不同生态区烟草的叶面腺毛基因表达

利用cDNA芯片技术,对同一烟草品种（Nicotina tabacum L cv. K326）、不同生态区(典型的浓香型烟区-河南平顶山和典型的清香型烟区-云南玉溪)、同一叶位(中部叶)、相同叶龄（60天）的叶片腺毛基因表达谱进行了比较研究,共发现有效差异表达基因445个。其中,与光合色素代谢、光合作用以及类萜生物合成相关基因在河南烟叶腺毛中高表达,与碳水化合物、蛋白质分解代谢过程相关的基因在云南烟叶腺毛中高表达;防御过程相关基因在两生态区均有高表达。该结果表明, 烟草叶面腺毛的基因表达水平受到生态因素的调控,生态因素有可能通过影响腺毛的物质代谢和分泌,从而对烟叶风格形成产生一定的影响。     

不同生态烟区烤烟δ~(13)C值与生理及品质特征的比较研究

以烤烟品种K326为试验材料,在云南、福建和河南三个生态烟区大田种植,自烟叶生理成熟期起至工艺成熟期,分4次采集中部(第11叶位)烟样,对烟叶的δ13 C值、总碳、全氮、光合色素等进行测定,比较不同生态烟区烤烟δ13 C值的分布、生理生态适应性及品质特征。结果表明:(1)三个生态烟区烟叶δ13 C值、总碳、碳氮比、比叶重、叶绿素a、总叶绿素含量均表现为云南福建河南,全氮含量则为河南云南福建,叶绿素b含量为云南河南福建,类胡萝卜素含量为河南福建云南,其中的δ13 C值、总碳、碳氮比、类胡萝卜素含量在福建和云南烟区间较为接近。(2)烟叶δ13 C值与总碳含量在云南呈正相关,在福建、河南呈负相关关系;三个生态烟区烟叶δ13 C值与全氮含量均呈负相关关系;δ13 C值与光合色素含量在云南、河南烟区均呈正相关关系,在福建烟区均呈负相关关系;δ13 C值与烟碱、氮、钾、氯呈负相关关系,与总糖、还原糖呈正相关关系。(3)云南烤烟香韵丰富,刺激性中等,化学成分协调性最好;河南烤烟香气量较高,刺激性较大;福建烤烟在香气量和化学成分协调性方面表现较差。研究发现,烟叶δ13 C值与烟叶的生理特征、品质特征存在紧密联系,用烟叶的δ13 C值、生理指标、化学品质可区分不同生态烟区烤烟香气风格和品质特征。     

不同生态区烤烟叶片稳定碳同位素组成特征

在河南、福建和云南3个生态区大田种植烤烟K326,采集不同叶位生理成熟的烟叶,测定δ13C值、总碳、全氮及比叶重,探讨了不同生态区烟叶δ13C值组成特征。结果表明:河南烟叶δ13C值分布范围为-31.2%—-27.3‰,平均值-29.7‰;福建烟叶δ13C值分布范围为-28.6%—-24.6‰,平均值-26.3‰;云南烟叶δ13C值分布在-27.1—-24.0‰之间,平均值-25.6‰;河南烟叶全氮含量最高,其总碳、碳氮比和比叶重均显著低于其余两地;福建和云南烟叶各生理指标值较为接近;各生态区烟叶δ13C值均与全氮含量呈负相关,与碳氮比及比叶重呈正相关。河南与其余两地烤烟生理特征的差异性,以及福建与云南烤烟生理特征的相似性表明,δ13C值可能与烤烟的品质存在关联。     

滤减UV B辐射对烟叶可溶性蛋白、〖HJ*3〗〖HJ*2〗光合色素和类黄酮的影响

在云南玉溪烟区种植烤烟海拔最高（1 806.0 m）的通海县,通过盆栽烤烟K326试验,研究了在滤减自然的太阳UV B辐射强度25%、50%和65%条件下,UV B辐射对烟叶发育过程中可溶性蛋白、光合色素和类黄酮的影响。结果表明：随叶龄增加,可溶性蛋白含量下降,光合色素降解,类黄酮在老叶中积累,蛋白质在生理成熟期对UV B辐射最敏感。与对照相比,减弱UV B辐射处理降低了烟叶类黄酮和可溶性蛋白含量,但光合色素含量上升;较低的UV B辐射降低了叶绿素的降解速度。结果从一侧面说明UV B辐射对烟叶蛋白质的合成是有益的,类黄酮和叶绿素的变化是对UV B辐射变化的适应性反应,类黄酮与蛋白质之间可能存在一定的偶联关系。     

