Functional Analysis of ABC Transporter Genes From Botrytis cinerea Identifies BcatrB as a Transporter of Eugenol

The role of multiple ATP-binding cassette (ABC) and major facilitator superfamily (MFS) transporter genes from the plant pathogenic fungus Botrytis cinerea in protection against natural fungitoxic compounds was studied by expression analysis and phenotyping of gene-replacement mutants. The expression of 11 ABC (BcatrA–BcatrK) and three MFS genes (Bcmfs1, Bcmfs2 and Bcmfs4) was studied. All genes showed a low basal level of expression, but were differentially induced by treatment with cycloheximide and the plant defence compounds camptothecin, eugenol, psoralen, resveratrol and rishitin. The latter compounds induced expression of BcatrB at a high level. Eugenol was more toxic to BcatrB gene-replacement mutants than to the control isolates. Eugenol also caused an instantaneous increase in mycelial accumulation of the fungicide fludioxonil, a known substrate of BcatrB. However, there was no difference in virulence between the wild-type and BcatrB gene-replacement mutants on Ocimum basilicum, a plant known to contain eugenol. The results indicate that BcatrB is a transporter of lipophilic compounds, such as eugenol, but its role in virulence remains uncertain.     

Estimation of Genetic Variation Among Verticillium Isolates using AFLP Analysis

Amplified fragment length polymorphism (AFLP) analysis was used to study genetic variation among 76 isolates of Verticillium. A dendrogram based on the AFLP data revealed three main groups. One group consisted of 35 European isolates derived from Brassica napus together with five Californian isolates taken from B. oleracea. This group displayed a high degree of genetic similarity and included three isolates earlier classified as Verticillium longisporum, indicating that all isolates in this group probably should be regarded as members of V. longisporum. V. dahliae isolates constituted the second group while the third group contained four V. albo-atrum isolates. In addition to these three groups, a cluster of six V. nigrescens isolates was observed. However, the genetic distances between the isolates of V. nigrescens were much higher than those between members in the other groups and the bootstrap value for the V. nigrescens cluster was subsequently low. Four isolates classified as V. tricorpus were highly diverse and did not cluster together. Analysis of molecular variance revealed that the isolates of V. longisporum were separated into four subgroups, based on geographic origin. The study furthermore shows that AFLP is a suitable method for studying population structure in Verticillium.     

生物传感器在农药残留分析中的应用

本文简要介绍了几类生物传感器（酶传感器、微生物传感器、免疫传感器及压电传感器）在农药残留分析中的应用进展，并探讨了其发展趋势。     

Restoration of tuber production by exposure to unshaded daylight in Eleocharis kuroguwai Ohwi shaded during the early tuber formation period

The loss of final tuber weight of Eleocharis kuroguwai Ohwi by shading during the early tuber formation period (TFP) is overcome by exposure to unshaded daylight thereafter (late TFP). In the present study, the growth parameters that contribute to the dry matter increase (DMI) per day of tubers in the late TFP were examined. DMI of the tuber during the late TFP was determined by that of the whole plant and the ratio of the DMI of the tuber to that of the whole plant during this period. The ratio of the DMI of the tuber to that of the whole plant during the late TFP was significantly correlated with the DMI of the whole plant during the first 14 days of the late TFP. During the late TFP after the exposure to unshaded daylight, DMI of the whole plant correlated with the surface area of the stem (SAS) and net assimilation ratio (NAR), and the SAS correlated with the stem dry weight (DW) and specific stem-surface area (SSA). SSA negatively influenced NAR, but NAR was increased by unshading. During the late TFP after shading, the effect of the decrease of the stem DW due to shading on the DMI of the whole plant was mitigated by the large SAS and high NAR. These results indicate that the growth parameters that contribute to the DMI of tuber during the late TFP after exposure to unshaded daylight are SAS and NAR just after unshading, and SSA during this period.     

棉花黄萎病菌致病型的AFLP分析

 选用41个棉花黄萎病菌(Verticillium dahliae)代表菌系,在温室条件下,对4个棉花品种鄂荆1号(感)、中棉所12(耐)、文-5(抗)和唐棉2号(抗)进行致病性测定,结果可将供试菌系分为落叶型与非落叶型2类。选取8对AFLP引物PCR扩增的结果中,统计带型稳定、清晰且有多态性的条带,共169条作系统聚类分析,将上述菌系分为2大类,第一类为非落叶型菌系,包括10个非落叶型菌系和1个过渡菌系;第二类为30个落叶型菌系。根据聚类分析建立树状图,发现菌系与地理来源存在一定的相关性,而依据菌系致病力强弱分类则相关关系不大。选用25对EcoRⅠ和MseⅠ引物组合,对供试的41个V.dahliae进行AFLP扩增,筛选到2对引物E64(GACTGCGTACCAATTCGAC)、M53(GATGAGTCCTGAGTAACCG)和E49(GACTGCGTACCAATTCCAG)、M65(GAT-GAGTCCTGAGTAAGAG),能分别扩增出433bp和110bp2条仅为V.dahliae非落叶型菌系独有的特异片段,可将落叶型与非落叶型菌系分开,这2条特异片段被命名为EM₄₃₃和EM₁₁₀。     

