戊唑醇对小麦纹枯菌超微结构的影响*

用光镜和电镜技术研究了新型三唑类杀菌剂戊唑醇对小麦纹枯病菌发育的组织学和超微结构的影响。光镜和电镜观察发现戊唑醇处理导致病菌发生一系列变化：菌丝呈现念珠状异常膨大和缢缩,菌丝原生质体内液泡和电子致密体增多,菌丝细胞壁不规则加厚,部分菌丝的隔膜发育受阻成畸形,不能形成正常的桶孔隔膜和桶孔覆垫。有些菌丝原生质体内出现菌丝内套菌丝的现象。这些变化最终导致菌丝细胞解体死亡。     

戊唑醇对小麦纹枯菌超微结构的影响

用光镜和电镜技术研究了新型三唑类杀菌剂戊唑醇对小麦纹枯病菌发育的组织学和超微结构的影响,光镜和电镜观察发现戊唑醇处理导致病菌发生一系列变化：菌丝呈现念珠状异常膨大和缢缩,菌丝原生质体内液泡和电子致密体增多,菌丝细胞壁不规则加厚,部分菌丝的隔膜发育受阻成畸形,不能形成正常的桶孔隔膜和桶孔覆垫。有些菌丝原生质体内出现菌丝内套菌丝的现象,这些变化最终导致菌丝细胞解体死亡。     

小麦条锈菌胞间菌丝的超微结构和细胞化学研究

本文应用电镜技术和细胞化学方法,对小麦条锈菌寄主胞间菌丝的超微结构进行了研究。观察发现:小麦条锈菌胞间菌丝有两种类型,即具隔膜菌丝和无隔膜菌丝。在胞间菌丝中,多核现象极为普遍。常规染色和细胞化学染色结果表明:胞间菌丝的细胞壁由四层组成,隔膜由三层构成,细胞壁的内层与隔膜的外层相连,细胞壁和隔膜中含有蛋白质和多糖物质。隔膜的发育可分三个阶段,即隔膜突的形成,隔膜壁的延伸和隔膜孔结构的形成。本研究中还观察到胞间菌丝间的融合现象。本文的研究结果表明:小麦条锈菌胞间菌丝的一些特征显然不同于其它锈菌。     

内吸杀菌剂甲霜灵对向日葵霜霉菌发育的影响

利用电镜技术研究了内吸杀菌剂甲霜灵对向日葵霜霉菌在寄主向日葵上发育的影响。观察发现甲霜灵引起病菌发生一系列的变化。病菌胞间菌丝细胞内液泡、电子致密体增加,线粒体肿胀畸形;菌丝细胞壁不规则地加厚,部分极度加厚的细胞壁构成了假隔膜,并且在菌丝上还有不规则的突起形成。病菌吸器外间质明显不规则地变宽,其中充满大量电子致密度高的物质：部分吸器体不能正常扩张膨大,发育受阻而成畸形。向日葵霜霉菌的以上变化导致菌丝细胞和吸器的坏死,从而抑制了病菌的进一步发育。     

玉蜀黍丝核菌的鉴定特征

在研究四川省玉米纹枯病菌系时,获得２３个ＲｈｉｚｏｃｔｏｎｉｚｅａｅＶｏｏｒｈｅｅｓ菌株,该菌菌丝形态与Ｐ．ｓｏｌａｎｉＫｕｈｎ相似;菌丝细胞含３个以上椭圆形细胞核,隔膜具桶孔,桶孔周围的隔膜肿体（ｓｅｐｔａｌｓｗｅｌｌｉｎｇ）与隔膜板垂直,两端不再膨大外延;菌丝生长较快,在ＰＤＡ平板２６℃下培养,每２４小时菌落直径增长３３．２～３９．１ｍｍ;菌丝与苯酚相互作用后,着色较深,呈深褐色,菌核初期为白     

