红豆杉细胞稳定生产紫杉醇中的同步化协同诱导作用

同步化协同诱导可以稳定提高曼地亚红豆杉细胞培养物中紫杉醇含量。细胞培养周期第8d的低温同步化处理可促使细胞达到最高同步化率（20.4%）,而茉莉酸甲酯（MJ）的协同诱导可提高紫杉醇产量,使紫杉醇产量最高值达到54.7mg·L^-1。在细胞生长周期第8d,未经低温同步化的红豆杉细胞中的关键酶基因DXR、HMGR、GGPPS和DBAT的表达量在MJ诱导24h后均迅速下降,但在低温同步化的细胞中紫杉醇表达量下降缓慢,60h后仍维持较高的水平。低温同步化和MJ协同诱导的红豆杉细胞中,紫杉醇合成关键酶基因高效稳定的表达可能是引起紫杉醇产量稳定提高的原因之一。     

树干注射印楝素的动态研究

本文主要以雪橙Citrus sinensis（um）Osbeckcv．Xue Cheng为研究对象,研究了成年雪橙树对印楝素药液的吸收速率,印楝素注射雪橙树后在雪橙叶片中的富集和消解动态,结果显示：采用药水吸收系数评价雪橙树对印楝索水溶液的吸收速率和日平均吸药量的试验结果表明,雪橙树对印楝素水溶液的吸收速度在不同时间存在明显差异,以14时至15时为吸药量最高点,吸收值为35．8mL／h,该时段也是试验当天最高温度时段。日平均累积吸药量为221．8mL,药水系数为2．98。不同浓度印楝素注射雪橙树后,在叶片中的富集量和消解动态一致。印楝素富集量在注射后9一lO天达到了峰值,注射后56天各注射浓度的雪橙树叶片中仍能检测到印楝素,而印楝素喷雾处理的雪橙树植株的叶片在药后7天即无检测量。     

A bioanalytical method for the proteome wide display and analysis of protein complexes from whole plant cell lysates

While protein interaction studies and protein network modeling come to the forefront, the isolation and identification of protein complexes in a cellular context remains a major challenge for plant science. To this end, a nondenaturing extraction procedure was optimized for plant whole cell matrices and the combined use of gel filtration and BN‐PAGE for the separation of protein complexes was studied. Hyphenation to denaturing electrophoresis and mass spectrometric analysis allows for the simultaneous identification of multiple (previously unidentified) protein interactions in single samples. The reliability and efficacy of the technique was confirmed (i) by the identification of well‐studied plant protein complexes, (ii) by the presence of nonplant interologs for several of the novel complexes (iii) by presenting physical evidence of previously hypothetical plant protein interactions and (iv) by the confirmation of found interactions using co‐IP. Furthermore practical issues concerning the use of this 2‐D BN/SDS‐PAGE display method for the analysis of protein–protein interactions are discussed.     

Effect of Pedoclimatic Conditions on the Chemical Composition of the Sigoise Olive Cultivar

The present work focused on the quality and the chemical composition of monovarietal virgin olive oil from the Sigoise variety grown in two different locations in Tunisia, viz., a sub‐humid zone (Béjaoua, Tunis) and an arid zone (Boughrara, Sfax). In addition to the quality characteristics (acidity, peroxide value, and the spectrophotometric indices K232 and K270) and the chemical composition (content of fatty acids, antioxidants, and volatile compounds) of the oil, the fruit characteristics of the olives were studied. Except for the content of the majority of the fatty acids, there were significant differences observed in the oil composition of olives that were cultivated in different locations. The content of total phenols and lipoxygenase (LOX) oxidation products was higher for olives grown at the higher altitude, whereas that of α‐tocopherol, carotenes, and chlorophylls was higher for olives from the Boughrara region (lower altitude). Moreover, olives produced at the higher altitude showed a higher ripeness index and oil content than those cultivated at the lower altitude.     

