Culture of Differentiated Adult Rabbit Auricular Chondrocytes     

V. Villar  B. Colao  I. Calles‐Venal  J. G. Fernndez‐lvarez  M. Fernndez‐Caso  J. M. Villar 《Anatomia, histologia, embryologia》2005,34(Z1):54-54

Chondrocytes dedifferentiate to a fibroblast‐like phenotype on plastic surfaces. Dedifferentiation is reversible if these cells are then cultured embedded in gels as alginate, agarose or collagen. Chondrocytes cultured in suspension on a non‐adherent surface are also known to form aggregates of differentiated cells. The knowledge of chondrocyte behavior in culture is relevant for tissue engineering purposes. In this report we describe a simple method to culture differentiated or redifferentiated rabbit auricular chondrocytes on plastic surfaces with a stable phenotype. When chondrocyte aggregates formed in suspension are next seeded on plastic surfaces, most of them attach to the plastic as round or polygonal cells, and this morphological differentiation, confirmed by the presence of type II collagen, is stable for long culture periods. We also report that the addition of aggregates to monolayer cultures of dedifferentiated chondrocytes results in their redifferentiation, as is shown by their morphological changes and the synthesis of type II collagen. Therefore, this simple method can be useful for the study of chondrocyte behavior on plastic surfaces and for redifferentiating previously proliferated chondrocytes in tissue engineering techniques. Furthermore, these results demonstrate that, in addition to culture conditions such as cell isolation method or cell‐density, chondrocyte behavior on plastic depends on the presence or absence of aggregates resulting from the dissociation process.  相似文献   

The effects of agricultural fields and human settlements on the use of rivers by wildlife in the mid-Zambezi valley,Zimbabwe     

Fritz  Hervé  Saïd  Sonia  Renaud  Pierre-Cyril  Mutake  Snoden  Coid  Craig  Monicat  François 《Landscape Ecology》2003,18(3):293-302

After the eradication of the Tse-Tse fly in the Mid-Zambezi valley, human settlements and fields extended mainly along the main rivers. In order to investigate the consequences of this human development on wildlife diversity we monitored three rivers of the Mid-Zambezi valley in Zimbabwe: Angwa, Manyame and Kadzi. The rivers were divided in segments of 200 m which were checked for spoors in order to assess the number of species and the number of individuals that used the segments. Human settlements were also recorded. We used a GIS to define the spatial characteristics of the fields present along the rivers, and related them to the distribution and abundance of wild species spoors in the river beds and banks. Our results show that the number of species in one segment of the river decreased with the increasing size of the field area bordering the segment. For all the major ungulate species, the numbers of individuals recorded per segment decreased with increasing field area. A similar trend was observed for small and medium-sized carnivores, though they were in lower numbers when present. Our analyses thus confirm that the extension of human agriculture in wildlife areas has an impact on most wild species, but we also define some threshold value of field size above which there seem to be an acceleration of the decrease in wildlife density and diversity: 3.2 ha for medium and small herbivores and carnivores; only the elephant seem to tolerate larger field area with a threshold value of 32 ha.This revised version was published online in May 2005 with corrections to the Cover Date.  相似文献   

Detection of the Defoliating Pathotype of Verticillium dahliae in Infected Olive Plants by Nested PCR   总被引：2，自引：0，他引：2  

Jesús Mercado-Blanco  Dolores Rodríguez-Jurado  Encarnación Pérez-Artés  Rafael M. Jiménez-Díaz 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(1):1-13

Spread of Verticillium wilt into newly established olive orchards in Andalucía, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants.  相似文献   

Gene Flow Analysis of Phytophthora porri Reveals a New Species: Phytophthora brassicae Sp. Nov.     

Willem A. Man in 't Veld  Arthur W.A.M. de Cock  Elena Ilieva  C. André Lévesque 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(1):51-62

