青杨叶锈病菌(Melampsora larici-populina Kleb.)侵染过程的超微结构研究

 采用电子显微镜技术对青杨叶锈病菌(Melampsora larici-populina Kleb.)的侵染过程进行了研究。发现该菌夏孢子萌发产生1~3个芽管,且具较多的树杈状分枝。芽管由气孔侵入,侵入前不形成明显的附着胞或仅个别芽管形成附着胞。芽管侵入气孔后在气孔腔内形成气孔下囊,再分化出圆形的膨大体而产生1~2支初生菌丝。初生菌丝在寄主细胞间扩展,与叶肉细胞壁接触后分化出吸器母细胞,吸器母细胞中的细胞器与胞间菌丝相同,双核。吸器母细胞产生侵入钉侵入叶肉细胞内部形成吸器,成熟吸器由细长具颈环的管状颈部和膨大的吸器体组成,此时胞间菌丝在吸器母细胞处分化出次生菌丝,在叶肉细胞间扩展形成次生菌落,产生孢子堆。病菌在寄主细胞间隙或沿寄主细胞壁延伸时,寄主细胞仍保持正常状态。     

多堆柄锈菌侵染不同抗性玉米的组织病理学研究

［目的］ 了解不同抗感玉米自交系对南方锈病的组织病理学反应,为今后筛选不同类型抗病自交系提供参考。［方法］ 采用曲利苯蓝透明染色法,以4个玉米自交系为材料,研究了玉米南方锈病致病菌—多堆柄锈菌在不同抗性材料上侵染过程的组织学特征。［结果］ 多堆柄锈菌侵入和定殖可以分为5个阶段：孢子萌发与芽管形成、附着胞形成、侵入细胞、胞内吸器产生、菌丝在细胞间扩展。在不同抗性的玉米材料上,病菌孢子萌发和芽管形成差别不明显,但侵入后病菌在不同抗性材料内的发育进程和发育程度具有显著差异。在抗病玉米材料上,病菌初生菌丝、吸器母细胞、次生菌丝的形成时间推迟,胞内吸器少,菌丝分枝少,菌丝生长缓慢。［结论］ 这些抗性特征与田间表现出的细胞过敏性坏死、叶片上夏孢子堆少的特征具有一致性。     

小麦叶锈菌在感病寄主上发育的组织病理学和超微结构研究

 应用荧光显微技术、微分干涉技术和生物电镜技术,系统地研究了小麦叶锈菌在感病寄主上的发育过程及其超微结构特征。小麦叶锈菌在感病品种上的发育过程可分为几个明显的阶段,即孢子的萌发、附着胞的形成、气孔下囊的分化、初生菌丝和次生菌丝的形成和生长、吸器母细胞和吸器的形成、夏孢子床和夏孢子堆的产生以及夏孢子的形成。小麦叶锈菌的胞间菌丝呈丝状,生长和分枝通常沿寄主细胞壁进行。胞间菌丝与寄主细胞的接触诱导了吸器母细胞的分化,吸器母细胞在与寄主细胞壁的接触部位发育形成入侵栓,穿透寄主细胞壁后于细胞内形成吸器。胞间菌丝和吸器母细胞均含有双核,而成熟吸器则含有单核。经常规染色后,胞间菌丝和吸器母细胞的壁与隔膜均可分辨出由多层构成。     

小麦新抗源一粒葡抗条锈病的组织学和超微结构研究

 采用荧光显微镜、微分干涉显微镜和电子显微镜技术,系统研究了小麦新抗源一粒葡抗小麦条锈病的组织学和超微结构特征。结果表明:相对于感病品种铭贤169,一粒葡对条锈菌的侵染,在组织学和超微结构上均表现出明显的抗性特征。在组织学水平,表现为菌丝生长受抑,菌落发育延迟或败育,吸器母细胞和吸器数目明显减少;同时,侵染点的寄主细胞表现出不同程度的过敏性坏死症状。电镜观察发现,在一粒葡和感病品种中,条锈菌均可由芽管顶端直接进入或通过形成附着胞进入小麦气孔。其后,在一粒葡上,病菌胞间菌丝、吸器母细胞、吸器在细胞和亚细胞水平均发生了一系列异常变化,表现为原生质染色逐渐加深,液泡增多变大,逐渐消解原生质;胞间菌丝、吸器母细胞细胞壁不规则增厚;胞间菌丝线粒体肿胀,数目增多,逐渐解体;吸器母细胞细胞质逐渐空泡化后丧失其生理功能;吸器外质膜皱褶;吸器外间质加宽并有丝状或颗粒状物质形成,吸器体壁逐渐消解出现孔洞,吸器体最终畸形坏死。同时,寄主细胞产生一系列显著的结构防卫反应:形成胞壁沉积物、乳突、吸器鞘等结构,以及发生坏死,阻碍并抑制病菌的发育及扩展。     