九台晚李PGIP基因的克隆及生物信息学分析

以九台晚李叶片基因组为模板,PGIP基因保守序列设计引物,PCR扩增到1条全长1192bp的目的片段(GenBank登录号:GU068978)。该基因包含有1个完整的开放阅读框,由2个外显子和1个内含子构成,外显子总长990bp,编码330个氨基酸,其编码的氨基酸序列中含有一段典型的亮氨酸重复序列。序列分析表明:该基因与中国李、杏、桃、马哈利樱桃、梅等李属植物的PGIP基因序列一致度达95%~99%。系统进化分析显示出属内亲缘关系较近、属间亲缘关系较远的特点。该序列为植物分子抗病育种提供了1条新的基因资源。     

黄山群体种自然杂交后代儿茶素组分的变异特性

对黄山群体种自然杂交后代种质资源表型性状进行调查,从中筛选出204份种质作为试材,对其儿茶素6种组分进行测定并进行遗传多样性分析。结果表明,黄山群体种自然杂交后代在儿茶素组成成分上存在丰富的多样性和变异,遗传多样性指数平均为1.78,变异系数平均为31.2%。根据儿茶素品质指数和儿茶素总量分别对204份种质进行聚类分析,筛选出适制绿茶的种质材料15份、适制乌龙茶的种质6份、适制红茶的种质7份。从中筛选出一批特异的资源,儿茶素总量、酯型儿茶素含量、酯型儿茶素/儿茶素总量之比均较高的种质材料4份,品质指数高的特异资源5份。     

根癌农杆菌介导转化川草二号老芒麦胚性愈伤组织

以川草二号老芒麦成熟种子为外植体,经过对培养基的筛选和培养条件优化,建立了愈伤组织再生系统。转化载体为pCAMBIA1304质粒,其T—DNA上携有潮霉素抗性基因（hptII）和类产碱假单胞菌杀虫蛋白基因（ppIP）,经根癌农杆菌EHA105介导转化结构致窑、颗粒状、黄白色的胚性愈伤组织。通过潮霉素筛选和对抗性植株进行分子检测,获得了转基因植株。同时优化了农杆菌遗传基因转化的参数,建立了农杆菌介导的川草二号老芒麦程序化转基因方案。     

Putrescine Influences Growth and Production of Coumarins in Hairy Root Cultures of Witloof Chicory (Cichorium intybus L. cv. Lucknow Local)

The effect of putrescine (Put) on the growth and production of two coumarins, esculin and esculetin, in hairy roots of chicory (Cichorium intybus L. cv. Lucknow local) was examined. To study the role of Put on growth and production of coumarins, polyamine inhibitors, namely α-dl-difluromethylornithine and α-dl-difluromethylarginine were used at 1 mM concentration. Put treatment at 1.5 mM produced a 1.9-fold increase in the growth of hairy roots, as well as the production of esculin and esculetin. The treatments with polyamine (PA) inhibitors resulted in much lower growth and production of coumarins compared with both 1.5-mM Put treatment and the control. Both free and conjugated PAs were studied over the whole culture period, and conjugates of all three PAs, namely Put, spermidine, and spermine, were higher than free PAs throughout the culture period. The treatments with PA inhibitors showed lower levels of endogenous PAs compared with Put-treated samples. The treatment with 1.5 mM Put showed maximum accumulation of endogenous conjugated Put (2,098 ± 157 nmoles gm⁻¹ fresh weight). The production of esculin and esculetin was strictly correlated with growth in all treatments. Put at 1.5 mM resulted in greater length of primary root (18.3 ± 1.4 cm) as compared with the control (11 ± 0.9 cm) and larger numbers of secondary and tertiary roots. Received July 14, 1999; accepted October 5, 1999     

Mechanism of dusky reddish-brown "kaki" color development of Japanese morning glory,Ipomoea nil cv. Danjuro

The mechanism of dusky reddish-brown "kaki" color development of morning glory, Ipomoea nil cv. Danjuro, was studied. Three major known anthocyanins were isolated as glucosylated pelargonidin derivatives. Measurement of the vacuolar pH with proton-selective microelectrodes revealed the vacuolar pH of the colored cell of open flowers to be 6.8, while that of buds was 5.8. Mixing of the three anthocyanins according to the composition ratio in petals at pH 6.8 allowed the identical color to that of petals to be reproduced. The typical "kaki" color development was mostly caused by 5-OH free acylated anthocyanins, which have two lambdamax around 435 and 535 nm in the visible region.