不同温湿度组合对安徽虫瘟霉 诱发桃蚜病害的影响

用孢子浴方法,对42批次桃蚜Myzus persicae（30～60头/批）接种大剂量（孢子79～90个/ mm²）安徽虫瘟霉Zoophthora anhuiensis的分生孢子,在20℃下保湿24 h后转入不同温度（10℃、15℃、20℃、25℃、30℃及自然变温1.5～16.6℃和8.5～20.2℃）和湿度(50%、65%、80%、90%、95%及100% RH)的组合条件下观察桃蚜的反应。结果表明,各组合条件下的桃蚜均能发病死亡,而且累计死亡率的显著差异存在于不同温度（F=7.46, P<0.01)和湿度间（F=12.54, P<0.01）。最适发病的温度为恒温20℃和变温8.5～20.2℃（日均温12.4℃）,死亡率随湿度升高而增大。在10～25℃和100% RH的组合中,温度的变化几乎不影响桃蚜的累计死亡率,但影响发病速率,在10℃、15℃、20℃和25℃下的致死时间LT₅₀值分别为8.4 天、7.1 天 、4.0 天和3.4 天。回归分析表明, 在100% RH下安徽虫瘟霉诱发桃蚜发病的起始温度为1.65℃。在10～15℃及自然变温下,病死蚜尸顺利产孢的湿度为80% RH;而在20～30℃下,蚜尸产孢的湿度为90% RH。在所有温湿组合的蚜尸中未见安徽虫瘟霉的休眠孢子发生。     

块状耳霉的分离、鉴定、培养和寄主范围

1981年自感染真菌的扁豆蚜(Aphis craccivora Koch)虫尸上分离到一株虫毒菌,经鉴定是耳霉属的块状耳霉(couidiobolus thromboides Drechsler)。该菌易于分离和培养,在培养基上可形成大量休眠孢子,休眠孢子容易萌发,六天中萌发率达53%。经试验该种虫霉可以感染多种蚜虫,是一种有希望制成杀蚜菌剂的真菌。     

飞虱虫疠霉分生孢子在桃蚜体壁上的附着与入侵

用飞虱虫疠霉（Pandora delphacis)初级分生孢子接种桃蚜（Myzus persicae),24h内定时取样,在扫描电镜下观察孢子的萌发及其对寄主体壁的入侵。结果表明,附着到蚜体表面的孢子在4h内有30－40％已萌发产生芽管,少数是呈叉状分枝的营养生长型芽管。且侵染性芽管的孢子部分孢入体壁蜡质层中,显示孢子有分泌物产生并作用于寄主体壁。接种10h内,侵染性芽管通过顶端膨大的附着胞或直接穿透入寄主体壁。到24h时,产生侵染性芽管的孢子全部侵入寄主体内,寄主体表仅留下少数未萌发的孢子或营养生长型芽管。初级分生孢子在蚜体表面很少产生次级分生孢子,说明桃蚜是适合该菌侵染的寄主。陷入寄主体壁的孢子不因若蚜蜕皮而被去掉,表明该菌对成蚜和若蚜都具有侵染力。     

环境因子对努利虫疠霉产孢和孢子萌发的影响

探讨努利虫疠霉Pandoranouryi(Remaudi埁re&Hennebert)H櫣mber在田间蚜虫种群中发生与流行的规律,研究外界环境因子对感菌虫尸产孢和孢子萌发的影响。结果表明,处于水琼脂培养平板上感染努利虫疠霉的桃蚜虫尸在8~25℃的温度范围内均能产生大量的初生分生孢子,在30℃下,仅弹射极少量孢子。8℃下,孢子弹射可以持续120h,当温度高于15℃,大部分的孢子会在48h内完成弹射。相对湿度小于95%,虫尸停止产孢。20℃下,光照条件不会影响虫尸弹射孢子的总量。在8℃和30℃时,24h后处于水琼脂培养平板上的孢子萌发率分别为45.23%和61.74%,显著低于15~25℃温度范围内的孢子萌发率(95%以上)。处于叶片上的真菌孢子,当相对湿度大于74%时出现萌发,但在盖玻片的表面,当湿度低于100%时未发现孢子萌发。     