三唑酮拌种对小麦条锈病菌叶部侵染过程的影响

 供试小麦种子用干拌法施药,在一叶期接种小麦条锈菌(Puccinia striiformis West)毒性小种,用整叶透明染色方法研究三唑酮拌种对条锈菌叶部侵染过程的影响.结果表明,由初生吸器形成开始,三唑酮对吸器产生和菌落扩展表现出多方面的影响.拌种植株叶片中条锈菌侵染菌丝的生长和分枝严重受阻,吸器母细胞和吸器形成显著减少,菌落亦不再进一步分化,多数侵染点还出现寄主细胞坏死。这些组织病理学变化与抗病品种的过敏性坏死反应很相似.     

小麦叶锈菌侵入方式的观测简报

小麦叶锈菌侵入方式的观测简报小麦叶锈菌的侵入方式，以往文献报道为萌发的芽管遇气孔后，形成附着胞覆盖在气孔上，长出侵入丝钻入气孔，并在气孔下腔内形成泡囊，再长出侵染丝侵入细胞内形成吸器，完成侵入过程。我们经３年的重复观察，发现除上述方式外，还有大量的锈...     

种衣剂17号包衣对小麦条锈菌影响的组织病理学研究

种衣剂１７号内含三唑醇等杀菌剂和其它化学成分，小麦种子用种衣剂包衣，出苗后一叶期接咱小麦条锈菌毒性小种，采用荧光染色技术和整叶透明技术，研究了种衣剂１７号对条锈菌在寄主内发育的影响。观察结果表明，该种衣剂对对叶表的夏孢子萌发和芽管入侵无明显影响，但可使菌丝扩展和菌丝分枝严重受抑，吸器母细胞和吸器形成的数目减少，每一侵染点的吸器一般不超过２个，并且对吸器具有致畸作用。在多数侵染点内可观察到寄主细胞坏     

小麦与条锈病菌不亲和互作的超微结构

 采用条锈菌同一小种的野生型菌系和弱毒突变菌系,分别接种同一小麦品种的方法,研究了不亲和互作的超微结构特征。在不亲和互作中,条锈菌的胞间菌丝、吸器母细胞和吸器明显受抑。吸器可以在发育早期受抑坏死,也可迟滞至吸器体形成之后坏死。吸器外质膜严重皱褶,并出现孔洞,吸器外间质加宽,沉积大量电子致密物质。侵染位点的小麦叶肉细胞表现与过敏性坏死反应相关联的一系列变化。细胞壁内侧还出现乳突状或颗粒状沉积物等防御结构或物质。     

菜豆锈病菌侵染对寄主超微结构的作用及菜豆抗锈病的细胞学表现

 在电镜下观察发现,菜豆锈病菌侵染菜豆后,逐步对其超微结构产生影响:寄主细胞发生质壁分离;叶绿体变形;叶绿体的片层结构排列零乱;线粒体脊模糊不清,直至叶绿体解体;线粒体空泡化;少数细胞的细胞壁分解;不同细胞的细胞器堆积在一起。同时,病原菌的侵染激发了寄主抗病性的细胞学表现:供试的抗感菜豆品种都表现为在病原菌侵入位点的寄主细胞壁内侧有高电子致密物质沉积;与吸器母细胞接触的寄主细胞壁加厚以及在吸器颈周围有电子不透明物质形成。只是这3种反应在抗病品种中表现得更加强烈。此外,抗病品种中还有一些特有的抗性特征,如被侵染细胞及其相邻细胞的快速坏死,吸器母细胞侵入位点的寄主细胞壁外侧也有一种高电子致密物质沉积,抗病品种中真菌吸器周围聚集含大量线粒体的寄主细胞的细胞质,且吸器外基质比感病品种中的宽。     

