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1.
Nature ; 626(7997): 194-206, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38096902

RESUMO

The LINE-1 (L1) retrotransposon is an ancient genetic parasite that has written around one-third of the human genome through a 'copy and paste' mechanism catalysed by its multifunctional enzyme, open reading frame 2 protein (ORF2p)1. ORF2p reverse transcriptase (RT) and endonuclease activities have been implicated in the pathophysiology of cancer2,3, autoimmunity4,5 and ageing6,7, making ORF2p a potential therapeutic target. However, a lack of structural and mechanistic knowledge has hampered efforts to rationally exploit it. We report structures of the human ORF2p 'core' (residues 238-1061, including the RT domain) by X-ray crystallography and cryo-electron microscopy in several conformational states. Our analyses identified two previously undescribed folded domains, extensive contacts to RNA templates and associated adaptations that contribute to unique aspects of the L1 replication cycle. Computed integrative structural models of full-length ORF2p show a dynamic closed-ring conformation that appears to open during retrotransposition. We characterize ORF2p RT inhibition and reveal its underlying structural basis. Imaging and biochemistry show that non-canonical cytosolic ORF2p RT activity can produce RNA:DNA hybrids, activating innate immune signalling through cGAS/STING and resulting in interferon production6-8. In contrast to retroviral RTs, L1 RT is efficiently primed by short RNAs and hairpins, which probably explains cytosolic priming. Other biochemical activities including processivity, DNA-directed polymerization, non-templated base addition and template switching together allow us to propose a revised L1 insertion model. Finally, our evolutionary analysis demonstrates structural conservation between ORF2p and other RNA- and DNA-dependent polymerases. We therefore provide key mechanistic insights into L1 polymerization and insertion, shed light on the evolutionary history of L1 and enable rational drug development targeting L1.


Assuntos
Endonucleases , Elementos Nucleotídeos Longos e Dispersos , DNA Polimerase Dirigida por RNA , Transcrição Reversa , Humanos , Microscopia Crioeletrônica , Endonucleases/química , Endonucleases/genética , Endonucleases/metabolismo , Elementos Nucleotídeos Longos e Dispersos/genética , RNA/genética , DNA Polimerase Dirigida por RNA/química , DNA Polimerase Dirigida por RNA/genética , DNA Polimerase Dirigida por RNA/metabolismo , Cristalografia por Raios X , DNA/biossíntese , DNA/genética , Imunidade Inata , Interferons/biossíntese
2.
Plant J ; 112(4): 881-896, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36164819

RESUMO

Narrow odd dwarf (nod) and Liguleless narrow (Lgn) are pleiotropic maize mutants that both encode plasma membrane proteins, cause similar developmental patterning defects, and constitutively induce stress signaling pathways. To investigate how these mutants coordinate maize development and physiology, we screened for protein interactors of NOD by affinity purification. LGN was identified by this screen as a strong candidate interactor, and we confirmed the NOD-LGN molecular interaction through orthogonal experiments. We further demonstrated that LGN, a receptor-like kinase, can phosphorylate NOD in vitro, hinting that they could act in intersecting signal transduction pathways. To test this hypothesis, we generated Lgn-R;nod mutants in two backgrounds (B73 and A619), and found that these mutations enhance each other, causing more severe developmental defects than either single mutation on its own, with phenotypes including very narrow leaves, increased tillering, and failure of the main shoot. Transcriptomic and metabolomic analyses of the single and double mutants in the two genetic backgrounds revealed widespread induction of pathogen defense genes and a shift in resource allocation away from primary metabolism in favor of specialized metabolism. These effects were similar in each single mutant and heightened in the double mutant, leading us to conclude that NOD and LGN act cumulatively in overlapping signaling pathways to coordinate growth-defense tradeoffs in maize.


Assuntos
Proteínas de Plantas , Zea mays , Zea mays/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Folhas de Planta/metabolismo , Fenótipo , Mutação , Regulação da Expressão Gênica de Plantas
3.
Plant J ; 112(1): 207-220, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35960639

