5-Aminolevulinic acid alleviates herbicide-induced physiological and ultrastructural changes in Brassica napus

It is well known that application of 5-aminolevuJinic acid （ALA） could promote the plant growth under abiotic stress in oilseed rape （Brassica napus L.）. However, the specifics of its physiological and ultrastructural regulation under herbi- cide stress conditions are poorly understood. In the present study, alleviating role of ALA in B. napus was investigated under four levels of herbicide propyl 4-（2-（4,6-dimethoxypyrimidin-2-yloxy） benzylamino） benzoate （ZJ0273） （0, 100, 200 and 500 mg L-1） with or without 1 mg L-1 ALA treated for 48 or 72 h. Results showed that after 48 h of herbicide stress, the growth of rape seedlings was significantly inhibited with the successive increases of the ZJ0273 concentrations from 0 to 500 mg L-1, but this inhibition was obviously alleviated by exogenous application of ALA. However, when treatment time prolonged to 72 h, the recovery effects of ALA could not be evaluated due to the death of plants treated with the highest concentration of ZJ0273 （500 mg L-1）. Further, the root oxidizability and activities of antioxidant enzymes （superoxide dismutase, perox- idase and ascorbate peroxidase） were dramatically enhanced by the application of 1 mg L-1 ALA under herbicide stress. Therefore, plants treated with ALA dynamically modulated their antioxidant defenses to reduce reactive oxygen species （ROS） accumulation and malondialdehyde （MDA） content induced by herbicide stress. Additionally, exogenously applied ALA improved the ultrastructure＇s of chloroplast, mitochondria and nucleus, and induced the production of stress proteins. Our results suggest that ALA could be considered as a potential plant growth regulator for the improvement of herbicide tolerance through alleviation of the physiological and ultrastructural changes induced by the herbicide in crop production.

Altered expression of nuclear matrix proteins in etoposide induced apoptosis in HL-60 cells

The events of cell death and the expression of nuclear matrix protein (NMP) have been investigated in a promyelocytic leukemic cell line HL-60 induced with etoposide. By means of TUNEL assay, the nuclei displayed a characteristic morphology change, and the amount of apoptotic cells increased early and reached maximun about 39% after treatment with etoposide for 2 h. Nucleosomal DNA fragmentation was observed after treatment for 4 h. The morphological change of HL-60 cells, thus, occurred earlier than the appearance of DNA ladder. Total nuclear matrix proteins were analyzed by 2-dimensional gel electrophoresis. Differential expression of 59 nuclear matrix proteins was found in 4 h etoposide treated cells. Western blotting was then performed on three nuclear matrix acssociated proteins, PML, HSC70 and NuMA. The expression of the suppressor PML protein and heat shock protein HSC70 were significantly upregulated after etoposide treatment, while NuMA, a nuclear mitotic apparatus protein, was down regulated. These results demonstrate that significant biochemical alterations in nuclear matrix proteins take place during the apoptotic process.

Effect of high salinity on cell growth and protein production of Antarctic ice microalgae Chlamydomonas sp. ICE-L

Antarctic ice microalgae Chlamydomonas sp.ICE-L can survive and thrive in Antarctic sea ice.In this study,Chlamydomonas sp.ICE-L could survive at the salinity of 132‰ NaCl.SDS-PAGE showed that the density of 2 bands（26 and 36 kD） decreased obviously at the salinity of 99‰ NaCl compared to at the salinity of 33‰ NaCl.The soluble proteins in Chlamydomonas sp.ICE-L grown under salinity of 33‰ and 99% NaCl were compared by 2-D gel electrophoresis.After shocking with high salinity,8 protein spots were found to disappear,and the density of 28 protein spots decreased.In addition,19 protein spots were enhanced or induced,including one new peptide（51 kD）.The changes of proteins might be correlated with the resistance for Chlamydomonas sp.ICE-L to high salinity.

