isolate EP155 was transformed by cDNA from a mild hypovirus mutant Cys(72), launching virus infection, and mutagenized by random plasmid insertion with pHygR conferring hygromycin resistance. We now report an extension of the study to characterize different mutant strains, with different phenotypes than their parental strain TCys(72)-1. TCys(72)-1 is moderately reduced in pigmentation and sporulation compared to the uninfected wild-type strain EP155. Mutants sfb1, sfb2 and k202 were characterized biologically and molecularly in comparison to the previously isolated mutant (namA) and the parental strain. These mutants harbored one (sfb1) or more copies (sfb2 and k202) of the mutagenic plasmid, pHygR. The three mutants had similar biological attributes; that is, vegetative growth rate, conidiation and virulence (assay on apples) was reduced on potato dextrose agar media, relative to the parental strain and pigmentation was the same or slightly increased. Interestingly, viral dsRNA accumulation levels were apparently unaltered in these mutants. The screening method was efficient for mining fungal mutants with unusual hypovirus symptoms. Further, characterization of the mutants provides interesting insights into symptom induction by the hypovirus.     

西兰花花梗褐心病研究

西兰花花梗褐心病的主要症状为病花梗表面和内部有小褐斑,髓部水渍状并可扩展到花球主轴髓部,花球中部分花梗和花蕾的发育受阻。病花球保湿2～3d后,其切口可形成大量白色霜霉状物。经鉴定,该病害是由寄生霜霉(Peronospora parasitica)引起的新症状,病菌主要在幼蕾期侵染。在西兰花幼蕾期喷施58%甲霜·锰锌可湿性粉剂500倍液有很好的防治效果。     

适用于板栗疫病菌分子相互作用研究的双分子荧光互补系统

双分子荧光互补技术近年来已广泛应用于研究体内蛋白质相互作用。为建立适合于板栗疫病菌的体内蛋白质相互作用检测体系,本研究以质粒pCPXHY2和pCPXG418为骨架,构建了由板栗疫病菌pgd启动子控制的、以增强型黄色荧光蛋白EYFP为报告基因和以潮霉素和G418抗性为选择标记的双质粒系统pCPXHY2N155和pCPXG418C156。质粒pCPXHY2N155携带EYFP的1~155位氨基酸残基的编码序列,pCPXG418C156携带EYFP基因的156~239位氨基酸残基的编码序列。为验证该质粒系统的有效性,将板栗疫病菌异质三联体G蛋白的β和γ亚基基因分别与pCPXHY2N155上的N155和pCPXG418C156上的C156相连,获得重组质粒pCPXHY2N155β和pCPXG418C156γ。将这两个重组质粒共转化板栗疫病菌野生型菌株EP155,在转化株中观察到明显的黄色荧光,表明Gβ和Gγ在细胞中发生了相互作用。本研究所构建的双分子荧光互补系统,为今后研究板栗疫病菌分子相互作用提供了新的技术手段。     

烟草黑胫病发生动态与黑胫病菌全基因组(DNA)遗传分化关系的研究

 为了检测烟草黑胫病发生动态与寄生疫霉 (Phytophthora parasitica var. nicotianae )全基因组 (DNA )遗传分化的关系 ,从 12 0个 RAPD(Random Am plified Polym orphic DNAs)随机引物中筛选出启动性强的 14个引物 ,借助于 PCR技术对不同年份同一烟草品种上分离获得的 30个菌株 (10个 /年 )和 5个不同烟草品种上分离获得的 15个菌株 (3个 /品种 )进行全基因组 DNA扩增片段指纹图谱及其遗传分化分析。利用 UPGMA(Unweigthted Pair- groupMethod with Arithm etic Average)软件对受试两组菌株的遗传分化构建遗传进化树 ,结果表明大多数不同年份烟草黑胫病菌株的遗传分化与黑胫病发生危害程度成正相关 ,不同烟草品种上的黑胫病菌株遗传分化与其寄主品种无相关性     

烟草黑胫病菌对不同杀菌剂的抗药性初探（英文）

为了解烟草黑胫病菌对不同杀菌剂产生的抗药性,在室内培养条件下,采用菌落生长测定法,对从黔东南州镇远县分离的烟草黑胫病菌进行了抗性测定。结果表明:烟草黑胫病菌对不同杀菌剂均存在不同程度的抗药性,其中对烯酰吗啉的抗药性最强,EC50值为1.19μg/ml。因此,在镇远烟区,长期单一使用烯酰吗啉防治黑胫病存在抗性风险。