Isozyme analysis and sequence analysis of the internal transcribed spacer regions (ITS-1 and ITS-2) and the 5.8S subunit of the ribosomal DNA gene repeat were used to examine whether isolates of Phytophthora porri from Allium and Brassica represent a single homogeneous species. Twenty-six strains of P. porri, 16 strains isolated from the genus Allium, and 10 strains isolated from the genus Brassica, were analyzed using malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH) and lactate dehydrogenase (LDH), represented altogether by four putative loci (Mdh-2, Idh-1, Idh-2, and Ldh-2). Isozyme analysis revealed that strains isolated from Allium contained five private alleles at three isozyme loci (Ldh-2 ⁸³, Ldh-2 ¹⁰⁴, Idh-1 ¹⁰⁸, Idh-1 ¹¹², and Idh-2 ⁹⁸), whereas six different alleles were observed at four isozyme loci (Ldh-2 ⁸⁵, Ldh-2 ¹⁰⁰, Ldh-2 ¹¹⁴, Idh-1 ¹⁰⁰, Idh-2 ¹⁰⁰, and Mdh-2 ¹¹¹) in strains obtained from Brassica. The heterozygosity at the Ldh-2 locus, differing in allele composition, however, between strains from Allium and Brassica, was present in all strains, indicating that it is probably fixed. Sequence analysis of the ITS regions and the 5.8S subunit showed consistent differences between isolates from Allium and isolates from Brassica. Based on isozyme data, ITS sequence analysis and formerly published differences in restriction enzyme patterns of mitochondrial DNA, morphology and pathogenicity, it was concluded that the isolates of P. porri Foister did not represent a homogeneous species. Isolates from Brassica constitute a distinct species which is described here as P. brassicae sp. nov. It was inferred from isozyme patterns, which were in no case intermediate between the two species, that P. porri and P. brassicae do not hybridize and are reproductively isolated by barriers to gene flow.  相似文献   

Induction of an antioxidant enzyme system and other oxidative stress markers associated with compatible and incompatible interactions between chickpea (Cicer arietinum L.) and Fusarium oxysporum f. sp.ciceris   总被引：1，自引：0，他引：1  

Carmen García-Limones  Ana Hervs  Juan A. Navas-Corts  Rafael M. Jimnez-Díaz  Manuel Tena 《Physiological and Molecular Plant Pathology》2002,61(6):325

To ascertain if active oxygen species play a role in fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris, the degree of lipid peroxidation (malondialdehyde formation) and the activity levels of diamine oxidase (DAO), an apoplastic H₂O₂-forming oxidase, and several antioxidant enzymes, namely ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guaiacol-dependent peroxidase (GPX) and superoxide dismutase (SOD), were determined spectrophotometrically in roots and stems of ‘WR315’ (resistant) and ‘JG62’ (susceptible) chickpea cultivars inoculated with the highly virulent race 5 of the pathogen. Moreover, APX, CAT, GPX and SOD were also analysed in roots and stems by gel electrophoresis and activity staining; and the protein levels of APX and SOD in roots were determined by Western blotting. In roots, infection by the pathogen increased lipid peroxidation and CAT and SOD activities, although such responses occurred earlier in the incompatible compared with the compatible interactions. APX, GPX and GR activities were also increased in infected roots, but only in the compatible interaction. In stems, infection by the pathogen increased lipid peroxidation and APX, CAT, SOD and GPX activities only in the compatible interaction, and DAO activity only in the incompatible one. In general, electrophoregrams agreed with the activity levels determined spectrophotometrically and did not reveal any differences in isoenzyme patterns between cultivars or between infected and non-infected plants. Further, Western blots revealed an increase in the root protein levels of APX in the compatible interaction and in those of SOD in both compatible and incompatible interactions. In conclusion, whereas enhanced DAO activity in stems, and earlier increases in lipid peroxidation and CAT and SOD activities in roots, can be associated with resistance to fusarium wilt in chickpea, the induction of the latter three parameters in roots and stems along with that of APX, GR (only in roots) and GPX (only in stems) activities are rather more associated with the establishment of the compatible interaction.  相似文献   

细胞骨架解聚药物对小麦与叶锈菌互作诱发的细胞过敏性反应的影响   总被引：5，自引：0，他引：5  

侯春燕  王冬梅  李小娟  韩胜芳  刘娟  王智炘 《植物病理学报》2002,32(2):147-152

 小麦(Triticum aestivum)品种洛夫林10和叶锈菌小种366组成不亲和组合,小麦叶片发生过敏性坏死反应(HR)是小麦抵抗叶锈菌侵染的重要因素。在接种前给小麦叶片分别预注射微管解聚药物磺草硝(oryzalin)和微丝解聚药物细胞松弛素D (cytochalasin D,CD),结果表明2种药物注射使得寄主因叶锈菌侵染诱导的细胞过敏性坏死数目明显减少,并且注射药物的浓度越大,寄主细胞发生HR的数量越少。说明肌动蛋白和微管蛋白的聚合状态是诱发小麦叶片发生HR防卫反应所必需的,细胞骨架在小麦抵抗叶锈菌侵染过程中可能起着重要作用。  相似文献   

