绿僵菌属的三个新种

作者从广东、贵州、北京等地收集的虫生真菌中,发现在我国具有绿僵菌属特征的真菌可分为四个种。根据菌落颜色、分生孢子团块大小、紧密和牢固程度、孢子链连接方式、分生孢子形状等,将我们收集的属于本属的菌株分为四个种。本文报道绿僵菌属三个新种的描述。金龟子绿僵菌Metarhizium anisopliae(Metsch.)Sorokin为最常见的甘蔗金龟子致病菌,是本属的模式种。平沙绿僵菌Metarhizium pingshaense Chen et Guo sp.nov.、柱孢绿僵菌Metarhizium cylindrosporae Chen et Guo sp.nov.和贵州绿僵菌Metarhizium guizhouense Chen et Guo sp.nov.是三个新种。     

一株绿僵菌的分离研究

利用马铃薯葡萄糖琼脂培养基,对一株绿僵菌进行了分离培养。对不同生长发育天数的菌落特征即颜色、大小、孢子堆的形成等特点进行了描述。在光学显微镜下观察了菌丝的形态,成熟孢子及分生孢子梗的形状、颜色及连接状况。利用扫描电镜观察了分生孢子和孢子梗的超微形态。经培养特征和形态学鉴定,确定该昆虫病原真菌是金龟子绿僵菌。     

虫草一新种及其绿僵菌无性型

从贵州省都匀茶场采得一虫草新种。为缅怀戴芳澜教授对我国真菌学的贡献,我们命名这个新种为戴氏虫草(Cordyceps taii Liang et Liu)。在虫草属于囊壳倾斜埋生群(Cre-mastocarpon)的肉质亚群(Subsect．Carnosae)中,戴氏虫草与已知种的主要鉴别特征是具有较长的次生子囊孢子(21—29μm)。通过分离培养和于囊孢子的微循环产孢(microcycle-conidiation)已基本证明,戴氏虫草的无性型是戴氏绿僵菌新种(Metarhizium taii Lianget Liu)。这种绿僵菌的主要鉴别特征是形成孢梗束并偶有双细胞分生孢子。     

7株绿僵菌防治德国小蠊的毒力筛选

本文用7个绿僵菌(Metarhizium)菌株对德国小蠊Blattella germanica(Linnaeus)进行注射生测,筛选出了一个致病菌株进行五代复壮选育.该株绿僵菌Metarhizium anisopli-ae(Metsch)Sorokin复壮前对雌雄成虫注射生测LD50依次为13.2与8.9天;复壮后对雌雄成虫背部点滴生测12天的LD50值均为1.3×105孢子/虫.通过CP包埋剂制作石蜡切片对僵虫体内菌丝核和体表孢子堆进行了观察.     

竹林金针虫对绿僵菌及其代谢产物的行为反应

绿僵菌(Metarhizium spp.)是地下害虫重要的生防真菌,为了明确绿僵菌对金针虫的驱避作用,本文以平沙绿僵菌WP08菌株为研究对象,通过Y型管等嗅觉生测方法测定了筛胸梳爪叩甲幼虫对绿僵菌分生孢子、非挥发物和挥发性物的行为反应。结果表明:金针虫的选择行为与土壤中平沙绿僵菌分生孢子浓度密切相关,当土壤中绿僵菌孢子浓度达到5×10~8个·g~(-1)干土时,绿僵菌对金针虫表现出极显著的驱避作用(P0.001),随着浓度的降低,驱避作用减弱,当浓度达到5×10~6个·g~(-1)干土时,驱避作用消失;平沙绿僵菌孢子发酵液(浓度为1×10~8个·mL~(-1))对金针虫驱避作用极显著(P0.001),而挥发物对金针虫驱避作用不明显(P0.05)。本研究结果为揭示绿僵菌驱虫的内在机制提供了重要的基础信息。     

