基于高通量测序及分离培养初步研究美国大豆中真菌多样性

中国是全球最大的大豆进口国,进境大豆所携带的病原真菌传入我国的风险极高。基于高通量测序技术对6批美国大豆真菌多样性进行分析,同时采用分离培养获得单一菌株,依据菌落形态、显微结构及分子技术进行鉴定。高通量测序结果显示,大豆中所有真菌共计5门15纲35目63科112属155种,主要为链格孢属(Alternaria)、球腔菌科(Mycosphaerellaceae)、小戴卫霉科(Davidiellaceae)、小囊菌科(Plectosphaerellaceae)、赤霉属(Gibberella)、附球霉属(Epicoccum)、间座壳属(Diaporthe)、隐球菌属(Cryptococcus)。分离培养结果显示,得到真菌共计40株10种,分别是大豆北方茎溃疡病菌(Diaporthe phaseolorum var.caulivora)、大豆南方茎溃疡病菌(Diaporthe phaseolorum var.meridionalis)、大豆拟茎点种腐病菌(Diaporthe longicolla/Phomopsis longicolla)、大豆炭腐病菌(Macrophomina phaseolina)、尖孢镰刀菌(Fusarium oxysporum)、菌核菌(Sclerotinia sclerotiorum)、镰刀菌(Fusarium sp.)、附球菌(Epicoccum sp.)、交链孢菌(Alternaria sp.)、小双胞腔菌(Didymella sp.)。     

马尾松叶表微生物多样性

为发掘对林木生长发育有利的优良微生物资源,并筛选适合叶表微生物的收集方法,以马尾松针叶为试验材料,分别用悬摇法和超声波法收集马尾松叶表微生物,用扩增子高通量测序技术、MUSCLE和Qiime软件研究马尾松叶表微生物的多样性。结果表明:扫描电镜观测结果显示,马尾松针叶表面定殖有大量微生物,包括真菌(菌丝及孢子)和细菌。扩增子高通量测序结果表明,马尾松叶表微生物物种丰富,包含细菌运算分类单位(OTUs)490个,真菌OTUs 1273个。马尾松叶表细菌以未分类的蓝细菌属(unidentified_Cyanobacteria)(36.53%)、未分类的拜叶林克氏菌属(unidentified_Beijerinckia)(28.60%)、鞘氨醇单胞菌属(Sphingomonas)(2.35%)为优势属;叶表真菌以枝孢属(Cladosporium)(2.45%)、拟盘多毛孢属(Pestalotiopsis)(0.92%)、无头孢菌属(Capnobotryella)(0.91%)为优势属。针对叶表细菌多样性的研究表明,悬摇法和超声波法均有较高的物种检出度;在叶表真菌多样性的研究中超声波法优于悬摇法,但超声波法样品间数据变异性较大,测定结果不稳定。     

Australian Uromyces viciae-fabae: Host and nonhost interaction among cultivated grain legumes

Uromyces viciae-fabae, rust of faba bean, parasitizes other legume crops such as lentils (Lens culinaris) and field peas (Pisum sativum) in some environments. In this study we examined the host range of two Australian isolates of U. viciae-fabae collected and purified from a faba bean crop and classified as U. viciae-fabae ex V. faba. Field pea (P. sativum), chickpea (Cicer arientinum), lupin (Lupinus spp.), lentil (L. culinaris), and mung bean (Vigna radiata) genotypes were tested with these isolates, as well as resistant and susceptible genotypes of the faba bean host. Race specificity for these two pathogen isolates was observed on Vicia faba, with two faba bean genotypes showing partial resistance. Both U. viciae-fabae isolates also colonized field pea seedlings and successfully produced uredinia under glasshouse conditions, despite this fungus not being known as a pathogen of Australian field pea crops. No sporulation of either isolate of U. viciae-fabae ex V. faba was observed on any of the remaining legume species tested. However, obvious differences in fungal growth were observed, ranging from small infection sites with very rare haustorium formation in mung bean to more extensive growth and the development of potential uredinial structures in chickpea. These observations are discussed in relation to the phylogenetic relationship of these host and nonhost species.     

