植物表达载体pGMAR-BADH的构建

将甜菜碱醛脱氢酶基因BADH和植物表达载体pGMAR通过内切酶BamHI和KpnI双酶切,T4连接酶进行连接反应。重组质粒在大肠杆菌菌株DH5α内扩增,提取并纯化质粒。经鉴定,基因BADH已被完整、正确的插入到pGMAR载体中,并成功将BADH插入到载体pGMAR的两个MAR序列之间。     

用小麦芽有效成分消除豆腥味的研究

利用小麦芽粉中的醛脱氢酶(ALDH)对大豆进行脱腥处理,使大豆中的腥味物质转变为酸,从而有效消除大豆中的豆腥味。通过测定总醛含量(以己醛计),结合感官评定,研究了作用温度、作用时间、ALDH添加量对豆腥味的影响。结果表明,小麦芽最佳作用温度为45℃;最佳作用时间为2.5h;最佳添加量为大豆干质量的2%,对豆腥味及适口性影响的因素大小依次为:作用温度>作用时间>添加量。     

Generation and analysis of expressed sequence tag sequences from a soft‐seeded pomegranate cDNA library

The cultivation of soft‐seeded pomegranate is an important direction in pomegranate breeding. To comprehensively understand the molecular mechanisms involved in the formation of soft‐seeded pomegranate (Punica granatum. var. Hongmanaozi), we established an expressed sequence tag (EST) resource. Two thousand valid sequences were generated, from which 907 unigenes were obtained after initial assembly using the clustalx program. Among these unigenes, 51 showed no similarity to any protein in the public databases, 433 matched with proteins of unknown function, and 423 matched with proteins of known or putative functions. The 423 unigenes were further classified into 13 categories. Among these categories, protein synthesis, cell structure, protein destination and storage, secondary metabolism, signal transduction and transporters accounted for 8%, 8%, 4%, 7%, 6% and 17%, respectively. We also successfully developed 10 highly polymorphic expressed sequence tag‐simple sequence repeat (EST‐SSR) markers for pomegranate. The results provide a new tool for future activities in pomegranate breeding.     

低温胁迫对番茄幼苗叶片中脯氨酸降解酶的活性及其基因表达量的影响

为了探讨脯氨酸代谢与番茄抗寒性之间的关系,采用常规方法测定了低温胁迫不同时间下番茄幼苗叶片中游离脯氨酸含量和脯氨酸脱氢酶活性,采用实时荧光定量PCR法检测了不同处理时间下脯氨酸脱氢酶基因的表达量,并分析了各项指标的动态变化。结果表明,在低温逆境下,耐寒性品种O-33-1中脯氨酸积累量总体呈增加的趋势,而冷敏感品种‘耐运2000’游离脯氨酸积累量下降,说明脯氨酸积累量的变化在一定程度上可以体现番茄抗寒性的强弱;ProDH活性在O-33-1中呈上升趋势,在‘耐运2000’中呈下降趋势,说明脯氨酸的积累受P5CS与ProDH相反方向的共同调节,使植物体内脯氨酸含量保持在适当的水平;2个品种在低温胁迫下ProDH基因表达量都显著降低,说明低温逆境抑制了该基因的表达,使脯氨酸的降解在转录水平上受到调节。     

土壤微生物产电信号评价芘污染毒性的研究

通过向土壤中加入葡萄糖促进微生物产电过程,研究了芘污染条件下土壤产电变化规律。利用双室微生物燃料电池(Microbial fuel cells,MFCs),实时、连续记录芘污染土壤产电电压。产电110 h后结束MFCs运行,采用循环伏安法检测芘对土壤微生物电化学活性的影响;结合PCR-DGGE及测序技术,分析芘对MFCs阳极表面细菌群落结构的影响。结果显示,MFCs产电电量随芘浓度增加显著降低。循环伏安检测显示芘降低了土壤微生物的电化学活性。DNA序列分析表明,阳极细菌与已报道的产电细菌高度相似,包括Sporolactobacillus、Clostridium、Enterobacter、Bacillus及Ethanoligenens。芘降低了Bacillus丰度。     

