Ubiquitination-mediated protein degradation and modification： an emerging theme in plant-microbe interactions

Post-translational modification is central to protein stability and to the naodulation of protein activity.Various types ofprotein modification,such as phosphorylation,methylation,acetylation,myristoylation,glycosylation,and ubiquitina-tion,have been reported.Among them,ubiquitination distinguishes itself from others in that most of the ubiquitinatedproteins are targeted to the 26S proteasome for degradation.The ubiquitin/26S proteasome system constitutes the majorprotein degradation pathway in the cell.In recent years,the importance of the ubiquitination machinery in the controlof numerous eukaryotic cellular functions has been increasingly appreciated.Increasing number of E3 ubiquitin ligasesand their substrates,including a variety of essential cellular regulators have been identified.Studies in the past severalyears have revealed that the ubiquitination system is important for a broad range of plant developmental processes andresponses to abiotic and biotic stresses.This review discusses recent advances in the functional analysis of ubiquitina-tion-associated proteins from plants and pathogens that play important roles in plant-microbe interactions.     

21世纪植物化学生态学前沿领域

植物和其它有机体通过次生物质为媒介的化学作用关系近年引入注目,其中植物的诱导化学防御,植物的化学通讯,植物次生物质和进化的关系,植物与人类的化学关系和海洋植物化学生态学是21世纪植物化学生态学值得关注的前沿领域。植物化学生态学前沿领域的进展将为实现21世纪的持续,发展是在生态安全条件下提高农业产量并达到对病虫草害的有效控制方面具有重要意义。     

植物芥子酶研究进展

芥子酶防御系统为白花菜目植物特有．由芥子酶及其底物硫代葡萄糖苷组成．芥子酶和硫代葡糖苷分别储藏在不同的细胞中,在受到病虫侵袭时,底物和酶相遇,硫代葡糖苷被降为有毒化合物,起防御作用．对植物芥子酶防御系统研究进展进行了综述,包括基因家族的结构、基因的表达调控、芥子酶的细胞定位、植物以外其它生物的芥子酶、硫代葡糖苷／芥子酶系统起源进化以及其可能功能等．     

云南哀牢山地区森林附生维管植物多样性及区系特征

附生植物作为山地森林生态系统中重要的结构性成分,在维持森林生态系统生物多样性格局、水分和养分循环等方面发挥着重要作用。本文通过野外调查、标本查阅并结合相关文献,对云南哀牢山地区附生维管植物物种组成及分布进行了系统研究。结果显示,哀牢山地区附生维管植物共有23科83属218种,其中附生蕨类和兰科植物最丰富。附生蕨类有34属93种,以附生-石生蕨类生活型占优势,其中水龙骨科17属62种,占附生蕨类的66.67%,瓦韦属(Lepisorus)和石韦属(Pyrrosia)分别有13种和10种。附生兰科植物有26属65种,其中石斛属(Dendrobium)和石豆兰属(Bulbophyllum)分别有12种和8种。该地区附生维管植物属的分布具有明显的热带性质并以热带亚洲分布居多。附生植物生长于生境因子变化剧烈、资源有限的林冠,对环境变化敏感,极易遭受破坏且破坏后难以恢复,不少附生植物具有很高的药用、观赏等价值。因此,应加强对附生维管植物这一特殊类群的保护。     

Mapping the eosinophil cationic protein antimicrobial activity by chemical and enzymatic cleavage

The eosinophil cationic protein (ECP) is a human antimicrobial protein involved in the host immune defense that belongs to the pancreatic RNase A family. ECP displays a wide range of antipathogen activities. The protein is highly cationic and its bactericidal activity is dependant on both cationic and hydrophobic surface exposed residues. Previous studies on ECP by site-directed mutagenesis indicated that the RNase activity is not essential for its bactericidal activity. To further understand the ECP bactericidal mechanism, we have applied enzymatic and chemical limited cleavage to search for active sequence determinants.Following a search for potential peptidases we selected the Lys-endoproteinase, which cleaves the ECP polypeptide at the carboxyl side of its unique Lys residue, releasing the N-terminal fragment (0-38).Chemical digestion using cyanogen bromide released several complementary peptides at the protein N-terminus. Interestingly, ECP treatment with cyanogen bromide represents a new example of selective chemical cleavage at the carboxyl side of not only Met but also Trp residues. Recombinant ECP was denatured and carboxyamidomethylated prior to enzymatic and chemical cleavage. Irreversible denaturation abolishes the protein bactericidal activity.The characterization of the digestion products by both enzymatic and chemical approaches identifies a region at the protein N-terminus, from residues 11 to 35, that retains the bactericidal activity. The most active fragment, ECP(0-38), is further compared to ECP derived synthetic peptides. The region includes previously identified stretches related to lipopolysaccharide binding and bacteria agglutination. The results contribute to define the shortest ECP minimized version that would retain its antimicrobial properties. The data suggest that the antimicrobial RNase can provide a scaffold for the selective release of cytotoxic peptides.     

