BackgroundHuanglongbing (HLB) is a highly destructive citrus disease which threatens citrus production worldwide and ‘Candidatus Liberibacter asiaticus’ (Las), a non-culturable phloem-limited bacterium, is an associated causal agent of the disease. To better understand the physiological and molecular processes involved in host responses to Las, 2-DE and mass spectrometry analyses, as well as ICP spectroscopy analysis were employed to elucidate the global protein expression profiles and nutrient concentrations in leaves of Las-infected grapefruit plants at pre-symptomatic or symptomatic stages for HLB.ResultsThis study identified 123 protein spots out of 191 spots that showed significant changes in the leaves of grapefruit plants in response to Las infection and all identified spots matched to 69 unique proteins/peptides. A down-regulation of 56 proteins including those associated with photosynthesis, protein synthesis, and metabolism was correlated with significant reductions in the concentrations of Ca, Mg, Fe, Zn, Mn, and Cu in leaves of grapefruit plants in response to Las infection, particularly in symptomatic plants. Oxygen-evolving enhancer (OEE) proteins, a PSI 9 kDa protein, and a Btf3-like protein were among a small group of proteins that were down-regulated in both pre-symptomatic and symptomatic plants in response to Las infection. Furthermore, a Las-mediated up-regulation of 13 grapefruit proteins was detected, which included Cu/Zn superoxide dismutase, chitinases, lectin-related proteins, miraculin-like proteins, peroxiredoxins and a CAP 160 protein. Interestingly, a Las-mediated up-regulation of granule-bound starch synthase was correlated with an increase in the K concentrations of pre-symptomatic and symptomatic plants.ConclusionsThis study constitutes the first attempt to characterize the interrelationships between protein expression and nutritional status of Las-infected pre-symptomatic or symptomatic grapefruit plants and sheds light on the physiological and molecular mechanisms associated with HLB disease development.

Iron is an essential nutrient that plays a role in bacterial differential gene expression and protein production. Accordingly, the comparative analysis of total lysate and outer membrane fractions isolated from A. baumannii ATCC 19606 T cells cultured under iron-rich and -chelated conditions using 2-D gel electrophoresis-mass spectrometry resulted in the identification of 58 protein spots differentially produced. While 19 and 35 of them represent iron-repressed and iron-induced protein spots, respectively, four other spots represent a metal chelation response unrelated to iron. Most of the iron-repressed protein spots represent outer membrane siderophore receptors, some of which could be involved in the utilization of siderophores produced by other bacteria. The ironinduced protein spots represent a wide range of proteins including those involved in iron storage, such as Bfr, metabolic and energy processes, such as AcnA, AcnB, GlyA, SdhA, and SodB, as well as lipid biosynthesis. The detection of an iron-regulated Hfq ortholog indicates that iron regulation in this bacterium could be mediated by Fur and small RNAs as described in other bacteria. The iron-induced production of OmpA suggests this protein plays a role in iron metabolism as shown by the diminished ability of an OmpA isogenic deficient derivative to grow under iron-chelated conditions.

