Importance of Cell Wall Degrading Enzymes Produced by Fusarium graminearum during Infection of Wheat Heads

Cytological studies were carried out to elucidate the importance of cell wall degrading enzymes (CWDE) during infection of wheat spikes by Fusarium graminearum. Scanning electron micrographs revealed that at 6–24 hours after inoculation (hai) of single spikelets with macroconidia of F. graminearum, the fungus germinated by forming several germ tubes and developed a dense hyphal network in the cavity of the spikelet. At 24–36hai, the fungus formed infection hyphae which invaded the ovary and inner surface of the lemma and palea. Transmission electron microscopical studies revealed that the fungus extended inter- and intracellularly in the ovary, lemma and rachis and caused considerable damage and alterations to the host cell walls. In different tissues of healthy and F. graminearum-infected wheat spikes the cell wall components cellulose, xylan and pectin were localized by means of enzyme-gold and immuno-gold labelling techniques. Localization of cellulose, xylan and pectin showed that host cell walls which were in direct contact with the pathogen surface had reduced gold labelling compared to considerable higher labelling densities of walls distant from the pathogen–host interface or in non-colonized tissues. The reduced gold labelling densities in the infected host cell walls indicate that these polysaccharide degrading enzymes might be important pathogenicity factors of F. graminearum during infection of wheat spikes. The results revealed that, infection and colonization of wheat spikes by F. graminearum and reactions of infected host tissue were similar to those reported for F. culmorum.     

Effect of phytoplasmal infection on photosystem II efficiency and thylakoid membrane protein changes in field grown apple (Malus pumila) leaves

We have studied the effect of the apple proliferation phytoplasmal infection on some features of the thylakoids from field grown apple (Malus pumila) leaves. Changes in photosynthetic pigments, soluble proteins, ribulose-1,5-bisphosphate carboxylase, nitrate reductase, photosynthetic activities and thylakoid membrane proteins were investigated. The level of total chlorophyll and carotenoids were reduced in phytoplasma-infected leaves. Similar results were also observed for soluble proteins and ribulose- 1,5-bisphosphate carboxylase activity. The in vivo nitrate reductase activity was significantly reduced in infected leaves. When various photosynthetic activities were followed in isolated thylakoids, phytoplasmal infection caused marked inhibition of whole chain and photosystem II activity while the inhibition of photosystem I activity was only marginal. The artificial exogenous electron donors, diphenyl carbazide and hydroxylamine significantly restored the loss of photosystem II activity in infected leaves. The same results were obtained when F_v/F_m was evaluated by chlorophyll fluorescence measurements. The marked loss of photosystem II activity in infected leaves could be due to the loss of 47, 33, 28–25, 23 and 17 kDa polypeptides. It is concluded that phytoplasmal infection inactivates the donor side of photosystem II. This conclusion was confirmed by immunological studies showing that the content of the 33 kDa protein of the water-splitting complex was diminished significantly in infected leaves.     

几种芸香科植物花粉形态观察

应用扫描电子显微镜对佛手、代代、柚、柑橘和金橘等芸香科植物花粉进行形态观察。发现这几种植物的花粉在形态、大小、外壁纹饰、网孔等方面均有差异,其中3个佛手品种的花粉具网状纹饰,白衣秀士佛手和青衣童子佛手的网脊连续,而赤金王子佛手的网脊不连续,其他4种芸香科花粉具穴状纹饰,代代和柚的花粉比柑橘和金橘的花粉大,而代代和金橘的花粉表面孔穴密度比柚和柑橘的花粉表面孔穴密度大。这些花粉表面微观形态的差异将为上述品种鉴定提供依据。     

黄瓜幼叶细胞中钙调素的免疫胶体金定位

 采用免疫胶体金电镜技术对黄瓜幼叶叶肉细胞中的钙调素(CaM)进行定位。结果表明：适温下生长的黄瓜幼苗叶肉细胞中的CaM主要分布于细胞核和叶绿体内，细胞核中CaM主要存在于染色质和核仁上，叶绿体中CaM主要存在于类囊体膜上；线粒体中也有一定量的CaM分布；细胞质、液泡、液泡膜及质膜上只有少量CaM存在；而细胞壁和细胞间隙却很难发现显示CaM存在的金颗粒。     

侧沟茧蜂触角感觉器的扫描电镜观察

扫描电镜观察表明：侧沟茧蜂Microplitis sp．的触角上存在5种感觉器，分别为毛状感觉器A型、B型、C型，槽状感觉器，锥形乳头状感觉器，刺形感觉器和钟形感觉器。对触角感觉器的形态和分布进行了描述，并对两性间的差别进行了讨论，为研究该蜂寻找生境和寄生行为提供依据。     

鸭疫里氏杆菌形态特征的电镜观察

自鸭传染性浆膜炎的病死鸭中分离鉴定的鸭疫里氏杆菌(RA)Ⅰ、Ⅱ型两个菌株，用负染法和超薄切片法制备样品，于透射电镜下进行观察。结果观察到约占1／3的RA具有特殊的形态结构：在细菌的顶端或侧面有1～2个与菌体相连的芽状赘生物，有的也能从菌体上脱落下来。它在超薄切片中的大小约为菌体的1／3～1／5，能观察到其内部也具有类似菌体内部的结构。且具有类似菌体的“细胞壁”，类似于核体部分的结构位于“细胞壁”的一侧。而用相同的方法制作的鸭大肠杆菌(E．cozi)对照样品，却见不到这种特殊的形态结构。说明这种形态特征可能是RA特有的。     

Spread of potato virus Y in potato cultivars (Solanum tuberosum L.) with different levels of sensitivity