茉莉酸对棉花单宁含量和抗虫相关酶活性的诱导效应

以植物生长调节物茉莉酸(Jasmonic acid,JA)为诱导子,以常规棉为研究对象,探讨了外源茉莉酸对棉花幼苗单宁和蛋白酶抑制素以及其它抗虫相关酶活性诱导的浓度依赖性和持久性,讨论了棉花抗虫相关物质的抗虫效果.结果表明,0.01、0.1和1.0 mmol/L茉莉酸都能在2周内诱导棉花单宁和胰蛋白酶抑制素(Proteinase inhibitors,PIs)含量增加,诱导多酚氧化酶(Polyphenol oxidase,PPO)、苯丙氨酸解氨酶(Phenylalanine ammonia-lyase,PAL)、过氧化物酶(Peroxidase,POD)和过氧化氢酶(Catalase,CAT)活性升高.对3种浓度茉莉酸的诱导效应进行分析表明,0.1 mmol/L茉莉酸对于诱导PIs、PPO、POD和CAT最有效,0.1和1.0 mmol/L茉莉酸对于诱导棉花单宁和苯丙氨酸解氨酶等效,二者的诱导效应均高于0.01 mmol/L.对茉莉酸诱导抗性的持久性进行分析表明,最佳诱导效应发生的时间各不相同:POD活性在JA处理后第1天最高,随后呈下降趋势,PIs和单宁含量分别在JA处理后第7天和第14天达最大值;JA处理后第1天和第7天的PPO活性无明显差异,但明显高于第14天;JA处理后第7天和第14天的PAL活性无明显差异,但明显高于第1天;JA处理后第1、7和14天棉花叶片的CAT活性均无明显差异.以上结果表明,茉莉酸可通过增加棉叶单宁和PIs含量、提高棉叶PAL、PPO、POD和CAT活性等增强棉花幼苗的抗虫性.     

MicroID2: A Novel Biotin Ligase Enables Rapid Proximity-Dependent Proteomics

Identifying protein–protein and other proximal interactions is central to dissecting signaling and regulatory processes in cells. BioID is a proximity-dependent biotinylation method that uses an “abortive” biotin ligase to detect proximal interactions in cells in a highly reproducible manner. Recent advancements in proximity-dependent biotinylation tools have improved efficiency and timing of labeling, allowing for measurement of interactions on a cellular timescale. However, issues of size, stability, and background labeling of these constructs persist. Here we modified the structure of BioID2, derived from Aquifex aeolicus BirA, to create a smaller, highly active, biotin ligase that we named MicroID2. Truncation of the C terrminus of BioID2 and addition of mutations to alleviate blockage of biotin/ATP binding at the active site of BioID2 resulted in a smaller and highly active construct with lower background labeling. Several additional point mutations improved the function of our modified MicroID2 construct compared with BioID2 and other biotin ligases, including TurboID and miniTurbo. MicroID2 is the smallest biotin ligase reported so far (180 amino acids [AAs] for MicroID2 versus 257 AAs for miniTurbo and 338 AAs for TurboID), yet it demonstrates only slightly less labeling activity than TurboID and outperforms miniTurbo. MicroID2 also had lower background labeling than TurboID. For experiments where precise temporal control of labeling is essential, we in addition developed a MicroID2 mutant, termed lbMicroID2 (low background MicroID2), that has lower labeling efficiency but significantly reduced biotin scavenging compared with BioID2. Finally, we demonstrate utility of MicroID2 in mass spectrometry experiments by localizing MicroID2 constructs to subcellular organelles and measuring proximal interactions.     

