一种简便易行的栗疫病菌单孢分离方法

介绍了一种分离栗疫病菌单分生孢子的简便易行的方法。此方法可以排除菌丝段的干扰,也适用于分生孢子器中产生微小分生孢子的其他真菌。     

一种简易单孢子分离技术

一种简易单孢子分离技术孙广宇，张天宇（西北农业大学植保系陕西杨陵７１２１００）单孢子分离是植病研究中一项常用的基本技术。单孢子分离的方法很多，这里介绍我们在研究工作中摸索出来的一种方法──直接挑取法，即在实体解剖镜下直接挑取单个孢子进行分离纯化的方法...     

介绍一种单孢直接挑取获得单孢菌株的方法

单孢分离是植物病原真菌研究中的一项基础技术,常用的有琼胶平板稀释纯化法、琼胶平板表面单孢挑取法、微块单孢分离法、印痕移取单孢分离法等,各有利弊,但均需预先制备琼胶平板。为简化单孢分离过程,参照国内外使用的方法,并根据我们的实验条件,     

马铃薯晚疫病菌单孢分离技术的改进

单孢分离是植物病理学的基本技术 ,它可纯化病原菌 ,是进行病原鉴定、生理生化、病菌遗传变异等方面研究的基础。单孢分离的方法很多 ,常采用ＰＤＡ平板划线法。但这种方法不太适用于马铃薯晚疫病菌 ,因为 :(1)市售琼脂杂质较多 ,经高压灭菌后仍可保留许多不规则的小块 ,在显微     

小麦矮腥黑穗病菌的单孢接种侵染

采用进口美麦中截获的单个、经萌发的Ｔｉｌｌｅｔｉａ ｃｏｎｔｒｏｖｅｒｓａ Ｋｕｈｎ（ＴＣＫ）冬孢子人工接种创伤和未创伤的美国硬红冬小麦的胚芽鞘,每个胚芽鞘接种一个萌发ＴＣＫ冬孢子,接种后麦苗在５℃、１２ｈ光照和黑暗交替条件下接头２１或２９天,然后移植到离网室中的塑料周转箱中作常栽培管理,结果均得了ＴＣＫ病穗。这是单个萌发ＴＣ冬孢子引起小麦发病的首次报道。本试验结果表明,在自然条件下,当寄主和环境     

非无菌操作下分离尖孢镰刀菌的培养基

 尖孢镰刀菌(F.oxysporum)是一类非常重要的土传病原真菌,寄主范围广,可引起100多种植物的萎蔫型病害。     

大连海关全国首次检出单孢共头霉菌和一种篮状菌属真菌

2021年11月,大连周水子机场海关在查验多哥入境快件时,发现疑似植物繁殖材料,样品经大连海关技术中心鉴定,其中有象牙椰子(Phytelephas macrocarpa)果实,经分离培养、多点基因测序分析后,在其上鉴定出3种病原真菌,包括单孢共头霉(Syncephalastrum monosporum)、蓝状菌属真菌1...     

禾长蠕孢菌和尖角突脐孢菌防治稗草的研究

 病原真菌禾长蠕孢菌稗草专化型(Helminthosporium gramineum Rabenh f.sp.echinochloae,HGE)和尖角突脐孢菌(Exserohilum monoceras,EM)在稗汁葡萄糖中的发酵滤液对稗草种子的发芽有明显的抑制效果,发芽抑制率分别为30.9%和13.5%.HGE菌在改良Fries、稗汁葡萄糖,EM菌在改良Fries中的发酵滤液对稗草根和芽的生长有明显抑制作用.HGE菌发酵滤液与其孢子混合使用比单独使用对稗草的防效明显提高,稗草感病株率、致死率分别达86.3%和69.5%,病情指数为78.7;发酵滤液与孢子结合(先后喷雾)使用后稗草的感病株率、致死率分别为83.9%和67.9%,病情指数为72.8.HGE与弯孢菌(Curvularia lunata)孢子混合喷雾接种对稗草防效明显高于2种菌孢子单独使用.     

一种简单快速鉴定面粉中孢子的方法

面粉常携带多种检疫性病虫,特别是易携带一类危险性病原菌小麦矮腥黑穗病菌TCK和小麦印度腥黑穗病菌TIM.因此,对面粉的检验主要是针对上述两种病原菌.2003年我们共检验面粉样品177份,占全局船舶检疫送检样品的52%,其中,截获TCK4批次,TIM10批次,较去年提高了250%.     