松毛虫赤眼蜂携带球孢白僵菌防治亚洲玉米螟技术研究与应用

高效的载菌技术是提高载菌赤眼蜂防治效果的重要基础。本研究对比了不同吸附助剂对松毛虫赤眼蜂羽化时间、羽化量、存活时间及载菌量影响，证明了不同助剂及吸附球孢白僵菌分生孢子后对赤眼蜂无不利影响。试验结果表明，0.1%（w/v）淀粉溶液作为吸附助剂可显著提高赤眼蜂的载菌量，达3.6×10⁴孢子/蜂以上，并应用该助剂建立了赤眼蜂高效携带白僵菌方法。从被害株数、蛀孔数、活虫数和僵虫数4个方面测定了载菌赤眼蜂田间防治玉米螟效果。结果表明，载菌赤眼蜂较单一使用赤眼蜂防治效果显著提高，被害株数、蛀孔数和活虫数均显著降低，分别降低了28.1%、22.8%和24.5%，并且载菌赤眼蜂处理组中僵虫数量显著高于单独使用赤眼蜂处理组，说明载菌赤眼蜂对玉米螟具有协同防控作用。利用该方法进行玉米螟田间防治，能够降低防治成本，提高防治效率，节约劳动力，具有巨大的应用和推广价值。     

基于高通量测序技术分析2种菌草根际土壤真菌群落多样性

为了解福州菌草基地巨菌草和绿洲一号2种菌草的根际土壤真菌群落多样性,采用Illumina Miseq高通量测序技术和生物信息学方法,对菌草根际土真菌的多样性及群落结构进行分析。结果表明：从3份土壤样本中获得525 292条ITS序列,在97%序列相似性基础上可划分为7267个可操作分类单元（OTU）。真菌群落的丰富度指数（ACE）以菌草非根际土最低,绿洲一号根际土略高于巨菌草根际土;而多样性指数（Shannon-Wiener和Simpson）菌草根际土高于非根际土,绿洲一号根际土略高于巨菌草根际土。3份样品的优势菌门为子囊菌门（Ascomycota）、担子菌门（Basidiomycota）、接合菌门（Zygomycota）、壶菌门（Chytridiomycota）、球囊菌门（Glomeromycota）;优势菌纲为散囊菌纲（Eurotiomycetes）、粪壳菌纲（ Sordariomycetes）、座囊菌纲（Dothideomycetes）、伞菌纲（Agaricomycetes）、锤舌菌纲（Leotiomycetes）、圆盘菌纲（Orbiliomycetes）、银耳纲（Tremellomycetes）、粘膜菌纲（Wallemiomycetes）;优势菌属为篮状菌属（Talaromyces）、深黄伞形霉属（Umbelopsis）、枝孢菌属（Cladosporium）、暗双孢菌属（Cordana）、镰刀菌属（Fusarium）、隐球菌属（Cryptococcus）、淡紫紫霉属（Purpureocillium）、赤霉菌属 （Gibberella）、灵芝属（Ganoderma）。研究结果表明,菌草根际土真菌群落多样性高于非根际土,为进一步更好地利用菌草提供理论参考依据。     

