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1.
用组织培养技术将富士苹果茎尖外植体建立在MS培养基中,形成试管苗。取4—6周龄的茎与愈伤组织,徒手切取约1mm厚的横切面,印迹在硝化纤维膜上。印迹组织经封闭后,与CLSV碱性磷酸酶标抗体反应。对反应结果进行显色。感染CLSV的印迹组织呈紫色反应,正常组织无显色反应。结果表明,该方法十分容易检测印迹组织中的CLSV。带毒愈伤组织较茎的显色反应敏感,显色反应时间仅为茎的一半(15min)。CLSV在愈伤组织中有两种分布情况:一是所有组织均有显色反应;二是呈不均匀分布。CLSV在茎组织中有三种分布情况:第一,除髓部外,表皮、皮层及维管系统中均有分布;第二,不均匀分布,CLSV集中分布于表度组织中,在皮层及韧皮部仅有少量分布;第三,“嵌合”分布,即同一种组织中部分组织带毒,部分为正常组织。 相似文献
2.
Wanyoike Mary Wanjiru Kang Zhensheng Heinrich Buchenauer 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(8):803-810
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. 相似文献
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Martin M.T. Glie B. 《European journal of plant pathology / European Foundation for Plant Pathology》1997,103(5):427-431
Plum pox potyvirus (PPV) induces in infected Nicotiana clevelandii cells characteristic crystalline inclusions known as nuclear inclusions (NI) when located in the nucleus and as dense material (Dm) when located in the cytoplasm. Crystalline inclusions contain protease (NIa) and RNA-dependent RNA polymerase (NIb) proteins. It is now well established for all potyviruses that cylindrical inclusions contain CI helicase ATPase protein (Martin et al., 1992). The intracellular location of other non-structural PPV proteins remains unknown. Using Escherichia coli expression vectors, specific antibodies were obtained against P1, P3, 6K2 and NIb PPV proteins for which antibodies were not yet available. As expected, NIb antiserum labelled crystalline inclusions. P1, P3 and 6K2 proteins were present in both types of crystalline inclusions found in the nucleus and in the cytoplasm of PPV-infected leaves of N. clevelandii, suggesting that nuclear inclusions and dense material were composed of the same proteins. This composition is discussed. 相似文献
6.
ZHU Ying-yuan WANG Zi-neng ZENG Yi-ke ZHENG Pei-e XU Jian-ping GUO Zu-wen TANG Fu-xing 《园艺学报》2004,20(6):1038-1041
AIM: To obverse the expression and localization of urocortin on ultrathin cryosections of syncytiotrophoblast of human term placenta with immunocytochemistry technique under transmission electron microscope. METHODS: The human term placenta tissue from Cesarean delivery and normal labor were fixed in 4% paraformaldehyde, and then divided into two parts. One part was for regular immunocytochemistry under microscope, and the other part was used to prepare ultrathin cryosections for immunocytochemistry under transmission electron microscope. RESULTS: 1.Uroncortin mainly distributed in cytoplasm of syncytiotrophoblast of human term placenta under microscope. Urocortin also appeared in cytoplasm in some stromal cells. 2. Under transmission electron microscope, the anti-urocortin gold particles were observed in cytoplasm of syncytioptrophoblast ultrathin cryosections and sited on rough-surfaced endoplasmic reticulum. The anti-urocortin gold particles also appeared on nucleus and nuclear membrane of syncytiotrophoblast. CONCLUSION: Syncytiotrophoblast of human term placenta synthesized and secreted urocortin. The internalization of urocortin within syncytiotrophoblast nuclear indicates that urocortin may act as intracrine. 相似文献
7.
