盐生植物海滨锦葵幼苗盐胁迫下基因差异表达分析

利用cDNA-AFLP技术对海滨锦葵幼苗盐胁迫下叶片和根部的基因差异表达模式进行分析和比较, 并对部分盐胁迫应答的转录衍生片段进行了回收、测序和功能推测, 以从转录水平分析海滨锦葵的耐盐分子机制。结果显示：(1) 盐胁迫下海滨锦葵幼苗叶片和根部的基因差异表达多以量的变化为主, 包括盐胁迫下基因表达上调、下调或随盐处理浓度高低和胁迫时间长短而波动的差异表达模式; 只有少量基因的差异表达表现出质的变化, 如盐胁迫下基因沉默或诱导表达; (2) 仅在盐胁迫处理2 h的海滨锦葵幼苗根部, 基因的差异表达以质的变化为主的类型比例略高于量的变化类型比例; (3) 盐胁迫应答基因在不同组织中上调、下调、诱导或沉默的比例随胁迫处理时段而动态变化, 在刚胁迫时基因表达的差异加剧, 而后随胁迫处理时段的延长而渐趋稳定。结果预示, 从基因表达水平探讨植物的耐盐分子机理, 尽管有一定的规律可循, 但由于不同组织对盐胁迫的应答是动态变化的过程, 海滨锦葵不同组织在盐胁迫不同阶段的基因时、空、序表达特征并没有固定的程式。对部分盐胁迫下上调或诱导表达的转录衍生片段(Trivially distributed file system, TDFs)进行的序列分析和功能推测表明, 苗期海滨锦葵在盐胁迫下应答基因至少涉及3类：(1) 离子平衡重建或减少胁迫损伤相关基因(特别是运转蛋白类); (2) 恢复盐胁迫下植物生长和发育相关基因：如参与能量合成和激素调节途径相关基因等; (3)信号转导相关基因及功能未确定的新基因。文章并对盐胁迫应答基因的差异表达模式与海滨锦葵的耐盐性关系进行了讨论。     

Annotation of Differential Gene Expression in Small Yellow Follicles of a Broiler-Type Strain of Taiwan Country Chickens in Response to Acute Heat Stress

This study investigated global gene expression in the small yellow follicles (6–8 mm diameter) of broiler-type B strain Taiwan country chickens (TCCs) in response to acute heat stress. Twelve 30-wk-old TCC hens were divided into four groups: control hens maintained at 25°C and hens subjected to 38°C acute heat stress for 2 h without recovery (H2R0), with 2-h recovery (H2R2), and with 6-h recovery (H2R6). Small yellow follicles were collected for RNA isolation and microarray analysis at the end of each time point. Results showed that 69, 51, and 76 genes were upregulated and 58, 15, 56 genes were downregulated after heat treatment of H2R0, H2R2, and H2R6, respectively, using a cutoff value of two-fold or higher. Gene ontology analysis revealed that these differentially expressed genes are associated with the biological processes of cell communication, developmental process, protein metabolic process, immune system process, and response to stimuli. Upregulation of heat shock protein 25, interleukin 6, metallopeptidase 1, and metalloproteinase 13, and downregulation of type II alpha 1 collagen, discoidin domain receptor tyrosine kinase 2, and Kruppel-like factor 2 suggested that acute heat stress induces proteolytic disintegration of the structural matrix and inflamed damage and adaptive responses of gene expression in the follicle cells. These suggestions were validated through gene expression, using quantitative real-time polymerase chain reaction. Functional annotation clarified that interleukin 6-related pathways play a critical role in regulating acute heat stress responses in the small yellow follicles of TCC hens.     

利用RNA-seq技术分析淹水胁迫下转BnERF拟南芥差异表达基因

为探究淹水胁迫下BnERF调节的耐淹防御相关途径,应用RNA-seq技术,对淹水6小时后的拟南芥(Arabidopsis thaliana)野生型(WT)和转BnERF株系(E33)幼苗进行基因表达分析。结果表明,淹水3天后,E33表现出较强的耐淹性,地上部生长状况和根系发育均明显强于野生型。E33幼苗未淹水处理时相对于野生型单独上调的基因有9个,4个为膜结合蛋白,其中2个参与MAPK级联途径,其它5个参与氧化胁迫及水分调节途径;与未淹水野生型相比,无论是未淹水处理还是淹水6小时后的E33幼苗中缺氧响应、抗氧化防护及细胞、器官发育相关基因的表达量均上调。另外,淹水6小时后E33的差异基因并未完全覆盖淹水6小时后野生型的差异基因;E33幼苗中缺氧响应、氧化胁迫响应、能量的产生与转变、乙醇代谢途径中的基因以及乙烯响应因子基因的表达量都明显高于野生型。上述结果表明,BnERF直接或间接调节植物的淹水胁迫相关生理代谢途径,参与淹水胁迫的防御过程。     

Analysis of Proteins Expressed by an Abiotic Stress Tolerant Pseudomonas putida (NBAII-RPF9) Isolate Under Saline and High Temperature Conditions

Pseudomonas putida (NBAII-RPF9) was identified as an abiotic stress tolerant bacterium capable of growing at 45 °C as well as in 1 M NaCl. The proteins expressed by this bacterium when subjected to these two stresses were analyzed by 2D gel and MALDI-TOF/MS. Two parameters viz., heat/saline shock (20 min at 45 °C/1 M solid NaCl added at mid log phase and incubated for 1 h) and heat/saline tolerance (24 h growth at 45 °C/in 1 M NaCl) were studied. Under heat shock 13 upregulated proteins and 1 downregulated protein were identified and under tolerance 6 upregulated proteins were identified. GroES and GroEL proteins were expressed under both tolerance and shock. Under saline shock 11 upregulated proteins were identified whereas under saline tolerance 6 upregulated proteins were identified and all these proteins had pI between 3 and 10 with molecular weights ranging from 14.3 to 97 kDa. Aspartate carbamoyltransferase was common under both the saline conditions studied. The analysis revealed involvement of heat stress responsive molecular chaperones and membrane proteins during heat stress. During salt stress, proteins involved in metabolic processes were found to be upregulated to favor growth and adaptation of the bacterium. Heat shock chaperones viz., DnaK and DnaJ were expressed under both saline and heat stress. This is the first report of protein profile obtained from a single bacterium under saline and heat stress and the studies reveal the complex mechanisms adapted by the organism to survive under high temperature or saline conditions.