肾阳虚证候的人血清比较蛋白质组学分析

利用双向电泳（2-DE）优化分离了除去高丰度蛋白（白蛋白和IgG）的老年体虚肾阳虚患者（以健康组为参照）的血清样本,对比分析pH 4～7范围的2-DE谱图,肾阳虚患者和参照组的平均蛋白点分别为(393±32)和(455±19)个. 其中肾阳虚证候表达量上调2倍（P<0.05）以上的蛋白点有26个,下调2倍以上的有33个. 用质谱获得上述差异蛋白的肽质量指纹图谱,并经数据库检索共鉴定出了49种差异蛋白质,其中有10种在肾阳虚血清中特异表达,有6种在健康组血清中特异表达. 蛋白功能分析发现,其中33种蛋白质的差异表达与肾阳虚证密切相关. 蛋白质TCRβ及transthyretin的蛋白印迹实验验证了2-DE 的结果. 该研究结果为阐明中医肾阳虚证的机理提供了一条新途径.     

湖南不同烟区烤烟烟碱含量差异的生态原因

于2007年在湖南省3个典型植烟生态区代表地(湘西北桑植县、湘中浏阳市、湘南永州市)分别同时进行大田本土栽培、客土与本土单独盆栽试验,检测烤烟K326收获期各处理中部烟叶杀青样品的烟碱含量,研究湖南不同烟区烤烟烟碱含量差异的主要生态原因.结果表明:湖南不同烟区大田本土正常栽培烤烟的平均烟碱含量存在显著差异,其中以桑植烟区所产烟叶烟碱含量最高,其次是浏阳烟区,永州烟区最低.盆栽试验结果表明,气候对湖南烟区烤烟烟碱含量有显著影响,土壤及其与气候的互作对湖南烟区烤烟烟碱含量的影响有限.气候、土壤及两者互作对湖南烟区烤烟烟碱含量变异的贡献率依次为60.0%、12.8%和27.2%.与湖南烟区烤烟烟碱含量关联度较大的主要亚生态因子依次为:成熟期的云量和伸根期的相对湿度、日照时数、昼夜温差、降雨量以及旺长期的平均气温.综合分析表明,气候是导致湖南烟区烤烟烟碱含量差异的主要生态因素.     

苦瓜性别分化的特异蛋白质研究

对苦瓜（ＭｏｍｏｒｄｉｃａｃｈａｒａｎｔｉａＬ．）的两性期和雌、雄花早期发育过程中的３个时期花蕾,用毛细管电泳法进行可溶性蛋白质分析,发现了一些与性别分化有关的特异蛋白质。其中１１ｋＤ的蛋白质从幼雌花时期开始出现后,在雌花发育的３个典型时期都存在,很可能是雌花分化程序表达中的一种“关键蛋白”。与之类似,３０ｋＤ的蛋白质很可能是雄花程序表达中的一种“关键蛋白”     

滤减UV B辐射对烟叶可溶性蛋白、〖HJ*3〗〖HJ*2〗光合色素和类黄酮的影响

在云南玉溪烟区种植烤烟海拔最高(1806.0m)的通海县,通过盆栽烤烟K326试验,研究了在滤减自然的太阳UV-B辐射强度25%、50%和65%条件下,UV-B辐射对烟叶发育过程中可溶性蛋白、光合色素和类黄酮的影响。结果表明:随叶龄增加,可溶性蛋白含量下降,光合色素降解,类黄酮在老叶中积累,蛋白质在生理成熟期对UV-B辐射最敏感。与对照相比,减弱UV-B辐射处理降低了烟叶类黄酮和可溶性蛋白含量,但光合色素含量上升;较低的UV-B辐射降低了叶绿素的降解速度。结果从一侧面说明UV-B辐射对烟叶蛋白质的合成是有益的,类黄酮和叶绿素的变化是对UV-B辐射变化的适应性反应,类黄酮与蛋白质之间可能存在一定的偶联关系。     

滤减UV-B辐射对烟叶可溶性蛋白、光合色素和类黄酮的影响

在云南玉溪烟区种植烤烟海拔最高(1806.0m)的通海县,通过盆栽烤烟K326试验,研究了在滤减自然的太阳UV-B辐射强度25%、50%和65%条件下,UV-B辐射对烟叶发育过程中可溶性蛋白、光合色素和类黄酮的影响。结果表明:随叶龄增加,可溶性蛋白含量下降,光合色素降解,类黄酮在老叶中积累,蛋白质在生理成熟期对UV-B辐射最敏感。与对照相比,减弱UV-B辐射处理降低了烟叶类黄酮和可溶性蛋白含量,但光合色素含量上升;较低的UV-B辐射降低了叶绿素的降解速度。结果从一侧面说明UV-B辐射对烟叶蛋白质的合成是有益的,类黄酮和叶绿素的变化是对UV-B辐射变化的适应性反应,类黄酮与蛋白质之间可能存在一定的偶联关系。     

