Analysis of Subcellular Metabolite Distributions Within Arabidopsis thaliana Leaf Tissue: A Primer for Subcellular Metabolomics

Author(s):  
Stephan Krueger ◽  
Dirk Steinhauser ◽  
Jan Lisec ◽  
Patrick Giavalisco
2005 ◽  
Vol 85 (1) ◽  
pp. 225-235 ◽  
Author(s):  
Rebecca H. Hallett ◽  
Heather Ray ◽  
Jennifer Holowachuk ◽  
Juliana J. Soroka ◽  
Margaret Y. Gruber

A bioassay arena and a laboratory screening protocol were developed for assessing lines of Arabidopsis thaliana L. (Heynh.) for feeding damage by the adult crucifer flea beetle, Phyllotreta cruciferae (Goeze). The arena consists of a 96-well microtitre plate with a modified top to contain flea beetles and allow ventilation. Eight lines of A. thaliana, arranged in an 8 × 8 Latin square design, were screened simultaneously in each arena using 50 starved flea beetles. Two cotyledons and the first pair of true leaves per plant were rated visually under a dissecting microscope using a visual damage rating scale. The protocol was used to screen 29 wild ecotypes, eight mutant lines and a single transgenic line of A. thaliana. Discrimination between both cotyledon and leaf tissue was apparent for young beetles that were both non-reproductive or reproductive, but not for old reproductive beetles. Differences were observed between Asian and European ecotypes of A. thaliana, suggesting that geographic origin may play a role in susceptibility of Arabidopsis ecotypes to flea beetle feeding. The transparent testa regulatory gene mutants (lines 82, 111, 164) were most susceptible to flea beetle feeding, possibly indicating a role for anthocyanins and/or flavonoids in governing flea beetle susceptibility. Significant variation in damage levels indicates that expression of flea beetle resistance in the A rabidopsis genome is plastic, and that potential exists to use the wide array of publicly available Arabidopsis germplasm as tools in the transfer of resistance to agronomically important host plants. Key words: Seedling bioassay, Arabidopsis thaliana, wild ecotypes and mutants, crucifer flea beetle, Phyllotreta cruciferae, host plant resistance


2001 ◽  
Vol 14 (2) ◽  
pp. 234-241 ◽  
Author(s):  
Wenqi Hu ◽  
Jing Yuan ◽  
Qiao-Ling Jin ◽  
Patrick Hart ◽  
Sheng Yang He

Hypersensitive reaction and pathogenicity (hrp) genes are required for Pseudomonas syringae pv. tomato (Pst) DC3000 to cause disease in susceptible tomato and Arabidopsis thaliana plants and to elicit the hypersensitive response in resistant plants. The hrp genes encode a type III protein secretion system known as the Hrp system, which in Pst DC3000 secretes HrpA, HrpZ, HrpW, and AvrPto and assembles a surface appendage, named the Hrp pilus, in hrp-gene-inducing minimal medium. HrpA has been suggested to be the Hrp pilus structural protein on the basis of copurification and mutational analyses. In this study, we show that an antibody against HrpA efficiently labeled Hrp pili, whereas antibodies against HrpW and HrpZ did not. Immunogold labeling of bacteria-infected Arabidopsis thaliana leaf tissue with an Hrp pilus antibody revealed a characteristic lineup of gold particles around bacteria and/or at the bacterium-plant contact site. These results confirm that HrpA is the major structural protein of the Hrp pilus and provide evidence that Hrp pili are assembled in vitro and in planta.


Genetics ◽  
1995 ◽  
Vol 140 (2) ◽  
pp. 725-732 ◽  
Author(s):  
M E Jenkins ◽  
G R Harlow ◽  
Z Liu ◽  
M A Shotwell ◽  
J Ma ◽  
...  

Abstract Five Arabidopsis mutants have been isolated on the basis of hypersensitivity of leaf tissue to UV light. For each mutant, the UV-hypersensitive phenotype (uvh) was inherited as a single recessive Mendelian trait. In addition, each uvh mutant represented a separate complementation group. Three of the mutations producing the UV hypersensitive phenotype have been mapped relative to either genetic markers or physical microsatellite polymorphisms. Locus UVH1 is linked to nga76 on chromosome 5, UVH3 to GL1 on chromosome three, and UVH6 to nga59 on chromosome 1. Each uvh mutant has a characteristic pattern of sensitivity based on UV sensitivity of leaf tissue, UV sensitivity of root tissue, and ionizing radiation sensitivity of seeds. On the basis of these patterns, possible molecular defects in these mutants are discussed.


