中国野生葡萄醛脱氢酶基因在拟南芥中的过量表达

将质粒pWR306中包含ED35s启动子、Omega元件及TNOS终止子的一段核苷酸序列定向克隆到质粒pCAMBIA1300,构建了植物中间表达载体pWR-II;将重组质粒pGEM-T-ALDH的醛脱氢酶(ALDH)基因片段定向克隆到pWR-II,构建了葡萄ALDH基因的植物过量表达载体pWR-II-ALDH。采用改进冻融法将其导入农杆菌EHA105,采用花序浸蘸法转化拟南芥,获得了潮霉素抗性的转化拟南芥植株。PCR检测证明外源基因已整合进拟南芥基因组,Real-timePCR结果表明ALDH基因在转化植株的表达量远高于野生种。转化植株和野生植株在形态上有一定的差异,说明葡萄ALDH基因在拟南芥中表达对其生长发育有一定影响。     

中国野生葡萄抗黑痘病基因RAPD标记的克隆、序列分析及辅助育种应用

对中国野生葡萄抗黑痘病基因RAPD标记OPS03-1300进行了克隆、测序及序列分析,结果表明该标记实际长度是1354bp,因此更名为OPS03-1354,其GenBank登录号为DQ350885。该标记序列与99条来自欧洲葡萄的EST有同源性,其中与1条来自赤霞珠叶片感染葡萄皮尔斯病病原菌后获得的EST同源性为82%,与2条来自赤霞珠果实在不同发育期获得的EST同源性分别为80%和85%。该序列与1条来自中国野生华东葡萄白河-35-1叶片接种霜霉病病原菌后获得的EST同源性为67%。该序列编码葡萄的一种假定蛋白。此外,应用该标记对中国野生葡萄与欧洲葡萄的种间杂交广西-1×京可晶F1代339株、白河-35-1×佳利酿F2代207株进行了辅助选择。     

山葡萄‘双红’、‘双优’在戈壁滩上栽培的表现

报道我国西部栽培的‘双红’和‘双优’两个山葡萄品种不埋土能正常越冬,调查、测定和分析这两个山葡萄品种在冬季寒冷、冬春又极干旱的西部戈壁滩上栽培的生产表现。两个山葡萄品种根系耐低温能力比欧亚种酿酒葡萄品种强,枝条保水能力更强,因此,在干旱荒漠的戈壁石砾地上不埋土能安全越冬和抗御春日多风和干旱。一年生枝条长势好,花芽能正常形成,但在开花季节花序干枯和脱落现象严重。‘双红’、‘双优’山葡萄表现生产性能差、果穗少、果穗小、穗重轻、果粒小,故产量低,仅为原产地的1／3,但其品质好,可溶性固形物高出3．9～4．6个百分点。为使‘双红’、‘双优’在河西戈壁滩上栽培,既不埋土又有较高的产量,提出一些解决措施。     

葡萄β型液泡加工酶基因(VvβVPE)的原核表达、纯化与多克隆抗体制备

液泡加工酶(VPE)基因是植物内启动和执行细胞程序性死亡的关键基因,β型VPE是种子特异表达并且为种子蛋白成熟所必须的基因.本研究以黑比诺葡萄(Vitis vinifera cv.Pinot Noir)花后8个不同时期胚珠的cDNA混合样为模板,通过RT-PCR扩增得到葡萄β型液泡加工酶基因(暂命名VvβVPE,GenBank登录号:KC136352),开放阅读框1 485 bp.进一步将VvβVPE克隆到pGEX-6P-1原核表达载体并转入大肠杆菌(Escherichia coli)BL21中,经IPTG诱导,获得约81.3 kD的包涵体融合蛋白GST-VvβVPE.将诱导条件优化后,在37℃、0.05 mmol/L IPTG诱导5h后融合蛋白GST-VvβVPE的表达量最大,采用电透析与盐酸胍处理后透析复性两种方法获得纯化包涵体蛋白,电透析纯化法获得纯化包涵体蛋白的浓度可达到免疫要求.使用纯化蛋白免疫新西兰大白兔制备多克隆抗血清,经Western blot检测,该抗血清(稀释到1∶10 000)与GST-VvβVPE融合蛋白识别反应良好,获得的抗血清可用于以后的VvβVPE的酶活性分析、免疫亚细胞定位分析以及转基因植株蛋白水平的鉴定等,为进一步明确VvβVPE在葡萄胚珠发育中的作用提供基础数据.     