间作木薯对橡胶树根际土壤真菌群落结构的影响

研究间作木薯对橡胶树根际土壤真菌群落多样性变化规律,为判断胶园间作木薯的可行性提供理论支撑。以橡胶与木薯间作为处理,橡胶单作为对照,采用传统方法和高通量测序技术分析橡胶树根际土壤理化性状及真菌群落结构。结果表明：（1）间作模式下橡胶树根际土壤的碱解氮、速效磷、速效钾、有机质含量和真菌Shannon指数显著降低,真菌Simpson指数显著提高,pH和Chao1指数虽有降低,但差异未达到显著水平;（2）门分类水平上,2个处理的橡胶树根际土壤真菌群落结构组成相近,共有的3大优势菌门均为Ascomycota、Zygomycota和Basidiomycota。间作木薯降低了橡胶树根际土壤中Zygomycota和Basidiomycota的相对丰度,但提高了Ascomycota相对丰度,达到91.54%;（3）属分类水平上,2个处理中相对丰度比例前10的优势菌属占所有检测出菌属的61.30%~65.59%,间作木薯提高了橡胶根际土壤中Phyllosticta、Gibberella和Lecythophora的相对丰度;（4）土壤碱解N、速效P和速效K含量对橡胶树根际土壤中真菌群落α-多样性产生影响,土壤真菌多样性变化与土壤有机质含量有关。因此,间作木薯增加了植物对土壤矿质营养元素的吸收,导致橡胶树根际土壤肥力下降,进一步降低了橡胶树根际土壤真菌的群落多样性,同时增加了病原真菌的丰度。     

Identification of resistance to Aspergillus flavus infection in cotton germplasm

Natural resistance in cottonseed to Aspergillus flavus infection has not been explored to date. A green fluorescent protein (GFP) expressing A. flavus strain was used to assess the resistance of seed from 35 cotton varieties from Gossypium arboreum, G. barbadense, and G. hirsutum. Mature cotyledons devoid of seed coat were wounded, inoculated, and assessed for innate resistance to A. flavus infection. Of the initial 35 varieties tested, we observed a range of resistance to infection in representatives of all three species. A subset of 15 representatives was further analyzed. Within this group, G. arboreum cultivar A2 186 and G. hirsutum cultivar SA 1582 were most resistant to fungal infection. The most susceptible cultivar in this group was G. hirsutum SA 1595. The remaining 12 representatives tested in the secondary screen (3 G. arboreum, 3 G. barbadense, and 6 G. hirsutum lines) exhibited intermediate resistance. We did not observe any relationship between species and resistance. Within each species, there was a range of responses to fungal infection. Future studies using this methodology to screen additional diploid and tetraploid cotton lines may enable us to identify naturally resistant germplasm that can be used to develop cotton with enhanced resistance to fungal infection.     

First report on the Dutch elm disease pathogen Ophiostoma novo‐ulmi from Latvia

During 2017, Dutch elm disease was investigated on 67 Ulmus glabra and 29 Ulmus laevis samples from 28 sites in Latvia. The presence of Ophiostoma novo‐ulmi was detected in 61.2% of the samples from U. glabra and in 34.5% of the samples from U. laevis. In 11% of cases, hybrids between subsp. novo‐ulmi and subsp. americana were identified, particularly in the sites in the southern part of Latvia. This study presents the first verified record of Ophiostoma novo‐ulmi and its subspecies in Latvia.     

局地尺度高寒草原土壤真菌多样性对地上植物多样性与生产力关系的调控作用

为研究局地尺度高寒草原土壤真菌多样性对地上植物多样性与生产力关系的调控作用，本研究以青海省湟源县局地尺度高寒草原为研究对象，对不同植物多样性梯度的地上生产力、土壤理化性状和土壤真菌多样性等指标进行调查、采样和分析。通过偏回归分析（partial least squares regression，PLSR）、方差分解分析（variance partitioning analysis，VPA）、构建结构方程模型（structural equation model，SEM）等分析手段，分析了土壤真菌多样性对地上植物多样性与生产力关系的调控作用。结果表明，高寒草原植物多样性与地上生产力存在显著线性正相关关系；土壤真菌多样性分别与植物多样性和地上生产力呈显著正相关关系；通过PLSR，VPA，SEM等方法控制土壤非生物因子效应后，真菌多样性与二者的显著正相关关系仍存在。综上所述，局地尺度下青海省湟源县高寒草原土壤真菌多样性是调控地上植物多样性与生产力正向关系的关键生物因子。