不同温湿度条件下飞虱虫疠霉对桃蚜生殖力和内禀增长力的影响

对7日龄桃蚜 Myzus persicae用高剂量飞虱虫疠霉Pandora delphacis接种后,饲养于不同温(10～30℃)、湿(74%～100%RH)度组合条件下观察其生殖力及内禀增长力(r_m) 的变化。结果表明,被接种个体生殖力比对照显著降低,且下降幅度受温度、湿度及温湿互作的影响。在各处理组合中,尤其以20～30℃和95%～100%RH的条件下下降更为明显。各处理r_m在不同温度间差异显著,均随温度呈抛物线形变化,25℃下达最高,但不同湿度间差异不显著。与对照相比,被接种蚜的r_m>除10℃与各湿度组合间无显著差异外,其余温湿组合下均显著低于对照。上述分析表明,飞虱虫疠霉的侵染能显著影响桃蚜生殖力及r_m,尤其是在桃蚜繁殖的最适温度范围内效果最为明显。     

努利虫疠霉感染桃蚜过程的扫描电镜观察

利用扫描电镜观察了努利虫疠霉Pandora nouryi (Remaudière & Hennebert) Humber初生分生孢子接种桃蚜Myzus persicae (Sulzer)后孢子萌发、入侵以及菌体突破虫体的整个侵染过程。结果表明：附着于虫体表面的初生分生孢子在3～5 h后即有60%以上的萌发率,萌发的孢子形成芽管或产生球形或棍棒状的附着胞;12 h后大部分孢子均已萌发,并成功入侵寄主虫体;接种60 h后,呈掌状分枝的假根首先从桃蚜胸部的腹面突破虫体长出体外,明显区别于新蚜虫疠霉Pandora neoaphidis (Remaudière & Hennebert) Humber假根突破虫体的位置;假囊状体不多见,且仅分布于蚜虫身体两侧,这可在一定程度上解释努利虫疠霉产孢对湿度条件要求较高的生物学现象。     

安徽虫瘟霉的黍米培养及其对桃蚜的侵染力

安徽虫瘟霉Zoophthora anhuiensis (Li) Humber是较难人工培养的蚜虫专化性病原真菌。将灭菌并适度熟化的黍米Panicum miliaceum L.作为固体基质与挑碎的安徽虫瘟霉平板菌落混合,在20℃和12L∶12D的温光条件下静止固体培养,获得了产孢潜能大、杀蚜活性强的米粒培养物。培养7天的黍米的产孢量达13.0×10⁴个孢子/粒,产孢持续时间长达6天。用此黍米培养物弹射的孢子对桃蚜Myzus persicae (Sulzer) 若蚜进行7.9～134.9个孢子/mm²共9个剂量的孢子浴接种,所获数据很好拟合时间 剂量 死亡率模型。接种后第5～7天各天的LC₅₀依次为59.8, 39.5和33.5个孢子/mm²,LC₉₀依次为354,234和198个孢子/mm²。在57.7～134.9个孢子/mm²的接种剂量范围内,致死中时LT₅₀从5.1天下降到4.3天。由此表明,安徽虫温霉的黍米培养不仅简单易行,而且菌种的产孢和侵染生物学特性在培养物中被充分体现,每颗米粒如同自然罹病而死的蚜尸,值得进一步研究开发和利用。     

飞虱虫疠霉继发性感染对桃蚜数量增长的控制作用

用飞虱虫疠霉（Pandora delphaics)“孢子浴”接种的桃蚜(Myzus persicae)无翅成蚜在离体甘蓝菜叶片（65cm^2)上建立蚜群,在不同温度（10－30℃）和湿度（74％－100％RH）的组合条件下任其繁衍,发病和交互感染,以评价该菌的控蚜效果。在25个温,湿度组合处理（8次重复,每重复含3头接种成蚜)中,蚜群均不同程度的发病死亡,在历时30d的观察中,以高温（20－30℃）,高湿（95％RH）组合条件下的蚜群发病快且死亡率高,蚜尸上产生的孢子有效地引起若蚜继发性感染。与相同温度下不带菌的对照蚜群相比,次于30℃下,各湿度除个别例外,第8d的控蚜率达30％以上,第20d达80％以上。在10℃和15℃下,控蚜效果一般不如上述较高温度下,且与湿度的关联程度相对较低,但最大控蚜效果均发生在100％RH处理中。结果表明,飞虱虫疠霉用于蚜虫防治的潜力很大,值得深入研究和开发利用。     