甘蓝型油菜蛋白质双向电泳体系的建立

选用油菜品种08127,采用TCA-丙酮提取方法,pH4～7胶条和改进的IEF聚焦程序,得到较好的蛋白质双向电泳结果,建立了一种适合甘蓝型油菜的双向电泳体系。利用其他甘蓝型油菜品种的幼苗、叶片和茎中的蛋白检验该实验体系,都能得到较好的蛋白质双向电泳结果。利用该实验体系比较甘蓝型油菜幼苗不同发育时期蛋白质双向电泳图,找到二者之间的差异点,选取部分点成功进行了质谱分析。

2个油菜CMS系统的酯酶和过氧化物酶同工酶分析

以甘蓝型油菜细胞质雄性不育系(CMS)Polima A及其保持系Polima B、陕2A及其保持系陕2B 4个材料初花期的叶片、叶柄以及花蕾组织为材料,采用聚丙烯酰胺垂直板凝胶电泳比较它们在酯酶(EST)和过氧化物酶(POD)同工酶酶谱的差异。结果表明,甘蓝型油菜不育系与其对应保持系的叶片、叶柄的EST同工酶谱均无明显差异,但两系花蕾的EST同工酶谱有一定的差异;Polima A与陕2A对应器官的EST同工酶谱均表现出明显差异。不育系与其对应保持系的叶片、叶柄的POD同工酶谱差异不明显,而两系花蕾的POD同工酶谱有明显的差异;两个不育系之间的POD同工酶谱明显不同。花蕾的EST和POD同工酶的条带数目明显多于相应材料的叶片和叶柄,且EST同工酶的条带数目明显多于相应的POD同工酶。因此,甘蓝型油菜CMS系统Polima A和陕2A有着不同的遗传背景。

甘蓝型油菜叶片蛋白质双向电泳体系优化

比较了甘蓝型油菜叶片总蛋白质提取的2种不同方法,探讨了叶片蛋白质分离的双向凝胶电泳技术优化方案。结果表明:采用TCA/丙酮法较Tris-Cl法更适合蛋白质提取,选择pH为4～7的固相胶条和分离胶浓度为12%的聚丙烯酰胺凝胶(SDS-PAGE)能够使蛋白质得到充分分离,采用考马斯亮蓝G250染色剂进行2次染色,可提高2-DE分辨率和实验解析度,优化后的双向电泳体系能够分离出2 000个以上的蛋白点,并且重复稳定性好。

Differential Proteomic Analysis of Carbon Ion Radiation in Sheep Sperm

This study is first to investigate proteomic changes in sheep sperm induced by carbon ion radiation using two-dimensional electrophoresis （2-DE） analysis in the project of breeding a new variety of sheep. Differential expression proteins were detected using the PDQuest 8.0 software after staining with Coomassie blue. Valid spots were then analyzed through liquid chromatography tandem mass spectrometry （LC-MS/MS）. Among the 480 total protein spots displayed in 2-D gels, 6 specific protein spots were observed in sperm gels. A search against protein sequences in the National Center for Biotechnology Information databases （NCBI） indicated that differentially expressed proteins correspond to two proteins, identified to be enolase and transcription factor AP-2-alpha （TFAP-2c0. The two proteins were up-regulated in the irradiated sperm. To the best of our knowledge, this study is the first to identify proteomic changes induced by carbon ion radiation in sheep sperm. The analysis of differential expression protein may be useful in identifying new breeding markers in sheep reproduction and in clarifying the mechanisms involved in irradiation or space breeding.

Proteomic Alterations of Antarctic Ice Microalga Chlamydomonas sp.Under Low-Temperature Stress

Antarctic ice microalga can survive and thrive in cold channels or pores in the Antarctic ice layer. In order to understand the adaptive mechanisms to low temperature, in the present study we compared two-dimensional polyacrylamide gel electrophoresis （2-DE） profiles of normal and low temperature-stressed Antarctic ice microalga Chlamydomonas sp. cells. In addition, new protein spots induced by low temperature were identified with peptide mass fingerprinting based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS） and database searching. Well-resolved and reproducible 2-DE patterns of both normal and low temperature-stressed cells were acquired. A total of 626 spots was detected in control cells and 652 spots were detected in the corresponding low temperature-stressed cells. A total of 598 spots was matched between normal and stressed cells. Two newly synthesized proteins （a and b） in low temperature-stressed cells were characterized. Protein spot A （53 kDa, pl 6.0） was similar to isopropylmalate/homocitrate/citramalate synthases, which act in the transport and metabolism of amino acids. Protein spot b （25 kDa, pl 8.0） was related to glutathione S-transferase, which functions as a scavenger of active oxygen, free radicals, and noxious metabolites. The present study is valuable for the application of ice microalgae, establishing an ice microalga Chlamydomonas sp. proteome database, and screening molecular biomarkers for further studies.