RESUMO

Zea mays (maize) makes phytoalexins such as sesquiterpenoid zealexins, to combat invading pathogens. Zealexins are produced from farnesyl diphosphate in microgram per gram fresh weight quantities. As farnesyl diphosphate is also a precursor for many compounds essential for plant growth, the question arises as to how Z. mays produces high levels of zealexins without negatively affecting vital plant systems. To examine if specific pools of farnesyl diphosphate are made for zealexin synthesis we made CRISPR/Cas9 knockouts of each of the three farnesyl diphosphate synthases (FPS) in Z. mays and examined the resultant impacts on different farnesyl diphosphate-derived metabolites. We found that FPS3 (GRMZM2G098569) produced most of the farnesyl diphosphate for zealexins, while FPS1 (GRMZM2G168681) made most of the farnesyl diphosphate for the vital respiratory co-factor ubiquinone. Indeed, fps1 mutants had strong developmental phenotypes such as reduced stature and development of chlorosis. The replication and evolution of the fps gene family in Z. mays enabled it to produce dedicated FPSs for developmentally related ubiquinone production (FPS1) or defense-related zealexin production (FPS3). This partitioning of farnesyl diphosphate production between growth and defense could contribute to the ability of Z. mays to produce high levels of phytoalexins without negatively impacting its growth.


Assuntos
Geraniltranstransferase , Sesquiterpenos , Geraniltranstransferase/genética , Geraniltranstransferase/metabolismo , Fosfatos de Poli-Isoprenil , Sesquiterpenos/metabolismo , Terpenos/metabolismo , Ubiquinona/metabolismo , Zea mays/genética , Zea mays/metabolismo , Fitoalexinas
4.
Bioessays ; 42(1): e1900154, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31815300

RESUMO

Linear chromosomes shorten in every round of replication. In Drosophila, telomere-specialized long interspersed retrotransposable elements (LINEs) belonging to the jockey clade offset this shortening by forming head-to-tail arrays at Drosophila telomere ends. As such, these telomeric LINEs have been considered adaptive symbionts of the genome, protecting it from premature decay, particularly as Drosophila lacks a conventional telomerase holoenzyme. However, as reviewed here, recent work reveals a high degree of variation and turnover in the telomere-specialized LINE lineages across Drosophila. There appears to be no absolute requirement for LINE activity to maintain telomeres in flies, hence the suggestion that the telomere-specialized LINEs may instead be neutral or in conflict with the host, rather than adaptive.


Assuntos
Drosophila/genética , Genoma de Inseto , Retroelementos/genética , Telômero/genética , Animais , Elementos Nucleotídeos Longos e Dispersos , Simbiose , Telomerase/genética , Telomerase/metabolismo , Telômero/metabolismo
5.
Mol Plant Microbe Interact ; 34(10): 1157-1166, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34165327

RESUMO

Seed maceration and contamination with mycotoxin fumonisin inflicted by Fusarium verticillioides is a major disease concern for maize producers worldwide. Meta-analyses of quantitative trait loci for Fusarium ear rot resistance uncovered several ethylene (ET) biosynthesis and signaling genes within them, implicating ET in maize interactions with F. verticillioides. We tested this hypothesis using maize knockout mutants of the 1-aminocyclopropane-1-carboxylate (ACC) synthases ZmACS2 and ZmACS6. Infected wild-type seed emitted five-fold higher ET levels compared with controls, whereas ET was abolished in the acs2 and acs6 single and double mutants. The mutants supported reduced fungal biomass, conidia, and fumonisin content. Normal susceptibility was restored in the acs6 mutant with exogenous treatment of ET precursor ACC. Subsequently, we showed that fungal G-protein signaling is required for virulence via induction of maize-produced ET. F. verticillioides Gß subunit and two regulators of G-protein signaling mutants displayed reduced seed colonization and decreased ET levels. These defects were rescued by exogenous application of ACC. We concluded that pathogen-induced ET facilitates F. verticillioides colonization of seed, and, in turn, host ET production is manipulated via G-protein signaling of F. verticillioides to facilitate pathogenesis.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Assuntos
Fumonisinas , Fusarium , Etilenos , Proteínas de Ligação ao GTP , Virulência , Zea mays
6.
Planta ; 254(4): 73, 2021 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-34529190

RESUMO

MAIN CONCLUSION: A maize receptor kinase controls defense response to fungal pathogens by regulating jasmonic acid and antimicrobial phytoalexin production. Plants use a range of pattern recognition receptors to detect and respond to biotic threats. Some of these receptors contain leucine-rich repeat (LRR) domains that recognize microbial proteins or peptides. Maize (Zea mays) has 226 LRR-receptor like kinases, making it challenging to identify those important for pathogen recognition. In this study, co-expression analysis with genes for jasmonic acid and phytoalexin biosynthesis was used to identify a fungal induced-receptor like protein kinase (FI-RLPK) likely involved in the response to fungal pathogens. Loss-of-function mutants in fi-rlpk displayed enhanced susceptibility to the necrotrophic fungal pathogen Cochliobolus heterostrophus and reduced accumulation of jasmonic acid and the anti-microbial phytoalexins -kauralexins and zealexins- in infected tissues. In contrast, fi-rlpk mutants displayed increased resistance to stem inoculation with the hemibiotrophic fungal pathogen Fusarium graminearum. These data indicate that FI-RLPK is important for fungal recognition and activation of defenses, and that F. graminearum may be able to exploit FI-RLPK function to increase its virulence.