红火蚁Toll受体家族免疫响应绿僵菌表达模式的研究

为探究红火蚁Solenopsis invicta Buren Toll受体家族基因如何免疫响应绿僵菌Metarhizium的侵染。本研究采用浸渍法测定了不同浓度下绿僵菌对红火蚁大中小3种不同大小工蚁的致病力,并用显微镜观察了绿僵菌侵染后红火蚁3种工蚁的表型变化。利用生物信息学筛选Toll受体基因,并对Toll受体家族基因的理化性质、结构域、染色体位置、系统进化进行分析鉴定。运用实时荧光定量PCR（RT-qPCR）检测了红火蚁大中小型工蚁中Toll受体家族的发育历期和绿僵菌侵染后的表达模式。结果表明,绿僵菌在96 h对红火蚁大中小型工蚁的致死中浓度（LC50）分别为5.8×10^7孢子/mL、3.1×10^7孢子/mL、1.5×10^7孢子/mL;绿僵菌侵染红火蚁第3天,显微镜下观察到红火蚁体壁细小的菌丝,虫体僵硬,寄主死亡后菌丝迅速蔓延并逐渐长成橄榄绿色分生孢子;RT-qPCR结果表明Toll受体基因在红火蚁不同发育阶段中的mRNA表达水平存在显著差异,且Toll受体基因在雌性生殖蚁表达水平显著高于雄性生殖蚁;RT-qPCR结果表明红火蚁大中小型工蚁Toll受体免疫响应绿僵菌不一样。在大型工蚁中,绿僵菌侵染后,Toll受体家族基因能显著上调表达,6 h是一个诱导高峰期,Toll2-1和LRR转录水平最高,响应绿僵菌最强;在中型工蚁中,绿僵菌不能刺激Toll2-2基因的表达,却能强烈诱导Toll1,Toll2-1,Toll6,Toll7和LRR基因的上调表达,Toll1和Toll2-1响应绿僵菌最强。在小型工蚁中,绿僵菌能显著诱导Toll受体家族基因基因的上调表达,在24 h时,LRR基因表达量最高,相比于对照,LRR基因表达提高25倍,LRR在6 h和24 h响应绿僵菌最强。以上研究表明Toll受体可以免疫响应入侵的绿僵菌,且不同的Toll对绿僵菌可能具有不同的响应机制。本研究为进一步阐明Toll受体的功能奠定基础,为进一步利用绿僵菌控制红火蚁提供技术指导。     

响应面设计优化绿僵菌固体发酵条件

【目的】为了提高绿僵菌分生孢子产量及孢子质量,应用响应面设计对金龟子绿僵菌菌株CY-1(Metarhizium anisopliae)进行固体发酵培养基的优化。【方法】单因素试验基础上,采用响应面试验设计方法优化培养基组分。【结果】添加了碳、氮营养的最佳固体发酵培养基为玉米粉:稻壳=8:2,料水比1:0.8,葡萄糖0.8%,硫酸铵2.5%,磷酸二氢钾0.8%;在固体培养基上的理论产孢量为7.45×10~9个/g;验证后实际为6.94×10~9个/g。【结论】运用响应面法对绿僵菌固体发酵的培养基成分进行优化,得到了绿僵菌孢子粉,为孢子粉进行地下害虫防治和制剂加工的研究奠定了基础。     

绿僵菌对小麦纹枯病菌的抑制作用研究

在实验室条件下,研究了金龟子绿僵菌（Metarhizium anisopliae）对小麦纹枯病菌（Rhizoctonia cerealis）的拮抗作用及其机理。结果表明,金龟子绿僵菌与小麦纹枯病菌对峙培养以及在培养基中加入金龟子绿僵菌孢子悬浮液,对小麦纹枯病菌菌丝生长均有较好的抑制作用。测定了培养不同天数的金龟子绿僵菌Ma55发酵液对小麦纹枯病菌菌丝生长、菌核产生量及菌核萌发率的影响。结果表明,液体振荡培养25 d的金龟子绿僵菌Ma55发酵液对小麦纹枯病菌的菌丝生长、菌核产生量及菌核萌发具有显著的抑制作用,且Ma55发酵液中的抑菌活性物质具有较好的热稳定性。在光学显微镜下,未观察到Ma55对小麦纹枯病菌的重寄生现象,但发现金龟子绿僵菌与小麦纹枯病菌对峙培养处小麦纹枯病菌营养菌丝的细胞质变稀薄、菌丝部分消解或断裂。上述结果显示,金龟子绿僵菌对小麦纹枯病菌的拮抗机制主要是营养竞争、空间竞争及抗生作用等。     

