茶树小分子量热激蛋白基因CsHSP17.2的克隆与表达分析

[目的]进一步了解茶树小分子量热激蛋白基因CsHSP17.2在逆境胁迫条件下的分子生物学功能.[方法]利用RT-PCR技术从茶树‘迎霜’中克隆得到CsHSP1 7.2基因,运用生物信息学软件分析其核苷酸和编码蛋白,通过Real-timePCR分析其表达模式.[结果]该基因开放阅读框长度为453 bp,编码150个氨基酸,蛋白质相对分子质量为17.2×10a,理论等电点5.56;无信号肽位点,属于非分泌型蛋白;被定位于细胞质中.系统发育树分析表明,茶树CsHSP1 7.2与水稻(GenBank登录号:P27777)和花生(ABC41131)的进化关系较近,属于小分子量热激蛋白基因家族第Ⅰ亚族.qRT-PCR分析发现,茶树CsHSP17.2属于组成型基因;高温(38℃)处理1h能显著提高CsHSP17.2 mRNA的相对表达量(P＜0.05);干旱(100 g·L-1 PEG 6000)、高盐(200 mmol· L-1 NaCl)和外源脱落酸(200 mg· L-1 ABA)处理条件下,该基因的转录水平均出现不同程度的上调.[结论]克隆得到茶树‘迎霜’小分子量热激蛋白基因CsHSP17.2,其在花中表达量最高,且响应高温、干旱、高盐和外源脱落酸胁迫.     

分子蒸馏技术纯化柠檬精油工艺研究

采用分子蒸馏技术对柠檬精油进行浓缩精制,分别考察了蒸馏温度、刮膜转速和蒸馏压力对浓缩精油中柠檬醛浓度的影响。在单因素试验基础上,通过L9(34)正交设计试验确定了柠檬精油分子蒸馏的最佳工艺参数。结果表明,柠檬精油分子蒸馏的最佳工艺参数为:进料速度1 mL/min,分子蒸馏温度45℃,刮膜转速240 r/min,蒸馏压力40 Pa。浓缩精油经过气相色谱仪(GC)测定,柠檬醛浓度由1.41 mg/mL上升至11.17 mg/mL,浓缩倍数达7.9。经过分子蒸馏后的浓缩精油呈深棕色,质地浓稠,气味浓郁、醇厚。     

格木寄生线虫中国新记录种——卡勒萨隐皮孢囊线虫Cryphodera kalesari

在对广东省主要珍贵树种上的线虫进行调查时从格木Erythrophleum fordii上分离到一种隐皮孢囊线虫,经形态特征观察和测量数据分析,将其鉴定为卡勒萨隐皮孢囊线虫Cryphodera kalesari。其诊断特征为:雌虫椭圆形至近球形,头部具有1个唇盘和2～3个唇环,口针长33.6～37.8μm,阴门唇突出,阴门与肛门之间区域凹陷,肛阴距为42～72μm;2龄幼虫头部具1个明显的唇盘和3个唇后环纹,口针长24.8～29.5μm,口针基部球前缘凹陷,侧区3条侧线,尾长圆锥形,长38～53.5μm,尾末端细圆,透明尾长17.1～25.1μm,侧尾腺孔位于肛门后2～5环;雄虫未发现。本研究首次获得了卡勒萨隐皮孢囊线虫的rDNA(LSU D2D3和ITS)序列,为此线虫的鉴定提供了可靠的分子数据。本研究还分析了卡勒萨隐皮孢囊线虫与本属其他种类的系统进化关系。卡勒萨隐皮孢囊线虫为中国的地理新记录种。格木为隐皮孢囊线虫的新寄主。     

Assessment of molecular detection of Fusarium circinatum in insects and passive spore traps in Pinus radiata plantations

Fusarium circinatum is the causal agent of pitch canker, a destructive disease that threatens natural and planted pine forests around the world. Although pitch canker has caused problems in Spain and Portugal, concerning Europe as a whole, the fungus is not established across the pine distribution area. Its dispersion by wind and/or insect vectors could nevertheless play a role in the colonization of currently uninfected stands. It is therefore crucial to develop monitoring tools for its detection. To this end, we assessed the molecular detection of the pathogen in environmental samples of bark beetles and passive spore traps, collected in two infected Pinus radiata plantations in Basque country, Spain. The spread pattern of F. circinatum was assessed by an experimental design that included insect and spore traps installed at the centre, at the edge and outside the plots. Our results showed that F. circinatum was detected in both types of samples, at almost all collection dates. In both type of samples, positive detections were mainly found at the centre of the plots, a lower proportion at the edge, and very few outside. This suggests that long‐distance dispersion of Fusarium circinatum does not rely on wind spore dispersal neither on insect flight. Our study also shows that molecular methods are a powerful tool to monitor the pathogen in environmental samples.     