The aim of this work was to correlate the appearance of the symptoms, multiplication and spread of virus after mechanical inoculation of potato (Solanum tuberosum L.) cultivars showing different levels of susceptibility and sensitivity to Potato virus Y^NTN (PVY^NTN). The potato cultivars used were the resistant cultivar Sante and susceptible cultivars Igor, Pentland squire and Désirée. The spread of the virus PVY^NTN in infected plants was monitored using different methods: DAS-ELISA, tissue printing, immuno-serological electron microscopy and real-time PCR. In all three susceptible cultivars, the virus was detected in the inoculated leaves 4–5 days after inoculation. From there virus spread rapidly, first into the stem, then more or less simultaneously to the upper leaves and roots. Real-time PCR was shown to be very sensitive and enabled viral RNA to be detected in non-inoculated leaves of susceptible cultivar Igor earlier than other methods. Therefore, for exact studies of plant–virus interaction, a combination of methods which detect viruses on the basis of their different properties (coat protein, morphology or RNA) should be used to monitor the spread of viruses.     

Footsteps from Insect Larvae Damage Leaf Surfaces and Initiate Rapid Responses

Plant resistance to insect herbivory involves gene expression in response to wounding and the detection of insect elicitors in oral secretions (Kessler and Baldwin, 2002, Ann. Rev. Plant/ Biol. 53: 299–328). However, crawling insect larvae stimulate the synthesis of 4-aminobutyrate within minutes and imprints of larval footsteps can be visualized within seconds through superoxide production or transient increases in chlorophyll fluorescence (Bown et al., 2002, Plant Physiol. 129: 1430–1434). Here cryo-scanning electron microscopy was used to demonstrate that larval feet, which are equipped with a perimeter row of hook-like crochets, damage leaf tissue and result in larval footprints. Staining for cell death shows that areas of wounding correspond to footsteps detected through increased chlorophyll fluorescence. Superoxide production in response to footsteps was inhibited by diphenyleneiodonium, an inhibitor of the plasma membrane NADPH oxidase enzyme. Inhibition of superoxide production, however, did not eliminate the detection of cell death. The results demonstrate that larval footsteps damage leaf tissue, and initiate rapid local responses which are not dependent on herbivory or oral secretions. It is proposed that superoxide production at the wound site prevents opportunistic pathogen infection.     

More evidence that anaerobic oxidation of methane is prevalent in soils: Is it time to upgrade our biogeochemical models?

Estimating future fluxes of CH₄ between land and atmosphere requires well-conceived process-based biogeochemical models. Current models do not represent the anaerobic oxidation of methane (AOM) in land surface soils, in spite of increasing evidence that this process is widespread. Our objective was to determine whether AOM, or potential AOM, commonly occurs in 20 hydromorphic soils spanning a wide range of chemical properties. Bulk soil samples were collected under shallow water near the shoreline of 15 recently drained fish ponds in southern Bohemia (Czech Republic), as well as from below the water table at 3 peatland locations in northeast Scotland and 2 acid sulfate soils on the southern coast of Finland. Each soil slurry was incubated under both oxic and anoxic conditions, with or without the addition of alternative electron acceptors (SO₄²⁻ and NO₃⁻) or H₂PO₄⁻. Here, “oxic” and “anoxic” conditions refer to anoxic soil respectively incubated in a headspace containing air or argon. Using the isotope dilution method, we determined the gross production and oxidation rates of CH₄ after 2 days incubation under oxic headspace conditions, and after 2, 21 and 60 days incubation under anoxic conditions. Large differences in net CH₄ fluxes were observed between soil types and between incubation conditions. AOM was detected in each of the 20 bulk soil samples, which spanned >6 pH units and 2 orders of magnitude in organic C content. Significant positive relationships were found between AOM and gross CH₄ production rates under anoxic conditions, resulting in AOM rates that were sometimes higher than CH₄ oxidation rates under oxic headspace conditions. There was no relationship between net and gross CH₄ production rates, such that 2 soil types could display similar low net rates, yet conceal very large differences in gross rates. The effects of alternative electron acceptors on AOM were idiosyncratic and resulted in no net trend. We did find, however, a negative effect of SO₄²⁻ and H₂PO₄⁻ on gross CH₄ production rates under anoxic and oxic conditions respectively. Under oxic headspace conditions, CH₄ oxidation was related to soil organic C content. Taken collectively, our results suggest that AOM, or potential AOM, is prevalent over a wide range of soil types, that AOM may contribute substantially to CH₄ oxidation in soils, and that AOM in soils should be integrated to current process-based CH₄ cycling models.     

不同固定条件对几种植物样品超微结构的影响

为比较几种固定条件对不同植物材料样品电镜超微结构图像清晰度的影响。以3个不同科属4种植物[油棕(Elaeis gineansis Jacq.)、拟南芥(Arabidopsis thaliana.)、花生(Arachis hypogaea Linn.)、柱花草(Stylosanthes guianensias Sw.)]为研究主体代表,探讨了2.5%戊二醛与4%戊二醛固定过夜、锇酸固定2 h及过夜的制备方法对其叶片超微结构的影响,并成功观察了柱花草接种胶苞炭疽野生型菌株CH008后的病菌侵入过程。结果表明:油棕、拟南芥、花生及柱花草叶片经4%戊二醛与1%锇酸固定液固定过夜,其组织成像的效果均较为理想,各种细胞器整体信息丰富、结构组织完好、线条清晰。表明,此制备方法能够较好的保存植物叶片和细胞组织,提高制样的成功率及观察分辨率。