响应面设计优化莱氏绿僵菌SZCY固体发酵培养条件及致病力测定

【背景】莱氏绿僵菌(Metarhizium rileyi)对新入侵我国的草地贪夜蛾(Spodoptera frugiperda)具有较强的致病力和田间流行性,因此具备深入开发的价值。【目的】优化莱氏绿僵菌SZCY固态发酵培养条件,测定所产分生孢子对草地贪夜蛾幼虫的毒力,为提高该菌株分生孢子规模化生产奠定基础。【方法】采用单因素试验确定了相对适宜的固态培养基,利用Box-Behnken响应面法优化该菌株的固态培养基和发酵参数,同时评价不同条件下该菌所产分生孢子对草地贪夜蛾幼虫的毒力。【结果】去颖稻谷(rice)为莱氏绿僵菌SZCY菌株固相产孢最佳载体。培养温度、光周期及酵母浸粉含量是影响莱氏绿僵菌SZCY固态发酵产孢量的主要因素。莱氏绿僵菌SZCY固态发酵最佳工艺参数为温度22.83℃、光周期18.68 h L:5.32 h D、酵母浸粉4.98 g/100 g,在此条件下,莱氏绿僵菌在去颖稻谷固态培养基上的产孢量为5.65×10¹⁰孢子/g,用其制备浓度为10⁷孢子/mL的孢子悬浮液,对草地贪夜蛾3龄幼虫的LT₅₀为3....     

草地贪夜蛾高致病力绿僵菌菌株的筛选与鉴定

为获取对草地贪夜蛾Spodoptera frugiperda具有高致病力的生防真菌,从福建省不同地区分离得到8株寄主为鳞翅目和半翅目幼虫僵虫的绿僵菌Metarhizium,采用浸渍法测定了其对草地贪夜蛾2龄幼虫和蛹的致病力,并根据形态学和分子生物学方法对高致病力菌株进行种类鉴定。结果表明,8株绿僵菌菌株对草地贪夜蛾2龄幼虫和蛹均表现出不同程度的致病力,其中菌株FJMR2和FJXY7表现出较强的致病力。在5×107个/mL孢子浓度下,FJMR2和FJXY7对草地贪夜蛾2龄幼虫的致死率分别为88.76%和82.13%,对蛹的致死率分别为86.57%和84.00%;对草地贪夜蛾2龄幼虫的LT50分别为4.81 d和4.93 d,对蛹的LT50分别为4.94 d和4.83 d。经鉴定菌株FJMR2和FJXY7均为莱氏绿僵菌Metarhizium rileyi。本研究获得2株对草地贪夜蛾2龄幼虫和蛹具有高致病力的莱氏绿僵菌菌株,在草地贪夜蛾的生物防治中具有较大应用潜力。     

Susceptibility of Oryctes rhinoceros adults to Metarrhizium anisopliae

The susceptibility of Oryctes rhinoceros adults to infections due to entomopathogenic fungi (Beauveria bassiana, Beauveria tenella, Metarrhizium anisopliae, Paecilomyces fumoso-roseus, and Spicaria rileyi) was studied by spraying titrated spores suspensions on the insect integument. The results show a definite susceptibility of the adults to Metarrhizium anisopliae strains of the major type only.     

Submerse cultivation of the fungusMetarrhizium anisopliae (Metsch.)

