不同辣椒炭疽病菌对唑菌酯的敏感性差异

辣椒炭疽病是辣椒生产上的重要病害,严重制约辣椒生产,唑菌酯防治辣椒炭疽病的相关研究未见报道。为此,本研究利用菌丝生长速率法测定了4种不同炭疽病菌对唑菌酯的敏感性,并对其作用靶标基因细胞色素b基因(Cytb)进行比较分析。结果表明:唑菌酯对Colletotrichum brevisporum YYGXZ07,C.truncatum CZHP03,C.acutatum HHBY48和C.gloeosporioides CSLL11的EC50分别为0.098、3.363、10.156和31.982μg/mL,而且供试菌株在含药培养基上的生长速率排序为C.truncatum CZHP03C.brevisporum YYGXZ07C.acutatum HHBY48C.gloeosporioides CSLL11;不同菌株的作用靶标基因细胞色素b氨基酸比对分析发现C.acutatum HHBY48、C.truncatum CZHP03和C.brevisporum YYGXZ07与C.gloeosporioides CSLL11的Cytb氨基酸序列同源性分别为82.98%、88.30%和87.77%,推测6个氨基酸位点可能与菌株对唑菌酯的敏感性存在关联。     

Required number of location-years for estimating functional lower developmental thresholds and required thermal summations of insects: The first emergence of adult Apthona nigriscutis Foudras as an example

Sampling theory was combined with resampling techniques to determine the number of location-years needed for estimating an insect's required thermal summation and lower developmental threshold from field-based studies. This was done by analysing 19 location-years of field observations on the first emergence of Apthona nigriscutis Foudras, an important biological control agent of the noxious weed, leafy spurge (Euphorbia esula L.). Results indicated that 5 - 7 location-years were needed to estimate that insect's mean required thermal summation and mean lower developmental threshold for predicting first adult emergence. However, if the wish was to estimate the median required thermal summation and median lower developmental threshold for that purpose, then 8 - 11 location-years were needed to obtain acceptable estimates of those parameters. The implications of these and other findings are discussed.     

Management of strawberry anthracnose by choice of irrigation system,mulching material and host resistance

We studied, by means of field experiments, the combined effects of irrigation system, mulching material and genetic resistance on the incidence of strawberry anthracnose caused by Colletotrichum acutatum. All trials involved artificial inoculation with fruits colonized by the pathogen. One experiment examined the effects of drip irrigation, overhead sprinkler irrigation, and localized low-pressure tape irrigation, and the effects of grass (Brachiaria sp.) and plastic (polyethylene) mulches, in a split plot, randomized complete block design with four replicates. Lower disease incidence, indicating lower pathogen dispersal, was verified with low-pressure tape and drip irrigation. Flower blight and fruit rot incidence in drip-irrigated plots was minimal (ca. 2% of the disease levels in the plots otherwise irrigated). Grass mulch reduced flower blight in the sprinkler system (up to 80%), but had no significant effect when disease levels were already low due to use of other irrigation systems. A second experiment compared the effects of grass mulch, pine (Pinus elliotti) mulch, and plastic mulch with overhead sprinkler irrigation on strawberry cultivars partially resistant (‘Dover’) and susceptible (‘Campinas’) to the disease in a factorial completely randomized design (CRD). Both organic mulches reduced flower blight and diseased fruit incidence by up to 76% compared to the plastic mulch treatment, especially in the early stages of the epidemics. ‘Dover’ had 70% lower incidence of flower blight than ‘Campinas’, and the effects of organic mulches were more pronounced in ‘Campinas’. A third experiment, with all three irrigation systems combined with inoculated and non-inoculated plots, estimated yield effects due to disease and irrigation system. All plots were covered with plastic mulch in a CRD with six treatments. This experiment clearly confirmed the higher flower blight and fruit rot incidences when sprinkler irrigation was used. In inoculated treatments, fruit yield was significantly reduced in tape-irrigated plots, and more so in sprinkler-irrigated plots. In drip-irrigated plots yield was reduced by only 1%. Overall, our results confirm the importance of cultural and genetic factors as valuable means of strawberry anthracnose management. Adoption of localized irrigation systems, the use of organic mulches and choice of a partially resistant cultivar significantly reduced disease levels in field plots. The importance of water splash for C. acutatum dispersal and its dependency on the soil mulch characteristics are the likely causes for the results observed.     