Assuntos
Ascomicetos , Zea mays , Bipolaris , Fusarium , Leucina , Doenças das Plantas , Proteínas Quinases , Zea mays/genética
7.
Metabolomics ; 17(1): 6, 2021 01 05.
Artigo em Inglês | MEDLINE | ID: mdl-33400019

RESUMO

INTRODUCTION: Studies investigating crop resistance to abiotic and biotic stress have largely focused on plant responses to singular forms of stress and individual biochemical pathways that only partially represent stress responses. Thus, combined abiotic and biotic stress treatments and the global assessment of their elicited metabolic expression remains largely unexplored. In this study, we employed targeted and untargeted metabolomics to investigate the molecular responses of maize (Zea mays) to abiotic, biotic, and combinatorial stress. OBJECTIVE: We compared the inducible metabolomes of heat-stressed (abiotic) and C. heterostrophus-infected (biotic) maize and examined the effects of heat stress on the ability of maize to defend itself against C. heterostrophus. METHODS: Ultra-high-performance liquid chromatography-high-resolution mass spectrometry was performed on plants grown under control conditions (28 °C), heat stress (38 °C), Cochliobolus heterostrophus infection, or combinatorial stress [heat (38 °C) + C. heterostrophus infection]. RESULTS: Multivariate analyses revealed differential metabolite expression between heat stress, C. heterostrophus infection, and their respective controls. In combinatorial experiments, treatment with heat stress prior to fungal inoculation negatively impacted maize disease resistance against C. heterostrophus, and distinct metabolome separation between combinatorial stressed plants and the non-heat-stressed infected controls was observed. Targeted analysis revealed inducible primary and secondary metabolite responses to abiotic/biotic stress, and combinatorial experiments indicated that deficiency in the hydroxycinnamic acid, p-coumaric acid, may contribute to the heat-induced susceptibility of maize to C. heterostrophus. CONCLUSION: These findings demonstrate that abiotic stress can predispose crops to more severe disease symptoms, underlining the increasing need to investigate defense chemistry in plants under combinatorial stress.


Assuntos
Resposta ao Choque Térmico , Metaboloma , Metabolômica , Doenças das Plantas/imunologia , Zea mays/imunologia , Zea mays/metabolismo , Cromatografia Líquida de Alta Pressão , Interações Hospedeiro-Patógeno , Espectrometria de Massas , Metabolômica/métodos , Doenças das Plantas/microbiologia , Zea mays/microbiologia
8.
Nucleic Acids Res ; 47(16): 8708-8719, 2019 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-31392993

RESUMO

Long Interspersed Elements (LINEs), also known as non-LTR retrotransposons, encode a multifunctional protein that reverse transcribes its mRNA into DNA at the site of insertion by target primed reverse transcription. The second half of the integration reaction remains very poorly understood. Second-strand DNA cleavage and second-strand DNA synthesis were investigated in vitro using purified components from a site-specific restriction-like endonuclease (RLE) bearing LINE. DNA structure was shown to be a critical component of second-strand DNA cleavage. A hitherto unknown and unexplored integration intermediate, an open '4-way' DNA junction, was recognized by the element protein and cleaved in a Holliday junction resolvase-like reaction. Cleavage of the 4-way junction resulted in a natural primer-template pairing used for second-strand DNA synthesis. A new model for RLE LINE integration is presented.


Assuntos
Enzimas de Restrição do DNA/genética , DNA Cruciforme/genética , Elementos Nucleotídeos Longos e Dispersos , RNA Mensageiro/genética , DNA Polimerase Dirigida por RNA/genética , Transcrição Reversa , Animais , Bombyx/genética , Bombyx/metabolismo , DNA/química , DNA/genética , DNA/metabolismo , Clivagem do DNA , Primers do DNA/genética , Primers do DNA/metabolismo , Enzimas de Restrição do DNA/metabolismo , DNA Cruciforme/química , DNA Cruciforme/metabolismo , Resolvases de Junção Holliday/genética , Resolvases de Junção Holliday/metabolismo , Conformação de Ácido Nucleico , RNA Mensageiro/metabolismo , DNA Polimerase Dirigida por RNA/metabolismo
9.
Planta ; 252(4): 62, 2020 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-32965567

