Differential expression of heat shock protein genes in preconditioning for photosynthetic acclimation in water-stressed loblolly pine

Heat shock proteins (HSPs) are induced not only under heat stress conditions but also under other environmental stresses such as water stress. In plants, HSPs families are larger than those of other eukaryotes. In order to elucidate a possible connection between HSP expression and photosynthetic acclimation or conditioning, we conducted a water stress experiment in loblolly pine (Pinus taeda L.) seedlings involving progressive treatment consisting of one cycle of mild stress (?1 MPa) followed by two cycles of severe stress (?1.7 MPa). Net photosynthesis was measured at each stress level. Photosynthetic acclimation occurred in the progressive treatment after the first cycle, but not in the severe treatment, suggesting that a cycle of mild stress conditioned the trees to adapt to a more severe stress. Real time results indicated specific patterns in needles in the expression of HSP70, HSP90 and sHSP genes for each treatment, both at maximum stress and at recovery. We identified a pine homolog to GRP94 (ER resident HSP90) that was induced after rehydration coincident with acclimation. Further analysis of the promoter region of the pine GRP94 showed putative cis-elements associated with water stress and rehydration, corresponding to the expression pattern observed in our experiment.     

Differentially expressed genes and proteins upon drought acclimation in tolerant and sensitive genotypes of Coffea canephora

The aim of this study was to investigate the molecular mechanisms underlying drought acclimation in coffee plants by the identification of candidate genes (CGs) using different approaches. The first approach used the data generated during the Brazilian Coffee expressed sequence tag (EST) project to select 13 CGs by an in silico analysis (electronic northern). The second approach was based on screening macroarrays spotted with plasmid DNA (coffee ESTs) with separate hybridizations using leaf cDNA probes from drought-tolerant and susceptible clones of Coffea canephora var. Conilon, grown under different water regimes. This allowed the isolation of seven additional CGs. The third approach used two-dimensional gel electrophoresis to identify proteins displaying differential accumulation in leaves of drought-tolerant and susceptible clones of C. canephora. Six of them were characterized by MALDI-TOF-MS/MS (matrix-assisted laser desorption-time of flight-tandem mass spectrometry) and the corresponding proteins were identified. Finally, additional CGs were selected from the literature, and quantitative real-time polymerase chain reaction (qPCR) was performed to analyse the expression of all identified CGs. Altogether, >40 genes presenting differential gene expression during drought acclimation were identified, some of them showing different expression profiles between drought-tolerant and susceptible clones. Based on the obtained results, it can be concluded that factors involved a complex network of responses probably involving the abscisic signalling pathway and nitric oxide are major molecular determinants that might explain the better efficiency in controlling stomata closure and transpiration displayed by drought-tolerant clones of C. canephora.     

Photosynthetic gene expression in black willow under various soil moisture regimes

This study was the first attempt to extract RNA from black willow (Salix nigra Marshall) that contains numerous secondary products and to examine the photosynthetic gene expression of black willow under a wide range of soil moisture regimes. Black willow cuttings were grown under control, continuous flooding, periodic flooding and periodic drought for 42 d. A modified lithium chloride precipitation method was used for RNA extraction. Results of real-time polymerase chain reaction showed reduced gene expression of oxygen evolving complex, large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenasse and ferredoxin on day 7 as well as the latter two on day 14 in response to flooding. Therefore, decreased expression of these three genes may have contributed to the observed reduced photosynthetic capacity in response to flooding.     

Expression pattern of the psbO gene and its involvement in acclimation of the photosynthetic apparatus during abiotic stresses in Festuca arundinacea and F. pratensis

PsbO, the manganese-stabilizing protein, plays a crucial role in oxygen-evolving complex functioning and stabilization, by maintaining optimal manganese, calcium and chloride concentrations at the active state of PSII. In this paper we present a study focused on recognizing the relationship between psbO gene activity and acclimation of the photosynthetic apparatus under abiotic stresses in the grasses Festuca arundinacea and F. pratensis. PsbO expression was compared between two distinct genotypes within each species which differed in their levels of stress tolerance (drought and frost, respectively) during drought treatment (F. arundinacea) and cold acclimation (F. pratensis). The research involved: (1) the analyses of psbO gene expression profiles using real-time PCR, and (2) the analyses of PsbO protein accumulation profiles using protein gel blot hybridization. The results indicate that PsbO plays a protective function with respect to the photosynthetic apparatus during abiotic stresses. In cold-treated F. pratensis plants the accumulation of PsbO seems to be responsible for differences in the PSII photochemical efficiency. Higher stability of PSII during drought, observed in the high-drought tolerant F. arundinacea genotype, is not associated with PsbO accumulation, although the degradation of this protein affects destabilization of the oxygen-evolving complex in drought.     

Studying the functional genomics of stress responses in loblolly pine with the Expresso microarray experiment management system

Conception, design, and implementation of cDNA microarray experiments present a variety of bioinformatics challenges for biologists and computational scientists. The multiple stages of data acquisition and analysis have motivated the design of Expresso, a system for microarray experiment management. Salient aspects of Expresso include support for clone replication and randomized placement; automatic gridding, extraction of expression data from each spot, and quality monitoring; flexible methods of combining data from individual spots into information about clones and functional categories; and the use of inductive logic programming for higher-level data analysis and mining. The development of Expresso is occurring in parallel with several generations of microarray experiments aimed at elucidating genomic responses to drought stress in loblolly pine seedlings. The current experimental design incorporates 384 pine cDNAs replicated and randomly placed in two specific microarray layouts. We describe the design of Expresso as well as results of analysis with Expresso that suggest the importance of molecular chaperones and membrane transport proteins in mechanisms conferring successful adaptation to long-term drought stress.     

干旱胁迫对华北绣线菊和金山绣线菊光合能力的影响

分别采取轻度、中度、重度干旱胁迫和复水处理,研究华北绣线菊和金山绣线菊的光合能力动态变化;利用二维双向电泳与质谱鉴定等技术,分析鉴定干旱胁迫前后2种绣线菊蛋白质的差异表达,以及引起其光合能力改变的生理机制.结果表明:干旱胁迫处理显著影响了2种绣线菊的光合能力,最大光合速率、光补偿点和光饱和点逐渐下降,其干旱胁迫反应为渐进效应.轻度和中度干旱胁迫后,2种绣线菊的恢复能力较强;而重度干旱胁迫后的恢复能力较弱.经干旱胁迫诱导后,抗旱能力弱的金山绣线菊有6处蛋白点消失、11处新增蛋白点、13处蛋白点上调表达、4处蛋白点下调表达,均为低分子量酸性蛋白;其中由干旱诱导表达的3种差异蛋白分别为放氧增强蛋白因子1、2和1,5-二磷酸核酮糖羧化酶/加氧酶大亚基的降解片断.绣线菊抗旱能力的差异与干旱胁迫期间光合能力的变化有关.