淡黄绒泡菌和全白绒泡菌原生质流向的观察

显型原质团是绒泡菌目黏菌的营养生长阶段,其最明显的现象是往返原生质流,但一直并不清楚原生质流反向流动的原因。观察研究了淡黄绒泡菌和全白绒泡菌原质团中的原生质流,结果表明：由于菌脉中堵塞或是在原质团前缘尚未分化通道引起反向原生质流,从而引起原质团多方向生长使原质团前缘呈现扇面状。原生质流总的方向是扇面端,并完成原质团运动。     

淡黄绒泡菌和全白绒泡菌孢囊形成过程显微观察的染色差异

为了研究黏菌孢囊形成过程中显微结构的变化,文中探讨了番红-固绿和铁帆-苏木精染色条件下淡黄绒泡菌和全白绒泡菌孢囊不同发育阶段显微结构的差异显示效果。结果表明：在幼孢囊中原质团有种水平的割裂,大割裂和微割裂,这些割裂和孢丝及孢子的形成有关;全白绒泡菌囊轴表现了和孢囊柄不一致的状态和染色结果;番红-固绿染色下,淡黄绒泡菌在孢囊形成前期原质团被染成淡红色,可以分辨出大量游离存在的细胞核;孢囊壁及囊轴被染成绿色,孢子灰绿色;全白绒泡菌原质团被染成绿色,初期可见较厚孢囊壁,囊轴绿色。铁帆-苏木精染色下,淡黄绒泡菌和全白绒泡菌原质团均被染成灰色,囊壁不明显,成熟孢子发生皱缩。     

三种绒泡菌属原质团的培养和孢子果的诱导发生

在黏菌个体发育研究的培养过程中,经常发生原质团不能转变为孢子果的情况,使个体发育停留在营养生长阶段,利用液体发酵和有饲培养相结合的方法获得的绒泡菌属黏菌原质团进行了孢子果的诱导。在饥饿条件下,通过对光照和温度的调节,获得了绒泡菌属黏菌在不同培养条件下形成孢子果和菌核的最佳条件。在固体培养基和液体培养基中获得全白绒泡菌孢子果的最佳培养条件分别为：24℃、6,000lx光照和20℃、6,000lx光照;获得扁绒泡菌孢子果的最佳培养条件分别为：26℃、6,000lx光照和20℃、6,000lx光照。淡黄绒泡菌Physarum melleum在固体培养基上既可以获得孢子果也可以获得菌核,最佳培养条件分别为：26℃、6,000lx光照和22℃、3,000lx光照;在液体培养基中只能形成菌核,条件为：22℃、6,000lx光照。     

细弱绒泡菌的生活史

利用燕麦-琼脂培养、基物培养及扫描电镜技术研究了细弱绒泡菌的个体发育过程,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,细弱绒泡菌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,表面具细小疣点。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。成熟原质团橘黄色。原质团类型为显型,具有扇形网络状菌脉。成熟原质团可形成多个孢囊。琼脂培养基上获得的细弱绒泡菌孢子与野生型相似,并具有可育性。     

实验室培养条件下绒泡菌科四种黏菌个体发育比较研究

本文应用悬滴培养、燕麦-琼脂培养等方法及显微成像技术对绒泡菌科Physaraceae 4种黏菌（黄头绒泡菌Physarum flavicomum、淡黄绒泡菌Physarum melleum、垂头绒泡菌Physarum album、针箍菌Physarella oblonga）的个体发育特征进行比较研究,首次实现了垂头绒泡菌在实验室条件下的完整生活循环。4种黏菌孢子萌发均为V-型开裂,但萌发时间有所不同,黄头绒泡菌萌发所需时间最短,仅需5h,而针箍菌萌发所需时间最长,需要2d;针箍菌和淡黄绒泡菌形成的黏变形体均可转变为游动胞,而相同条件下黄头绒泡菌和垂头绒泡菌并未有游动胞形成;4种黏菌原生质团的生长速率不同,在光照刺激下针箍菌、淡黄绒泡菌、黄头绒泡菌和垂头绒泡菌的原生质团分别经过2-3d、2-3d、7d和13d,发育成子实体;完成孢子到孢子的整个生活史,针箍菌和淡黄绒泡菌需要25-30d,黄头绒泡菌和垂头绒泡菌需要40-45d。     

扁绒泡菌的生活史

黏菌的生活史对于研究其营养方式的多样性以及系统发育等具有重要价值,目前国内外有关黏菌生活史的报导很少。利用燕麦-琼脂培养、基物培养及扫描电镜技术研究了扁绒泡菌的个体发育过程,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,扁绒泡菌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,表面具细小疣点。孢子萌发为裂式,释放1黏变形体。原质团类型为显型。成熟原质团乳白色,可形成多个孢囊。琼脂培养基上获得的扁绒泡菌孢子与野生型相似,并具有可育性。     