飞虱虫疠霉分生孢子在桃蚜体壁上的附着与入侵*

用飞虱虫疠霉(Pandora delphacis)初级分生孢子接种桃蚜(Myzus persicae),24 h内定时取样,在扫描电镜下观察孢子的萌发及其对寄主体壁的入侵。结果表明,附着到蚜体表面的孢子在4 h内有30~40%已萌发产生芽管,其中多数为侵染性芽管,少数是呈叉状分枝的营养生长型芽管。具侵染性芽管的孢子部分陷入体壁蜡质层中,显示孢子有分泌物产生并作用于寄主体壁。接种10 h内,侵染性芽管通过顶端膨大的附着胞或直接穿透侵入寄主体壁。到24 h时,产生侵染性芽管的孢子全部侵入寄主体内,寄主体表仅留下少数未萌发的孢子或营养生长型芽管。初级分生孢子在蚜体表面很少产生次级分生孢子,说明桃蚜是适合该菌侵染的寄主。陷入寄主体壁的孢子不因若蚜蜕皮而被去掉,表明该菌对成蚜和若蚜都具有侵染力。     

飞虱虫疠霉液培菌丝的凝胶化及其杀蚜活性*

用丙烯酰胺淀粉凝胶包埋飞虱虫疠霉(Pandora delphacis)液培菌丝,制成含营养和不含营养的虫霉凝胶,并测试其产孢能力及对桃蚜(Myzus persicae)的侵染力。经持续9 d产孢观察,相同菌丝量条件下,含营养的虫霉凝胶的累计产孢量(1910.9 个/mm2）为不含营养的虫霉凝胶的3.14倍（608.5个/mm2）和对照（纯液培菌丝）的4.69倍（387.5个/mm2）。用虫霉凝胶的“孢子浴”接种桃蚜居群（成蚜与不同龄期若蚜的混合）,在2.23个/mm2的剂量下,接种后第7 d的死亡率达82.3%。结果表明,丙烯酰胺淀粉凝胶作为载体材料,与飞虱虫疠霉的基本生物学特性相容,对虫霉的剂型化具有重要意义     

The phylogenetic placement of Ostropales within Lecanoromycetes (Ascomycota) revisited

Results of molecular studies regarding the phylogenetic placement of the order Ostropales and related taxa within Lecanoromycetes were thus far inconclusive. Some analyses placed the order as sister to the rest of Lecanoromycetes, while others inferred a position nested within Lecanoromycetes. We assembled a data set of 101 species including sequences from nuLSU rDNA, mtSSU rDNA, and the nuclear protein-coding RPB1 for each species to examine the cause of incongruencies in previously published phylogenies. MP, minimum evolution, and Bayesian analyses were performed using the combined three-region data set and the single-gene data sets. The position of Ostropales nested in Lecanoromycetes is confirmed in all single-gene and concatenated analyses, and a placement as sister to the rest of Lecanoromycetes is significantly rejected using two independent methods of alternative topology testing. Acarosporales and related taxa (Acarosporaceae group) are basal in Lecanoromycetes. However, if the these basal taxa are excluded from the analyses, Ostropales appear to be sister to the rest of Lecanoromycetes, suggesting different ingroup rooting as the cause for deviating topologies in previously published phylogenies.     

Genome Analysis inNeurospora crassa;Cloning of Four Lociarginine-1(arg-1),methionine-6(met-6),unknown-7(un-7), andribosome production-1(rip-1) and Associated Chromosome Walking

We have cloned fourNeurospora crassagenes by complementation analysis. Cloned genes include thearginine-1(arg-1),methionine-6(met-6),unknown-7(un-7), andribosome production-1(rip-1) loci. Chromosome walks were initiated in ordered cosmid libraries from the cloned loci. A total of about 700 kb of theNeurosporagenome is covered in these walks.     