Screening of Human CYP1A2 and CYP3A4 Inhibitors from Seaweed In Silico and In Vitro

Phenolic compounds and carotenoids are potential inhibitors of cytochrome P450s. Sixteen known compounds, phenolic compounds and carotenoids from seaweed were examined for potential inhibitory capacity against CYP1A2 and CYP3A4 in silico and in vitro. Morin, quercetin, and fucoxanthin inhibited the enzyme activity of CYP1A2 and CYP3A4 in a dose-dependent manner. The IC₅₀ values of morin, quercetin, and fucoxanthin were 41.8, 22.5, and 30.3 μM for CYP1A2 and 86.6, 16.1, and 24.4 μM for CYP3A4, respectively. Siphonaxanthin and hesperidin did not show any significant effect on CYP1A2, but they slightly inhibited CYP3A4 activity at high concentrations. In silico modeling of CYP’s binding site revealed that the potential inhibitors bound in the cavity located above the distal surface of the heme prosthetic group through the 2a or 2f channel of CYPs. This study presents an approach for quickly predicting CYP inhibitory activity and shows the potential interactions of compounds and CYPs through in silico modeling.     

甘肃华山松溃疡病病原真菌类群的初步鉴定

近年来甘肃陇南地区华山松溃疡病发生严重，导致华山松大面积死亡，但有关病害发生的原因等尚不清楚。采用组织分离法从感病华山松的健康部位、病健交界处、子实体及病组织处分离获得97个真菌菌株，结合形态学及系统学对分离到的菌株进行初步分析。结果显示，所得菌株分为9个种，其中Neocosmospora silvicola占14.17%，Fusicolla violacea占1.67%，Microcera sp.占8.33%，Neonectria neomacrospora占11.67%，Dermea pinicola占8.33%，Nematogonum ferrugineum占9.17%，Pezicula sporulosa占4.17%，Phaeoacremonium fraxinopennsylvanicum占14.17%，Sarea resinae占9.17%。以丛赤壳科为优势菌群，占总菌株数量的44.33%；初步推断Neocosmospora silvicola为疑似病原菌。     

外来植物土荆芥入侵的化学基础探讨

土荆芥为藜科藜属一年生或多年生直立草本植物,1864年首次发现于中国台湾省台北淡水,2010年1月7日被列入中国《第二批外来入侵植物种名单》。目前对土荆芥的入侵学研究主要集中在生物学、生态学、化学成分、他感作用、逆境抗性等方面,因此其入侵机制主要是化学武器假说,如次生代谢旺盛,具化感潜力,抗逆性强等。但并未从分子学角度进行相关研究,如土荆芥的遗传进化特点、土荆芥入侵的分子标记、土荆芥的代谢组学等。尤其是土荆芥入侵之后,其与本土植物的种子萌发速度或生长速度比较(优先效应),目前尚未有报道。因此,未来在上述研究的基础上,必须对其分子生物学开展深入研究,尤其应重点研究土荆芥入侵的优先效应机制。     

肾上腺皮质肿瘤良恶性鉴别方法探究

肾上腺皮质肿瘤（Adrenocortical Tumor,ACT）是一种常见的肿瘤,存在于人体的内分泌系统中,并且是引发肾上腺皮质癌（Adrcnocortical Carcinoma,ACC）进而导致死亡的主要原因.经典的＂Weiss系统＂在诊断明确性上存在种种无法解决的问题,但随着科技水平的发展,人们开始探索新的途径来鉴别ACT的良恶性,于是开始有了影像学和分子生物学技术等特异性的检查方法,从而进一步增强了诊断的明确性.现通过对比的方式,探究肾上腺皮质肿瘤（Adrenocortical Tumor,ACT）影像学检查和分子生物学技术检查的结果,从而能更准确地鉴别其良恶性.