Annual bluegrass (Poa annua L.) control in bentgrass (Agrostis palustris Huds.) green with sequential application of bispyribac‐sodium combined with dinitroanilines*     

NAM‐IL PARK  YUKO SUTO  YUTAKA MIURA  NOBUO NAKATANI  SHIN‐ICHI IORI  MASARU OGASAWARA 《Weed Biology and Management》2002,2(3):159-162

  相似文献   

红肉脐橙果肉中主要色素的定性及色素含量的变化   总被引：23，自引：3，他引：23  

徐娟  邓秀新 《园艺学报》2002,29(3):203-208

 运用薄层层析法及二极管阵列检测器与高效液相色谱联用的方法, 确定红肉脐橙果肉中含有番茄红素和β- 胡萝卜素两种主要色素, 它们在液相色谱图中占总峰面积(计溶剂峰) 的73. 57 %～77. 78 % , 番茄红素占55. 36 %～58. 49 % , β- 胡萝卜素占18. 21 %～19. 93 %。利用反相高效液相色谱仪以及番茄红素和β- 胡萝卜素标样制作标准曲线, 测定果实发育及室温贮存过程中色素的变化动态, 结果表明, 该脐橙果肉中的番茄红素在成熟期含量最高, 达109. 67μg·g^-1DM, β- 胡萝卜素此时也达到最高值(15. 52μg·g^-1DM) ; 番茄红素的含量在采收后基本呈下降趋势(但采后30 d 有显著升高) , 说明果肉中存在类胡萝卜素采后合成的现象; 贮存4 个月后, 番茄红素含量急剧降低至最高时的1/ 10。  相似文献   

红肉脐橙(Citrus sinensis L.)果肉中特征色素提取方法探索   总被引：4，自引：0，他引：4  

徐娟  邓秀新 《果树学报》2002,19(4):223-226

红肉脐橙是目前为止惟一果肉呈粉红色或红色的脐橙,其呈色色素尚未见报道。为深入研究其果肉中特征色素的构成及在果实生长发育和贮存过程中色素含量的变化,本研究在对该色素初步定性的基础上对提取方法进行了探索。在分析红肉脐橙果肉中色素萃取液的紫外可见吸收光谱时发现,该色素有类胡萝卜素的特征吸收峰,将萃取液进行薄层层析则进一步证实其内含有番茄红素。在色素提取的前处理方式上,本研究认为冷冻干燥样品的浸提效果较真空干燥的样品稳定(变异系数小),且浸提率相差无几,所以选择冷冻干燥作为合适的样品前处理方式。色素浸提条件的正交实验结果表明,最佳浸提条件为50℃、1.5h内浸提4次,浸提的料液比为1:60。此外,浸提温度、浸提时间、浸提料液比和浸提次数对浸提率都有显著影响,影响最大的因素是浸提温度,最佳温度为50℃,随温度降低浸提率显著降低。其次是侵提时间的长短,最佳浸提时间为1.5h,延长或缩短0.5h都会使浸提率下降。浸提次数对浸提率的影响是最小的。  相似文献   

柑橘类果实汁胞的红色现象及其呈色色素   总被引：10，自引：0，他引：10  

徐娟  邓秀新 《果树学报》2002,19(5):307-313

柑橘类果实中不乏汁胞为红色类型者,如红肉葡萄柚(Citrus paradisii Macf.)的粉红色是番茄红素所致,血橙(C.sinensis L.)的血红色来源于花青素,此外,还有红色的红江橙(C.sinensis L.)、橙红色的元红(C.reticulata Blan-co)和紫红色的五布红心柚(C.grandis L.)等,它们因色泽鲜艳而具备特殊的市场潜力,但呈色色素尚未见报道。在符合审美观的同时,红色汁胞品种还具有特殊的营养价值,如类黄酮、类胡萝卜素等色素富集可软化血管、抗癌、抗氧化及消除体内自由基等。综合有关文献,首次将国内外主要红色汁胞柑橘品种依宽皮柑橘、甜橙、柚、葡萄柚及杂柑等分类进行综述,分析可能的呈色色素,阐明导致红色性状产生的两类色素—花青素、类胡萝卜素生物合成途径  相似文献