马铃薯晚疫病菌单孢分离物生物学特性的初步研究*

研究了马铃薯晚疫病菌单游动孢子分离的方法,并对其单游动孢子分离物菌落生长直径、产孢量以及对甲霜灵的敏感性进行了初步研究。结果表明同一菌株的不同单游动孢子菌落生长直径和产孢量有显著差异,但不同游动孢子分离物对甲霜灵敏感性没有显著差异。     

马铃薯晚疫病菌单孢分离物生物学特性的初步研究

研究了马铃薯晚疫病菌单游动孢子分离的方法,并对其单游动孢子分离物菌落生长直径、产孢量以及对甲霜灵的敏感性进行了初步研究。结果表明同一菌株的不同单游动孢子菌落生长直径和产孢量有显著差异,但不同游动孢子分离物对甲霜灵敏感性没有显著差异。     

大豆疫霉菌ITS分子检测程序的建立及其应用

依据GenBank中登录的大豆疫霉菌(Phytophthora sojae)、近缘种及相似种rDNA的ITS区序列差异,进行多重比较后设计合成一对大豆疫霉菌特异引物,并在PCR反应体系和扩增条件优化的基础上,对包括大豆疫霉菌在内的共140个菌株进行PCR检测。结果表明,电泳后只有大豆疫霉菌扩增出一条288bp的特异性条带。运用设计的大豆疫霉菌专用引物(专利申请号200610089105.4)及建立的检测程序对大豆疫霉菌纯培养游动孢子、接种于土壤中的游动孢子和卵孢子以及接种发病的大豆染病组织进行了检测应用,结果显示该检测程序对接种于土壤中的大豆疫霉菌游动孢子和卵孢子的检测理论精度分别达0.3和0.06个孢子,对染病组织检测也表现出了较高的灵敏度。     

大豆疫霉菌抗甲霜灵特性研究

大豆疫霉Phytophthorasojae易对甲霜灵产生抗性,从大豆疫霉野生型菌株可诱变筛选到对甲霜灵有抗性的菌株Mtr。Mtr抗性菌株的抗性水平可达野生型单游动孢子菌株的870倍以上。Mtr性状在无性后代稳定遗传,在游动孢子后代连续三代未发生抗药性分离。大豆疫霉Mtr性状的保持对甲霜灵没有表现依赖性。Mtr单游动孢子菌株在不含甲霜灵的胡萝卜培养基(CA)平板上培养30d后对甲霜灵的抗性没有下降,其单游动孢子后代也未出现抗药性分离。     

大豆疫霉对寄主大豆和非寄主菜豆根及根分泌物的响应与其对寄主选择性的关系

【背景】大豆疫霉根腐病作为大豆生产上的一种毁灭性病害已被美国、加拿大等多国报道,其病原菌大豆疫霉(Phtophthora sojae Kaufmann and Gerdemann)为典型的土传病原菌。近年来,土传病原菌与植物根系的互作成为研究土传病原菌寄主选择机制的主要方向。【目的】探究寄主大豆和非寄主菜豆根及根分泌物对大豆疫霉的不同影响,阐明这种影响与大豆疫霉对寄主选择的关系。【方法】应用原位土培法种植大豆疫霉感病品种Sloan、抗病品种Williams82和非寄主菜豆一点红,测定了单个大豆疫霉游动孢子对寄主大豆和非寄主菜豆幼根的侵染行为,收集了寄主及非寄主根分泌物,测定了根分泌物对大豆疫霉游动孢子的趋化作用,包括吸引游动孢子的能力以及对游动孢子成囊、孢囊萌发和芽管生长的影响。【结果】大豆疫霉单个游动孢子对寄主大豆幼根表现强烈趋向性,沿着根面进行多次试探性接触后在根尖伸长区快速成囊并萌发,产生的芽管顶端贴附在幼根表面,在感病大豆品种根面上的芽管比抗病大豆品种上的短且粗,而对非寄主菜豆幼根则不具有趋向性,接触一次后即远离,最终在距离幼根75μm的位置成囊萌发,且芽管生长不具有方向性。此外,大豆疫霉游动孢子对抗病、感病大豆和非寄主菜豆幼根的侵染行为差异完全在根分泌物试验中重现,即寄主大豆根分泌物对大豆疫霉游动孢子具有较强的趋向作用,能够有效吸引游动孢子,促进游动孢子快速成囊及萌发,抑制芽管的伸长,而非寄主菜豆根分泌物不具有上述作用。【结论】大豆疫霉对寄主的选择性与根分泌物有关,为进一步了解大豆疫霉的寄主选择机制提供理论依据。     