基于rDNA ITS序列对绒泡菌目黏菌系统发育的探讨

绒泡菌目Physarida是黏菌纲Myxogastria最大的一个目,对其系统发育关系的研究一直是根据形态特征。为了从分子水平探讨绒泡菌目乃至黏菌纲的系统发育关系,以黏菌r DNA ITS通用引物对绒泡菌目5属8种黏菌的r DNA ITS进行扩增和测序,结合Gen Bank中已有的黏菌r DNA ITS序列,利用贝叶斯推断法(Bayesian inference,BI)和最大似然法(Maximum likelihood,ML)构建系统发育树。结果表明:绒泡菌目不同物种的r DNA ITS区在碱基组成和长度上差异明显,长度为777–1 445bp,G+C mol%在53.4%–61.9%之间。绒泡菌目与发网菌目Stemonitida聚类为两个明显的分支,在绒泡菌目分支上,绒泡菌科Physaraceae和钙皮菌科Didymiaceae各聚为一支,支持了形态学上以孢丝是否具有石灰质为依据区分这两个科的观点。由多份不同地理来源的鳞钙皮菌Didymium squamulosum材料组成的钙皮菌科又形成3个分支,证实了这个形态种是由地域来源广泛、繁殖亲和性各异和遗传变异较大的不同生物种组成的复合体。     

黑发菌核糖体DNA小亚基片段的序列测定

用Rusketal 于 1995年设计的引物SMNUR10 1和Whiteetal 1990年设计的引物NS4对黑发菌 (Co matrichanigra)的rDNA小亚基片段进行了PCR扩增 ,并将扩增产物克隆到大肠杆菌JM10 9中进行测序 ,测得的序列长度为 115 4bp。同时将其与多头绒泡菌的相应序列进行了比较 ,其序列同源性为 6 6 %。     

梭绒盘菌(子囊菌门:锤舌菌纲)对寄主植物Medeola virginiana的系统性侵染(英文)

设计了用于扩增梭绒盘菌ITS rDNA片段的特异性引物。研究发现,梭绒盘菌不仅存在于被侵染植物的茎部病斑中,而且存在于寄主Medeola virginiana未被感染的叶片、茎及根部。由于寄主植物通过克隆繁殖,因此病害发生与营养体的系统性侵染有关。文中评述了寄主植物的生长模式以及该菌在其寄主群体中长期生存的状况。     

小绒泡菌生活史及其形态建成期显微结构的观察

在完成小绒泡菌Physaum pusillum生活史的同时,观察了小绒泡菌P. pusillum生活史不同阶段显微结构的形成。生活史研究结果表明：小绒泡菌P. pusillum的孢子萌发方式为裂式,黏变形体具两根鞭毛,较短一根不易观察到,原质团类型为显型原质团,子实体形态建成过程约需12-13h。显微结构观察结果表明：石灰结和孢丝结构在原质团隆起约6h后开始形成,孢子在原质团隆起约9h后开始形成,石灰结和孢丝先于孢子形成。     

基于核糖体基因的鳞钙皮菌种内分子关系分析

为探讨不同地域的鳞钙皮菌Didymium squamulosum种内分子亲缘关系,通过PCR扩增鳞钙皮菌子实体及原质团DNA,得到SSU、ITS1-5.8S-ITS2 rRNA基因区域,并以SSU、5.8S rRNA基因片段构建NJ亲缘关系树。     

一种快速高效提取病原真菌DNA作为PCR模板的方法

真菌rDNA-ITS序列分析适合于较高等级水平的生物群体间的系统分析。真菌DNA的提取采用传统的方法,步骤繁琐,需要较长时间。采用Chelex-100法提取真菌DNA,使用PCR扩增rDNA-ITS序列评价提取核酸的质量。结果显示,该方法具有经济、简便、快速、高效的特点,是一种比较理想的提取真菌基因组DNA作为PCR模板的方法。     

An immunological approach to the isolation of factors with mitotic activity from the plasmodial stage of the myxomycete Physarum polycephalum

Nuclear divisions in plasmodia of Physarum polycephalum were advanced by applying immunologically purified plasmodial extracts of late G2 phase on the surface of plasmodia which were 1.5 h before the third mitosis. The purification of G2 extracts was achieved by interaction of antibodies prepared against the antigens of early S phase plasmodia with the antigens of late G2 plasmodia. There was no advancement of mitosis by extracts prepared from early S phase plasmodia. Untreated G2 extracts did not accelerate mitosis with the same effectiveness as did antibody purified G2 extracts.     