不同浓度增效剂对金龟子绿僵菌、黄绿绿僵菌和球孢白僵菌产孢的影响

生防菌作为一种重要的生物杀虫剂,被开发成多种制剂应用于农林害虫的防治;微生物制剂的生产成本高、耗时长,极大影响其生产效率。本试验通过液固两相发酵法筛选出提高金龟子绿僵菌Metarhizium anisopliae、黄绿绿僵菌Metarhizium flavoviride和球孢白僵菌Beauveria bassiana产孢量的最优增效剂浓度。结果表明0.1%浓度增效剂对金龟子绿僵菌的促产孢作用达最大化,浓度达4.48×10⁹孢子/g;黄绿绿僵菌的最适增效剂浓度为0.05%,浓度达2.75×10⁹孢子/g;当增效剂浓度小于0.1%时利于黄绿绿僵菌的产孢,增效剂高于0.1%时抑制产孢;而球孢白僵菌的产孢量与增效剂浓度正相关,最大浓度达7.41×10⁹孢子/g。在真菌液体发酵过程中添加产孢增效剂,有效提高了真菌产孢量,缩短了真菌培养周期,为提高真菌制剂生产效率、节约成本提供理论支持。     

一种快速简便的植物病原真菌基因组DNA提取方法

 尿素提取法是最新报道的一种植物拟南芥基因组DNA的提取方法,但是还没有应用于植物病原真菌DNA的提取。本研究采用改进的4种不同的方法(尿素提取法、CTAB提取法、氯化苄提取法以及SDS-CTAB提取法),分别对5种分类地位不同的植物病原真菌(草莓疫霉病菌、西瓜蔓枯病菌、草莓炭疽病菌、西瓜枯萎病菌和水稻纹枯病菌)的基因组DNA进行提取和比较。结果表明,尿素提取法和SDS-CTAB法均能提取到所有5种真菌的基因组DNA,而尿素提取法提取DNA的效率更高、质量更好,操作步骤更为快速简单。     

新疆棉花枯、黄萎病的发生现状及其快速分离技术

为明确新疆不同地区造成棉花萎蔫,维管束变褐的病原菌种类及其发生现状,本研究对新疆47个主要植棉区有萎蔫症状且维管束变褐的棉花病株进行了分离鉴定,并对其快速分离鉴定方法进行了改进。结果表明:南疆20个地点,得到358个纯化菌株,黄萎病菌占81.8%,枯萎病菌占18.2%;北疆27个地点,得到495个纯化菌株,黄萎病菌占93.9%,枯萎病菌占6.1%。南北疆均以黄萎病为主,枯萎病南疆重于北疆。维管束病害的快速分离方法每分离一个样品只需2 min,污染率仅为6.7%。     

Production,characterization and interaction of single-spore isolates ofPlasmodiophora brassicae

Out of 164 plants of clubroot-susceptible Chinese cabbage inoculated with single resting spores ofPlasmodiophora brassicae, two plants developed clubroot symptoms. The two single-spore isolates (SSIs) extracted from these plants gave an identical reaction pattern on the European Clubroot Differential set (ECD) and seven doubled-haploid lines (DH-lines). Their reaction pattern differed from that of the original field isolate on four hosts: ECD hosts 06 and 07 were susceptible to the field isolate but resistant to both SSIs, while for DH-lines Bi and Pt the reverse was true. DH-line Pt was significantly less diseased by mixed inocula consisting of the field isolate and SSI-1 than by SSI-1 alone. It was concluded that the SSI-1 pathotype was a minor component of the field isolate, although it was isolated twice. The results also suggest that the alleviating effect of the field isolate in mixed inoculations with SSI-1 on DH-line Pt was due to induced resistance, rather than to competitive interactions.Abbreviations cv cultivar - DH-line doubled haploid line - ECD European Clubroot Differential set - SSI single-spore isolate     

The development of a rapid PCR assay for detection of Fusarium moniliforme

The fungus Fusarium moniliforme infects a wide range of crops throughout the world. In maize (Zea mays L.) it causes seedling blight and root, stalk, and ear rots. A simple procedure that can be used to detect infection by F. moliliforme from infected plant tissues has been developed. A F. moniliforme genomic library was prepared and used to identify the recombinant clones containing fungal DNA sequences not hybridizing with the DNA of the host plant, maize. Based on the nucleotide sequence information obtained from the F. moniliforme pUCF2 genomic clone, specific oligonucleotides were designed and used as primers for in vitro DNA amplification by the polymerase chain reaction. An amplification product was obtained with F. moniliforme DNA preparations whereas no amplified DNA was detected with DNAs from other fungal pathogens, including various Fusarium species, or from the host plant. This PCR analysis was successfully employed to identify F. moniliforme directly from the mycelia that develop from naturally infected maize seeds, with no need to obtain pure fungal cultures for reliable diagnosis. The protocol can be used for the diagnosis of infected plants and soils in epidemiological studies of Fusarium diseases, for seed health testing, and for evaluation of susceptibility to colonization in commercial maize hybrids.     

