马铃薯叶片特异性启动子克隆分析及其驱动的人白细胞介素-12表达载体构建

利用植物生物反应器表达具有应用价值的药用蛋白是近年来生物技术研究热点。本研究利用PCR技术从马铃薯基因组中克隆其叶片组织特异性启动子Prbcs后,利用PLACE等数据库对序列进行启动子元件分析,接着通过重叠PCR、限制性内切酶酶切、体外连接、转化等技术,以双元载体pCambia-2301为骨架,构建Prbcs驱动的药用蛋白人白细胞介素12（hIL12）表达载体。结果显示,本研究成功从马铃薯基因组中扩增得到了623 bp大小的启动子片段,序列分析表明该片段含有典型的启动子元件如TATA-box,以及一些组织特异性的相关响应元件,符合组织特异性启动子的特征;成功获得了基于pCambia-2301的Prbcs驱动的hIL12植物表达载体。本研究获得的Ppatatin启动子及其驱动的hIL12表达载体,为提高hIL12在植物生物反应器中表达量以及优化马铃薯生物反应器打下了基础。     

小麦质不育系花药及雌蕊邻苯二酚氧化酶同工酶分析

小麦细胞质雄性不育系、保持系花药三个不同发育时期的邻苯二酚氧化酶同工酶存在明显差异,不育系中的同工酶谱带数多于保持系.考虑到小麦细胞质雄性不育系的细胞核与保持系相同,但具有与保持系不相同的不育细胞质,故邻苯二酚氧化酶同工酶可能与植物的雄性不育特性有某种联系.雌蕊中的邻苯二酚氧化酶同工酶在不育系与保持系间基本相同,这可能与雌蕊中不育系、保持系间的核质组成相同有关,这一事实似乎从另一侧面说明了邻苯二酚氧化酶与不育特性存在着某种联系.     

Molecular phylogeny of the apterygotan insects based on nuclear and mitochondrial genes

Basal hexapodan orders (Apterygota) are traditionally divided in two well defined taxonomic groups, Entognatha and Ectognatha. Entognathy (the enclosed mouthparts condition) is considered the most distinctive character joining the Ellipura (Protura+ Collembola) with the order Diplura, whereas the presence of exposed mouthparts (ectognathy) is the feature shared by Microcoryphia and Zygentoma with the pterygote insects. In spite of the growing interest for the evolutionary history of the Apterygota, there is no complete agreement among the general phylogenetic hypotheses based on the study of morphological characters. In this study we analyzed the DNA sequence of segments of the nuclear Elongation Factor-1α(EF-1α) and of the mitochondrial 12S rRNA genes, and used different methods of phylogenetic reconstruction. The 12S gene seems to be more suited than the EF-1α to resolve some of the most outstanding systematic disputes, whereas the lack of resolution at the deeper nodes does not allow to assess the phylogenetic relationships within Microcoryphia and between ectognathan orders. We have obtained a fairly high support for the monophyly of the orders Diplura and Zygentoma. In the 12S analysis, the Ellipura and the Entognatha form monophyletic assemblages. In addition, the study of the distribution of introns in the EF-1α suggests a relationship between Collembola and Diplura.     

莲藕的酶促褐变及其贮藏中褐变的控制

为有效控制莲藕褐变,试验研究了温度、pH值、底物浓度以及抑制剂对莲藕多酚氧化酶（PPO）的影响。结果表明：莲藕PPO最适pH值为7.5,最适温度为35℃,以邻苯二酚为底物,米氏常数Km为0.0273 mol/L。亚硫酸钠、抗坏血酸（VC）对莲藕PPO活性具有较好抑制作用,乙酸、柠檬酸、植酸、木瓜蛋白酶、半胱氨酸也对莲藕PPO活性具有一定抑制作用。0.2%VC+2%乙酸+0.03%亚硫酸钠抑制剂处理对莲藕褐变的抑制效果最佳。可以通过浸泡亚硫酸钠和抗坏血酸（VC）、调节pH值、低温贮藏等方法来抑制莲藕PPO活性,控制莲藕褐变。     

种衣剂对玉米幼苗某些酶活性及苗体生长的影响

实验用三种种衣剂包衣玉米种子,研究了玉米幼苗中某些与植物抗逆代谢有关的酶活性变化。三种种衣剂均使超氧化物歧化酶、过氧化氢酶和过氧化物酶活性升高,苗体干鲜重增加。表明玉米种衣剂可通过调节玉米幼苗中某些酶的活性,增强植株的抗逆性。     