RESUMO

MAIN CONCLUSION: Linolenic acid produced by the ω-3 fatty acid desaturase MSD3 in sorghum is used for insect-induced jasmonic acid production and is important for resistance against Spodoptera frugiperda. Jasmonic acid (JA) is a phytohormone that regulates both plant development and stress responses. In sorghum (Sorghum bicolor), the ω-3 fatty acid desaturase Multiseeded3 (MSD3) and the 13-lipoxygenase Multiseeded2 (MSD2) are important for producing JA to regulate panicle development and spikelet fertility, but their function in plant defense remains unknown. In this study, we examined whether these genes are important for the production of JA in response to herbivory by the insect pest Spodoptera frugiperda. Compared to wild-type controls, the msd3 mutant accumulated less JA in leaves of both infested and uninfested plants, revealing that MSD3 is involved in stress-induced JA production. In contrast, herbivore-induced JA production in the msd2 mutant was indistinguishable from wild type, indicating that MSD2 does not function in herbivore-induced JA production. An increase of S. frugiperda growth was observed on both the msd3 and msd2 mutants, hinting at roles for both JA and additional oxylipins in sorghum's defense responses.


Assuntos
Ácidos Graxos Dessaturases , Lipoxigenase , Defesa das Plantas contra Herbivoria , Sorghum , Spodoptera , Animais , Ácidos Graxos Dessaturases/metabolismo , Herbivoria , Lipoxigenase/genética , Lipoxigenase/metabolismo , Mutação , Oxilipinas/metabolismo , Defesa das Plantas contra Herbivoria/genética , Sorghum/enzimologia , Sorghum/genética , Sorghum/parasitologia , Spodoptera/fisiologia
10.
Plant Cell Environ ; 43(1): 223-234, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31411732

RESUMO

To grow and thrive plants must be able to adapt to both adverse environmental conditions and attack by a variety of pests. Elucidating the sophisticated mechanisms plants have developed to achieve this has been the focus of many studies. What is less well understood is how plants respond when faced with multiple stressors simultaneously. In this study, we assess the response of Zea mays (maize) to the combinatorial stress of flooding and infestation with the insect pest Spodoptera frugiperda (fall armyworm). This combined stress leads to elevated production of the defence hormone salicylic acid, which does not occur in the individual stresses, and the resultant salicylic acid-dependent increase in S. frugiperda resistance. Remodelling of phenylpropanoid pathways also occurs in response to this combinatorial stress leading to increased production of the anti-insect C-glycosyl flavones (maysins) and the herbivore-induced volatile phenolics, benzyl acetate, and phenethyl acetate. Furthermore, changes in cellular redox status also occur, as indicated by reductions in peroxidase and polyphenol oxidase activity. These data suggest that metabolite changes important for flooding tolerance and anti-insect defence may act both additively and synergistically to provide extra protection to the plant.


Assuntos
Resistência à Doença/fisiologia , Inundações , Insetos/fisiologia , Zea mays/metabolismo , Animais , Catecol Oxidase/metabolismo , Regulação da Expressão Gênica de Plantas , Herbivoria/fisiologia , Larva/fisiologia , Peroxidase/metabolismo , Doenças das Plantas , Ácido Salicílico/metabolismo , Spodoptera/fisiologia
11.
Anal Biochem ; 603: 113809, 2020 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-32511965

RESUMO

Long interspersed elements (LINEs) replicate by target primed reverse transcription (TPRT). Insertion involves two half reactions. Each half reaction involves DNA cleavage followed by DNA synthesis. The linker region, located just beyond the reverse transcriptase in the LINE open reading frame, contains a set of predicted helices that may form an α-finger, followed by a gag-like zinc-knuckle. Point mutations of moderately conserved amino-acid residues in the presumptive α-finger severely impair the DNA endonuclease and reverse transcriptase activities of the integration reaction during both half reactions. Mutations in the gag-like zinc-knuckle also impair DNA cleavage and DNA synthesis in some instances. Mutations in core residues that presumably disrupt the protein structure of the presumptive α-finger and the gag-like zinc-knuckle lead to a promiscuous DNA endonuclease and protein-nucleic acid complexes that get stuck in the well during analysis. The linker region appears to function as a protein, DNA, and RNA conformational switching area. The linker is used to properly position nucleic acid substrates into the active sites of the reverse transcriptase and of the DNA endonuclease.