种衣剂17号包衣对小麦条锈菌发育影响的超微结构研究

本研究采用电镜种衣剂１７号泽小麦条锈菌发育的影响。观察结果表明,该种衣剂引病菌和寄主细胞内发生了一系列变化。病菌菌丝和吸器内脂肪粒和液泡明显增加;菌丝壁和吸器壁呈不规则加厚;菌丝分枝处玩隔膜产生或隔膜畸形;有的吸器母细胞产生的畸形人侵栓,大都不能穿管寄主细胞壁,初生吸器外音质同内沉积有染色较深的物质,次生吸器可产生多个不规则分枝,但不能扩张膨大,菌丝外渗的物质可能引起寄主细胞的坏死;大多数受侵寄主     

戊唑醇对赤霉病菌生长发育影响的细胞学和免疫细胞化学研究

本文采用电子显微镜和免疫细胞化学技术研究了三唑类杀菌剂戊唑醇 (Tebuconazole) 对小麦赤霉病菌Fusarium graminearum菌丝的形态结构、细胞壁成份和毒素产生的影响。结果表明,戊唑醇不但强烈抑制了培养基上菌丝的生长,而且可引起菌丝形态和结构的明显畸形。电镜观察发现,药剂处理后菌丝呈现不规则的肿胀、过度分枝;菌丝细胞壁不规则加厚,尤其是菌丝顶端部位加厚明显;菌丝形成的不完整隔膜增多,且隔膜壁不规则增厚;菌丝细胞内液泡增加、脂肪粒累积,细胞器排列紊乱,原生质最终坏死。有时在坏死菌丝内可发现新的子菌丝,但子菌丝细胞壁不规则加厚、细胞质坏死、也呈不正常状态,并可再度形成新的菌丝。免疫细胞化学标记表明,药剂处理后菌丝细胞中毒素的标记密度明显低于对照菌丝,表明毒素的产生受到了抑制;而菌丝细胞壁的主要成份几丁质和-1,3-葡聚糖的标记密度明显高于对照处理,表明药剂处理可引起菌丝细胞壁中几丁质和-1,3-葡聚糖的过度累积。     

立枯丝核菌菌丝隔膜超微结构的研究*

通过生长两天的菌丝超微结构的观察,我们首次发现在生长两天的菌丝中内含体堵塞隔膜孔帽的孔口和隔膜孔通道中折叠的细胞膜挤压通道,使通道成扁半圆形。因此,推测内含体和隔膜孔通道中折叠的细胞膜在菌丝隔膜形成后的最初阶段调控相邻细胞间物质的流量和物质交流,维持细胞间的动态平衡。通过生长两天的菌核内隔膜超微结构的观察,隔膜孔通道已被完全堵塞,菌丝细胞显示出了老化的结构特征。     

立枯丝核菌菌丝隔膜超微结构的研究

通过生长两天的菌丝超微结构的观察,我们首次发现在生长两天的菌丝中内含体堵塞隔膜孔帽的孔口和隔膜孔通道中折叠的细胞膜挤压通道,使通道成扁半圆形。因此,推测内含体和隔膜孔通道中折叠的细胞膜在菌丝隔膜形成后的最初阶段调控相邻细胞问物质的流量和物质交流,维持细胞问的动态平衡。通过生长两天的菌核内隔膜超微结构的观察,隔膜孔通道已被完全堵塞,菌丝细胞显示出了老化的结构特征。     

玉蜀黍丝核菌的鉴定特征

在研究四川省玉米纹枯病菌系时,获得23个RhizoctoniazeaeVoorhees菌株。该菌菌丝形态与R.solaniKuhn相似;菌丝细胞含3个以上椭圆形细胞核;隔膜具桶孔,桶孔周围的隔膜肿体（septalswelling）与隔膜板垂直,两端不再膨大外延;菌丝生长较快,在PDA平板26℃下培养,每24小时菌落直径增长33．2～39．1mm;菌丝与苯酚相互作用后,着色较深,呈深褐色。菌核初期为白色或奶油色,成熟后为红褐色;常散生于基质内,少数着生在基质表面和培养皿的内壁上;球形或近球形,直径0.25～1．0mm;结构致密,无菌环和菌髓分化,表面较光滑。223个R.zeae菌株均与Oniki等建立的WaiteacircinataWAG－Z标准菌株发生融合,但与R.solani的融合群标准菌株均无融合反应。R.zeae对玉米的致病性较强,稍弱于玉米纹枯病主菌系R.solaniAG－1－IA,其病斑多半不连续,颜色较深,也可0侵染果穗引致穗腐并在苞叶上产生大量红褐至黑褐色的小菌核。     