小麦条锈菌夏孢子阶段核相状况的研究

 本文应用电镜和荧光染色技术对小麦条锈菌夏孢子阶段的细胞核相状况进行了系统研究。观察发现夏孢子通常为双核体,夏孢子萌发的芽管可为双核、三核或四核,但仍以双核芽管为主。尽管在胞间菌丝和吸器母细胞中可观察到典型的双核细胞,但胞间菌丝和吸器母细胞中的多核现象极为普遍。经连续切片观察证实,吸器的细胞核可为双核、三核、四核、五核和六核,一般以四核为主。夏孢子堆中的不同类型的细胞均为双核。上述研究结果表明,小麦条锈菌的细胞核相状况较为复杂,并且显然不向于其它锈菌。关于小麦条锈菌多核现象的生物学意义值得进一步研究。     

Scanning electron microscopy study of infection structure formation of Puccinia graminis f.sp. tritici in host and non-host cereal species

The development of uredospore-derived infection structures of Puccinia graminis f.sp. tritici in wheat, barley, sorghum and maize was examined by scanning electron microscopy (SEM). Germ tubes grew over the leaf surface until a stoma was located. An appressorium formed over the stoma and the leaf was penetrated by an infection peg. Within the substomatal chamber of all species the infection peg developed a substomatal vesicle by 6 h post-inoculation (hpi). from which a primary infection hypha developed parallel to the long axis of the leaf. In wheat, barley and maize, when a primary infection hypha abutted onto a host cell, a septum was laid down between the tip of the hypha and the substomatal vesicle, delimiting a haustorial mother cell by 12 hpi; haustorial mother cells did not form in sorghum. Secondary infection hyphae arose on the substomatal vesicle side of the septum; infection did not progress further in maize, but in wheat and barley secondary infection hyphae branched, and proliferated intercellularly forming the fungal thallus. A haustorial mother cell was delimited when an intercellular hypha abutted onto a host cell. Infection sites with haustorial mother cells were observed at 12 hpi in barley and 24 hpi in wheat. In all four plant species, some atypical substomatal vesicle initials, substomatal vesicles and primary infection hyphae were observed.     

禾顶囊壳小麦变种对小麦种子根的侵染过程

 光镜和电镜观察表明,禾顶囊壳小麦变种(Gaeumannomyces graminis var.tritici,小麦全蚀病菌)对小麦种子根的侵染过程可分为侵入前、侵入表皮层、进入皮层和进入中柱等4个连续阶段。麦根接菌后在15℃下培养,48 h后侵入表皮层细胞,60 h后进入皮层,120 h后进入中柱。病原菌主要以侵染菌丝直接侵入表皮层,表皮细胞间隙和根毛基细胞是主要侵入部位,少数由附着枝侵入。菌丝穿透细胞壁有明显的酶解作用特征,菌丝先端前方胞壁上还产生电子密物质。皮层细胞是病原菌定殖和发展的主要场所,病原菌还能离解胞间层,形成胞外空间,特别有利于菌丝和菌丝束的扩展。在侵入位点的寄主细胞壁和质膜之间,形成多种形状的木质管,其数量与侵入菌丝的数目相对应,但木质管不能阻止菌丝进入细胞。菌丝进入中柱后,可阻塞导管和筛管。小麦细胞发生退行性病变,尤以细胞壁膨大崩坏和早期质壁分离最明显,细胞间隙还产生性质不明的黄色物质。     

荸荠茎点霉秆枯病菌侵染过程的超微观察

<正>荸荠(Eleocharis dulcis),又称马蹄,为莎草科多年生草本植物,是一种具有食用和药用价值的水生蔬菜。近年来,随着荸荠在我国种植面积的不断扩大,病害发生也呈逐年上升趋势。荸荠茎点霉秆枯病是2009年在湖北省荸荠产区发现的一种新病害,由Phoma bellidis侵染引起,该病在湖北省团风地区发生尤为严重,对荸荠的产量和品质造成严重影响;病害一般在8~12月发生,发病初期在荸荠茎秆上产生圆形或梭形红褐色小斑,随后病斑沿茎     

Nonhost versus host resistance to the grapevine downy mildew, Plasmopara viticola, studied at the tissue level