Assuntos
DNA/química , DNA/metabolismo , Elementos Nucleotídeos Longos e Dispersos/fisiologia , Motivos de Aminoácidos , Sítios de Ligação , Sequência Conservada , DNA/biossíntese , Clivagem do DNA , Desoxirribonuclease I/metabolismo , Proteínas de Insetos , Mutação Puntual , Polimerização , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , DNA Polimerase Dirigida por RNA/metabolismo
12.
Plant J ; 93(5): 799-813, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29315977

RESUMO

Maize white seedling 3 (w3) has been used to study carotenoid deficiency for almost 100 years, although the molecular basis of the mutation has remained unknown. Here we show that the w3 phenotype is caused by disruption of the maize gene for homogentisate solanesyl transferase (HST), which catalyzes the first and committed step in plastoquinone-9 (PQ-9) biosynthesis in the plastid. The resulting PQ-9 deficiency prohibits photosynthetic electron transfer and eliminates PQ-9 as an oxidant in the enzymatic desaturation of phytoene during carotenoid synthesis. As a result, light-grown w3 seedlings are albino, deficient in colored carotenoids and accumulate high levels of phytoene. However, despite the absence of PQ-9 for phytoene desaturation, dark-grown w3 seedlings can produce abscisic acid (ABA) and homozygous w3 kernels accumulate sufficient carotenoids to generate ABA needed for seed maturation. The presence of ABA and low levels of carotenoids in w3 nulls indicates that phytoene desaturase is able to use an alternate oxidant cofactor, albeit less efficiently than PQ-9. The observation that tocopherols and tocotrienols are modestly affected in w3 embryos and unaffected in w3 endosperm indicates that, unlike leaves, grain tissues deficient in PQ-9 are not subject to severe photo-oxidative stress. In addition to identifying the molecular basis for the maize w3 mutant, we: (1) show that low levels of phytoene desaturation can occur in w3 seedlings in the absence of PQ-9; and (2) demonstrate that PQ-9 and carotenoids are not required for vitamin E accumulation.


Assuntos
Alquil e Aril Transferases/metabolismo , Proteínas de Plantas/metabolismo , Plastoquinona/metabolismo , Tocoferóis/metabolismo , Zea mays/metabolismo , Ácido Abscísico/metabolismo , Alquil e Aril Transferases/genética , Carotenoides/genética , Carotenoides/metabolismo , Mutação , Oxirredutases/genética , Oxirredutases/metabolismo , Fenótipo , Fotossíntese , Filogenia , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Plastídeos/genética , Plastídeos/metabolismo , Sementes/genética , Sementes/metabolismo , Vitamina E/genética , Vitamina E/metabolismo , Zea mays/genética
13.
BMC Plant Biol ; 19(1): 310, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31307401

RESUMO

BACKGROUND: The hypersensitive defense response (HR) in plants is a fast, localized necrotic response around the point of pathogen ingress. HR is usually triggered by a pathogen recognition event mediated by a nucleotide-binding site, leucine-rich repeat (NLR) protein. The autoactive maize NLR gene Rp1-D21 confers a spontaneous HR response in the absence of pathogen recognition. Previous work identified a set of loci associated with variation in the strength of Rp1-D21-induced HR. A polygalacturonase gene homolog, here termed ZmPGH1, was identified as a possible causal gene at one of these loci on chromosome 7. RESULTS: Expression of ZmPGH1 inhibited the HR-inducing activity of both Rp1-D21 and that of another autoactive NLR, RPM1(D505V), in a Nicotiana benthamiana transient expression assay system. Overexpression of ZmPGH1 in a transposon insertion line of maize was associated with suppression of chemically-induced programmed cell death and with suppression of HR induced by Rp1-D21 in maize plants grown in the field. CONCLUSIONS: ZmPGH1 functions as a suppressor of programmed cell death induced by at least two autoactive NLR proteins and by two chemical inducers. These findings deepen our understanding of the control of the HR in plants.


Assuntos
Apoptose/fisiologia , Proteínas de Plantas/fisiologia , Poligalacturonase/fisiologia , Zea mays/fisiologia , Apoptose/genética , Mapeamento Cromossômico , Cromossomos de Plantas , Genes de Plantas , Leucina , Proteínas de Plantas/química , Proteínas de Plantas/genética , Poligalacturonase/química , Poligalacturonase/genética , Recombinação Genética , Sequências Repetitivas de Aminoácidos , Nicotiana/genética , Zea mays/enzimologia , Zea mays/genética , Zea mays/imunologia
14.
Planta ; 249(1): 21-30, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30187155