离体细胞共培养中科间细胞共质体的形成

离体培养下选出的绿色胡萝卜（Ｄａｕｃｕｓ ｃａｒｏｔａ）细胞系和白色普通烟草（Ｎｉｃｏｔｉａｎａｔａｂａｃｕｍ ）细胞系,各自具有独特的细胞结构标志,在愈伤组织、光镜和电镜３ 个水平上均可区分。对两个细胞系进行分散、混合、Ｋ＋ 液低渗处理后在固体培养基上共培养,１０—１５ ｄ 后可观察到两种细胞的镶嵌生长。光镜和电镜下均观察到烟草细胞和胡萝卜细胞之间隔离层的存在与消失。在隔离层消失的区域可见到异种细胞间次生胞间连丝的形成,从而将独立的两个共质体连成一个统一的共质体。对科间细胞共质联系的建立过程进行了讨论,认为细胞接触后首先非特异粘连——以隔离层形成并适度加厚为标志,然后特异的细胞识别在隔离层中启动,从而导致隔离层或消失而重新建立共质联系或加厚、木质化、木栓化     

细叶黄芪叶肉原生质体发育早期细胞壁再生的研究

采用透射电镜术、电镜多糖细胞化学染色、细胞壁荧光染色以及香豆素抑制细胞壁再生等方法,对细叶黄芪（Ａｓｔｒａｇａｌｕｓｍ ｅｌｉｌｏｔｏｉｄｅｓ ｖａｒ．ｔｅｎｕｉｓ）叶肉原生质体细胞壁的再生及其化学特点进行了研究。结果表明,离体培养２４ 小时的原生质体表面产生一些突起小泡,有时可见少量纤维组分的形成。培养３ 天时这种纤维组分明显增多。至５ 天时可清楚看到再生壁是由纤维和颗粒构成。六亚甲四胺银染色证明它们都是由多糖组分组成的。另外,培养３６ 小时的原生质体有相互粘连的现象。电镜观察、荧光染色及香豆素处理的研究表明粘连与再生壁的形成有关。根据上述观察结果,对原生质体再生壁的结构及其化学性质等问题进行了讨论     

Application of fluorescent probes to study structural changes in Aspergillus fumigatus exposed to amphotericin B, itraconazole, and voriconazole

The broad objective of this study was to document patterns of structural changes following antifungal treatment, and to determine any relationship with minimum inhibitory concentration (MIC) of an antifungal. Three clinical isolates of Aspergillus fumigatus, with high, intermediate, and low amphotericin B (AB), itraconazole (IZ), and voriconazole (VZ) MICs were studied in 24-well plates with cover slips. The fluorescent probes used were Calcofluor White (cell wall), propidium iodide (nucleus), and MitoTracker Green FM (mitochondria). Fluorescent microscopy as early as 3-h after exposure revealed that AB treated hyphae had intact cell wall with deformed mitochondria and nuclei while IZ and VZ treated hyphae revealed no intact cell wall, and deformation of mitochondria and nuclei. At 48 h, AB treated cells revealed rupture of hyphae and disintegration of mitochondria, and nuclei, IZ treated hyphae were swollen with disintegration of mitochondria, and nuclei while VZ treated hyphae showed rupture and disintegration of mitochondria and nuclei. The structural changes for the three strains studied were similar in fluorescent microscopy as long as the incubation time and their respective MICs were used. Thus, AB, IZ, and VZ induced gross organelle defects in A. fumigatus nuclei, mitochondria, and cell wall, which were consistent with respective MICs of antifungals used.     