A rapid and simple method for determining fungicide resistance in Botrytis

A simple test based on the germination of conidia of Botrytis on agar media augmented with various fungicides has been developed. Average concentrations causing a 50% reduction of germ-tube growth (EC₅₀) of highly sensitive isolates were determined on 1% malt extract agar (thiophanate-methyl 0.090 ppm; iprodione 0.566 ppm; fludioxonil 0.026 ppm; fenhexamid 0.144 ppm), 1% malt extract agar with 100 ppm salicyl hydroxamic acid (QoI fungicides, viz. trifloxystrobin 0.009 ppm; pyraclostrobin 0.013 ppm; azoxystrobin 0.087 ppm), 0.5% yeast extract agar (boscalid 0.069 ppm) and 0.5% sucrose agar (cyprodinil 0.053 ppm). In order to detect different levels of resistance against these various fungicides, two discriminatory concentrations were identified for each compound. A routine assay method was developed in which drops of a conidial suspension harvested directly from diseased plant material or sporulating cultures were incubated on each of 20 different agar media. Because of a very short time-span of 24–48 h between sample collection and evaluation of results, field-specific information on the occurrence, frequency and types of resistance of Botrytis against common botryticides in soft-fruit production may be generated prior to the main fungicide spray season at blossom time.

     

小麦白粉病纯化菌种保存方法

利用经初选确定的5种具有较好保鲜能力的配方,采用离体叶段滤纸保存法、离体叶段琼脂保存法和试管苗保存法对小麦白粉病菌纯化菌系保存效果进行了研究.结果表明,对小麦幼苗第一叶叶段保鲜效果最好的配方是沈保6号,当使用40～60μg/mL最适浓度时,第25天叶段绿色仍可达到90%以上.保存菌种的最适环境条件研究结果表明,在4℃冰箱中,以离体叶片沈保6号琼脂保存和离体叶沈保6号浸润滤纸保存效果最佳,在第40天叶片绿色仍分别达到78.0%和68.0%,孢子萌发率可达26.7%和26.8%.     

Patterns of Splash Dispersed Conidia of Fusarium poae and Fusarium culmorum

Splash dispersal of Fusarium culmorum and Fusarium poae spores was studied, using inoculated straw placed on tiles as the inoculum source to infect agar strips and artificially produced leaves. In addition, patterns of spread were studied with spores from inoculated artificial leaves onto agar strips. Observed patterns of spore dispersal for each species were indistinguishable, although F. culmorum produced fewer colonies than F. poae. Furthermore, spore dispersal from inoculated straw and artificial leaves were essentially identical, with one exception; colonies arose from single conidia when spread from artificial leaves, but consisted of clumps of conidia when derived from inoculated straw. Splash dispersal patterns of both species onto the upper- and undersides of artificial leaves were different. On the upperside of the leaf, most colonies were found at the tip, while on the underside of the leaf most colonies were found at the base of the leaf. This is the first time that artificially produced leaves have been used in splash dispersal experiments.     

云南罗平田间稻瘟菌株性状及无毒基因研究

稻瘟菌和水稻是研究禾本科作物病原-寄主互作机制的模式病理系统.云南罗平县不仅是云南省水稻主产区,栽培水稻品种多样,同时也是稻瘟病易发区,田间稻瘟菌群体组成复杂,信息流强度大.田间单孢菌株的分离和无毒基因的研究,是揭示稻瘟菌毒性变异机制和制定田间稻瘟病综合防控策略的重要基础.本研究通过单孢分离,从2017年云南罗平田间病...     

一种用玉米拟轮枝镰孢穗腐病病粒制备接种体的简易方法

制备接种体是玉米穗腐病抗性鉴定中的一个重要环节, 本研究提出了一种用玉米拟轮枝镰孢穗腐病病粒制备接种体的简易方法, 即将上年的拟轮枝镰孢穗腐病带菌病粒按一定重量加入无菌水洗脱制成接种体。田间接种试验结果表明, 采用本方法简易制备的接种体接种后可获得与常规制备接种体相当的接种效果, 且在-18℃冻存15 d仍可达到新鲜的常规制备接种体接种后的水平。使用简易制备接种体对33个玉米品种进行了田间穗腐病抗性鉴定, 21个品种表现为感病或高感。与实验室常规培养制备接种体的方法相比, 本制备方法简便易操作, 用时短, 成本低, 无需专业仪器设备, 能够一次制备大量接种体, 可广泛用于玉米种质材料和新品种的拟轮枝镰孢穗腐病田间抗性鉴定及抗性材料筛选等。