RESUMO

MAIN CONCLUSION: Maize produces an array of herbivore-induced terpene volatiles that attract parasitoids to infested plants and a suite of pathogen-induced non-volatile terpenoids with antimicrobial activity to defend against pests. Plants rely on complex blends of constitutive and dynamically produced specialized metabolites to mediate beneficial ecological interactions and protect against biotic attack. One such class of metabolites are terpenoids, a large and structurally diverse class of molecules shown to play significant defensive and developmental roles in numerous plant species. Despite this, terpenoids have only recently been recognized as significant contributors to pest resistance in maize (Zea mays), a globally important agricultural crop. The current review details recent advances in our understanding of biochemical structures, pathways and functional roles of maize terpenoids. Dependent upon the lines examined, maize can harbor more than 30 terpene synthases, underlying the inherent diversity of maize terpene defense systems. Part of this defensive arsenal is the inducible production of volatile bouquets that include monoterpenes, homoterpenes and sesquiterpenes, which often function in indirect defense by enabling the attraction of parasitoids and predators. More recently discovered are a subset of sesquiterpene and diterpene hydrocarbon olefins modified by cytochrome P450s to produce non-volatile end-products such kauralexins, zealexins, dolabralexins and ß-costic acid. These non-volatile terpenoid phytoalexins often provide effective defense against both microbial and insect pests via direct antimicrobial and anti-feedant activity. The diversity and promiscuity of maize terpene synthases, coupled with a variety of secondary modifications, results in elaborate defensive layers whose identities, regulation and precise functions are continuing to be elucidated.


Assuntos
Terpenos/metabolismo , Zea mays/metabolismo , Zea mays/parasitologia , Animais , Regulação da Expressão Gênica de Plantas , Insetos/patogenicidade , Doenças das Plantas/parasitologia , Sesquiterpenos/metabolismo , Fitoalexinas
15.
Int J Mol Sci ; 20(19)2019 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-31597271

RESUMO

As in other cereal crops, the panicles of sorghum (Sorghum bicolor (L.) Moench) comprise two types of floral spikelets (grass flowers). Only sessile spikelets (SSs) are capable of producing viable grains, whereas pedicellate spikelets (PSs) cease development after initiation and eventually abort. Consequently, grain number per panicle (GNP) is lower than the total number of flowers produced per panicle. The mechanism underlying this differential fertility is not well understood. To investigate this issue, we isolated a series of ethyl methane sulfonate (EMS)-induced multiseeded (msd) mutants that result in full spikelet fertility, effectively doubling GNP. Previously, we showed that MSD1 is a TCP (Teosinte branched/Cycloidea/PCF) transcription factor that regulates jasmonic acid (JA) biosynthesis, and ultimately floral sex organ development. Here, we show that MSD2 encodes a lipoxygenase (LOX) that catalyzes the first committed step of JA biosynthesis. Further, we demonstrate that MSD1 binds to the promoters of MSD2 and other JA pathway genes. Together, these results show that a JA-induced module regulates sorghum panicle development and spikelet fertility. The findings advance our understanding of inflorescence development and could lead to new strategies for increasing GNP and grain yield in sorghum and other cereal crops.


Assuntos
Ciclopentanos/metabolismo , Fertilidade , Oxilipinas/metabolismo , Desenvolvimento Vegetal , Sorghum/fisiologia , Sequência de Aminoácidos , Sítios de Ligação , Grão Comestível , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Redes e Vias Metabólicas , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regiões Promotoras Genéticas , Ligação Proteica , Sorghum/classificação , Fatores de Transcrição/metabolismo
16.
Int J Mol Sci ; 20(21)2019 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-31661847

RESUMO

Grain number per panicle is an important component of grain yield in sorghum (Sorghum bicolor (L.)) and other cereal crops. Previously, we reported that mutations in multi-seeded 1 (MSD1) and MSD2 genes result in a two-fold increase in grain number per panicle due to the restoration of the fertility of the pedicellate spikelets, which invariably abort in natural sorghum accessions. Here, we report the identification of another gene, MSD3, which is also involved in the regulation of grain numbers in sorghum. Four bulked F2 populations from crosses between BTx623 and each of the independent msd mutants p6, p14, p21, and p24 were sequenced to 20× coverage of the whole genome on a HiSeq 2000 system. Bioinformatic analyses of the sequence data showed that one gene, Sorbi_3001G407600, harbored homozygous mutations in all four populations. This gene encodes a plastidial ω-3 fatty acid desaturase that catalyzes the conversion of linoleic acid (18:2) to linolenic acid (18:3), a substrate for jasmonic acid (JA) biosynthesis. The msd3 mutants had reduced levels of linolenic acid in both leaves and developing panicles that in turn decreased the levels of JA. Furthermore, the msd3 panicle phenotype was reversed by treatment with methyl-JA (MeJA). Our characterization of MSD1, MSD2, and now MSD3 demonstrates that JA-regulated processes are critical to the msd phenotype. The identification of the MSD3 gene reveals a new target that could be manipulated to increase grain number per panicle in sorghum, and potentially other cereal crops, through the genomic editing of MSD3 functional orthologs.