不同光照强度对兴安落叶松几种主要防御蛋白活力的影响

林木组成抗性对抵御害虫危害发挥着重要作用。为了研究环境因子对其组成抗虫性的影响,以兴安落叶松幼苗为试验材料,通过搭建遮阳棚设置3个光照强度(自然光照作为对照、50%和25%自然光照强度),模拟森林幼苗生长的林缘、林窗和林下3种不同的光环境,分析不同光照强度对其针叶内与植物抗虫性有关的几种主要防御蛋白活力的影响。研究结果表明,不同光照强度对针叶内主要防御蛋白活力有显著影响(P<0.05)。50%和25%光照强度下,POD、SOD、PAL、PPO和CI的活性显著高于自然光照下的(P<0.05),POD和CI的活性在50%光照强度下最高,SOD、PAL和PPO的活性在25%光照强度下最高;自然光照条件下CAT的活性显著高于50%和25%光照强度下的(P<0.05);遮荫处理对TI的活性有显著影响(P<0.05),但在不同月份中波动较大,规律性不明显。相同的光照强度条件下,主要防御蛋白活力在3个月中也有显著变化(P<0.05)。7月和8月份保护性酶POD和CAT的活性显著高于6月(P<0.05);防御性酶PAL、PPO以及蛋白酶抑制剂CI的活性在6月份最高。综合以上研究结果,光照可影响兴安落叶松针叶中几种主要防御蛋白活力,这也说明在较荫蔽的光照环境下,兴安落叶松有较强的抗性。其中SOD、CAT、PAL、PPO和POD是植物次生物质生成过程中的关键酶,其活性的改变在一定程度上表征着植物防御体系在变化。研究环境因子与植物防御之间的关系具有重要的理论和实际应用价值。     

The relationship between the reduction of nonstructural carbohydrate induced by defoliator and the growth and mortality of trees

Large scale herbivorous insect outbreaks can cause death of regional forests, and the events are expected to be exacerbated with climate change. Mortality of forest and woodland plants would cause a series of serious consequences, such as decrease in vegetation production, shifts in ecosystem structure and function, and transformation of forest function from a net carbon sink into a net carbon source. There is thus a need to better understand the impact of insects on trees. Defoliation by insect pests mainly reduces photosynthesis (source decrease) and increases carbon consumption (sink increase), and hence causes reduction of nonstructural carbohydrate (NSC). When the reduction in NSC reaches to a certain level, trees would die of carbon starvation. External environment and internal compensatory mechanisms can also positively or negatively influence the process of tree death. At present, the research of carbon starvation is a hotspot because the increase of tree mortality globally with climate change, and carbon starvation is considered as one of the dominating physiological mechanisms for explaining tree death. In this study, we reviewed the definition of carbon starvation, and the relationships between the reduction of NSC induced by defoliation and the growth and death of trees, and the relationships among insect outbreaks, leaf loss and climate change. We also presented the potential directions of future studies on insect-caused defoliation and tree mortality.     

Pair territoriality in the longnose filefish,oxymonacanthus longirostris

The longnose filefish,Oxymonacanthus longirostris, usually lives in heterosexual pairs, the male and female swimming together and sharing the same territory. Pair territoriality in the species was examined in detail in relation to sexual differences in territorial defense activities. Rigorous pair territoriality was maintained only during the breeding season, although pairs used their home ranges exclusively to a certain extent, during the non-breeding season. The frequency of aggression against other conspecific pairs in the breeding season was higher than in the non-breeding season. Agonistic interactions appear to be over both mates and food resources, the strict pair territoriality in the breeding season possibly being due to mutual mate guarding. In intraspecific aggressive interactions, males usually led their partner females when attacking intruders. The feeding frequency of males was much lower than that of females in the breeding season. Mate removal experiments indicated that females could not defend their original territories solitarily and their feeding frequency decreased. Conversely, males could defend territories solitarily without a decrease in feeding frequency. These results suggest that males contribute most to the defense of the pair territory, with females benefiting from territorial pair-swimming with their partner males.