大鼠海马的表达蛋白质组学实验研究

目的：用蛋白质组学方法初步分析大鼠海马蛋白质的表达。方法：提取大鼠海马蛋白质样品后,用双向凝胶电泳对其分离,经考马斯亮蓝染色后,产生大鼠海马蛋白质双向凝胶电泳图谱。从凝胶上切割分离的蛋白质,经胰蛋白酶胶内酶解,通过基质辅助激光解吸／电离飞行时间质谱（MALDI-TOF-MS）对酶解后的肽段进行分析。根据肽段质谱数据,经数据库（NCBI）检索,对蛋白质进行鉴定。结果：鉴定了37种具有明确功能的蛋白质,它们分别属于代谢酶、细胞骨架蛋白、热休克蛋白、抗氧化蛋白、信号传导蛋白、蛋白酶体相关蛋白、神经元特异蛋白及神经胶质蛋白。另外,鉴定了3种未知功能蛋白。结论：为建立大鼠海马蛋白质组数据库提供必要的资料,为在大鼠模型上研究神经疾病发病机理奠定基础。     

Low nitrogen-induced expression of cyclophilin in Nicotiana tabacum

Leaf morphology and the leaf protein expression profiles of flue-cured tobacco grown in central Henan province of China under low nitrogen (low-N) and normal nitrogen (normal-N) nutrition were examined. The leaf length and width were measured at 50, 60, and 70 days after transplanting. Leaves grown under low-N conditions were shorter and more narrow than those grown under normal-N conditions. The protein expression profiles of tobacco leaves harvested at 70 days after transplanting were analyzed by 2-dimensional electrophoresis, and five differentially expressed proteins including a putative protein were identified. Except for the MCM protein-like protein, the other three differentially expressed proteins of cyclophilin-like protein, vacuolar invertase INV2, MAR-binding protein and the one putative protein showed increased expression in the low-N nutrition group. Among these proteins, the cyclophilin-like protein, which is a stress-responsive signal protein, may play pivotal roles in regulating leaf development under stress conditions. Real-time quantitative PCR analysis showed that the mRNA expression level of the cyclophilin-like protein at day 50, 60 and 70 under low-N conditions was 0.90, 1.43 and 6.9-fold higher than that under normal-N conditions, indicating that the gene expression of cyclophilin-like protein was strongly induced by low-N conditions.     

萎凋处理对乌龙茶风味品质形成的转录组分析

乌龙茶是一种高香型半发酵茶,因其具有的花果香和鲜醇浓厚口感而广受消费者青睐.在鸟龙茶加工过程中,萎凋是促进乌龙茶风味品质形成的第一道工序.然而,乌龙茶萎凋过程中影响风味品质形成的分子机制尚不明确.利用转录组测序对乌龙茶鲜叶、室内萎凋叶和日光萎凋叶进行分析.结果 表明,从3个样品中共鉴定出10793个差异表达基因.KEG...     

Leaf proteome characterization in the context of physiological and morphological changes in response to copper stress in sorghum

Copper (Cu) is an essential micronutrient required for normal growth and development of plants; however, at elevated concentrations in soil, copper is also generally considered to be one of the most toxic metals to plant cells due to its inhibitory effects against many physiological and biochemical processes. In spite of its potential physiological and economical significance, molecular mechanisms under Cu stress has so far been grossly overlooked in sorghum. To explore the molecular alterations that occur in response to copper stress, the present study was performed in ten-day-old Cu-exposed leaves of sorghum seedlings. The growth characteristics were markedly inhibited, and ionic alterations were prominently observed in the leaves when the seedlings were exposed to different concentrations (0, 100, and 150 µM) of CuSO₄. Using two-dimensional gels with silver staining, 643 differentially expressed protein spots (≥1.5-fold) were identified as either significantly increased or reduced in abundance. Of these spots, a total of 24 protein spots (≥1.5-fold) from Cu-exposed sorghum leaves were successfully analyzed by MALDI-TOF-TOF mass spectrometry. Of the 24 differentially expressed proteins from Cu-exposed sorghum leaves, 13 proteins were up-regulated, and 11 proteins were down-regulated. The abundance of most identified protein species, which function in carbohydrate metabolism, stress defense and protein translation, was significantly enhanced, while that of another protein species involved in energy metabolism, photosynthesis and growth and development were severely reduced. The resulting differences in protein expression patterns together with related morpho-physiological processes suggested that these results could help to elucidate plant adaptation to Cu stress and provide insights into the molecular mechanisms of Cu responses in C₄ plants.     