Assuntos
Produtos Agrícolas/enzimologia , Ciclopentanos/metabolismo , Ácidos Graxos Dessaturases/genética , Ácidos Graxos Dessaturases/metabolismo , Oxilipinas/metabolismo , Sorghum/enzimologia , Alelos , Produtos Agrícolas/efeitos dos fármacos , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Ciclopentanos/farmacologia , Grão Comestível/efeitos dos fármacos , Grão Comestível/genética , Grão Comestível/crescimento & desenvolvimento , Sequenciamento de Nucleotídeos em Larga Escala , Ácido Linoleico/química , Ácido Linoleico/metabolismo , Mutação , Oxilipinas/farmacologia , Fenótipo , Sementes/efeitos dos fármacos , Sementes/genética , Sementes/crescimento & desenvolvimento , Sorghum/genética , Sorghum/metabolismo , Ácido alfa-Linolênico/biossíntese , Ácido alfa-Linolênico/química
17.
Plant J ; 89(4): 746-763, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27862526

RESUMO

We used a systems genetics approach to elucidate the molecular mechanisms of the responses of maize to grey leaf spot (GLS) disease caused by Cercospora zeina, a threat to maize production globally. Expression analysis of earleaf samples in a subtropical maize recombinant inbred line population (CML444 × SC Malawi) subjected in the field to C. zeina infection allowed detection of 20 206 expression quantitative trait loci (eQTLs). Four trans-eQTL hotspots coincided with GLS disease QTLs mapped in the same field experiment. Co-expression network analysis identified three expression modules correlated with GLS disease scores. The module (GY-s) most highly correlated with susceptibility (r = 0.71; 179 genes) was enriched for the glyoxylate pathway, lipid metabolism, diterpenoid biosynthesis and responses to pathogen molecules such as chitin. The GY-s module was enriched for genes with trans-eQTLs in hotspots on chromosomes 9 and 10, which also coincided with phenotypic QTLs for susceptibility to GLS. This transcriptional network has significant overlap with the GLS susceptibility response of maize line B73, and may reflect pathogen manipulation for nutrient acquisition and/or unsuccessful defence responses, such as kauralexin production by the diterpenoid biosynthesis pathway. The co-expression module that correlated best with resistance (TQ-r; 1498 genes) was enriched for genes with trans-eQTLs in hotspots coinciding with GLS resistance QTLs on chromosome 9. Jasmonate responses were implicated in resistance to GLS through co-expression of COI1 and enrichment of genes with the Gene Ontology term 'cullin-RING ubiquitin ligase complex' in the TQ-r module. Consistent with this, JAZ repressor expression was highly correlated with the severity of GLS disease in the GY-s susceptibility network.


Assuntos
Folhas de Planta/genética , Folhas de Planta/microbiologia , Zea mays/genética , Zea mays/microbiologia , Ascomicetos/patogenicidade , Cromossomos de Plantas/genética , Redes Reguladoras de Genes/genética , Redes Reguladoras de Genes/fisiologia , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Locos de Características Quantitativas/genética
18.
Planta ; 248(1): 105-116, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29616394

RESUMO

MAIN CONCLUSION: The maize inbred line W22 has lower herbivore-induced volatile production than B73 but both fall armyworm larvae and the wasps that parasitize them prefer W22 over B73. Maize inbred line W22 is an important resource for genetic studies due to the availability of the UniformMu mutant population and a complete genome sequence. In this study, we assessed the suitability of W22 as a model for tritrophic interactions between maize, Spodoptera frugiperda (fall armyworm) and the parasitoid wasp Cotesia marginiventris. W22 was found to be a good model for studying the interaction as S. frugiperda prefers W22 over B73 and a higher parasitism rate by C. marginiventris was observed on W22 compared to the inbred line B73. W22 also produced lower amounts of many herbivore-induced volatile terpenes and indole emission upon treatment with S. frugiperda oral secretions. We propose that some of the major herbivore-induced terpene volatiles are perhaps impeding S. frugiperda and C. marginiventris preference and that as yet unidentified compounds are produced at low abundance may be positively impacting these interactions.