The effect of silicon (Si) nutrition on lowlevel cadmium (Cd) toxicity symptoms was investigated in hydroponically-grown rice seedlings (Oryza sativa L.). Silicon (0.0, 0.2, or 0.6 mM) was added when seedlings were 6 or 20 days old representing early (Si E ) or late (Si L ) Si treatment, respectively. Cadmium (0.0 or 2.5 μM) was added when seedlings were 6 days old. Measurements included generation of CO 2 and light response curves; chlorophyll fluorescence analysis; growth; and tissue-element content analysis. Our results showed that low-level Cd treatment generally inhibited growth and photosynthesis. However, the addition of 0.2 or 0.6 mM Si E or Si L significantly reduced root-and leaf-Cd content. Consequently, the addition of 0.6 mM Si L significantly alleviated lowlevel Cd-induced inhibition of growth. Furthermore, 0.2 mM Si treatment significantly reduced g s compared to 0.0 or 0.6 mM Si without inhibiting A, especially in +Cd plants, suggesting an increase in instantaneous water-use-efficiency (IWUE). Additionally, in +Cd plants, the addition of 0.6 mM Si E significantly reduced F o but increased F v /F m , while treatment with 0.2 mM Si L significantly increased q P , suggesting an increase in light-use-efficiency. We thus, propose that 0.6 mM Si L treatment is required for the alleviation of low-level Cd-mediated growth inhibition. Furthermore, we suggest that 0.2 mM Si concentration might be close to the optimum requirement for maximum Siinduced increase in IWUE in rice plants, especially when under low-level Cd-stress. Our results also suggest that Si alleviates low-level Cd toxicity by improving light-use-efficiency. Keywords Chlorophyll fluorescence . Instantaneous water-use-efficiency . Low-level cadmium . Silicon . Stomatal conductance Abbreviations A net CO 2 assimilation rate A max maximum net CO 2 assimilation rate C a ambient CO 2 concentration C E carboxylation efficiency C i intercellular CO 2 concentration E transpiration rate F m maximum chlorophyll fluorescence yield in a dark-adapted state F o minimum chlorophyll fluorescence yield in a dark-adapted state F o /F m basal quantum yield of non-photochemical processes in PS2 in a dark-adapted state F v maximum variable fluorescence yield in a dark-adapted state F v /F m quantum efficiency of open PS2 centers in a dark-adapted state Plant Soil (2008) 311:73-86

The best known silicon (Si)-accumulating plant, rice (Oryza sativa L.), stores most of its Si in leaves, but the importance of Si has been limited to a mechanical role. Our initial studies showed that Si-induced cadmium (Cd) tolerance is mediated by the enhancement of instantaneous water-use-efficiency, carboxylation efficiency of ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO), and light-use-efficiency in leaves of rice plants. In this study, we investigated changes in the rice leaf proteome in order to identify molecular mechanisms involved in Si-induced Cd tolerance. Our study identified 60 protein spots that were differentially regulated due to Cd and/or Si treatments. Among them, 50 were significantly regulated by Si, including proteins associated with photosynthesis, redox homeostasis, regulation/protein synthesis, pathogen response and chaperone activity. Interestingly, we observed a Si-induced up-regulation of a class III peroxidase and a thaumatin-like protein irrespective of Cd treatment, in addition to a Cd-induced up-regulation of protein disulfide isomerase, a HSP70 homologue, a NADH-ubiquinone oxidoreductase, and a putative phosphogluconate dehydrogenase, especially in the presence of Si. Taken together, our study sheds light on molecular mechanisms involved in Si-induced Cd tolerance in rice leaves and suggests a more active involvement of Si in plant physiological processes than previously proposed.

Citrus huanglongbing (HLB) is a highly destructive disease of citrus presumably caused by ‘ Candidatus Liberibacter asiaticus ’ (Las), a gram-negative, insect-transmitted, phloem-limited α-proteobacterium. Although almost all citrus plants are susceptible to HLB, reports have shown reduced susceptibility to Las infection in lemon ( Citrus limon ) plants. The aim of this study is to identify intra-species specific molecular mechanisms associated with Las-induced responses in lemon plants. To achieve this, comparative 2-DE and mass spectrometry, in addition to Inductively Coupled Plasma Spectroscopy (ICPS) analyses, were applied to investigate differences in protein accumulation and the concentrations of cationic elements in leaves of healthy and Las-infected lemon plants. Results showed a differential accumulation of 27 proteins, including an increase in accumulation of starch synthase but decrease in the production of photosynthesis-related proteins in Las-infected lemon plants compared to healthy plants. Furthermore, there was a 6% increase (P > 0.05) in K concentration in leaves of lemon plants upon Las infection, which support results from previous studies and might represent a common response pattern of citrus plants to Las infection. Interestingly, contrary to reports from prior studies, this study showed a general reduction in the production of defense-related pathogen-response proteins but a 128% increase in Zn concentration in lemon plants in response to Las infection. Taken together, this study sheds light on general and intra-species specific responses associated with the response of citrus plants to Las.