生物酶和微生物技术改善烟叶香气的研究进展

生物酶和微生物在烟叶醇化发酵过程中发挥着重要作用。目前,利用生物酶和微生物技术提高烟叶品质、改善烟叶香气,已成为烟草行业关注的热点。利用酶制剂处理烟叶可以降解烟叶的蛋白质、果胶、纤维素等生物大分子,以达到提高烟叶品质和改善烟叶香气的作用。利用微生物对烟叶进行发酵可以有效调整和改善烟叶内部化学组分的比例,增加烟叶中的香气物质。综述了生物酶和微生物技术在烟叶产香发酵中的研究进展及其在烟叶发酵机理及增香技术中的应用,重点阐述了微生物和生物酶提高烟叶香气、改善烟叶品质以及降解烟叶中蛋白质、淀粉、果胶、纤维素等大分子物质的研究现状,分析了目前微生物和生物酶在实际应用中存在的问题,以期为今后使用生物酶和微生物技术改善烟叶香气提供理论依据。     

A new multigene family inducible by tobacco mosaic virus or salicylic acid in tobacco.

A previously undescribed cDNA family was isolated from tobacco challenged with tobacco mosaic virus (TMV). A cDNA library was constructed with mRNA from upper leaves of Xanthi nc tobacco plants that had been inoculated with TMV on the lower leaves 11 days previously. The library was screened differentially with radiolabeled cDNA synthesized with mRNA from upper, uninoculated leaves of either TMV-inoculated or mock-inoculated tobacco plants. The new cDNA family, designated SAR8.2, had at least five expressed members, one or more of which were inducible by TMV inoculation and by salicylic acid treatment. The cDNAs encoded small, highly basic proteins containing N-terminal hydrophobic signal peptides and highly conserved cysteine-rich C-terminal domains. One of the SAR8.2 family members contained a direct repeat of the C-terminal domain in tandem. Hybridization of SAR8.2 cDNA to tobacco genomic DNAs indicated a gene family of 10-12 members.     

iTRAQ-based comparative proteomic analysis reveals high temperature accelerated leaf senescence of tobacco (Nicotiana tabacum L.) during flue-curing

Tobacco (Nicotiana tabacum) is extensively cultivated all over the world for its economic value. During curing and storage, senescence occurs, which is associated with physiological and biochemical changes in postharvest plant organs. However, the molecular mechanisms involved in accelerated senescence due to high temperatures in tobacco leaves during curing need further elaboration. We studied molecular mechanisms of senescence in tobacco leaves exposed to high temperature during curing (Fresh, 38 °C and 42 °C), revealed by isobaric tags for relative and absolute quantification (iTRAQ) for the proteomic profiles of cultivar Bi’na1. In total, 8903 proteins were identified, and 2034 (1150 up-regulated and 1074 down-regulated) differentially abundant proteins (DAPs) were obtained from tobacco leaf samples. These DAPs were mainly involved in posttranslational modification, protein turnover, energy production and conversion. Sugar- and energy-related metabolic biological processes and pathways might be critical regulators of tobacco leaves exposed to high temperature during senescence. High-temperature stress accelerated tobacco leaf senescence mainly by down-regulating photosynthesis-related pathways and degrading cellular constituents to maintain cell viability and nutrient recycling. Our findings provide a valuable inventory of novel proteins involved in senescence physiology and elucidate the protein regulatory network in postharvest organs exposed to high temperatures during flue-curing.     

Proteome analysis of tobacco leaves under salt stress

The mechanisms responsible for the effects of salt stress on tobacco plants were examined by means of proteomic analysis. Tobacco plants were exposed to 0, 150, 250, 300, or 400 mM NaCl. At 150 mM NaCl or above, the plants showed a reduction in fresh weight and an increase in proline levels. Proteins extracted from the leaves of tobacco plants exposed to 150 mM NaCl were separated by 2-DE. Of 205 protein spots that were detected reproducibly in each gel, 18 were differentially expressed under NaCl treatment. Up-regulated proteins belonged to the photosynthesis category, whereas down-regulated proteins correspond to defense-related functions. Dose- and time-dependent studies showed that a stromal 70-kDa heat shock-related protein was markedly down-regulated by NaCl. Thus, down-regulation of the stromal 70-kDa heat shock protein in response to salt stress is likely the cause of failure to protect cells against salt stress of tobacco plants.