水稻Rubisco小亚基启动子的克隆及光诱导表达载体的构建

高效特异表达的启动子往往是转基因研究的关键因素。Rubisco小亚基启动子具有光诱导性、组织特异性和高表达的特性,可利用该启动子为转基因研究服务。本文以水稻（Oryza sativa）中花11叶片为材料,根据GenBank所报道的水稻Rubisco小亚基启动子序列,设计特异引物从水稻基因组DNA中扩增得到长度约1600bp的DNA片段,将该片段连接至T载体pMD18-TSimple,测序结果表明该片段序列与GenBank报道序列一致为100%。Plant CARE序列分析表明,该启动子具有12个与光诱导表达相关的元件。为构建光诱导表达载体,将该启动子和植物表达载体pCactF分别以KpnⅠ和XbaⅠ酶切后连接。光诱导表达载体的构建为进一步研究基因功能及利用光诱导表达载体改良作物品质奠定基础。     

干热胁迫下紫花苜蓿Rubisco羧化酶和活化酶活性变化及其基因表达的研究

为探讨核酮糖-1,5-二磷酸羧化酶(Rubisco)和Rubisco活化酶(Rubisco activase,RCA)活性变化及其基因表达,本研究以云南野生紫花苜蓿(Medicago sativa)和'阿尔冈金'紫花苜蓿为材料,测定其在干旱和高温条件下植株叶片Rubisco和RCA酶活性,并采用RT-qPCR分析Rub...     

海南杜鹃热诱导基因RhRCA1启动子的克隆与功能分析

为了阐述RhRCA1基因的调控机制,以海南杜鹃(Rhododendron hainanense)3年生扦插苗为材料,分析RhRCA1高温响应和组织表达特性;并克隆获得RhRCA1的启动子序列,将该启动子分别驱动荧光素酶报告基因在烟草中瞬时表达、GUS报告基因在拟南芥中稳定表达,分析该启动子活性、组织特异性和热诱导性。结果显示:(1)25℃条件下,RhRCA1表达量极低,37℃处理后其表达量显著升高,呈现典型的热诱导特性,并且RhRCA1在绿色组织中的表达量显著高于非绿色组织,呈现组织特异性;(2)获得的启动子长1 624 bp,其含有多个非生物胁迫响应、光响应、组织特异性等相关元件;(3)构建RhRCA1启动子和萤光素酶融合的植物表达载体,在烟草叶片中瞬时表达,荧光成像结果表明RhRCA1启动子能强烈响应高温胁迫;(4)构建RhRCA1启动子和GUS融合的植物表达载体,转化拟南芥植株筛选至T3代,GUS染色结果显示,高温能显著诱导RhRCA1启动子在拟南芥子叶、成熟叶、茎、萼片、果荚等绿色组织中的表达。研究结果表明RhRCA1启动子是1个兼具高温诱导型和组织特异性的启动子,可应用于植物...     

小麦叶片内1,5-二磷酸核酮糖羧化酶/加氧酶大亚基由53 000到50 000裂解反应的定位研究

研究了小麦叶片内1，5-二磷酸核酮糖羧化酶／加氧酶（Rubisco，EC 4．1．1．39）大亚基（LSU）由53000裂解为50000的反应。结果显示，成熟叶片的粗提液在pH5．5的条件下反应后能检测到50000的裂解产物，而暗诱导衰老叶片的粗提液在pH7．5的条件下也能发现LSU的这一降解。分别从成熟叶片和衰老叶片中提取叶绿体，以其裂解液为反应体系研究LSU由53000裂解为50000这步反应的细胞器定位。结果显示，衰老叶片中的叶绿体裂解液在pH7．5时反应1h后能检测到50000降解产物，而成熟叶片叶绿体裂解液在pH5．5和pH7．5的条件下反应后均未检测到LSU的50000裂解产物。上述结果表明：衰老叶片中ISU由53000部分裂解为50000的反应定位于叶绿体内，而成熟叶片中LSU由53000裂解为50000的反应可能定位于叶绿体外。     

雷竹光合特性的研究

对雷竹叶片光合速率及其主要影响因子的周年变化进行了研究 ,结果表明 :在晴到少云的天气条件下 ,雷竹光合速率日变化呈双峰曲线 .光强、温度、水分等外界环境因子对雷竹光合作用有显著影响 .强光、高温、低湿引起雷竹叶片光合“午睡” .雷竹光合速率年变化也呈双峰曲线型 .环境因子及 Rubisco,Rubisco活化酶 ,蛋白质 ,叶绿素等对光合速率都有显著影响 .雷竹幼叶的光合速率 ,光饱和点及相关影响因子较二龄叶高 ,具有较高的光合生理特性 .雷竹叶片的光饱和点约为 1 688～ 1 834μmol CO2 · m-2 · s-1;光补偿点约为 1 60～ 2 2 2μmol CO2 · m-2 · s-1.     