百合细胞中内生菌的发现

在健康的百合鳞茎、地上茎、叶和幼胚等组织的细胞中发现了“内生菌”。通过光学显微镜、电镜和组培等观察,“内生菌”不影响百合细胞的正常分裂和分化,再生植株健壮。这些细菌还能随着宿主细胞的分裂转移到子细胞。“内生菌”椭圆形或近圆球形。电镜切片菌壁厚,单层,均质,为典型革兰氏阳性菌壁结构。电镜观察结果与光学显微镜观察内生菌的革兰氏染色反应是一致的。     

吲(口朶)乙酸、激动素和单糖对苘麻愈伤组织细胞壁组分的影响

本实验采用 H~3标记糖饲喂示踪分析法。将苘麻愈伤组织分别用吲哚乙酸和激动素处理,并在培养基中供给 H~3葡萄糖或 H~3半乳糖。——实验表明,激素对细胞壁组成分影响,不仅与激素的种类有关,也与供给的外源单糖有关。而外源单糖(半乳糖、葡萄糖)单独加入或同时加入,使激素对苘麻愈伤组织诱导的影响又有所不同。在吲哚乙酸作用下,促进了 H~3葡萄糖的掺入;而半乳糖的加入又抑制了 H~3葡萄糖掺入到细胞壁各组分。在激动素作用下,促进H~3半乳糖的掺入,而葡萄糖的加入又抑制了 H~3半乳糖掺入到壁的各组分。     

Ultrastructural effects of date extract on Candida albicans

The effect of Berhi date extract on the ultrastructure of Candida albicans was studied by scanning and transmission electron microscopy. Exposure of yeast to 5% (w/v) date extract showed evidence of weakening in the cell wall with indications of cell distortion and partial collapse in some cases as seen by scanning electron microscopy. Increasing the concentration of date extract (20%, w/v) led to more drastic damage to the yeast with cell lysis and concurrent leakage of cytoplasmic material with eventual cell death. Ultrastructural investigation showed irregular shapes of cells treated with date extract, with prominent effects on cell wall layers. Cell membranes lost their integrity, aggregation of the cytoplasmic contents and large detachment of plasmalemma from cell wall was observed in the treated cells. These results suggest that date extract may have multiple effects on Candida with an increasing potential of using it for prophylaxis purposes. This revised version was published online in June 2006 with corrections to the Cover Date.     

An Ustilago maydis Mutant Partially Blocked in P45014DM Activity Is Hypersensitive to Azole Fungicides

An Ustilago maydis ergosterol biosynthesis mutant (A14) which is partially blocked in sterol 14alpha-demethylase (P45014DM) activity is described. This mutant accumulated the abnormal 14alpha-methyl sterols, eburicol, 14alpha-methylfecosterol, and obtusifoliol, along with significant amounts of ergosterol. Although the A14 mutant grew nearly as well as the wild type, it was impaired in cell extension growth, which indicated a dysfunction in apical cell wall synthesis. The mutant was also found to be hypersensitive to the azole fungicides penconazole and tebuconazole.     

3,4-Dehydroproline inhibits cell wall assembly and cell division in tobacco protoplasts.

We investigated the function of cell wall hydroxyproline-rich glycoproteins by observing the effects of a selective inhibitor of prolyl hydroxylase, 3,4-dehydro-L-proline (Dhp), on wall regeneration by Nicotiana tabacum mesophyll cell protoplasts. Protoplasts treated with micromolar concentrations of Dhp do not develop osmotic stability and do not initiate mitosis. The architecture of regenerated cell walls was examined using deep-etch, freeze-fracture electron microscopy of rapidly frozen tobacco cells. Untreated protoplasts assemble a dense fibrillar cell wall consisting of laterally associating subelementary fibrils. In contrast, treatment of protoplasts with Dhp alters the structure of the regenerated wall fibrils in several ways: first, the microfibrils are coated with globular knobs; second, some larger fiber bundles have an open ribbon-like appearance; and third, the smallest subelementary fibrils were not visible. Tobacco cells develop an abnormal morphology as a consequence of this abnormal cell wall structure. Thus, inhibition of prolyl hydroxylase results in the regeneration of a cell wall with abnormal structural and functional properties. These data provide experimental evidence that hydroxyproline-rich glycoproteins are important for the structural integrity of primary cell walls and for the correct assembly of other wall polymers, and that wall structure is an important regulator of cell division and cell morphology.