scholarly journals The regulation and catalytic mechanism of the NADP-malic enzyme from tobacco leaves

2009 ◽  
Vol 74 (8-9) ◽  
pp. 893-906 ◽  
Author(s):  
Veronika Doubnerová ◽  
Lucie Potůcková ◽  
Karel Müller ◽  
Helena Ryslavá

The non-photosynthetic NADP-malic enzyme EC 1.1.1.40 (NADP-ME), which catalyzes the oxidative decarboxylation of L-malate and NADP+ to produce pyruvate and NADPH, respectively, and which could be involved in plant defense responses, was isolated from Nicotiana tabacum L. leaves. The mechanism of the enzyme reaction was studied by the initial rate method and was found to be an ordered sequential one. Regulation possibilities of purified cytosolic NADP-ME by cell metabolites were tested. Intermediates of the citric acid cycle (?-ketoglutarate, succinate, fumarate), metabolites of glycolysis (pyruvate, phosphoenolpyruvate, glucose-6-phosphate), compounds connected with lipogenesis (coenzyme A, acetyl-CoA, palmitoyl-CoA) and some amino acids (glutamate, glutamine, aspartate) did not significantly affect the NADP-ME activity from tobacco leaves. In contrast, macroergic compounds (GTP, ATP and ADP) were strong inhibitors of NADP-ME; the type of inhibition and the inhibition constants were determined in the presence of the most effective cofactors (Mn2+ or Mg2+), required by NADP-ME. Predominantly non-competitive type of inhibitions of NADP-ME with respect to NADP+ and mixed type to L-malate were found.

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mari Kurokawa ◽  
Masataka Nakano ◽  
Nobutaka Kitahata ◽  
Kazuyuki Kuchitsu ◽  
Toshiki Furuya

AbstractMicroorganisms that activate plant immune responses have attracted considerable attention as potential biocontrol agents in agriculture because they could reduce agrochemical use. However, conventional methods to screen for such microorganisms using whole plants and pathogens are generally laborious and time consuming. Here, we describe a general strategy using cultured plant cells to identify microorganisms that activate plant defense responses based on plant–microbe interactions. Microbial cells were incubated with tobacco BY-2 cells, followed by treatment with cryptogein, a proteinaceous elicitor of tobacco immune responses secreted by an oomycete. Cryptogein-induced production of reactive oxygen species (ROS) in BY-2 cells served as a marker to evaluate the potential of microorganisms to activate plant defense responses. Twenty-nine bacterial strains isolated from the interior of Brassica rapa var. perviridis plants were screened, and 8 strains that enhanced cryptogein-induced ROS production in BY-2 cells were selected. Following application of these strains to the root tip of Arabidopsis seedlings, two strains, Delftia sp. BR1R-2 and Arthrobacter sp. BR2S-6, were found to induce whole-plant resistance to bacterial pathogens (Pseudomonas syringae pv. tomato DC3000 and Pectobacterium carotovora subsp. carotovora NBRC 14082). Pathogen-induced expression of plant defense-related genes (PR-1, PR-5, and PDF1.2) was enhanced by the pretreatment with strain BR1R-2. This cell–cell interaction-based platform is readily applicable to large-scale screening for microorganisms that enhance plant defense responses under various environmental conditions.


2021 ◽  
Vol 22 (8) ◽  
pp. 4214
Author(s):  
Gautam Anand ◽  
Meirav Leibman-Markus ◽  
Dorin Elkabetz ◽  
Maya Bar

Plants lack a circulating adaptive immune system to protect themselves against pathogens. Therefore, they have evolved an innate immune system based upon complicated and efficient defense mechanisms, either constitutive or inducible. Plant defense responses are triggered by elicitors such as microbe-associated molecular patterns (MAMPs). These components are recognized by pattern recognition receptors (PRRs) which include plant cell surface receptors. Upon recognition, PRRs trigger pattern-triggered immunity (PTI). Ethylene Inducing Xylanase (EIX) is a fungal MAMP protein from the plant-growth-promoting fungi (PGPF)–Trichoderma. It elicits plant defense responses in tobacco (Nicotiana tabacum) and tomato (Solanum lycopersicum), making it an excellent tool in the studies of plant immunity. Xylanases such as EIX are hydrolytic enzymes that act on xylan in hemicellulose. There are two types of xylanases: the endo-1, 4-β-xylanases that hydrolyze within the xylan structure, and the β-d-xylosidases that hydrolyze the ends of the xylan chain. Xylanases are mainly synthesized by fungi and bacteria. Filamentous fungi produce xylanases in high amounts and secrete them in liquid cultures, making them an ideal system for xylanase purification. Here, we describe a method for cost- and yield-effective xylanase production from Trichoderma using wheat bran as a growth substrate. Xylanase produced by this method possessed xylanase activity and immunogenic activity, effectively inducing a hypersensitive response, ethylene biosynthesis, and ROS burst.


2004 ◽  
Vol 85 (1) ◽  
pp. 33-37 ◽  
Author(s):  
Magali Merkx-Jacques ◽  
Jacqueline C. Bede

Abstract Plants exhibit remarkable plasticity in their ability to differentiate between herbivorous insect species and subtly adjust their defense responses to target distinct pests. One key mechanism used by plants to recognize herbivorous caterpillars is elicitors present in their oral secretions; however, these elicitors not only cause the induction of plant defenses but recent evidence suggests that they may also suppress plant responses. The absence of “expected changes” in induced defense responses of insect-infested plants has been attributed to hydrogen peroxide produced by caterpillar salivary glucose oxidase (GOX). Activity of this enzyme is variable among caterpillar species; it was detected in two generalist caterpillars, the beet armyworm (Spodoptera exigua) and the bertha armyworm (Mamestra configurata), but not in other generalist or specialist caterpillar species tested. In the beet armyworm, GOX activity fluctuated over larval development with high activity associated with the salivary glands of fourth instars. Larval salivary GOX activity of the beet armyworm and the bertha armyworm was observed to be significantly higher in caterpillars reared on artificial diet as compared with those reared on Medicago truncatula plants. This implies that a factor in the diet is involved in the regulation of caterpillar salivary enzyme activity. Therefore, plant diet may be regulating caterpillar oral elicitors that are involved in the regulation of plant defense responses: our goal is to understand these two processes.


1995 ◽  
Vol 92 (10) ◽  
pp. 4134-4137 ◽  
Author(s):  
Z. Chen ◽  
J. Malamy ◽  
J. Henning ◽  
U. Conrath ◽  
P. Sanchez-Casas ◽  
...  

1998 ◽  
Vol 39 (11) ◽  
pp. 1245-1249 ◽  
Author(s):  
A. Kiba ◽  
M. Sugimoto ◽  
K. Toyoda ◽  
Y. Ichinose ◽  
T. Yamada ◽  
...  

1990 ◽  
Vol 45 (6) ◽  
pp. 569-575 ◽  
Author(s):  
Dierk Scheel ◽  
Jane E. Parker

Abstract Plants defend themselves against pathogen attack by activating a whole set of defense responses, most of them relying on transcriptional activation of plant defense genes. The same responses are induced by treatment of plant cells with elicitors released from the pathogen or from the plant surface. Several plant/elicitor combinations have been used successfully as experimental systems to investigate the molecular basis of plant defense responses. Receptor-like structures on the plasma membrane of plant cells appear to bind the elicitors. Thereby, intracellular signal transduction chains are initiated which finally result in the activation of plant defense genes. A better understanding of the molecular mechanisms of early processes in plant defense responses, as provided by these studies, may in the long term help to develop environmentally safe plant protection methods for agriculture.


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