Selecting in situ conservation sites for grape genetic resources in the USA

Previously, there has not been any in situ conservation sites for crop germplasm within the United States Department of Agriculture (USDA), Agricultural Research Service (ARS), National Plant Germplasm System (NPGS). Using morphologic and molecular (SSR markers) techniques, we assessed the genetic variation present in populations of rock grape (Vitis rupestris Scheele), a native American grape species, throughout its range. We identified seven in situ conservation sites for rock grape using a strategy based on morphologic and molecular data, taxonomic information, population size and integrity, and landholder commitment. In collaboration with federal and state landholding agencies, we have established the first NPGS in situ conservation sites for American wild relatives of a crop.     

中国野生毛葡萄芪合酶基因表达及对葡萄抗白粉病的影响

[目的]克隆中国野生毛葡萄(Vitis quinquangularis)'丹凤-2'芪合酶(stilbene synthase)基因(STS)并研究其功能,为提高欧洲葡萄(V.vinifera)的白粉病抗性及品质提供依据.[方法]利用同源克隆法获得中国野生毛葡萄'丹凤-2'芪合酶基因VqSTS9和VqSTS21,构建植...     

不同葡萄品种愈伤组织诱导苗超氧化物歧化酶同工酶比较研究

对3个葡萄品种茎尖愈伤组织诱导苗的超氧化物歧化酶(SOD)同工酶分析,证明葡萄为Mn SOD,里扎玛特诱导苗存在1个类型的无性系变异,为体细胞无性系变异在品种改良中的应用提供了依据。     

欧亚种葡萄对白粉病与霜霉病的抗性研究

通过田间自然鉴定、田间人工接种鉴定及室内离体叶圆片接种鉴定的方法,研究了10个欧亚种葡萄品种(系)及一个欧山杂种对葡萄霜霉病及白粉病的抗性。结果表明,无论采取那种鉴定方法,爱格丽和北醇对两种病害的抗性都最强;赤霞珠、白羽和佳利酿3个品种对霜霉病的抗性最弱,88-02、88-03、88-04、梅鹿特、白克列海特和白诗南6个品种对霜霉病的抗性中等;赤霞珠和佳利酿2个品种对白粉病的抗性最弱,88-02、88-03、88-04、白羽、梅鹿特、白克列海特和白诗南7个品种对白粉病的抗性中等。另外,3种鉴定方法对两病的鉴定结果具有极显著的相关性,尤以田间接种鉴定效果为最佳。     

不同光环境对葡萄幼苗光合产物分配与转化的影响

 以一年生京玉葡萄幼苗 (VitisvineferaL .cv .Jingyu)为试材 ,利用光合测定仪和同位素示踪技术 ,比较研究了自然光环境和遮阳环境下生长的葡萄幼苗净光合速率日变化、生物量分配、光合产物分配与转化等方面的差异。与对照相比 ,遮阳环境中生长的葡萄幼苗净光合速率日变化曲线偏低 ;叶片和茎的生物量比较高 ,根和新稍的生物量比较低 ;其光合产物分配较多流向叶片和茎 ,而向根的分配随遮阳程度的加深明显降低 ;分配到各部位的光合产物的转化利用情况也发生了不同的变化。     

A 3-Year Adaptation Study of Three Distinct Grapevine Cultivars under Midwestern Field Conditions

Simultaneous responses of American (Concord), European (Pinot Noir), and American-European hybrid (Traminette) cultivars to seasonal and diurnal variation in field environmental conditions were assessed to propose suitable cultivars for potential use for selection and breeding in grapevines in dry areas. ‘Pinot Noir’ had higher gas exchange parameters, higher water potential, and chlorophyll fluorescence, while ‘Concord’ had the lowest values in all three seasons and experienced moderate stress by the end of each season. The dry conditions in 2005 caused water potential to decline sharply compared to values recorded in 2004 and 2006. ‘Pinot Noir’ and ‘Traminette’ exhibited a mild stress, while ‘Concord’ exhibited a moderate stress in 2006. Photosynthetic rate decreased slightly during the summer with a slight recovery at the beginning of September, and did not differ across the seasons among cultivars. Stomatal conductance followed the same pattern. However, in both 2004 and 2005, stomatal conductance dropped sharply late in the season because of frequently occurring high temperatures that were greater than 32 °C. Maximum morning values of Fv/Fm were within the optimal range. The study suggests the presence of marginal photoinhibition during mid-summer in Concord cultivars, because no full recovery of photochemical efficiency early in the evening was observed. The European cultivar (Pinot Noir) and the American-European hybrid (Traminette) are more adapted to field conditions in Kansas than Concord cultivars. However, high cold-mortality of Pinot Noir due to high sensitivity to cold winter temperature makes the hybrid cultivars the most suitable cultivar for Midwest regions with relatively cold winters.