砂培体系中甜菜多粘菌生物学特性的研究

在改进的砂培体系中,甜菜多粘菌（Polymyxabetae）完成生活循环只需7天。利用砂培体系研究了多粘菌在不同pH值、光照、接种材料和接种量条件下,对寄主的侵染以及在其中繁殖的情况,研究了多粘菌完成侵染所需时间,侵染的游动孢子最初释放时间,游动孢子体外存活期和休眠孢子对温度的敏感性等生物学特性。     

毛细管束法分离土壤稀有放线菌

描述了一种改进的从土壤中分离稀有放线菌的毛细管法。本法能在较低的稀释条件下,利用放线荫孢子和孢囊的疏水性、游动孢子在水中的运动性和趋化性(包括趋氧性)。从而大大提高稀有放线菌的检出率。     

黄柄钙皮菌的生活史

利用基物培养、燕麦-琼脂培养技术及扫描电镜技术研究了黄柄钙皮菌的个体发育过程,在燕麦琼脂培养基上完成了从孢子到孢子的生活史。结果表明,生活史包括单核的黏变形体或游动胞、多核的营养体原质团以及孢子形成阶段。孢子球形,表面具疣突。孢子萌发为裂式,释放1黏变形体。黏变形体行变形运动,在有水的条件下,可转变为游动胞并游动。可观察到1长具极性的鞭毛。合子形成原质团。成熟原质团棕色。原质团类型为显型,具有扇形网络状菌脉。琼脂培养基上获得的黄柄钙皮菌孢子与野生型相似,并具有可育性。     

EPA高产菌的繁殖体诱导条件初探*

研究了诱导介质、处理时间及温度等因素对高产EPA、AA等多不饱和脂肪酸的菌株———轮梗霉 (Diasporangiumsp.)产游动孢子能力的影响。结果表明 :胡萝卜片介质上、2 5℃诱导 6d ,可得到最大数量的孢子 ,同一条件下游动孢子量则在第 8d达到最高。进一步探讨孢子的萌发条件发现 :2 5℃、营养琼脂上孢子的萌发率最高。     

针箍菌的生活史

利用基物和燕麦-琼脂技术研究了针箍菌的个体发育过程,在燕麦琼脂培养基上完成了从孢子到孢子的生活史。结果表明,生活史包括单核的黏变形体或游动胞、多核的营养体原质团以及孢子形成阶段。琼脂培养基获得的针箍菌孢子与野生型相似,并具有可育性。     

Pattern formation of the coenobial algae Pediastrum biwae Negoro

It is suggested that flat colony patterns of the coenobial green algae Pediastrum biwae Negoro can be determined only by some properties of the zoospores without any other control system as a whole. These suggestions are made through observations of colony patterns and colony formation during asexual reproduction and digital electronic computer simulation. The zoospore can be regarded as a sphere which has two C-sites (presumptive sites for connection) and one H-site (presumptive site for horn formation) on its equator. This sphere swims rapidly at random in a transparent vesicle and undergoes a series of changes in its properties: (i) the regions along the equator might come to have a slight affinity for each other, which could cause the arrangement of the zoospores into a plane; (ii) the two C-sites might become connection sites and the zoospores, therefore, might form strings, leading to the characteristic pattern of the adult colony (e.g. a few concentric circular strings, a spiral string and so on); and finally, (iii) the H-site of the zoospore determines whether it grows to a horn or not, i.e. the zoospore grows into a horn cell or a triangular cell according to the absence or presence of other zoospores which prevent its growth.     

Studies on British chytrids XXVIII. Rhizophydium nobile sp. nov., parasitic on the resting spore of Ceratium hirundinella O.F.Müll. from the plankton

The life history of a new species of the Chytridiales Rhizophydium nobile is described. It occurs in the autumn on resting spores of the alga Ceratium hirundinella O.F.M. in Blelham Tarn, Windermere and Esthwaite Water, lakes in the English Lake District. The sporangium develops from the zoospore and possesses a branched rhizoidal system. The zoospores are fully formed in the sporangium. On dehiscence part of the sporangium content flows out surrounded by a vesicle which eventually bursts and liberates the zoospores. Only a few resting spores were seen associated with the sporangial stage. They were small spherical thick-walled bodies containing several globules. Further observations need to be made upon this stage.     

