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Seerangan K.,Bharathiar University | Seerangan K.,Institute of Plant and Microbial Biology | Thangavelu M.,Bharathiar University
Journal of Botany | Year: 2014

Investigations on the prevalence of arbuscular mycorrhizal (AM) and dark septate endophyte (DSE) fungal symbioses are limited for plants growing in tropical aquatic and wetland habitats compared to those growing on terrestrial moist or dry habitats. Therefore, we assessed the incidence of AM and DSE symbiosis in 8 hydrophytes and 50 wetland plants from four sites in south India. Of the 58 plant species examined, we found AM and DSE fungal symbiosis in 21 and five species, respectively. We reported for the first time AM and DSE fungal symbiosis in seven and five species, respectively. Intermediate-type AM morphology was common, and AM morphology is reported for the first time in 16 plant species. Both AM and DSE fungal colonization varied significantly across plant species and sites. Intact and identifiable AM fungal spores occurred in root zones of nine plant species, but AM fungal species richness was low. Though no clear relationship between AM and DSE fungal colonization was recognized, a significant negative correlation between AM colonization and spore numbers was established. Our study suggests that the occurrence of AM and DSE fungal symbiosis in plants growing in hydrophytic and wetland habitats is not as common as in terrestrial habitats. © 2014 Kumar Seerangan and Muthukumar Thangavelu. Source

Lovell J.T.,University of Texas at Austin | Lovell J.T.,Colorado State University | Mullen J.L.,Colorado State University | Lowry D.B.,Michigan State University | And 6 more authors.
Plant Cell | Year: 2015

Soil water availability represents one of the most important selective agents for plants in nature and the single greatest abiotic determinant of agricultural productivity, yet the genetic bases of drought acclimation responses remain poorly understood. Here, we developed a systems-genetic approach to characterize quantitative trait loci (QTLs), physiological traits and genes that affect responses to soil moisture deficit in the TSUxKAS mapping population of Arabidopsis thaliana. To determine the effects of candidate genes underlying QTLs, we analyzed gene expression as a covariate within the QTL model in an effort to mechanistically link markers, RNA expression, and the phenotype. This strategy produced ranked lists of candidate genes for several drought-associated traits, including water use efficiency, growth, abscisic acid concentration (ABA), and proline concentration. As a proof of concept, we recovered known causal loci for several QTLs. For other traits, including ABA, we identified novel loci not previously associated with drought. Furthermore, we documented natural variation at two key steps in proline metabolism and demonstrated that the mitochondrial genome differentially affects genomic QTLs to influence proline accumulation. These findings demonstrate that linking genome, transcriptome, and phenotype data holds great promise to extend the utility of genetic mapping, even when QTL effects are modest or complex. © 2015 American Society of Plant Biologists. All rights reserved. Source

Lin L.-C.,Academia Sinica, Taiwan | Chueh C.-M.,Academia Sinica, Taiwan | Wang L.-C.,Institute of Plant and Microbial Biology
Methods in Molecular Biology | Year: 2014

Conventional mutant screening in forward genetics research is indispensible to understand the biological operation behind any given phenotype. However, several issues, such as functional redundancy and lethality or sterility resulting from null mutations, frequently impede the functional characterization of genetic mutants. As an alternative approach, chemical screening with natural products or synthetic small molecules that act as conditional mutagens allows for identifying bioactive compounds as bioprobes to overcome the above-mentioned issues. Ethylene is the simplest olefin and is one of the major phytohormones playing crucial roles in plant physiology. Most of the current information on how ethylene works in plants came primarily from genetic studies of ethylene mutants identified by conventional genetic screening two decades ago. However, we lack a complete picture of functional interaction among components in the ethylene pathway and cross talk of ethylene with other phytohormones. Here, we describe our methodology for using chemical genetics to identify small molecules that interfere with the ethylene response. We set up a phenotype-based screening platform and a reporter gene-based system for verification of the hit compounds identified by chemical screening. We have successfully identified small molecules affecting the ethylene phenotype in etiolated seedlings and showed that a group of structurally similar compounds are novel inhibitors of ACC synthase, a rate-limiting enzyme in the ethylene biosynthesis pathway. © Springer Science+Business Media New York 2014. Source

Chen P.T.,Biotechnology Center | Shaw J.-F.,National Chung Hsing University | Chao Y.-P.,Feng Chia University | David Ho T.-H.,Institute of Plant and Microbial Biology | And 2 more authors.
Journal of Agricultural and Food Chemistry | Year: 2010

Bacillus subtilis is most commonly employed for secretion of recombinant proteins. To circumvent the problems caused by using plasmids, the T7 expression system known for its high efficiency was rebuilt in B. subtilis. Accordingly, a markerless and replicon-free method was developed for genomic insertion of DNAs. By the act of homologous recombination via the guide DNA, a suicidal vector carrying the gene of interest was integrated into genomic loci of bacteria. Removal of the inserted selection marker and replicon flanked by FRT sites was mediated by the FLP recombinase. By using the mentioned system, B. subtilis strain PT5 was constructed to harbor a genomic copy of the spac promoter-regulated T7 gene 1 located at wprA (encoding the cell wall-associated protease). Similarly, the T7 promoter-driven nattokinase or endoglucanase E1 of Thermomonospora fusca genes were also integrated into mpr (encoding an extracellular protease) of strain PT5. Consequently, the integrant PT5/Mmp-T7N or PT5/MT1-E1 resulted in a "clean" producer strain deprived of six proteases. After 24 h, the strain receiving induction was able to secret nattokinase and endoglucanase E1 with the volumetric activity reaching 10860 CU/mL and 8.4 U/mL, respectively. This result clearly indicates the great promise of the proposed approach for high secretion of recombinant proteins in B. subtilis. © 2010 American Chemical Society. Source

Chou Y.-L.,I - Shou University | Ko C.-Y.,Institute of Plant and Microbial Biology | Yen C.-C.,National Chung Hsing University | Chen L.-F.O.,Institute of Plant and Microbial Biology | And 2 more authors.
Journal of Agricultural and Food Chemistry | Year: 2015

Natural chlorophyll metabolites have exhibited physiological activity in vitro. In this study, a recombinant chlorophyllase1 gene from Chlamydomonas reinhardtii (CrCLH1) was isolated and characterized. Recombinant CrCLH1 can perform chlorophyll dephytylation and produce chlorophyllide and phytol. In a transient assay, the subcellular localization of CrCLH1-green fluorescent protein was determined to be outside the chloroplast. Biochemical analyses of the activity of recombinant CrCLH1 indicated that its optimal pH value and temperature are 6.0 and 40°C, respectively. Enzyme kinetic data revealed that the recombinant CrCLH1 had a higher catalytic efficiency for chlorophyll a than for chlorophyll b and bacteriochlorophyll a. According to high-performance liquid chromatography analysis of chlorophyll hydrolysis, recombinant CrCLH1 catalyzed the conversion of chlorophyll a to pheophorbide a at pH 5. Therefore, recombinant CrCLH1 can be used as a biocatalyst to produce chlorophyllide derivatives. © 2015 American Chemical Society. Source

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