Для глубинной культивации гриба Metarrhizium anisopliae (Metsch.) лучше всего подходит среда в составе 4% глюкозы, 3% кукурузного экстракта (corn steep) и 4% экстракта дрожжей при исходном pH 7,2. С шарикообразным ростом мицелия боролись, прибавляя Tween 80 в концентрации 0,2–0,8%. В этой концентрации он влиял благоприятно на рост гриба. В глубинной культуре имело место полное спорообразование. Образующиеся споры по своей форме сходны со спорами из мицелия, выращиваемого на воздухе, но формируются изолированно. Путем опытов заражения было установлено, что Metarrhizium anisopliae при применении указанной среды образует вирулентные споры, пригодные для борьбы с насекомыми.     

A COMPARISON OF EGG SIZE,CAPSULE SIZE,AND DEVELOPMENT PATTERNS IN THE ORDER ASCOGLOSSA (SACOGLOSSA) (MOLLUSCA: OPISTHOBRANCHIA)

Data on egg size, capsule size, development type, and the presence and nature of albumen are summarized for 32 species of Ascoglossa, including new data on 23 species from Florida and the Caribbean. Lower limits of egg sizes for Ascoglossa with lecithotrophic and capsular development were substantially lower than limits reported for other opisthobranchs, probably because of the use of albumen as nutrient reserves in species with extended development. Capsule size is a more accurate predictor of development type than egg size. The use of albumen as a nutrient resource is associated with a relatively high plasticity of development.     

座隔孢属一新种

座隔孢属(Toxosporiopsis Sutton & Sellar)为腔孢纲(Coelomycetes)之一小属,已描述2种。本文报道了该属产自中国的一个新种:中国座隔孢(Toxosporiopsis sinensisW.P.Wu)。新种生于胡桃(Juglans regia L.)枯枝上,它与已知种尖顶座隔孢(Toxosporio-psis capitata Sutton & Sellar)和大孢座隔孢(Toxosporiopsis macrosperma(Cavara)Sutton& Dyko)在子座大小,产孢细胞长度,分生孢子形态、大小,侧丝长度及其外围胶质鞘的有无等特征上差别极大,故另立新种。     

Ultrastructural pathology of the epidermis of molting elaterid larvae (Coleoptera) with a fungus and a bacterium in the ecdysial space

Toxins produced by the fungus Metarrhizium anisopliae and the bacterium Pseudomonas aeruginosa in the ecdysial space of a molting wireworm are absorbed through the thin new cuticle and ultrastructurally change the epidermal cells into two distinct types. One is a rounded, degenerative type characterized by a “light” cytoplasm with vesiculated rough endoplasmic reticulum, rounded mitochondria with degenerated cristae, little ground plasm, and a rounded nucleus with little nucleoplasm and large globules of condensed chromatin from which chromatin fibrils separate in loose folds or granulelike tight folds. The other type has very irregular outlines and is characterized by a “dark” cytoplasm with abundant, whorled laminae of rough endoplasmic reticulum and abundant free ribosomes in a dense ground plasm, large rounded clear vacuoles, and apparently normal mitochondria and nuclei. The fungal toxins are believed to be primarily responsible for the formation of the light cells, and the bacterial toxins, for the separation of the chromatin into fibrils in the light cells, the fusion of their nuclei into large nuclear bodies, and the changes in the cytoplasmic contents of the dark cells. The dark cells, although abnormal, appear to retain a limited secretory activity.     

Description of a new species of Thynnascaris Dollfus, 1933 (Nematoda: Anisakidae) from the stomach of the marine teleost fish Rachycentron canadus (Linnaeus)

A new species of the nematode genus Thynnascaris obtained from the teleost Rachycentron canadus (Linnaeus) is reported. A detailed examination of this specimen has allowed us to erect a new species Thynnascaris to accommodate the worm. It shows remarkable differences from other known species of Thynnascaris in respect of body size, length of the oesophageal appendix and intestinal caecum, location of nerve ring and excretory pore, on the papillae on the lips arrangement, type of the striations on the cuticle, vulva position, arrangement of caudal papillae and size of the spicules. The new species is designated as Thynnascaris shyamasundarii.     

