I.小麦非叶器官碳酸酐酶活性与定位分析II.镉毒害导致芦苇胚性细胞死亡的激素调控研究

第一部分： 通过生理测定和化学染色分析了冬小麦品种小堰54和京411的叶片和非叶片组织的碳酸酐 酶活性。叶片碳酸酐酶活性(CA)在挑旗时期达到最大值，之后减少到最小，而在饱粒期又呈 增加趋势。从灌浆期到饱粒期，颖片和内稃的CA活性均减少，而外稃和种皮的CA活性均增加。在饱粒期，小堰54的叶片、颖片、外稃和种皮CA活性均高于京411。组织化学染色表明，CA主要分布在旗叶的叶肉细胞叶绿体中，也分布在非叶片组织颖片、外稃和内稃的叶肉和维管束鞘细胞的胞质中。这些结果表明，小麦非叶组织叶肉和维管束鞘细胞的胞质中的CA可能对饱粒期冬小麦的C4光合途径起作用。饱粒期小堰54的C02传递到Rubisco酶速率和抗旱性较京411高。 第二部分： 以继代培养的芦苇胚性细胞为材料，利用台盼兰拒染法检测了悬浮细胞死亡过程，并利用石蜡切片法及苏木精染色法观察了不同浓度镉对芦苇细胞的毒害作用。1000μM的CdCl2迅速导致芦苇悬浮细胞死亡，200μM的CdClz在接种后第5天引起悬浮细胞死亡，100μM的CdCl2在接种后第7天引起悬浮细胞死亡，≤50μM的CdClz在接种后7天不引起悬浮细胞死亡。同时对不同浓度镉处理的芦苇胚性细胞的内源植物激素和可溶性蛋白质进行分析，≤50μM的镉浓度显著地降低胚性细胞内IAA、ZR、GA3和GA4的含量，却提高ABA的含量，抑制可溶性蛋白质的合成：≥100μM的镉浓度显著地提高IAA、ZR、GA3和GA4的含量，却降低ABA的含量，促进可溶性蛋白质的合成。这些结果表明，镉的毒害至少包括镉浓度决定的两种细胞死亡机制，高浓度的镉(1000μM)引起的细胞死亡应当为坏死，而100μM的镉引起植物悬浮细胞发生程序性死亡。在较高浓度(≥100μM)的镉处理下，芦苇细胞内内源IAA、ZR、GA3和GA4的浓度较高，可能调控可溶性蛋白质的合成而促进细胞发生程序化死亡。

Deletion of yeast CWP genes enhances cell permeability to genotoxic agents

We have previously reported the development of a novel genotoxic testing system based on the transcriptional response of the yeast RNR3-lacZ reporter gene to DNA damage. This system appears to be more sensitive than other similar tests in microorganisms, and is comparable with the Ames test. In an effort to further enhance detection sensitivity, we examined the effects of altering major cell wall components on cell permeability and subsequent RNR3-lacZ sensitivity to genotoxic agents. Although inactivation of single CWP genes encoding cell wall mannoproteins had little effect, the simultaneous inactivation of both CWP1 and CWP2 had profound effects on the cell wall structure and permeability. Consequently, the RNR3-lacZ detection sensitivity is markedly enhanced, especially to high molecular weight compounds such as 4-nitroquinoline-N-oxide (> sevenfold) and phleomycin (> 13-fold). In contrast, deletion of genes encoding representative membrane components or membrane transporters had minor effects on cell permeability. We conclude that the yeast cell wall mannoproteins constitute the major barrier to environmental genotoxic agents and that their removal will significantly enhance the sensitivity of RNR-lacZ as well as other yeast-based genotoxic tests.

土壤镉污染的遗传及生态毒性研究

针对蚕豆根尖微核试验方法现存的问题并结合土壤自身的特性，进行了蚕豆微核试验方法改进的试验研究。在此基础上，本文选择蚕豆幼苗次生根根尖为供试材料，以细胞有丝分裂指数（MI）、染色体畸变率（CAF）和微核率（MNF）为试验指标，开展了我国典型土壤（红壤、棕壤和黑土）中低剂量（0~10 mg•kg-1）镉污染遗传毒性剂量-效应关系研究。研究针对蚕豆根尖微核遗传毒性评价指标“污染指数”的缺陷，首次提出“毒性指数”。“毒性指数”将有丝分裂指数、染色体畸变率和微核率指标所得数据结合，评价土壤镉遗传毒性水平。与此同时，研究选择蚕豆幼苗叶片为供试材料，进行了以抗氧化酶活性（SOD、POD和CAT）和植物激素含量（ABA、GA3和Z&ZR）为指标的镉污染胁迫生态毒理试验研究，并比较了上述三类指标对土壤镉污染的毒性响应及敏感性。 蚕豆根尖微核方法改进试验结果表明，在三种备选试验材料中，蚕豆初生根、次生根根尖细胞和芽尖细胞均对镉胁迫响应显著。与芽尖细胞相比，根尖细胞MI、CAF和MNF之间具有较好的相关性；而次生根尖细胞比与初生根尖细胞对镉响应更敏感，遗传毒性效应显著相关。在上述方法学研究基础上，将蚕豆幼苗次生根根尖微核试验应用到典型土壤镉污染的研究中，结果表明，同一剂量下，红壤中镉的遗传毒性效应最强，棕壤次之，黑土最不显著。土壤pH值与有机质含量对镉的遗传毒性影响较大。 抗氧化防御酶活性试验结果表明，POD和SOD在0~10 mg•kg-1镉浓度范围内先升高后降低，对镉毒性的指示效果最明显；CAT反应与SOD、POD相反，其作用机理有待进一步探讨。植物激素试验结果表明，ABA和Z&ZR对镉胁迫响应敏感，其含量与镉浓度存在良好的剂量-效应关系；GA3随着镉浓度的增加表现出无规律性，表明它对污染物的响应具有选择性。 试验数据显示，不同指标对镉胁迫均具有响应，但响应的域值及敏感度不同。比较而言，蚕豆根尖微核遗传毒性指标指示效应更明显。将各指标联合使用可使土壤镉污染的遗传和生态毒性诊断更为全面、有效。本研究为土壤镉污染早期诊断敏感指标筛选及指标组合提供了实验依据，为土壤重金属污染早期诊断奠定试验和理论基础。

Effects of sodium dodecylbenzene sulfonate and sodium dodecyl sulfate on the Mytilus galloprovincialis biomarker system

The effects of in vivo exposure of Mytilus galloprovincialis to two anionic surfactants (SDBS and SDS) on the molecular biomarker system were studied. After continuous exposure for 72 days, activities/levels of GST, GPx and GSH were significantly higher than in corresponding control groups following exposure to 3.000 mg/L SDS and SDBS. Activities of SOD and CAT were significantly inhibited by experimental SDBS (except CAT in 0.100 mg/L group), but not by SDS. Statistical analysis of enzyme activities/levels suggested that there were significant positive relationships between GST and GPx, and negative relationships were found between GSH and CAT, GSH and SOD. Amplified fragment length polymorphism (AFLP) results showed that a greater genotoxic effect was observed for SDBS than for SDS. Based on the above results, the biomarker system of mussels can be affected by the two anionic surfactants (>= 3.000 mg/L); it was more easily affected by SDBS than by SDS. Crown Copyright (C) 2009 Published by Elsevier Inc. All rights reserved.