利用蛋白质组学研究水果采后成熟的分子机理

果实成熟是发生在高等植物中复杂而有序的过程,由于植物种类和环境条件的不同,果实从不成熟到发育成熟过程包括了众多阶段。近年来,植物本身的生理过程与农业产业息息相关,继而影响经济效益,因此果实成熟过程的研究日益重要。果实成熟过程中,伴随着不同色素水平变化和糖等物质含量增加,果实色泽发生变化,大量生理代谢过程例如颜色、质地、风味和香气等受到外因(光照、温度)和内因(发育相关基因和激素)调控而发生变化。因而利用差异蛋白质组学方法系统研究果实从未成熟到成熟的不同生理过程和阶段分子水平上的动态变化具有十分重要的意义。到目前为止,一些重要果实及其不同发育和成熟过程的蛋白质组数据库日益完善和更新。     

南方早熟砂梨缺镁黄叶症诊断与矫治

通过对福建省德化县早熟砂梨“黄叶病株”及“正常植株”叶片及土壤的营养诊断分析：黄叶病株叶片平均含镁量0.131%,极显著低于正常植株0.328%;黄叶症较重的果园土壤交换态镁含量低12.6～24.5㎎/㎏及土壤酸性强pH3.6～4.5,诊断结论砂梨叶片黄化现象为镁素缺乏所致。矫治试验效果,叶面喷施1%硝酸镁3次,可以叶色转绿;12月份土壤基施钙镁磷肥株1.3 kg,翌年5月份后有明显矫治效果。     

HXKs基因在香蕉果实后熟过程中功能初探

香蕉是世界主要水果之一,对乙烯非常敏感,属于呼吸跃变型果实。目前,香蕉果实后熟机理还未完全探明,后熟过程中己糖激酶(HXK)与乙烯的关系还不清楚。通过BLAST比对从香蕉基因组数据库中发现HXK基因家族的11个成员,经克隆获得这些基因的序列,并研究HXKs基因在香蕉果实自然后熟过程中的转录表达谱,重点研究乙烯利、甘露糖、NAG和1-MCP对HXKs基因表达、HXK酶活和内源乙烯生物合成的影响。结果表明:在香蕉果实后熟过程中HXKs基因呈现差异性表达,乙烯利上调大多数HXKs基因的表达和HXK酶活,1-MCP的作用正好相反,这表明外源乙烯正作用于HXK。另一方面,甘露糖加快内源乙烯生物合成,NAG却推迟内源乙烯生物合成,这说明HXK正作用于内源乙烯生物合成。所以,在香蕉果实后熟过程中乙烯和HXK之间存在相互促进的关系。这将为进一步阐明香蕉果实后熟机制提供理论依据,也将为香蕉果实保鲜新技术的挖掘提供新思路。     

葡萄果实转色过程中线粒体呼吸代谢变化

为明确线粒体呼吸代谢与果实成熟期间颜色形成的关系,本试验探究了3种葡萄果实(夏黑、巨峰和金手指)成熟期间颜色、总花色苷含量、呼吸强度、内源乙烯产生和线粒体呼吸代谢的变化规律。结果表明,夏黑和巨峰葡萄果实的果皮颜色指数(CIRG)值和总花色苷含量随着果实成熟呈上升趋势,而金手指葡萄果实CIRG值在整个成熟期间变化不显著;葡萄果实成熟期间呼吸强度和内源乙烯均呈先上升后下降趋势,在花后63d达到峰值。同时,葡萄果实成熟期间,线粒体膜通透性转换孔(MPTP)逐渐增加,线粒体膜流动性(MMF)持续下降;线粒体琥珀酸脱氢酶(SDH)活性随着成熟进程逐渐下降,而细胞色素氧化酶(CCO)活性在花后63d开始上升。表明盛花63d之后是果实色泽快速形成阶段,可能为葡萄果实色泽形成的关键时期。线粒体呼吸代谢增强促进了果实成熟期间呼吸强度增加和内源乙烯释放,从而使夏黑和巨峰果实中花色苷大量积累,加快了金手指葡萄果实叶绿素的降解和类胡萝素的合成。本研究结果为调控南方葡萄果实着色提供了理论依据。     

miR172超表达载体的构建及其在micro-Tom番茄中的遗传转化

为了进一步阐明miRNA在番茄不同生长发育阶段尤其是在果实成熟阶段的调控途径,利用聚合酶链式反应(PCR)从番茄(Solanum lycopersicum)的cDNA克隆出miR172基因核心片段,将该基因片段,以及pCAMBIA1300-221载体通过特异性的限制性内切酶双酶切,然后将miR172片段正向连接到载体上,转化大肠杆菌Trans5α,鉴定重组质粒正确后,利用冻融法将重组载体转入农杆菌EHA105中。再次回转大肠杆菌验证获得的重组载体,通过PCR以及双酶切鉴定是否符合预期结果,将测序结果与NCBI上的基因序列相比较,同源性高达100%。结果成功构建了适用于番茄农杆菌遗传转化的植物表达载体,接下里进行转基因植株的培育和鉴定。为通过miR172基因进一步研究果实成熟衰老机理奠定了物质基础。     