2021 ◽  
Author(s):  
Anna van Weringh ◽  
Asher Pasha ◽  
Eddi Esteban ◽  
Paul J. Gamueda ◽  
Nicholas J. Provart

Drought is an important environmental stress that limits crop production. Guard cells (GC) act to control the rate of water loss. To better understand how GCs change their gene expression during a progressive drought we generated guard cell-specific RNA-seq transcriptomes during mild, moderate, and severe drought stress. We additionally sampled re-watered plants that had experienced severe drought stress. These transcriptomes were generated using the INTACT system to capture the RNA from GC nuclei. We optimized the INTACT protocol for Arabidopsis thaliana leaf tissue, incorporating fixation to preserve RNA during nuclear isolation. To be able to identify gene expression changes unique to GCs, we additionally generated transcriptomes from all cell types, using a 35S viral promoter to capture the nuclei of all cell types in leaves. These data sets highlight shared and unique gene expression changes between GCs and the bulk leaf tissue. The timing of gene expression changes is different between GCs and other cell types: we found that only GCs had detectable gene expression changes at the earliest drought time point. The drought responsive GC and leaf RNA-seq transcriptomes are available in the Arabidopsis ePlant at the Bio-Analytic Resource for Plant Biology website.


2019 ◽  
Vol 20 (22) ◽  
pp. 5637 ◽  
Author(s):  
Lifang Zou ◽  
Bingwei Yu ◽  
Xing-Liang Ma ◽  
Bihao Cao ◽  
Guoju Chen ◽  
...  

Chinese kale (Brassica oleracea var. chinensis Lei) is an important vegetable crop in South China, valued for its nutritional content and taste. Nonetheless, the thermal tolerance of Chinese kale still needs improvement. Molecular characterization of Chinese kale’s heat stress response could provide a timely solution for developing a thermally tolerant Chinese kale variety. Here, we report the cloning of multi-protein bridging factor (MBF) 1c from Chinese kale (BocMBF1c), an ortholog to the key heat stress responsive gene MBF1c. Phylogenetic analysis showed that BocMBF1c is highly similar to the stress-response transcriptional coactivator MBF1c from Arabidopsis thaliana (AtMBF1c), and the BocMBF1c coding region conserves MBF1 and helix-turn-helix (HTH) domains. Moreover, the promoter region of BocMBF1c contains three heat shock elements (HSEs) and, thus, is highly responsive to heat treatment. This was verified in Nicotiana benthamiana leaf tissue using a green fluorescent protein (GFP) reporter. In addition, the expression of BocMBF1c can be induced by various abiotic stresses in Chinese kale which indicates the involvement of stress responses. The BocMBF1c-eGFP (enhanced green fluorescent protein) chimeric protein quickly translocated into the nucleus under high temperature treatment in Nicotiana benthamiana leaf tissue. Overexpression of BocMBF1c in Arabidopsis thaliana results in a larger size and enhanced thermal tolerance compared with the wild type. Our results provide valuable insight for the role of BocMBF1c during heat stress in Chinese kale.


1998 ◽  
Vol 11 (7) ◽  
pp. 706-709 ◽  
Author(s):  
Robert T. Lartey ◽  
Soumitra Ghoshroy ◽  
Vitaly Citovsky

Following inoculation, many plant viruses spread locally from cell to cell until they reach the vascular system, through which they then move to other parts of the plant, resulting in systemic infection. To isolate host genes involved in systemic transport of plant viruses, ethyl methanesulfonate-mutagenized Arabidopsis thaliana plants were screened for significant delays in the systemic movement of turnip vein clearing virus (TCVC). One such mutant, designated vsm1 (virus systemic movement), was identified. Unlike the wild-type plants, vsm1 did not develop viral disease and did not allow the systemic spread of the virus. The local viral movement within the inoculated vsm1 leaves, however, was not affected. TVCV systemic movement within the vsm1 plants was likely blocked at the step of viral entry into the host plant vasculature from the infected leaf tissue. vsm1 plants also restricted the systemic movement of another tobamovirus but not of an unrelated carmovirus.


2010 ◽  
pp. no-no ◽  
Author(s):  
DANA ZEIDLER ◽  
IAN A. DUBERY ◽  
PHILLIPPE SCHMITT-KOPPLIN ◽  
UTA VON RAD ◽  
JÖRG DURNER

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