动态水环境容量研究——以潇河流域为例

传统的水环境容量以90%设计枯水流量为计算条件,存在水文计算条件单一、水环境容量不易管控等缺点。考虑河道流量与入河污染物影响下的水环境容量逐月变化,提出了基于水文过程与污染源变化条件下的动态水环境容量与入河污染管控计算方法。以山西潇河流域晋中段为例,计算了研究流域两水功能区的逐月水环境容量及其余量,运用MIKE11模型计算了逐月允许的动态入河污染负荷。研究结果表明:(1)2019年水功能区A的COD和氨氮水环境容量阈值分别为138.37~216.54 t和6.38~14.19 t,7-10月水环境容量较大;水功能区B的COD和氨氮水环境容量阈值分别为417.67~821.05 t和9.32~29.49 t,5-8月份水环境容量较小。(2)在满足水功能区目标水质的基础上,2019年水功能区A允许的入河负荷均大于现状入河量,汛期COD和氨氮容量余量较多;水功能区B在1月、2月应削减氨氮0.003、0.086 t;削减COD 0.05、0.143 t,4-7月份允许的污染负荷大于实际入河量,余量较多。计算的动态水环境容量能够客观反映变化水文条件下的水环境容量及其余量实际,给出了满足河道水质目标前提下的入河污染物允许阈值,为流域水环境保护与管理提供重要参考和新思路。     

Effects of Boron Stress on copper enzyme activity in tomato

Moderate B deficiency in plants has been reported to enhance Cu deficiency by keeping these plants in the vegetative growth stage. In this study, ascorbate oxidase activity was used as an index of the effect of B stress on Cu activity. When T3238FER (B‐inefficient) and Rutgers (B‐efficient) tomatoes (Lycopersicon esculentum Mill.) were grown in nutrient solutions at various B levels, B‐stressed plants had higher ascorbic acid oxidase activity than B‐sufficient plants. This activity was significantly higher in T3238FER than in Rutgers. Ascorbic acid oxidase activity may be directly or indirectly related to the role of B in plant growth and may be responsible for the more efficient use of B by Rutgers than by T3238FER tomato.     

The effects of triacontanol on seedling growth and polyphenol oxidase activity in dark and light growth lettuce

The present study was designed to study the effects of various concentrations (0.00; 0.01, 0.10 and 1.00‐mg/ml) of triacontanol on root, stem and leaf growth and polyphenol oxidase activity in each respective tissue; in Grand Rapids and Great Lakes varieties of eleven‐day‐old dark and light grown lettuce seedlings.

Root, stem and leaf growth was less than in non‐triacontanol‐treated Grand Rapids seedlings in both TC‐treated dark and light exposed seedlings. With respect to Great Lakes seedlings, all dark‐grown roots exhibited greater growth than the corresponding untreated control. Light‐grown Great Lakes roots treated with 0.01‐mg/ml and 1.0‐mg/ml or triacontanol respectively, grew more than control or 0.1‐mg/ml triacontanol‐treated seedlings. Both dark and light‐grown triacontanol‐treated stem and leaf tissues of Great Lakes seedlings all produced less growth than the untreated controls.

The Grand Rapids variety had less polyphenol oxidase activity in both dark and light‐grown root and stem tissues than in untreated controls; however, both dark and light‐grown leaf tissue, treat ed with 0.01‐mg/ml and 1.0‐mg/ml of triacontanol respectively exhibited more polyphenol oxidase activity than 1.0‐mg/ml triacontanol ‐treated or untreated control tissues.

TC treatment of 0.1‐mg/ml caused no enhancement of PPO activity in dark or light‐grown root, stem and leaf tissues of Great Lakes tissue, however, seedlings treated with a concentration of 0.01‐mg/ml TC exhibited more PPO activity than non‐TC‐treated controls.     

Effect of aluminum uptake on growth and enzyme activity in Pisum sativum var. ‘Alaska’ and ‘Little Marvel’

It is usually assumed that plant tissue responses to nutritional elements are due to specific genetic differences that may exist either between inbred or closely related species. Little Marvel (dwarf) and Alaska (normal) varieties of 14‐day old pea seedlings were treated with four different concentrations of Al‐containing nutrient solution (0.0mM, 0.2mM, 0.6mM and distilled H₂O), prior to being exposed for 14 days to either DARK, LIGHT, or UV. Selected tissues (root tip, main root, main stem and proximal stem) were bioassayed for peroxidase and polyphenol oxidase enzyme activities, fresh wt vs. dry wt, water uptake and stem growth. The present study suggests that Little Marvel and Alaska pea tissue responds to high toxicity levels of Al by demonstrating an enhancement of enzymic activity. Tissue weight, growth and water uptake also show differential tissue specificity in both Little Marvel and Alaska tissue, in terms of Al toxicity response, given a particular external exposure.     

金银花多酚氧化酶特异性与抑制剂动力学研究

对金银花多酚氧化酶底物特异性和不同抑制剂的抑制效应进行了动力学分析,结果表明:金银花多酚氧化酶的最适作用底物为绿原酸,Km值为0.0059 mmol/L。维生素C、4-己基间苯二酚、L-半胱氨酸对多酚氧化酶的抑制均属可逆抑制,其抑制类型分别属于混合性、竞争性、反竞争性,对游离酶的抑制常数KI分别为1.620、4.587、0 mmol/L,对酶-底物络合物的抑制常数KIS分别为1.995、0、3.780 mmol/L;柠檬酸的抑制效应属不可逆抑制。维生素C、4-己基间苯二酚、L-半胱氨酸、柠檬酸对金银花多酚氧化酶半抑制浓度分别为0.062、0.053、0.140、0.048 mmol/L。