壮芽灵对烤烟种子的处理效应

对云烟85、K326和红花大金元3个品种进行不同成熟期种子活力测定及壮芽灵浸种处理。结果表明：完熟期的种子活力高于褐熟期和枯熟期；壮芽灵处理种子中，以稀释1000倍，浸种24小时的效果较好，可提高中、低等活力水平的烤烟种子的活力，而对高活力烤烟种子则有轻微的抑制作用。本法对提高中、低等活力水平的烤烟具有明显作用。     

灌浆成熟期温度对稻米品质及有关生理生化特性的影响

1987年在湖南农学院分早、中、晚三季研究灌浆成熟期温度对稻米品质及有关生理生化特性的影响,结果表明:a.茎、鞘、叶的总淀粉和粗蛋白质含量快速减少,籽粒总淀粉和蛋白质积累快,直链淀粉和蛋白质含量升高,淀粉酶和蛋白酶浓度提高,气温大于35℃时淀粉酶和蛋白酶的活性则更高,可溶性糖在灌浆成熟后期增加,温度在30～35℃范围内,Q酶活性最高,籽粒直链淀粉减少。b.灌浆成熟期温度提高,籽粒灌浆速率增大,时间缩短,谷粒物质致密度差,容重下降,垩白指数增大,胶稠度变硬,糊化温度上升。此外,还探讨了稻米品质形成的气候生态生理机制。     

Uneven ripening in ‘Bangalore blue’ grape (Vitis vinifera × Vitis labrusca) berries in India is linked to seed viability

Uneven ripening (UR) is a physiological disorder of unknown origin in ‘Bangalore blue’ grape (Vitis vinifera × Vitis labrusca) leading to wine of inferior quality. A preliminary study found wide variations in total dehydrogenase activity (TDH) of seeds from unevenly ripe berries. In our experiments, gibberellin (GA₃) applied to young grapes increased seed TDH activity and reduced the incidence of uneven ripening to 2% compared with 35% in the control. In contrast, paclobutrazol (PBZ) decreased TDH activity and increased the incidence of the disorder to 58%. GA₃-treated berries had higher concentrations of sucrose and TDH activity in seed representing mature seeds with high viability. In contrast, PBZ-treated and control berries had higher concentrations of glucose and lower TDH activity, indicating immature seeds with low viability. These results suggested that competition among developing berries can lead to differences in seed gibberellin content, seed viability and the rate of berry growth resulting in green, purple, and black berries at harvest. The study established the role of seed viability in uneven ripening and demonstrated that the incidence of the disorder is reduced by the application of GA₃ to immature berries.     

Postharvest UV-C treatment combined with 1-methylcyclopropene (1-MCP), followed by storage in continuous low-level ethylene atmosphere,improves the quality of tomatoes

Mature green tomatoes (Solanum lycopersicum L. cv Neang Pich) were exposed to 13.6 kJ m^?2 UV-C or 0.5 μL L^?1 1-MCP or combination of 13.6 kJ m^?2 UV-C and 0.5 μL L^?1 1-MCP, with appropriate untreated controls. After treatment, tomatoes were stored in air containing 0.1 μL L^?1 ethylene at 20°C and 100% RH. The untreated fruit ripened significantly faster than those of all other treatments. UV-C treatment alone was able to delay fruit ripening by up to 5 days longer compared to untreated fruits whilst the additional of 1-MCP further delayed fruit ripening. UV-C and 1-MCP treatments alone or in combination had significantly slower ethylene production rates throughout the storage period. The fruit treated with the combination of 1-MCP and UV-C was significantly firmer and had higher total phenolic content compared to that of the other treatments. However, there was no difference between treatments in soluble solids content/titratable acids ratio, chlorophyll content, lycopene content and total antioxidant activity. These results show that UV-C and 1-MCP treatment delay ripening and improve the quality of tomatoes in the presence of low-level ethylene during storage. This new treatment could be used to extend the shelf-life of mature green tomatoes through the supply chain without the use of refrigeration.