Assuntos
Herbivoria , Feromônios/metabolismo , Spodoptera , Compostos Orgânicos Voláteis/metabolismo , Vespas , Zea mays/metabolismo , Animais , Benzoxazinas/metabolismo , Perfilação da Expressão Gênica , Indóis/metabolismo , Modelos Biológicos , Spodoptera/parasitologia , Terpenos/metabolismo , Vespas/fisiologia , Zea mays/parasitologia
19.
Planta ; 247(4): 863-873, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29260396

RESUMO

MAIN CONCLUSION: Chemical isolation and NMR-based structure elucidation revealed a novel keto-acidic sesquiterpenoid, termed zealexin A4 (ZA4). ZA4 is elicited by pathogens and herbivory, but attenuated by heightened levels of CO 2 . The identification of the labdane-related diterpenoids, termed kauralexins and acidic sesquiterpenoids, termed zealexins, demonstrated the existence of at least ten novel stress-inducible maize metabolites with diverse antimicrobial activity. Despite these advances, the identity of co-occurring and predictably related analytes remains largely unexplored. In the current effort, we identify and characterize the first sesquiterpene keto acid derivative of ß-macrocarpene, named zealexin A4 (ZA4). Evaluation of diverse maize inbreds revealed that ZA4 is commonly produced in maize scutella during the first 14 days of seedling development; however, ZA4 production in the scutella was markedly reduced in seedlings grown in sterile soil. Elevated ZA4 production was observed in response to inoculation with adventitious fungal pathogens, such as Aspergillus flavus and Rhizopus microsporus, and a positive relationship between ZA4 production and expression of the predicted zealexin biosynthetic genes, terpene synthases 6 and 11 (Tps6 and Tps11), was observed. ZA4 exhibited significant antimicrobial activity against the mycotoxigenic pathogen A. flavus; however, ZA4 activity against R. microsporus was minimal, suggesting the potential of some fungi to detoxify ZA4. Significant induction of ZA4 production was also observed in response to infestation with the stem tunneling herbivore Ostrinia nubilalis. Examination of the interactive effects of elevated CO2 (E-CO2) on both fungal and herbivore-elicited ZA4 production revealed significantly reduced levels of inducible ZA4 accumulation, consistent with a negative role for E-CO2 on ZA4 production. Collectively, these results describe a novel ß-macrocarpene-derived antifungal defense in maize and expand the established diversity of zealexins that are differentially regulated in response to biotic/abiotic stress.


Assuntos
Sesquiterpenos/metabolismo , Zea mays/metabolismo , Alquil e Aril Transferases/metabolismo , Anti-Infecciosos/metabolismo , Aspergillus flavus/metabolismo , Dióxido de Carbono/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Imunidade Vegetal , Rhizopus/metabolismo , Plântula/metabolismo , Zea mays/efeitos dos fármacos , Zea mays/microbiologia
20.
J Exp Bot ; 69(5): 1235-1245, 2018 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-29301018

RESUMO

Reactive oxygen species (ROS) can be elicited by many forms of stress, including pathogen attack, abiotic stress, damage and insect infestation. Perception of microbe- or damage-associated elicitors triggers an ROS burst in many plant species; however, the impact of herbivore fatty-acid amides on ROS elicitation remains largely unexplored. In this study we show that the lepidopteran-derived fatty-acid amide elicitor N-linolenoyl-L-glutamine (GLN18:3) can induce a ROS burst in multiple plant species. Furthermore, in Arabidopsis this ROS burst is partially dependent on the plasma membrane localized NADPH oxidases RBOHD and RBOHF, and an Arabidopsis rbohD/F double mutant produces enhanced GLN18:3-induced jasmonic acid. Quantification of GLN18:3-induced ROS in phytohormone-deficient lines revealed that in Arabidopsis reduced levels of jasmonic acid resulted in a larger elicitor-induced ROS burst, while in tomato reduction of either jasmonic acid or salicylic acid led to higher induced ROS production. These data indicate that GLN18:3-induced ROS is antagonistic to jasmonic acid production in these species. In biological assays, rbohD/F mutant plants were more resistant to the generalist herbivores Spodoptera exigua and Trichoplusia ni but not to the specialist Plutella xylostella. Collectively, these results demonstrate that in Arabidopsis herbivore-induced ROS may negatively regulate plant defense responses to herbivory.


Assuntos
Amidas/metabolismo , Arabidopsis/fisiologia , Herbivoria , Mariposas , Espécies Reativas de Oxigênio/metabolismo , Solanum lycopersicum/fisiologia , Animais , Proteínas de Arabidopsis/metabolismo , Ácidos Graxos , Cadeia Alimentar , NADPH Oxidases/metabolismo , Spodoptera
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