116株芒果炭疽病菌对咪鲜胺的敏感性测定

研究芒果炭疽病菌对咪鲜胺的敏感性, 以期为该病的综合防治提供参考。从国内及国外芒果产区采集病样, 分离、纯化芒果炭疽菌菌株, 采用生长速率法测定芒果炭疽病菌对咪鲜胺的敏感性。结果表明, 分离获得的116株芒果炭疽病菌菌株对咪鲜胺的EC50值为0.000 8～0.073 9 μg/mL, 平均值为0.012 4 μg/mL, EC50最大值是最小值的92.4倍。不同采集地区和采集部位菌株对咪鲜胺的敏感性不同, 美国夏威夷的菌株敏感性最低, 广东的最高; 分离自叶片的菌株敏感性最高, 果实的最低。芒果炭疽病菌对咪鲜胺具有较高的敏感性, 生产上仍具有较高的使用价值, 但不同菌株的敏感性之间具有一定差异性, 存在潜在的抗性风险。     

A quantitative real‐time PCR assay for detection of Colletotrichum lindemuthianum in navy bean seeds

Bean anthracnose is a seedborne disease of common bean (Phaseolus vulgaris) caused by the fungal pathogen Colletotrichum lindemuthianum. Using seed that did not test positive for the pathogen has been proven to be an effective strategy for bean anthracnose control. To quantify the extent of anthracnose seed infection, a real‐time PCR‐based diagnostic assay was developed for detecting C. lindemuthianum in seeds of the commercial bean class navy bean. The ribosomal DNA (rDNA) region consisting of part of the18S rDNA, 5^.8S rDNA, internal transcribed spacers (ITS) 1, 2 and part of the 28S rDNA of seven races of C. lindemuthianum, 21 isolates of Colletotrichum species and nine other bean pathogens were sequenced with the universal primer set ITS5/ITS4. Based on the aligned sequence matrix, one primer set and a probe were designed for a SYBR Green dye assay and a TaqMan MGB (minor groove binder) assay. The primer set was demonstrated to be specific for C. lindemuthianum and showed a high sensitivity for the target pathogen. The detection limit of both assays was 5 fg of C. lindemuthianum genomic DNA. To explore the correlation between the lesion area and the DNA amount of C. lindemuthianum in bean seed, seeds of the navy bean cultivar Navigator with lesions of different sizes, as well as symptomless seeds, were used in both real‐time PCR assays.     

A new anthracnose disease of pyrethrum caused by Colletotrichum tanaceti sp. nov

A new pathogen of pyrethrum (Tanacetum cinerariifolium) causing anthracnose was described as Colletotrichum tanaceti based on morphological characteristics and a four‐gene phylogeny consisting of rDNA‐ITS, β‐tubulin (TUB2), glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) and actin (ACT) gene sequences. The fungus produced perithecia in culture, requiring an opposite mating type isolate in a heterothallic manner. The initial infection strategy on pyrethrum leaves involved the formation of appressoria followed by production of multilobed infection vesicles in the epidermal cells. Infection and colonization then proceeded through thinner secondary hyphae, which resulted in the initial production of water‐soaked lesions followed by black necrotic lesions. The infection process was suggestive of a hemibiotrophic infection strategy. Moreover, phylogenetic analysis clearly showed that C. destructivum, C. higginsianum and C. panacicola were separate species that also had similar intracellular hemibiotrophic infection strategies as C. tanaceti, which all clustered in the C. destructivum complex. Colletotrichum spp. were detected at 1% incidence in seed of 1 of 19 seed lines, indicating the potential for seed as a source of inoculum into crops. Colletotrichum tanaceti was detected in leaf lesions from 11 of 24 pyrethrum fields surveyed between April and July 2012, at a frequency of 1·3–25·0% of lesions. Anthracnose probably contributes to the complex of foliar diseases reducing green leaf area in pyrethrum fields in Australia.     