光暗处理对水稻叶片光合关键酶的效应

稻苗置暗中24h后,Rubisco含量及Rubisco与可溶性蛋白的比值随光强的增加而提高;经2h光照后再置黑暗中,Rubisco含量在最初20min下降迅速,说明苗期Rubisco蛋白在光暗交替下周转较快.Rubisco的初始活力在光照下迅速增加,其上升倍数大大高于量的增加,光强越大达到最大活力所需的时间越短.照光后把稻苗置暗中,Rubisco初始羧化活力很快下降,说明Rubisco的光下活化及暗中失活迅速.Rubisco活化酶的含量随光照增加而逐渐上升,光照处理后再置暗中Rubisco活化酶含量不断下降,但速率不如Rubisco.Rubisco活化酶的活化只依赖低光强,光照处理后再置黑暗中,Rubisco活化酶活力迅速下降.     

Photosynthetic and stomatal responses of potatoes grown under elevated CO2 and/or O3—results from the European CHIP-programme

The physiological effects of elevated CO₂ and/or O₃ on Solanum tuberosum cv. Bintje were examined in Open-Top Chambers during 1998 and 1999 at experimental sites across Europe as part of the EU ‘Changing Climate and Potential Impacts on Potato Yield and Quality’ programme (CHIP). At tuber initiation (≈20 days after emergence, DAE) elevated CO₂ (680 μl l⁻¹) induced a 40% increase in the light saturated photosynthetic rate (A_sat) of fully expanded leaves in the upper canopy. This was 16% less than expected from short-term exposures of plants grown under ambient CO₂ (360 μl l⁻¹) to elevated CO₂, indicating that photosynthetic acclimation began at an early stage of crop growth. This effect resulted from a combination of a 12% reduction in stomatal conductance (g_s) and a decline in photosynthetic capacity, as indicated by the significant reductions in the maximum carboxylation rate of Rubisco (Vc_max) and light-saturated rate of electron transport (J_max) under elevated CO₂. The seasonal decline in the promotion of photosynthesis by elevated CO₂ reflected the concurrent decrease in g_s. Vc_max and J_max were both reduced in plants grown under elevated CO₂ until shortly after maximum leaf area (MLA) was attained. Although non-photorespiratory mitochondrial respiration in the light (R_d) increased during the later stages of the season, net photosynthesis was consistently increased by elevated CO₂ during the main part of the season. Photosynthetic rate declined more rapidly in response to elevated O₃ under ambient CO₂, and the detrimental impact of O₃ was most obvious after MLA was attained (DAE 40–50). Several exposure indices were compared, with the objective of determining the critical ozone level required to induce physiological effects. The critical O₃ exposure above which a 5% reduction in light saturated photosynthetic rate may be expected (expressed in terms of cumulative exposure above 0 nl l⁻¹ O₃ between emergence and specific dates during the season (AOT0-cum)) was 11 μl l⁻¹ h; however this value should only be extrapolated beyond the CHIP dataset with caution. The interaction between O₃ and stomatal behaviour was more complex, as it was influenced by both long-term and daily exposure levels. Elevated CO₂ counteracted the adverse effect of O₃ on photosynthesis, perhaps because the observed reduction in stomatal conductance decreased O₃ fluxes into the leaves. The results are discussed in the context of nitrogen deficiency, carbohydrate accumulation and yield.     

Study on Variation of --Galactosidase Activity During Tea Processing

不同芽叶中β-半乳糖苷酶活性存在差异，嫩芽叶>老叶>嫩茎。芽叶中的β-半乳糖苷酶活性在摊放2h后上升到最高峰，为鲜叶酶活性的1.2倍；随着摊放时间的延长，酶活性又逐渐下降，8h后酶活性降至鲜叶酶活的85%。揉捻及发酵过程中β-半乳糖苷酶活性呈大幅度下降。微波杀青处理后，β-半乳糖苷酶快速失活，摊放冷却后没有复性。     

温室效应与植物光合作用——大气CO_2浓度升高对植物光合机理影响的分析

比较分析了植物光合同化 CO2 速率、Rubisco活性等性状对长期和短期高浓度 CO2 的反应 .结果表明 :所研究的植物在高浓度 CO2 下生长 ,可以长期保持高的 CO2 同化速率 ;长期在高浓度 CO2 下生长植物的光合速率增加有两个来源 ,其一 ,是 CO2 浓度增加而增加的底物浓度效应 (Δ Pc) ,它的大小随外界短期 CO2 浓度改变而改变 ,其二 ,是植物光合系统结构改变而提高光合能量转换或是电子传递效率所产生的光合速率增加 (ΔPs) ,它的大小不随外界短期 CO2 浓度变化而改变。植物光合速率对大气 CO2 浓度升高的增加量是上述两者之和     

温度对龙眼体胚发生早期POD活性及同工酶的影响

以龙眼品种红核子LC2胚性细胞系诱导获得的体胚发生早期过程中不同发育阶段的同步化胚性培养物为材料,2个温度梯度(20 ℃、30 ℃)处理,以25℃为对照,初步研究了POD活性及同工酶在龙眼体胚发生早期的变化规律.结果表明:不同温度处理的胚性培养物,随着胚性细胞的分化,POD活性总体呈下降趋势,且无论高温(30℃)或低温(20 ℃)处理后龙眼体胚发生早期各阶段胚性培养物的POD活性均明显大于对照(25 ℃);温度处理对龙眼体胚发生早期POD同工酶谱影响也较大,表现为新谱带的出现.推测POD活性与同工酶谱的规律变化与龙眼体胚发生早期细胞分化以及维持体胚正常发育有密切关系.