本研究从已构建的苦瓜果实均一化文库筛选得到一个EIN3-like同源EST序列,结合RACE技术,克隆得到EIN3基因c DNA全长序列,命名为Mc EIL2(Gen Bank登录号:KF595122)。该基因全长2 122 bp,开放阅读框1 890 bp,编码629个氨基酸,预测该蛋白的分子量为71.58 k D,与黄瓜Cus EIN3、甜瓜Cm EIL1、苜蓿Mt EIL1、绿豆Vr EIL1和番茄Le EIL1的蛋白同源性分别为89.61%、78.37%、71.23%、69.09%和65.66%。Mc EIL2基因的编码蛋白亚细胞定位于细胞核,荧光定量分析表明Mc EIL2基因表达量在幼果期先强后弱,绿熟期最低,破色期表达量大幅增加至最高随后下降,推测该基因参与苦瓜果实成熟软化调控过程。本研究初步明确了Mc EIL2基因在苦瓜成熟过程表达模式,可为今后进一步揭示苦瓜果实成熟软化的分子机制奠定基础。 相似文献
8.
本研究旨在克隆鸡(Gallus gallus)脂滴包被蛋白基因(Perilipin1)的cDNA序列,并检测其在鸡前脂肪细胞分化过程中的亚细胞定位.本研究以7周龄肉鸡腹部脂肪组织为材料,采用RT-PCR和RACE的方法扩增并克隆了鸡Perilipin1基因的5'UTR、CDS和3'UTR片段,分析了该基因的结构;以鸡原代前脂肪细胞为材料,利用免疫荧光及激光共聚焦技术,在诱导分化的不同时间点(12~120 h),检测了鸡Perilipin1在前脂肪细胞中的表达位置.结果表明,本研究获得的鸡Perilipin11基因5'UTR、CDS和YUTR序列总长度为2 379 bp,该基因由9个外最子、8个内含子组成(ATG位于第二外显子上);在鸡原代前脂肪细胞诱导分化过程中,Perilipin1始终包被在脂滴周围.综上,本研究成功克隆了鸡Perilipinl基因的完整cDNA序列并确定了Perilipin1在鸡前脂肪细胞分化过程中与脂滴的空间位置关系,该结果为深入开展鸡Perilipinl基因的功能研究,揭示鸡脂肪代谢的分子遗传机理提供了重要的理论依据. 相似文献
9.
M. Rokebul Anower M. D. Peel I. W. Mott Y. Wu 《Journal of Agronomy and Crop Science》2017,203(6):506-518
We previously reported an alfalfa half‐sib family, HS‐B, with improved salt tolerance, compared to parental plants, P‐B. In this study, we conducted additional experiments to address potential physiological mechanisms that may contribute to salt tolerance in HS‐B. Vegetatively propagated HS‐B and P‐B plants were treated with a nutrient solution (control) or a nutrient solution containing NaCl (EC = 12 dS/m). Shoots and roots were harvested at various time points after treatment for quantification of proline, soluble sugar, and H2O2 using spectrophotometers. Subcellular localization and quantification of Na in roots were conducted using a Na+‐specific dye under a confocal microscope. HS‐B produced 86 and 89% greater shoot and root dry biomass, respectively, compared to parental plants, P‐B, under salinity in the greenhouse. Under saline conditions the HS‐B shoots accumulated 115% and roots 55% more soluble sugars than P‐B counterparts. The non‐saline HS‐B shoots, however, showed 72% less soluble sugars than the non‐saline P‐B plants. Under saline conditions HS‐B accumulated 39% less proline in shoots, while roots accumulated 56% more proline, compared to their P‐B parents. HS‐B plants also showed a greater reduction of stomatal conductance under mild saline stress. HS‐B shoots and roots contained 3–4 times less reactive oxygen species (H2O2) after salt treatment compared to P‐B plants. Sodium localization and distribution analysis using Na+‐specific dye revealed HS‐B plants accumulated 88% and 48% less Na+ in stele and xylem vessels compared to P‐B. The study provides insights into the potential mechanisms that may contribute to salt tolerance in HS‐B: maintaining RWC by accumulating soluble sugars while reducing transpiration, maintaining healthy status by reducing oxidative stresses, and preventing salt toxicity by reducing accumulation of Na+ inside root cells and xylem. 相似文献
10.