New species of Phaeoramularia, Pseudocercospora and Stenella from Western Ghats of India

Four new species of the hyphomycete genera Phaeoramularia viz. Ph. caesalpinae, Pseudocercospora viz., Ps. tiliacearum, Stenella, viz. S. argyreiae and S. grewiae occurring on Caesalpinia bonducella Fleming (Caesalpiniaceae), Grewia sp. (Tiliaceae), Argyrea sp. Lour (Convolvulaceae) and Grewia sp. L. (Tiliaceae), respectively are described and illustrated here. All these fungi were collected from Western Ghats of India.     

The phylogenetic placement of the Leucogastrales, including Mycolevis siccigleba (Cribbeaceae), in the Albatrellaceae using morphological and molecular data

The morphology of many hypogeous fungi converges on a homogeneous reduced form, suggesting that disparate lineages are subject to a uniform selection pressure. The primary goal of this study was to evaluate the morphology and infer the phylogeny of the Leucogastrales with Mycolevis siccigleba using a Bayesian methodology. A comprehensive morphological assessment was used for an a priori phylogenetic inference to guide the sequencing effort. All structures except spore ornamentation pointed to the Albatrellaceae as the most likely sister taxon. Polyporoletus sublividus, a close relative of Albatrellus, produces ornamented basidiospores with a similar structure to M. siccigleba basidiospores. The ITS from 30 taxa was used for the molecular phylogenetic analysis. P. sublividus was found sister to Mycolevis. Leucophleps spinispora and L. magnata formed a group sister to the Polyporoletus/Mycolevis group, whereas Leucogaster was polyphyletic with respect to the core of the Leucogastrales and sister to A. caeruleoporus. This relationship was expected as previously undescribed chlamydospores produced by members of Albatrellus had a similar morphology to the basidiospores of L. rubescens.     

下延叉蕨和芽胞叉蕨配子体发育的研究

利用光学显微镜详细观察了叉蕨属(Tectaria)下延叉蕨(Tectaria decurrens)和芽胞叉蕨(T.fauriei)的配子体发育过程,记录了配子体各发育阶段的特征。结果表明:(1)下延叉蕨和芽胞叉蕨的孢子均为单裂缝,具周壁,由周壁形成纹饰,孢子极面观椭圆形,赤道面观豆形或肾形。(2)孢子萌发方式为向心型。(3)原叶体发育方式为三叉蕨型。(4)成熟原叶体心脏形,两翼向斜上方扩展。(5)均具单细胞和多细胞毛状体,在丝状体或片状体阶段出现。研究认为,从配子体发育角度看,叉蕨属是较进化的陆生真蕨类;毛状体的类型、位置和出现时间等特征在叉蕨属种间存在差异,可作为该属种间分类的特征。     

Characterisation of cell wall polysaccharides, arabinogalactans-proteins (AGPs) and phenolics of Cola nitida, Cola acuminata and Garcinia kola seeds

Many Cola plant species are endemic to West and Central Africa. Cola acuminata and Cola nitida are used as masticatory when fresh, while the dried nuts are used for beverages and pharmaceutical purposes in Europe and North America. Garcinia kola seeds, that serve as a substitute for the true kola nuts, are used in African traditional medicine for the treatment of various diseases, including colic, headache and liver cirrhosis. Seeds extracts of G. kola are also known for their anti-inflammatory, antimicrobial and antiviral properties. To gain information on the chemical properties of the kolas, we have isolated and analyzed cell wall polysaccharides, arabinogalactan-proteins and phenolic substances from the seeds of the three kola species. The sugar composition of cell wall material of C. acuminata, C. nitida and G. kola revealed that Gal (up to 30%), Ara, GalA and Glc as the predominant monosaccharides, representing approximately 90% by mol of the total hydrolysable sugar present in this material. In Ammonium oxalate cell wall fraction, GalA was found to be the major sugar present in all kola species. In the alkali-soluble fraction, there were significant differences in the level of Glc and Gal. The level of Glc was high in C. acuminata and C. nitida while the level of Gal and Xyl were high in C. nitida and G. cola. Isolation and quantification of arabinogalactan-proteins demonstrate that G. kola seeds contained four to eight times more of these proteoglycans than the seeds of the other two species. Finally, analysis of soluble phenolic substances shows that caffeine and catechin were largely represented in C. acumina and C. nitida seeds, with caffeine accounting for 50% of all soluble phenolics. These findings indicate that the three Kola seeds are highly enriched in pectins and proteoglycans and that C. acuminata and C. nitida can be used as a possible source of caffeine and catechin.     