Description and phylogenetic relationships of Rodentolepis gnoskei n. sp. (Cyclophyllidea: Hymenolepididae) from a shrew Suncus varilla minor in Malawi

Rodentolepis gnoskei n. sp. is described based on specimens obtained from a lesser dwarf shrew Suncus varilla minor collected in the Nyika National Park, Malawi. The new species is one of the smallest hymenolepidids known from African shrews and is morphologically closest to two other miniature hymenolepidids from African shrews, Staphylocystis loossi and S. khalili. The new species differs from both of them by a much smaller strobila size and fewer proglottids. The rostellar hooks in the new species are more numerous and smaller in size than in S. loossi. The rostellar hooks in R. gnoskei n. sp. are almost three times shorter than hooks in S. khalili. The hook shape in both S. loossi and S. khalili is substantially different from that in the new species. Molecular phylogenetic analysis indicates that the new species is close to Rodentolepis fraterna, a parasite of rodents. The new species clearly differs from R. fraterna by the much shorter strobila, larger hooks, relatively longer cirrus sac, rate of proglottid development, the number of eggs per proglottid and parasitism in shrews. Although the new species fits the current diagnosis of Rodentolepis, its generic allocation is provisional and will likely be revised in the future because the type species of Rodentolepis, R. straminea, belongs to a different well supported clade. Thus, a new genus needs to be established for the lineage that includes R. fraterna and R. gnoskei n. sp. However, this systematic rearrangement is not recommended until Staphylocystis pistillum, the type species of Staphylocystis, is included in future phylogenetic analyses. Rodentolepis gnoskei n. sp. is the first tapeworm species reported from shrews in Malawi and the first species of cestode reported from S. varilla minor and any member of Suncus in Africa.     

Penetration of the cuticular layers of elaterid larvae (Coleoptera) by the fungus Metarrhizium anisopliae, and notes on a bacterial invasion

The penetrant hyphae of Metarrhizium anisopliae in the exuvial cuticle of a molting wireworm can form secondary appressoria on the developing new cuticle. From these a new penetrant fungal apparatus can develop through the new cuticle toward the body cavity. The penetrant fungal apparatus in the ecdysial space of the host does not appear to be affected by the histolytic enzymes in the wireworm molting fluid. A mucoidlike substance that envelopes the fungus in the ecdysial space may be, in part, the protective mechanism involved. Bacteria from the soil often invade the ecdysial space of molting wireworms that have difficulty in shedding their exuvia. Pseudomonas aeruginosa can histolyze the proteinaceous exocuticle of the exuvium, the ecdysial membrane, and the dense inner epicuticle of the new cuticle, but not the epicuticle of the exuvium, when it invades the ecdysial space of a molting wireworm.     

中国广西贝尔蛛属一新种记述(蜘蛛目,幽灵蛛科)

记述了中国幽灵蛛科贝尔蛛属1新种:张氏贝尔蛛Belisana zhangi sp.nov..新种种名依照采集者、鱼类学家张春光教授的姓氏命名.模式标本保存于中国科学院动物研究所,北京.     

塔里木河中下游地区荒漠河岸林群落种间关系分析

采用 2× 2列联表, 应用Fisher精确检验法研究了新疆塔里木河中下游荒漠河岸林群落种间关系, 测定了16种植物、共 12 0个种对的种间联结性。研究结果表明 :1) 12 0个种对中有 17个种对分别在不同的样方尺度中表现出显著或极显著的种间联结, 约占总数的 14.2 % ;其中 13个种对为正关联, 4个种对为负关联 ;2 ) 不同取样面积对种间联结性分析的有效性有影响, 不同种对表现出种间联结的最小样方尺度不同 ;3) 随着样方面积的增大, 各种对自有不同的种间联结变化规律, 可归纳为 4种类型 ;4 ) 17个具种间联结的种对以灌木草本和草本草本的种对居多, 占总数的 76.5 % ;主要乔木树种胡杨 (Populuseuphratica) 与灌木之间、灌木和灌木之间趋向独立分布。