高浓度细胞分裂素诱导棉花悬浮细胞程序性死亡

棉花悬浮细胞经高浓度细胞分裂素（2mg／L KT或4mg／L KT）处理后第4天大量死亡，培养物呈褐黑色；抽提总DNA，进行琼脂糖胶电泳，结果表明：2mg／L KT和4mg／L KT两处理中有明显的大小为140～180bp及其倍增量片段的DNA“梯”。进行PI和FDA染色显微观察可以看到：2mg／L KT处理中大部分细胞的细胞核较小，核质浓缩，呈棒形或弯月形；胞质浓缩，出现了不对称的质壁分离，且丧失了FDA染色活性。这些结果表明，由高浓度细胞分裂素处理引起的棉花悬浮细胞大规模死亡是一种细胞程序性死亡（PCD）。不同的生理状态（UCSA和BAE）和不同生长时期（指数增长期和静止期）的棉花悬浮细胞对高浓度细胞分裂素的诱导反应存在差异。     

用葡萄球菌甲蛋白协同凝集鉴别猪痢疾密螺旋体的研究

应用猪痢疾蜜螺旋体（Ｔ．Ｈ．）免疫血清结合葡萄球菌甲蛋白（ＳＰＡ）与Ｈ１０，２－５，Ｘ２３，Ｃ１７等Ｔ．Ｈ．菌株进行协同凝集试验，均出现明显凝集反应。而免疫血清结合不含ＳＰＡ的菌株，无特定病原（ＳＰＦ）血清结合含ＳＰＡ的菌株均无凝集反应。免疫血清结合ＳＰＡ与７种肠道菌均无凝集反应。免疫血清用量以０．２ｍｌ为合适。最低用菌量为３０倍稀释，即每个视野Ｔ．Ｈ．约４０个菌体。     

Presence of Muscarinic Acetylcholine Receptors in the Cattle Tick Boophilus microplus and in Epithelial Tissue Culture Cells of Chironomus tentans

A muscarinic acetylcholine receptor (mAChR) has been demonstrated and partially characterized in larvae of the cattle tick Boophilus microplus. Its properties are compared with mAChR from an epithelial cell line from the dipteran insect Chironomus tentans. Competition studies with cholinergic ligands of different specificity revealed the muscarinic nature of the cholinergic receptors investigated in both species. In homogenates from tick larvae, specific binding sites for [³H]quinuclidinyl benzilate (QNB) with high affinity (1·2±(0·13) nM ; B_max 22·5 pmol mg protein⁻¹) were detected that do not bind nicotinic compounds specifically. The estimated IC₅₀ values for nicotine, imidacloprid and α-bungarotoxin were all in the mM range. Additionally, with tick larvae, high-affinity nicotinic binding sites were detected with [³H]nicotine which could be displaced by high concentrations of imidacloprid or QNB. The estimated IC₅₀ values for nicotine, α-bungarotoxin, imidacloprid and QNB were 43(±8) nM , 0·8(±0·2) μM , 2·8(±0·6) μM and 78(±1·9) μM , respectively. With homogenates of the non-neuronal insect cell line from C. tentans, only high-affinity binding sites for [³H]QNB were found. Muscarinic antagonists selectively displaced [³H]quinuclidinyl benzilate (QNB) binding to tick larvae homogenates. The mAChR of B. microplus preferred pirenzepine (IC₅₀ 2·13(±1·02) μM ) among different subtype-specific mAChR antagonists (4-DAMP had IC₅₀ 49·9(±9·13) μM and methoctramine had IC₅₀ 121(±14·2) μM ) indicating a type of binding site similar to the vertebrate M1 mAChR subtype. The tick muscarinic receptor seems to be a G-protein-coupled receptor, as concluded from the 4·8-fold reduction in receptor affinity for binding of the muscarinic agonist oxotremorine M upon treatment with the non-hydrolysable GTP-analogue γ-S-GTP. Binding data for the agonists oxotremorine M (IC₅₀ 71·3(±19·6) μM ) and carbachol (IC₅₀ 253(±87·1) μM ) parallel the biological efficacy of these compounds, in that, while oxotremorine M showed some activity against ticks, carbachol was ineffective.     

Caco-2细胞模型及其在营养素小肠吸收机理研究中的应用

Caco-2细胞源自人结肠癌细胞,体外培养时能自发地进行类似肠道细胞的形态学和生化学上的分化,获得许多小肠吸收细胞的特性,如形成微绒毛结构;在细胞表面形成良好的刷状缘;在细胞间形成紧密连接;分泌水解酶以及合成转运糖、氨基酸和药物等的载体转运系统。由培养在微孔滤膜上的Caco-2细胞构建的模型为研究营养素在小肠的吸收机理提供了一个有效且易于操作的实验手段。本文主要综述了Caco-2细胞模型的建立、特征、检测及其在氨基酸、维生素、核苷和微量元素等营养素小肠吸收机理研究中的应用。     