小麦苗枯病菌的ITS分析及PCR检测

 小麦苗枯病菌(Clavibacter fangii,Cf)是引起小麦细菌性苗枯病的病原,本研究用16S~23S rDNA间的内源转录间隔区(internally transcribed spacer,ITS)序列通用引物L1(5'-AGTCGTAACAAGGTAGCCGT-3')和L2(5'-GTGCCAAGGCATCCACC-3')扩增Cf和其它相关细菌的基因组DNA;并对其PCR产物进行回收、克隆和测序,将所获序列和其它已报道的细菌ITS序列进行多重比较后设计出Cf的特异性引物I1(5'-TGCCAAGTCACACTGAGACGA-3')和I2(5'-CAATGATCTACCACCCTCCGA-3')。此引物可以从Cf中扩增出351bp的特异性片段,而其余参试的21个细菌PCR反应结果均为阴性。该方法可以应用于小麦苗枯病菌的快速、可靠检测。此外,本研究对多种植物病原棒形杆菌的ITS序列进行比较研究,发现其具有一定的分类意义。     

新疆美洲斑潜蝇寄主种类及适合度的调查

通过乌鲁木齐、昌吉等地调查，发现新疆美洲斑潜蝇的寄主植物计有17科55种，其中蔬菜28种(变种)、大田作物8种，花卉和草坪植物13种、野生植物6种。以豆科、葫芦科植物受害最重。应用欧氏距离法的系统聚类分析，研究了美洲斑潜蝇对14种常见寄主植物的适合度；将其划分为5个类型，即最优适宜寄主、良好适宜寄主、适宜寄主、稍次适宜寄主和非适宜寄主。为害虫的防治决策提供了科学依据。     

中国栽培平菇的酯酶同工酶分析

采用电泳技术对我国大规模栽培的64个平菇商业菌株进行酯酶同工酶分析。结果表明，我国栽培平菇菌种多样性丰富，64个菌株有44个酶谱类型，不同菌株有各自特有的酯酶同工酶酶谱。聚类分析表明，供试菌株在60％相似水平上分为6大类：第一类包括以白黄侧耳为代表的21个菌株；第二类包括以德国引进菌株为代表的6个未定种名菌株；第三类包括7个白黄侧耳与佛罗里达侧耳杂交的杂交种及4个未定种名菌株；第四类包括以佛罗里达侧耳为代表的5个菌株；第五类包括以糙皮侧耳为代表的14个菌株；第六类包括以肺形侧耳为代表的7个菌株。     

Phylogenetic relationships between rice dwarf phytoreovirus isolates from five countries

Rice dwarf virus isolates were collected from several locations in Japan, the Philippines, China, Nepal and Korea. Genomic dsRNA segment profiles in polyacrylamide gel electrophoresis differed among the isolates. There were less differences in the profiles between isolates from Japan and Korea than in those between these two Countries and others. Nucleic acid hybridization was used to examine the extent of genomic variation. Full-length cDNAs to all genomic segments encoding non-structural proteins (S4, S6, S9, S10, S11 and S12) were synthesized from two Japanese isolates, and were used for dot-blot hybridization. Hybridizations using probes generated from the full-length cDNA clones failed to differentiate isolates from different geographical areas. However, cDNA probes covering a variable region of S12 were able to distinguish Japanese and Korean isolates from those of other countries. Phylogenetic tree analysis based on the amino acid sequence of P12 encoded by S12 grouped Japanese and Korean isolates together. The Chinese isolates from two different locations (Yunnan and Fujian) were closely related to each other, and were the most distantly related to Japanese and Korean isolates.     

Biological and molecular characterization of Tomato spotted wilt Virus in Israel

Received April 24, 1997; received in final form June 29, 1997. Symptoms resembling tomato spotted wilt virus (TSWV) infections were documented among ornamental and vegetable crops in commercial greenhouses and open fields in Israel. Plants exhibiting these symptoms were collected from January 1992 to December 1996. Among cultivated plants analyzed for TSWV by enzyme-linked immunosorbent assay (ELISA), 19 species representing five families were found to be infected; natural infection was also recorded in six plant species of weeds. Virus identity was characterized by host range, serology and electron microscopy. Serological reaction with the isolates, found in Israel, using antisera from different sources as well as the sequence analysis of the nucleocapsid gene, demonstrated that the Israeli isolates of TSWV are a member of tospovirus serogroup I, type I (BR-01 strain). No virus transmission was found in seeds collected from virus-infected vegetable and ornamental crops. A non-radioactive molecular probe derived from the cloned nucleocapsid isolate enables specific detection of the virus in crude sap from infected plants. The detection of TSWV in Israel constitutes a severe potential threat to the ornamental and vegetable industry.