There is remarkable conservation in the recognition of pathogen-associated molecular patterns (PAMPs) by innate immune responses of plants, insects and mammals. We developed an Arabidopsis thaliana leaf cell system based on the induction of early-defence gene transcription by flagellin, a highly conserved component of bacterial flagella that functions as a PAMP in plants and mammals. Here we identify a complete plant MAP kinase cascade (MEKK1, MKK4/MKK5 and MPK3/MPK6) and WRKY22/WRKY29 transcription factors that function downstream of the flagellin receptor FLS2, a leucine-rich-repeat (LRR) receptor kinase. Activation of this MAPK cascade confers resistance to both bacterial and fungal pathogens, suggesting that signalling events initiated by diverse pathogens converge into a conserved MAPK cascade.
Carotenoids are lipophilic isoprenoid compounds synthesized by all photosynthetic organisms and some non-photosynthetic prokaryotes and fungi. With some notable exceptions, animals (including humans) do not produce carotenoids de novo but take them in their diets. In photosynthetic systems carotenoids are essential for photoprotection against excess light and contribute to light harvesting, but perhaps they are best known for their properties as natural pigments in the yellow to red range. Carotenoids can be associated to fatty acids, sugars, proteins, or other compounds that can change their physical and chemical properties and influence their biological roles. Furthermore, oxidative cleavage of carotenoids produces smaller molecules such as apocarotenoids, some of which are important pigments and volatile (aroma) compounds. Enzymatic breakage of carotenoids can also produce biologically active molecules in both plants (hormones, retrograde signals) and animals (retinoids). Both carotenoids and their enzymatic cleavage products are associated with other processes positively impacting human health. Carotenoids are widely used in the industry as food ingredients, feed additives, and supplements. This review, contributed by scientists of complementary disciplines related to carotenoid research, covers recent advances and provides a perspective on future directions on the subjects of carotenoid metabolism, biotechnology, and nutritional and health benefits.
Flagellin, the main protein of the bacterial flagella, elicits defence responses and alters growth in Arabidopsis seedlings. Previously, we identified the FLS1 locus, which confers flagellin insensitivity in Ws-0. To identify additional components involved in flagellin perception, we screened for flagellin insensitivity mutants in the flagellin-sensitive accession La-er. Here, we describe the identification of a new locus, FLS2, by a map-based strategy. The FLS2 gene is ubiquitously expressed and encodes a putative receptor kinase. FLS2 shares structural and functional homologies with known plant resistance genes and with components involved in the innate immune system of mammals and insects.
Crocus sativus stigmas are the source of the saffron spice and accumulate the apocarotenoids crocetin, crocins, picrocrocin, and safranal, responsible for its color, taste, and aroma. Through deep transcriptome sequencing, we identified a novel dioxygenase, carotenoid cleavage dioxygenase 2 (CCD2), expressed early during stigma development and closely related to, but distinct from, the CCD1 dioxygenase family. CCD2 is the only identified member of a novel CCD clade, presents the structural features of a bona fide CCD, and is able to cleave zeaxanthin, the presumed precursor of saffron apocarotenoids, both in Escherichia coli and in maize endosperm. The cleavage products, identified through high-resolution mass spectrometry and comigration with authentic standards, are crocetin dialdehyde and crocetin, respectively. In vitro assays show that CCD2 cleaves sequentially the 7,8 and 7′,8′ double bonds adjacent to a 3-OH-β-ionone ring and that the conversion of zeaxanthin to crocetin dialdehyde proceeds via the C 30 intermediate 3-OH-β-apo-8′-carotenal. In contrast, zeaxanthin cleavage dioxygenase (ZCD), an enzyme previously claimed to mediate crocetin formation, did not cleave zeaxanthin or 3-OH-β-apo-8′-carotenal in the test systems used. Sequence comparison and structure prediction suggest that ZCD is an N-truncated CCD4 form, lacking one blade of the β-propeller structure conserved in all CCDs. These results constitute strong evidence that CCD2 catalyzes the first dedicated step in crocin biosynthesis. Similar to CCD1, CCD2 has a cytoplasmic localization, suggesting that it may cleave carotenoids localized in the chromoplast outer envelope.β-citraurin | symmetric carotenoid cleavage