苹果山梨醇脱氢酶(NAD-SDH)基因的克隆及其反义表达载体对农杆菌的转化

山梨醇是一种糖醇,是蔷薇科植物所特有的,是蔷薇科植物主要的光合产物、运输糖和贮藏物质(B ieleskiand Redg-w ell,1995),在其糖代谢中占有重要地位。另一方面,山梨醇能够提高植物抗环境胁迫的能力(徐迎春等,2001;Brow n etal.,1999)。N A D-山梨醇脱氢酶(N A D-SD H)是山梨醇     

Oxygen enrichment with magnesium peroxide for minimizing hypoxic stress of flooded corn

Flooding/waterlogging is a major factor responsible for hypoxic stress in agriculture. The aim of this study was to develop an effective oxygen buffer with magnesium peroxide (MgO₂) to generate hydrogen peroxide (H₂O₂) and release bioavailable oxygen. MgO₂ provided a relatively stable level (approx. 300 µM) of bioavailable oxygen. The oxygen‐buffer system is adjustable and controllable by adding Mg²⁺ or EDTA to the aqueous system. Regular H₂O₂ was also able to provide bioavailable oxygen but the system was poorly buffered with respect to oxygen release. The accessibility of plants to bioavailable oxygen was indicated by the activity of alcohol dehydrogenase (ADHase, EC 1.1.1.1), an anaerobically induced enzyme of flooded plants. The application of MgO₂ to flooded soil reduced ADHase activity in corn‐root tips by 91.3%. This application of MgO₂ presents a novel pathway to significantly (P < 5%) minimize adverse impacts of hypoxia on flooded corn seedlings. This finding may have broad implications for addressing hypoxicity problems in crop science and technology.     

谷子MDH基因非生物逆境响应特性研究

为揭示谷子MDH基因对逆境胁迫的响应,本研究通过序列比对得到2个谷子苹果酸脱氢酶(malate dehydrogenase,MDH)基因SiMDH1、SiMDH2,并对SiMDH1和SiMDH2的编码蛋白特征、基因结构、功能、进化等性状进行了系统的分析和预测,用荧光实时定量PCR(qPCR)检测了SiMDH1和SiMDH2在苗期不同逆境及关键生育期干旱胁迫和不同光照强度下的表达。蛋白特征参数和序列分析表明,SiMDH1和SiMDH2蛋白特征参数相似度很高,序列非常保守,都含有MDH基因的特征功能域苹果酸酶NAD结合域和苹果酸酶C端功能域。亚细胞定位预测显示,SiMDH1和SiMDH2主要定位于叶绿体、细胞质和线粒体。启动子区域顺式元件分析发现,在SiMDH1和SiMDH2启动子区域含有大量光应答、激素类和其他类应答元件。苗期逆境qPCR表达谱分析发现,SiMDH1和SiMDH2受脱落酸(ABA)、聚乙二醇(PEG)、高盐和低温不同程度诱导,揭示SiMDH1和SiMDH2参与了谷子苗期非生物逆境胁迫应答。进一步分析发现,SiMDH1和SiMDH2参与了谷子在拔节、抽穗、灌浆期的干旱应答,...     

Interactive effects of biochar and micronutrients on faba bean growth,symbiotic performance,and soil properties

Leguminous crops are significantly involved in the global symbiotic biological N₂ Fixation (BNF), an eco‐friendly process in the agriculture system. Biochar is considered as a vital amendment in improving growth and quality of crops and soils. Few investigations have been conducted to determine the combination effect of biochar with microelements on growth of legumes and soil properties. This study was designed to study the effect of soybean straw‐derived biochar (SSDB) with or without microelements on soil microbial and chemical properties, growth, yield, and seed chemical composition of faba bean (Vicia faba L.). Results revealed that dehydrogenase (DHA) and phosphatase (P‐ase) activities were markedly improved with the increase of SSDB rates under addition of microelements and their highest values were recorded after 90 d. Significant increases were noticed in nodulation activities, nodulation numbers (30.1–72.8), concentrations of N (1.62–1.93%), P (0.15–0.21%), and K (0.53–0.67%), and seed chemical constituents due to the addition of SSDB in the presence of microelements. Moreover, the combination of biochar with microelements caused significant changes in microbial counts. Overall, this investigation shows the potential and role of SSDB in enhancing the growth quality of faba bean seeds as well as an improvement of soil characteristics.