RELATIONSHIP OF TEMPERATURE TO ZOOSPORE PRODUCTION IN TETRASPORA GELATINOSA

A strain of Tetraspora gelatinosa was isolated from a field collection and a technique for measuring quantitatively the production of zoospores was devised. The method employed compound colonies grown on agar from zoospore suspensions. The effect of 3 temperatures on zoospore production was tested: no zoospores were produced by the colonies at 3 C; zoospores were produced at 11 and 23 C and the final yield of zoospores at the 2 temperatures was not significantly different. However, at 11 C the maximum density of zoospores produced, measured by cell count, occurred after IS hr, while the maximum density at 23 C occurred after 21 hr.     

Studies on Antagonistic Effects of Trichoderma Isolates Against Phytophthora cactorum

Bioassays were used to demonstrate the antibiotic effect of Trichoderma isolates on P. cactorum. When both fungi were grown on benomyl-containing PDA medium, the mycelial growth of Trichoderma was suppressed. However, the production of antibiotics by this fungus remained active, leading to inhibition of the mycelial growth of P. cactorum. The antibiotic effect of Trichoderma on zoospores and cysts was tested on a PDA substrate precultured with Trichoderma on cellophane sheets. On the substrate of some Trichoderma isolates, lysis of zoospores, formation of extracellular vesicles, and hypertrophy of the water expulsion vesicle did occur, both resulting in the death of the zoospores. Conidial suspensions of Trichoderma isolates also induced zoospore lysis. It is presumed that membrane-active peptide antibiotics (peptaibols) are involved in zoospore lysis. The peptaibol paracelsin caused lysins of zoospores at a concentration of 2.5 × 10^?4 M. The effect on cysts depended on the Trichoderma isolate tested and the age of Trichoderma preculture. Old cultures (after beginning of sporulation) affected cysts more severely than young cultures (before sporulation) which usually were not lethal to the cysts but induced preferably microsporangium formation, inhibition of cyst germination, and retardation of germ tube growth.     

A putative DEAD-box RNA-helicase is required for normal zoospore development in the late blight pathogen Phytophthora infestans

The asexual multinucleated sporangia of Phytophthora infestans can germinate directly through a germ tube or indirectly by releasing zoospores. The molecular mechanisms controlling sporangial cytokinesis or sporangial cleavage, and zoospore release are largely unknown. Sporangial cleavage is initiated by a cold shock that eventually compartmentalizes single nuclei within each zoospore. Comparison of EST representation in different cDNA libraries revealed a putative ATP-dependent DEAD-box RNA-helicase gene in P. infestans, Pi-RNH1, which has a 140-fold increased expression level in young zoospores compared to uncleaved sporangia. RNA interference was employed to determine the role of Pi-RNH1 in zoospore development. Silencing efficiencies of up to 99% were achieved in some transiently-silenced lines. These Pi-RNH1-silenced lines produced large aberrant zoospores that had undergone partial cleavage and often had multiple flagella on their surface. Transmission electron microscopy revealed that cytoplasmic vesicles fused in the silenced lines, resulting in the formation of large vesicles. The Pi-RNH1-silenced zoospores were also sensitive to osmotic pressure and often ruptured upon release from the sporangia. These findings indicate that Pi-RNH1 has a major function in zoospore development and its potential role in cytokinesis is discussed.     

德里腐霉游动孢子诱发试验的研究

本试验研究了不同培养基、菌龄大小、温度高低、诱饵种类和预处理时间长短对德里腐霉(Pythium.deliense)游动孢子产生的影响。结果显示,CMA、WA、PDA、TSA和LBA培养基均适宜于培养P.deliense并使其产生较多量的游动孢子,其中尤以CMA和WA效果为好;幼龄培养物的产孢能力比老龄的强,并以菌龄2天为最好;此菌在8℃以下和35℃以上不产孢,15—25℃为其产孢适宜温度;以马唐叶和胡萝卜为诱饵的处理,其孢子始见期和产孢量均明显比对照组早和高,马唐叶的刺激效应较胡萝卜的为强;随着预处理时间的增加,产孢量随之相应增多,但龄期重迭现象则越严重。     

Reproduction by zoospores inOedogonium

Summary Emergence of zoospores ofOedogonium and their subsequent developmental changes have been studied using live material and sections prepared for light and electron microscopy. Release commences with rupture of the cell wall at its pre-weakened site near the apical caps. The pliable protoplast of the zoospore becomes completely spherical once free of the wall; it is enclosed within the hyaline vesicle which expands continuously and then disappears. Meanwhile, as the flagella become active, the zoospore begins to elongate and its dome starts to protrude from a circular constriction where the flagella are inserted. Once free of the hyaline vesicle, it is actively motile for a variable period, during which elongation continues. The motile phase ceases when the zoospore begins to vibrate, whereupon the flagella are all violently shed. Soon after this, the constriction disappears from around the dome which becomes more pointed; the immobile cell now elongates further, increasing in volume. The cell periphery contains numerous contractile vacuoles. Zoospore elongation may be associated with a proliferation of longitudinal microtubules, and once the flagella are shed, the flagellar rootlet system disintegrates, probably releasing the rootlet microtubules. Mechanisms involved in the release of the zoospore are also discussed.     