Following inoculation of host and nonhost plants with Plasmopara viticola, the grapevine downy mildew, a histological survey was undertaken to identify the stage where its development is contained in nonhosts and in resistant host plants. Three herbaceous nonhost species, Beta vulgaris, Lactuca sativa, and Capsicum annuum, and three grapevine species displaying different level of resistance (Vitis vinifera [susceptible], Vitis riparia [partially resistant] and Muscadinia rotundifolia [totally resistant]) where inoculated by P. viticola using a controlled leaf disk inoculation bioassay. During the early steps of infection, defined as encystment of zoospores on stomata, penetration of the germ tube, and production of the vesicle with the primary hypha, there was no evidence of a clear-cut preference to grapevine tissues that could attest to host specificity. The main difference between host grapevine species and nonhosts was observed during the haustorium formation stage. In nonhost tissues, the infection was stopped by cell wall-associated defense responses before any mature haustorium could appear. Defense responses in resistant grapevines were triggered when haustoria were fully visible and corresponded to hypersensitive responses. These observations illustrate that, for P. viticola, haustorium formation is not only a key stage for the establishment of biotrophy but also for the host specificity and the recognition by grapevine resistance factors.     

The development of different pathotype groups of Mycosphaerella pinocles in susceptible and partially resistant pea leaves

The progress of infection by high- and low-virulence isolates of Mycosphaerella pinodes was examined in susceptible and partially resistant pea leaves. Conidia germinated with one or more germ tubes which frequently branched and formed appressorium-like structures on the leaf surface. Penetration occurred through the epidermal walls. A structure similar to an infection vesicle was formed, lying partly in the epidermal wail and partly in the cell lumen. From this structure, a penetration hypha was derived which initiated the development of the intra- and intercellularly-growing fungal colony. Infections led to rapid tissue collapse in both susceptible and resistant interactions. In resistant interactions, the formation of infection vesicles and penetration hyphae was reduced, and the development and spread of lesions was retarded.     

Leaf wax layer may prevent appressorium differentiation but does not influence orientation of the leaf rust fungus Puccinia hordei on Hordeum chilense leaves

The infection process of most rust fungi start with spore germination, directional growth of the germ tube towards a stoma, differentiation of an appressorium over the stoma, and penetration into the substomatal cavity. In the South American wild barley Hordeum chilense Roem. & Schult., wide variation occurs in the degree to which several rust fungal species are able to form appressoria over the stomata. Apparently, features of the plant may hamper early stages of the infection process. Such an early defence is called avoidance. In order to find out how germ tube growth is directed towards stomata, and whether the cuticular wax layer plays a role in this orientated growth and in appressorium differentiation, several orientation and differentiation parameters of Puccinia hordei germ tubes were measured on H. chilense leaves with and without the wax layer. Orientated growth of the germ tubes started upon contact with the epidermal cell junctions. The growth of lateral branches of the germ tube over the first epidermal cell junction that it meets, may help the germ tube to grow along the transverse axis of the leaf. No evidence was found of attraction of the germ tube to stomata. Removal of the cuticular wax layer did not result in loss of germ tube orientation. This suggests that the leaf wax layer has no role in the guidance of germ tubes. On high avoidance accessions, removal of the wax layer allowed appressoria to develop over stomata that would otherwise be overgrown. No effect of the cell widths in stomatal complexes was found on the chance that stomata were overgrown. This suggests that the overgrowth of stomata on H. chilense leaves by P. hordei germ tubes is mainly caused by the wax covering of the stomatal apparatus.     

Prepenetration stages in infection of clematis by Phoma clematidina

A detailed study of conidial germination, germ-tube growth and the formation of infection structures in Phoma clematidina , the causal agent of clematis wilt, is described for two clematis varieties differing in disease resistance. On both the resistant and susceptible varieties, the fungus entered leaves and stems by direct penetration of the cuticle, often, but not always, following the formation of infection structures. More germ tubes per conidium were formed on the susceptible host, but these germ tubes were on average shorter than on the resistant host. Although germ tubes regularly entered the plant via trichomes, stomata were not found to be sites of entry. Following penetration of the cuticle of resistant plants, germ-tube growth was sometimes restricted to the subcuticular region, and halo formation occurred at the sites where penetration was attempted. Subcuticular growth and halo formation were not observed on susceptible plants. These observations may partly explain the resistance of small-flowered clematis varieties to P. clematidina .