水稻不育系培矮64S的空间诱变效应及后代的SSR分析

对经过卫星搭载的培矮64S种子及后代进行了有关性状研究和SSR标记分析,结果表明:培矮64S空间诱变后代SP1代发芽率、存苗率、株高、抽穗期、株叶型的性状与对照相比没有明显变化;但SP2代在株高、粒型和育性等性状上发生了变异;从SP3代中获得柱头比培矮64S明显增大的变异株系,这对于改良培矮64S的异交结实率和提高制种产量具有重要意义。此外,对10个SP3变异株进行SSR分析结果表明,扩增片段数目变化约占55.21%,扩增片段分子量变化约占44.79%,空间诱变引起的变异可能是以DNA缺失-重复为主。SSR座位的变异频率平均为23.33%,变异座位在水稻基因组中是随机分布的。     

不同LED光质对培养火龙果胚性愈伤组织的影响

以“玫瑰红龙”火龙果成熟胚诱导的愈伤组织为材料,研究不同光质配比的LED光处理对火龙果愈伤组织生长的动态影响。结果表明1红1绿光质配比,12h/d照射,最有利于火龙果愈伤组织的生长,在28d至42d细胞增长最快且活性最高,为0.78-0.75。其次是白光>1红1蓝>1绿1蓝>1红1绿1蓝>红光,在70d培养周期内,第35d是较理想的收获时间。     

植物增殖细胞核抗原与DNA复制关系的研究现状

为了更深入的对植物增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)进行研究,本研究归纳了PCNA的研究现状,总结了PCNA的结构特征和包括PCNA参与DNA复制、DNA损伤修复和细胞周期调控等在内的PCNA的功能特点。目前有关PCNA的研究以医学和动物体为主,有关植物PCNA研究的报道很少且相对落后,但已有文献指出,PCNA调控DNA复制,参与DNA复制以确保植物体正常生长发育,因此植物PCNA的功能值得继续研究。     

Phenolic extract of Morchella angusticeps peck inhibited the proliferation of HepG2 cells in vitro by inducing the signal transduction pathway of p38/MAPK

Morchella angusticeps Peck, one of the most popular edible mushrooms, has attracted great attention due to its delicious taste and healthy properties. However, both its biological effects and the possible mechanism of action have not yet been known. We investigated the anti-proliferative activity of the phenolic extract derived from Morchella angusticeps Peck (MPE) against HepG2 human hepatocellular carcinoma cells. Results showed that MPE at non-cytotoxicity doses significantly inhibited the proliferation of HepG2 cells in a dose-dependent manner with inhibitory rates ranging from 18 to 90% (P<0.01). The possible mechanism might be that MPE induced apoptosis through initiating the mitochondrial death pathway by regulating Bax, Bcl-2 and cleaved caspase-3. On the other hand, MPE might trigger cell cycle arrest at G₀/G₁/S phases by managing p21, Cyclin D1, cyclin-dependent kinases-4 (CDK4) and proliferating cell nuclear antigen (PCNA). Additionally, MPE downregulated TRAF-2 and p-p53, while upregulated p-ASK1 and p-p38. Therefore, it could be inferred that MPE might induce the anti-proliferative function to HepG2 cells through the p38/MAPK signal transduction pathway.     

The M43 domain-containing metalloprotease RcMEP1 in Rhizoctonia cerealis is a pathogenicity factor during the fungus infection to wheat

Wheat(Triticum aestivum L.) is an important staple crop for global human. The necrotrophic fungus Rhizoctonia cerealis is the causal pathogen of sharp eyespot, a devastating disease of wheat. Herein, we identified RcMEP1, a zinc metalloproteaseencoding gene from R. cerealis genomic sequences, and characterized its pathogenesis function. RcMEP1 expressed at markedly-high levels during R. cerealis infection process to wheat. The predicted protein RcMEP1 comprises of 287 amino acid residues and contains a signal peptide and a M43 metalloprotease domain harboring the active site motif(HEVGHWLGLYH). The assays of Agrobacterium tumefaciens-mediated transient expression in Nicotiana benthamiana leaves indicated that RcMEP1 is an apoplastic elicitor of cell death, and that the predicted signal peptide functions and is required for secretion and cell death-induction. The purified RcMEP1 protein and its M43 domain peptide were individually able to induce plant cell death and H2 O2 accumulation, and to inhibit expression of host chitinases when infiltrated into wheat and N. benthamiana leaves, while the M43 domain-deleting peptide and negative control lacked the capacity. Moreover, compared with the control pretreatment, the purified RcMEP1 protein or its M43-domain peptide resulted in enhanced pathogenesis in the inoculated wheat, whereas the M43 domain-deleting peptide failed. These results suggest that RcMEP1 acted as an important pathogenicity factor during R. cerealis infection to wheat and that its signal peptide and M43 domain are required for the secretion and pathogenesis of RcMEP1. This study provides insights into pathogenesis role of M43 domain-containing metalloproteases during R. cerealis infection to wheat.