ALKALINE EARTH ELEMENTS AND ZOOSPORE RELEASE AND DEVELOPMENT IN PROTOSIPHON BOTRYOIDES

O'Kelley , Joseph C., and Walter R. Herndon . (U. Alabama, University.) Alkaline earth elements and zoospore release and development in Protosiphon botryoides . Amer. Jour. Bot. 48(9): 796–802. Illus. 1961.—Cells of Protosiphon botryoides Klebs from depleted nutrient medium containing Ca were washed and resuspended in fresh complete medium with Ca; or in media with a Sr, Ba or Na replacement, respectively, for Ca; or in an equivalent CaCl₂ solution or deionized water. Zoospore release was observed in these media upon illumination following a 12-hr dark period. Free zoospores were less abundant in Sr-, Ba- and Na-replacement media than in the Ca medium. Zoospore production and release also were depressed in solutions of only CaCl₂ and in deionized water. In the Sr and Ba media, zoospores were formed but not released from the parent cell, as a rule; some zoospores were released in mass within a gelatinous vesicle which did not liquefy and set the zoospores free; these zoospores lost motility and continued development in Sr, producing characteristic, spheroidal clusters of aplanospores. In the Na medium, protoplasmic cleavage preceding zoospore formation was severely inhibited. A study of the reversibility of Sr inhibition of the zoospore-release mechanism revealed evidence of reversion 12 hr after replacement of Sr by Ca. Walls of cells produced in Ca are rich in ruthenium red-positive materials, whereas cells produced under conditions of Sr replacement lack these materials. The significance of these findings in relation to the Ca requirement of other algal species is discussed.     

Zoospore adhesion and germination upon non-toxic substances in Pseudulvella sp. (Chlorophyta)

One of the most significant processes in the life history of an alga is the colonization of a new substratum. In the present study, we evaluate whether different organic compounds, such as agar, gelatine, chicken albumin, glycerine and polylysine, promote zoospore recruitment and germination in a periphytic, fresh-water green microalga of the genus Pseudulvella(Chlorophyta). Given the low adhesion capacity of its zoospores a series of experiments were conducted in order to find a substance and its optimal concentration that increases zoospore recruitment and allows us to follow the processes of settlement, attachment and germination of zoospores. Polylysine significantly increased the number of zoospores attached with no significant effect on the germination rate. The minimum effective concentration of polylysine for improving zoospore settlement was 0.1%. %     

Flow cytometric determination of zoospore DNA content and population DNA distribution in cultured Pfiesteria spp. (Pyrrhophyta)

The relative cellular DNA content from 23 different clonal cultures of Pfiesteria spp. zoospores was determined using a DNA fluorochrome and flow cytometry. Significant differences between Pfiesteria piscicida and P. shumwayae were detected, both in mean zoospore DNA content and population cell cycle DNA distribution. Intraspecific differences in DNA content were found between clonal zoospore cultures established from different geographical regions. Long-term cultures (years) of P. piscicida were available for testing, and a negative correlation was observed between zoospore DNA content and time in culture. Zoospore cell cycle-related DNA distributions were also markedly different between the two species in these clonal cultures. In most cultures tested, P. piscicida zoospores exhibited bimodal DNA flow histograms with G1-S-G2+M distributions, typical of eukaryotic asynchronously cycling cells. In contrast, cultures of P. shumwayae zoospores exhibited one DNA peak distribution, indicative of synchronized cells. The data are consistent with the hypothesis that P. shumwayae zoospores are interphasic cells, and mitosis in zoospore cultures of this species predominantly occurs as benthic or adherent non-motile division cysts. Light microscopy observations of the nuclear condition of electrostatically sorted zoospores of each Pfiesteria species also support this hypothesis. If highly conserved, this disparity in modes of vegetative reproduction would ramify the population dynamics of the two Pfiesteria species.