Evolutionary relationships among aquatic anamorphs and teleomorphs: Lemonniera, Margaritispora, and Goniopila

The hypothesis that similar conidial morphologies in aquatic hyphomycetes are a result of convergent evolution was tested using molecular sequence data. Cladistic analyses were performed on partial sequences of 28S rDNA of seven species of Lemonniera, one species of Margaritispora and one species of Goniopila. Lemonniera has tetraradiate conidia with long arms, whereas Margaritispora and Goniopila have typically globose (isodiametric) conidia, with short conical protuberances in a stellate or quadrangular arrangement. Lemonniera and Margaritispora have phialidic conidiogenesis and both produce dark, minute sclerotia in culture whereas Goniopila has holoblastic conidiogenesis and does not produce sclerotia in culture. Goniopila produces a microconidial phialidic synanamorph in culture. All three genera have schizolytic conidial secession. Molecular analyses demonstrate that Lemonniera species are placed in two distinct clades: one within Leotiomycetes; the other within Pleosporales, Dothideomycetes. Margaritispora is placed with Lemonniera species within Leotiomycetes. Goniopila and Lemonniera pseudofloscula are placed within Dothideomycetes. No morphological character was entirely congruent with the molecular derived phylogeny. This suggests that for the group of species studied, conidial shape is not a reliable indicator of phylogeny but more likely the result of convergent evolution in response to the aquatic environment.     

甘菊ClHSP70与ClHSP90基因的克隆及表达分析

该研究以甘菊(Chrysanthemum lavandulifolium)为实验材料,通过RT-PCR方法从甘菊转录组数据中分离出热激蛋白合成相关基因,命名为ClHSP70和ClHSP90。序列分析表明,ClHSP70基因ORF全长为2 559bp,编码852个氨基酸,蛋白功能区预测表明含有典型的HSP70蛋白NBD和SBD保守结构域;ClHSP90基因ORF全长为2 094bp,编码697个氨基酸,含有HATPase结构域和HSP90保守结构域。生物信息学分析表明,甘菊ClHSP70与大豆(Glycine max)和烟草(Nicotiana tomentosiformis)HSP70蛋白有较高的一致性,ClHSP90基因编码的氨基酸序列与紫茎泽兰(Ageratina adenophora)HSP90高度相似;实时荧光定量表达分析表明,在42℃处理不同时间,甘菊叶片中ClHSP70和ClHSP90基因表达均在0.5h时显著增加,1h达到最大值,2h后缓慢下降;不同组织表达分析表明,甘菊在42℃处理1h后,ClHSP70在成熟叶中的表达量显著高于嫩叶和根等其他组织;ClHSP90在成熟茎中的表达量最高。研究说明,ClHSP70和ClHSP90基因具有热激蛋白特征,参与了甘菊热胁迫应答过程,该研究结果为以后深入研究其基因功能奠定了基础。     

Molecular and morphological characterization of Leveillula (Ascomycota: Erysiphales) on monocotyledonous plants

Leveillula on monocotyledonous plants have been recorded as L. taurica by several authors, whereas the fungus on Allium has been described as an independent species, namely L. allii, by some authors. We sequenced ca 600 bp of the rDNA ITS region for two Leveillula specimens from Allium and Polianthes (both from monocotyledons) and compared them with several already published sequences from Leveillula isolates from dicotyledons. Pair-wise percentages of sequence divergences were calculated for all Leveillula isolates. The ITS sequence of the Polianthes isolate was identical to L. taurica on Helianthus and Vicia. The sequence of the Allium isolate was 99.5 % identical to L. taurica on Euphorbia, Haplophylum, Peganum, etc. These results suggest close relationships between monocot and dicot pathogenic Leveillula species. The identity between two monocot isolates was 98.4 %. Phylogenetic analysis revealed that the two monocot isolates do not group into a clade together. This result suggests that Leveillula acquired parasitism to monocots at least twice independently.