柑橘炭疽病高效杀菌剂的筛选及抗药性菌株的发现

采用生长速率法,用50%多菌灵可湿性粉剂(WP)、80%代森锰锌可湿性粉剂(WP)、10%苯醚甲环唑水分散剂(WG)、45%咪鲜胺水乳剂(EW)和25%嘧菌酯悬浮剂(FW)5种杀菌剂对来自广东不同地区和不同柑橘品种的10个柑橘炭疽病菌菌株进行了毒力测定.结果表明,咪鲜胺对柑橘炭疽病菌具有最好的抑制效果,其次是苯醚甲环唑;所有供试菌株对嘧菌酯和代森锰锌都具有一定程度的抗药性;不同地区的菌株对多菌灵的敏感性存在明显的差异,其中菌株CCMZ035、CCMZ004、CCST003和CCST026对多菌灵表现出很强的耐药性(抗药性).因此,咪鲜胺和苯醚甲环唑可作为防治柑橘炭疽病的首选药剂.     

进境棉籽中胶孢炭疽病菌的分离与鉴定

对一批澳大利亚进境的棉籽进行了病原菌分离,在棉籽上分离到1株疑似胶孢炭疽病菌的菌株CG-92963。通过对该菌株进行病原菌形态学观察、致病性测定并结合分子生物学方法检测。结果表明,菌株CG-92963在PDA上产生大量分生孢子,分生孢子呈长椭圆形或圆筒形;分别用真菌通用引物、特异引物对CG-92963进行扩增和测序,Blast分析结果表明其与GenBank中胶孢炭疽病菌ColletotrichumgZOP0sp0以0idPs序列同源性为100％;该菌不仅侵染棉花,还侵染芒果和梨。根据上述实验结果,将分离获得的菌株CG-92963鉴定为胶孢炭疽病菌ColletotrichumgtoeoSportoiaes（Penz）Sacc,     

与苦丁茶炭疽病抗性基因连锁的RAPD标记

为了研发出与苦丁茶炭疽病抗性基因紧密连锁的RAPD-SCAR特异标记,本研究中首先利用92条RAPD随机引物对苦丁茶冬青中已知对炭疽病高抗或高感的不同种质材料进行RAPD分析,并从中寻找到4个与苦丁茶炭疽病抗性基因紧密连锁的RAPD特异标记特异性片段S69-300,S227-300,S227-2000和S247-400。后续的研究可对这些RAPD特异片段进行回收,扩增和测序,并将测序结果作为开发与苦丁茶炭疽病抗性基因紧密连锁的SCAR标记的基础。     

黄瓜新炭疽病原的鉴定及生物学特性初探

明确引起黄瓜炭疽病的病原,旨在为该病的有效防治提供理论依据。通过形态学方法、柯赫氏法则验证,基于ITS、ACT和GDPH序列分析鉴定病原菌,并对该菌的生物学特性进行研究。结果表明,该菌回接黄瓜后的症状与田间症状一致。在PDA培养基上菌落为黑色,气生菌丝稀疏、灰白色;分生孢子长柱形,透明无隔;分生孢子盘着生黑色刚毛;孢子附着胞浅褐色,卵圆形,个别双生;菌丝附着胞深褐色,不规则形、浅裂或深裂。结合病原菌三基因联合建树的分析结果,可将其鉴定为Colletotrichum brevisporum。最适菌丝生长和产孢的温度25℃、pH 8,在PDA培养基上菌丝延伸最快,OA培养基上产孢最多,碳源中葡萄糖利于菌丝生长,可溶性淀粉和乳糖利于产孢,氮源中蛋白胨和酵母浸粉最适合菌丝生长,且后者产孢最多,致死温度为49℃。