Functional characterization of an unobtrusive protein, CkMT4, in re-establishing desiccation tolerance in germinating seeds.

Desiccation tolerance (DT) is gradually lost during seed germination, while it can be re-established by pre-treatment with polyethylene glycol (PEG) and/or abscisic acid (ABA). Increasing knowledge is available on several stress-related proteins in DT re-establishment in herb seeds, but limited information exists on novel proteins in wood seeds. This study aimed to investigate the role of metallothionein CkMT4, a protein species with the highest fold increase in abundance in Caragana korshinskii seeds on PEG treatment. The fluctuation in mRNA levels of CkMT4 during seed development was consistent with the changes in DT, and the expression of CkMT4 could be up-regulated by ABA. Besides metal-binding capacity, CkMT4 might supply Cu2+/Zn2+ to superoxide dismutase (SOD) under high redox potential provided by PEG treatment for excess reactive oxygen species (ROS) scavenging. The overexpression of CkMT4 in yeast results in enhanced oxidation resistance. Experimentally, this study demonstrated the overexpression of CkMT4 in Arabidopsis seeds benefited the re-establishment of DT and enhanced the activity of SOD. On the whole, these findings suggested that CkMT4 facilitated the re-establishment of DT in C. korshinskii seeds mainly through diminishing excess ROS, which put the mechanism underlying the re-establishment of DT in xerophytic wood seeds into a new perspective.

Comprehensive transcriptome profiling of Caragana microphylla in response to salt condition using de novo assembly.

OBJECTIVE: To investigate the response of Caragana microphylla in salt condition, transcriptome analysis, differentially expressed genes (DEGs) comparison with Arabidopsis thaliana, and the chlorophyll content analysis were performed. RESULTS: Gene Ontology (GO) term, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses of DEGs indicated that salt condition affected photosynthesis and chlorophyll in C. microphylla. The DEGs compared with salt responsive genes of A. thaliana indicated that C. microphylla's responses to salt differed greatly from those of the model plant and that the results also indicated up-regulated genes related to photosynthesis and chlorophyll in C. microphylla. Moreover, we confirmed that salt-treated C. microphylla increased chlorophyll content, and the genes of protoporphyrin IX downstream in chlorophyll biosynthesis were induced in the heatmap analysis. CONCLUSIONS: These results showed a similar pattern to some halophytes plants with increased chlorophyll at a certain salt concentration, and we assumed that C. microphylla also has a mechanism to adapt or tolerate moderate salt conditions.

Two NAC transcription factors from Caragana intermedia altered salt tolerance of the transgenic Arabidopsis.

BACKGROUND: Plants are continuously challenged by different environment stresses, and they vary widely in their adjustability. NAC (NAM, ATAF and CUC) transcription factors are known to be crucial in plants tolerance response to abiotic stresses, such as drought and salinity. ANAC019, ANAC055, and ANAC072, belong to the stress-NAC TFs, confer the Arabidopsis abiotic stress tolerance. RESULTS: Here we isolated two stress-responsive NACs, CiNAC3 and CiNAC4, from Caragana intermedia, which were induced by ABA and various abiotic stresses. Localization assays revealed that CiNAC3 and CiNAC4 localized in the nuclei, consistent with their roles as transcription factors. Histochemistry assay using Pro(CiNAC4)::GUS transgenic Arabidopsis showed that the expression of the GUS reporter was observed in many tissues of the transgenic plants, especially in the root vascular system. Overexpression of CiNAC3 and CiNAC4 reduced ABA sensitivity during seed germination, and enhanced salt tolerance of the transgenic Arabidopsis. CONCLUSIONS: We characterised CiNAC3 and CiNAC4 and found that they were induced by numerous abiotic stresses and ABA. GUS histochemical assay of CiNAC4 promoter suggested that root, flower and local damaged tissues were the strongest stained tissues. Overexpression assay revealed that CiNAC4 play essential roles not only in promoting lateral roots formation, but also in responding to salinity and ABA treatment of Arabidopsis.

Effects of elevated CO2, warming and precipitation change on plant growth, photosynthesis and peroxidation in dominant species from North China grassland.

Warming, watering and elevated atmospheric CO2-concentration effects have been extensively studied separately; however, their combined impact on plants is not well understood. In the current research, we examined plant growth and physiological responses of three dominant species from the Eurasian Steppe with different functional traits to a combination of elevated CO2, high temperature, and four simulated precipitation patterns. Elevated CO2 stimulated plant growth by 10.8-41.7 % for a C3 leguminous shrub, Caragana microphylla, and by 33.2-52.3 % for a C3 grass, Stipa grandis, across all temperature and watering treatments. Elevated CO2, however, did not affect plant biomass of a C4 grass, Cleistogenes squarrosa, under normal or increased precipitation, whereas a 20.0-69.7 % stimulation of growth occurred with elevated CO2 under drought conditions. Plant growth was enhanced in the C3 shrub and the C4 grass by warming under normal precipitation, but declined drastically with severe drought. The effects of elevated CO2 on leaf traits, biomass allocation and photosynthetic potential were remarkably species-dependent. Suppression of photosynthetic activity, and enhancement of cell peroxidation by a combination of warming and severe drought, were partly alleviated by elevated CO2. The relationships between plant functional traits and physiological activities and their responses to climate change were discussed. The present results suggested that the response to CO2 enrichment may strongly depend on the response of specific species under varying patterns of precipitation, with or without warming, highlighting that individual species and multifactor dependencies must be considered in a projection of terrestrial ecosystem response to climatic change.

Effects of exogenous GABA on gene expression of Caragana intermedia roots under NaCl stress: regulatory roles for H2O2 and ethylene production.

gamma-aminobutyric acid (GABA) is a four-carbon non-protein amino acid presented in a wide range of organisms. In this study, a suppression subtractive hybridization (SSH) library was constructed using roots of a legume shrub, Caragana intermedia, with the combined treatment of 300 mm NaCl and 300 mm NaCl + 10 mm GABA. We obtained 224 GABA-regulated unique expressed sequence tags (ESTs) including signal transduction, transcriptional regulation, hormone biosynthesis, reactive oxygen species (ROS) and polyamine metabolism, etc. The key H(2)O(2)-generated genes, NADPH oxidase (CaGR60), peroxidase (CaGR61) and amine oxidase (CaGR62), were regulated at the mRNA level by 10 mm GABA, which clearly inhibited H(2)O(2) accumulation brought about by NaCl stress in roots and leaves with the observation of 3,3'-diaminobenzidine (DAB) staining. Similarly, 10 mm GABA also regulated the expression of 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase (ACO) genes (CaGR30 and CaGR31) and ethylene production in NaCl-treated roots. Surprisingly, these H(2)O(2)-generated genes were enhanced at the mRNA level by a lower concentration of GABA, at 0.25 mm, but not other alternative nitrogen sources, and endogenous GABA accumulated largely just by the application of GABA at either concentration. Our results further proved that GABA, as a signal molecule, participates in regulating the expression of genes in plants under salt stress.