不同地理种群美洲斑潜蝇及近缘种的rDNA-ITS1序列分析和比较

【目的】通过对美洲斑潜蝇Liriomyza sativae Blanchard 不同地理种群及近缘种间的核糖体DNA第一内转录间隔区（rDNA-ITS1）进行比较,分析美洲斑潜蝇不同地理种群间的遗传分化情况,并为美洲斑潜蝇与近缘种间提供分子鉴别标记。【方法】用PCR产物直接测序法及克隆测序法对我国美洲斑潜蝇8个地理种群的rDNA-ITS1序列进行测序,并调用GenBank中3个近缘种的rDNA-ITS序列,运用软件MEGA3.1对美洲斑潜蝇不同地理种群及近缘种间的rDNA-ITS1序列进行分析。【结果】美洲斑潜蝇8个地理种群间的分化程度较低,只有8个变异位点,遗传距离都在0.02以下,但4个近缘种间的碱基差异显著,遗传距离为0.149～0.390,有126个变异位点,12个美洲斑潜蝇特异性识别位点。【结论】虽然基于rDNA-ITS1序列所显示的美洲斑潜蝇各地理种群之间的遗传分化很小,但是其分化趋势与地理分布基本相吻合;得到的12个特异性识别位点不仅可以作为美洲斑潜蝇与其近缘种间鉴别的分子标记,而且可为今后设计鉴别性PCR引物提供重要的参考依据。     

Evolution of Lycopodiaceae from results of sequencing spacer sequences of chloroplast rRNA genes

Nucleotide sequences of a chloroplast rDNA region including 8 bp from the 3' end of 23S rDNA-ITS2-4.5S rDNA-ITS3-5S rDNA-ITS4 (approximately 800 bp) were determined in 25 species of Lycopodiaceae and two species of the genus Isoetes. The rate of molecular evolution of spacers significantly varied in different Lycopsida taxa. A phylogenetic analysis by the neighbor-joining (NJ) method revealed that the family Lycopodiaceae is monophyletic. The topology of phylogenetic trees suggests the isolation of four or probably five genera in family Lycopodiaceae. For these genera, synapomorphic indels were detected. The obtained data were compared with the results of phylogenetic analysis of Lycopsida with regard to other sequences. The relationships of taxa within the family Lycopodiaceae is discussed.     

Advancing the onset of mitosis by cell free preparations of Physarum polycephalum

Mitotic divisions in the plasmodia of Physarum polycephalum were advanced by about 1 h by applying to the plasmodial surface extracts of other plasmodia. Advancement of mitosis was greatest when the extracts were prepared from plasmodia harvested at late G2. The activity in the extracts responsible for the advancement of mitosis was found to be heat labile and non-dialysable. It is suggested that this activity belongs to proteins responsible for the regulation of mitosis.     

Temperature-sensitive mutants of Physarum polycephalum: viability, growth, and nuclear replication.

Using a selfing strain of Physarum polycephalum that forms haploid plasmodia, we have isolated temperature-sensitive growth mutants in two ways. The negative selectant, netropsin, was used to enrich for temperature-sensitive mutants among a population of mutagenized amoebae, and, separately, a nonselective screening method was used to isolate plasmodial temperature-sensitive mutants among clonal plasmodia derived from mutagenized amoebae. Complementation in heterokaryons was used to sort the mutants into nine functional groups. When transferred to the restrictive temperature, two mutants immediately lysed, whereas the remainder slowed or stopped growing. Of the two lytic mutants, one affected both amoebae and plasmodia, and the other affected plasmodia alone. The growth-defective mutants were examined for protein and deoxyribonucleic acid synthesis and for aberrations in mitotic behavior. One mutant may be defective in both protein and deoxyribonucleic acid synthesis, and another only in deoxyribonucleic acid synthesis. The latter shows a striking reduction in the frequency of postmitotic reconstruction nuclei at the restrictive temperature. We believe that this mutant, MA67, is affected in a step in the nuclear replication cycle occurring late in G2. Execution of this step is necessary for both mitosis and chromosome replication.     

Patterns of oscillation during mitosis in plasmodia ofPhysarum polycephalum

Summary The rhythmic contraction pattern in plasmodia ofPhysarum polycephalum was studied to determine whether characteristic changes occur during the synchronized nuclear division. An electrical method that measures the contraction rhythm in situ during several cell cycles was used. Biopsies of the plasmodia were taken at 17 min intervals for precise determination of the cell cycle stages and were correlated with the simultaneously measured contraction rhythm. All measurements were performed in a temperature controlled environment (27 °C) at 100% relative humidity with the plasmodia (less than 24 h old) growing on a semi-defined agar medium. A total of 14 different plasmodia have been examined, and on one occasion the plasmodium was followed through 3 subsequent mitoses. The mitotic stages were identified with aceto-orcein coloring techniques and by fluorescence methods. Except for a few cases where a mitotic asynchrony of 2–3 min was observed, the mitotic events occurred simultaneously in the nuclei within a single plasmodium. Both the occurrence of the first mitosis after inoculation and the intermitotic times were highly variable. Our study indicates that the contraction rhythm in plasmodia ofPhysarum is unperturbed during the synchronized nuclear division. However, in 5 of the 17 examined mitoses an amplitude decay was observed. We discuss possible explanations for the obtained results with emphasis on the applied techniques, interpretation of the oscillation patterns, and possible restrictions in the cell itself.