Saffron, the processed stigma of Crocus sativus, is characterized by the presence of several apocarotenoids that contribute to the color, flavor, and aroma of the spice. However, little is known about the synthesis of aroma compounds during the development of the C. sativus stigma. The developing stigma is nearly odorless, but before and at anthesis, the aromatic compound -ionone becomes the principal norisoprenoid volatile in the stigma. In this study, four carotenoid cleavage dioxygenase (CCD) genes, CsCCD1a, CsCCD1b, CsCCD4a, and CsCCD4b, were isolated from C. sativus. Expression analysis showed that CsCCD1a was constitutively expressed, CsCCD1b was unique to the stigma tissue, but only CsCCD4a and -b had expression patterns consistent with the highest levels of -carotene and emission of -ionone derived during the stigma development. The CsCCD4 enzymes were localized in plastids and more specifically were present in the plastoglobules. The enzymatic activities of CsCCD1a, CsCCD1b, and CsCCD4 enzymes were determined by Escherichia coli expression, and subsequent analysis of the volatile products was generated by GC/MS. The four CCDs fell in two phylogenetically divergent dioxygenase classes, but all could cleave -carotene at the 9,10(9 ,10 ) positions to yield -ionone. The data obtained suggest that all four C. sativus CCD enzymes may contribute in different ways to the production of -ionone. In addition, the location and precise timing of -ionone synthesis, together with its known activity as a fragrance and insect attractant, suggest that this volatile may have a role in Crocus pollination.Crocus sativus is a triploid sterile plant characterized by its long red stigmas. These stigmas, when desiccated, constitute the spice known as saffron. Saffron is considered a complex mixture of volatile and non-volatile compounds that contribute to its overall aroma and flavor. The main components of saffron responsible for coloring strength are the derived carotenoids cis-and trans-crocins, picrocrocin, and its degradation product, the odor-active safranal that composes up to 70% of total volatiles (1). In addition to safranal, 150 volatile compounds are estimated to be present in the spice, and ϳ60 constituents have been identified (2). The structures of several of these reveal an isoprenoid-based origin like safranal, and are assumed to be the products of the oxidative cleavage of carotenoids.In recent years, a family of enzymes that cleave carotenoid substrates at different double bond positions have been described in plants. This family, the carotenoid cleavage dioxygenases (CCDs), 2 are specific for the location of the double bond in the molecule they cleave, but many are promiscuous in their carotenoid substrate choice (3, 4). The founding member of the CCD family was Vp14, a 9-cis-epoxycarotenoid dioxygenase (NCED). This and other closely related dioxygenases catalyze the cleavage at the 11,12 double bond of both 9-cis-violaxanthin and 9-cis-neoxanthin to produce xanthoxin (5-7), which is subsequently conv...
In Arabidopsis, activation of defense responses by flagellin is triggered by the specific recognition of the most conserved domain of flagellin, represented by the peptide flg22, in a process involving the FLS2 gene, which encodes a leucine-rich repeat serine/threonine protein kinase. We show here that the two fls2 mutant alleles, fls2-24 and fls2-17 , which were shown previously to confer insensitivity to flg22, also cause impaired flagellin binding. These features are rescued when a functional FLS2 gene is expressed as a transgene in each of the fls2 mutant plants, indicating that FLS2 is necessary for flagellin binding. The point mutation of the fls2-17 allele lies in the kinase domain. A kinase carrying this missense mutation lacked autophosphorylation activity when expressed in Escherichia coli . This indicates that kinase activity is required for binding and probably affects the stability of the flagellin receptor complex. We further show that overexpression of the kinase-associated protein phosphatase (KAPP) in Arabidopsis results in plants that are insensitive to flagellin treatment, and we show reduced flg22 binding in these plants. Furthermore, using the yeast two-hybrid system, we show physical interaction of KAPP with the kinase domain of FLS2. These results suggest that KAPP functions as a negative regulator of the FLS2 signal transduction pathway and that the phosphorylation of FLS2 is necessary for proper binding and signaling of the flagellin receptor complex. INTRODUCTIONMany biological recognition phenomena involve ligands and receptors. For example, the critical steps of pathogen recognition in plants appear to involve interactions of plant receptors with microbial ligands, the so-called elicitors, and the induction of a phosphorylation cascade that leads to defense responses (Boller, 1995;Yang et al., 1997). One of the most active bacterially derived elicitor molecules in plant cells is flagellin (Felix et al., 1999), which induces rapid defense responses (Gómez-Gómez et al., 1999) and triggers the activation of a phosphorylation cascade (Nühse et al., 2000) in Arabidopsis plants. We recently identified a gene involved in flagellin perception, FLS2 , by positional cloning (Gómez-Gómez and Boller, 2000). FLS2 encodes a receptor kinase with an extracellular domain composed of 28 tandem leucine-rich repeats (LRRs) and an intracellular serine/threonine protein kinase domain. Interestingly, the encoded receptor kinase is homologous with the rice resistance gene Xa21, which provides resistance to Xanthomonas oryzae (Song et al., 1995), and the extracellular LRR domain of FLS2 resembles resistance genes from the tomato Cf gene family that provide resistance to various strains of Cladosporium fulvum (Jones et al., 1994; Dixon et al., 1996), suggesting that the recognition pathway for general elicitors such as flagellin may resemble the pathway involved in the highly specific resistance defined by gene-for-gene interactions (Jones and Jones, 1996). In insects and mammals, members of the family of TOL...
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.