RESUMO
Active DNA demethylation is critical for controlling the DNA methylomes in plants and mammals. However, little is known about how DNA demethylases are recruited to target loci, and the involvement of chromatin marks in this process. Here, we identify 2 components of the SWR1 chromatin-remodeling complex, PIE1 and ARP6, as required for ROS1-mediated DNA demethylation, and discover 2 SWR1-associated bromodomain-containing proteins, AtMBD9 and nuclear protein X1 (NPX1). AtMBD9 and NPX1 recognize histone acetylation marks established by increased DNA methylation 1 (IDM1), a known regulator of DNA demethylation, redundantly facilitating H2A.Z deposition at IDM1 target loci. We show that at some genomic regions, H2A.Z and DNA methylation marks coexist, and H2A.Z physically interacts with ROS1 to regulate DNA demethylation and antisilencing. Our results unveil a mechanism through which DNA demethylases can be recruited to specific target loci exhibiting particular histone marks, providing a conceptual framework to understand how chromatin marks regulate DNA demethylation.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Desmetilação do DNA , Histonas/metabolismo , Complexos Multiproteicos/metabolismo , Acetilação , Cromatina/metabolismo , Inativação Gênica , Modelos Biológicos , Mutação/genética , Ligação Proteica , Subunidades Proteicas/metabolismoRESUMO
Plants generate reactive oxygen species (ROS) in the apoplast in response to pathogen attack, especially following resistance (R) gene-mediated pathogen recognition; however, the mechanisms activating ROS generation remain unknown. Here, we demonstrate that RKN (Meloidogyne incognita) infection rapidly induces ROS accumulation in the roots of tomato (Solanum lycopersicum) plants that contain the R gene Mi-1.2 but rarely induces ROS accumulation in the susceptible or Mi-1.2-silenced resistant genotypes. RNK also induces the hypersensitive response, a form of programmed cell death, in Mi-1.2 plants. RKN induces the expression of numerous class-A heat shock factor (HsfA) genes in resistant tomato plants. Silencing HsfA1a compromises Mi-1.2-mediated resistance, apoplastic H2O2 accumulation, and the transcription of whitefly induced 1 (Wfi1), which encodes a respiratory burst oxidase homolog. HsfA1a regulates Wfi1 transcription by binding to the Wfi1 promoter, and silencing of Wfi1 compromises Mi-1.2-mediated resistance. HsfA1a and Wfi1 are involved in Mi-1.2-triggered Hsp90 accumulation and basal defense in susceptible tomato. Thus, HsfA-1aWfi1-dependent ROS signaling functions as a crucial regulator of plant defense responses.
Assuntos
Fatores de Transcrição de Choque Térmico/metabolismo , Proteínas de Plantas/genética , Solanum lycopersicum/genética , Solanum lycopersicum/parasitologia , Tylenchoidea/patogenicidade , Animais , Regulação da Expressão Gênica de Plantas , Proteínas de Choque Térmico HSP90/genética , Proteínas de Choque Térmico HSP90/metabolismo , Fatores de Transcrição de Choque Térmico/genética , Interações Hospedeiro-Parasita/genética , Peróxido de Hidrogênio/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Raízes de Plantas/parasitologia , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas , Espécies Reativas de Oxigênio/metabolismoRESUMO
Strigolactones (SLs) are carotenoid-derived phytohormones that are known to influence various aspects of plant growth and development. As root-derived signals, SLs can enhance symbiosis between plants and arbuscular mycorrhizal fungi (AMF). However, little is known about the roles of SLs in plant defense against soil-borne pathogens. Here, we determined that infection with root-knot nematodes (RKNs; Meloidogyne incognita) induced SL biosynthesis in roots of tomato (Solanum lycopersicum). Silencing of SL biosynthesis genes compromised plant defense against RKNs, whilst application of the SL analog racGR24 enhanced it. Accumulation of endogenous jasmonic acid (JA) and abscisic acid (ABA) in the roots in response to RKN infection was enhanced by silencing of SL biosynthetic genes and was suppressed by application of racGR24. Genetic evidence showed that JA was a positive regulator of defense against RKNs while ABA was a negative regulator. In addition, racGR24 enhanced the defense against nematode in a JA-deficient mutant but not in an ABA-deficient mutant. Silencing of SL biosynthetic genes resulted in up-regulation of MYC2, which negatively regulated defense against RKNs. Our results demonstrate that SLs play a positive role in nematode defense in tomato and that MYC2 negatively regulates this defense, potentially by mediating hormone crosstalk among SLs, ABA and JA.
Assuntos
Lactonas/metabolismo , Doenças das Plantas/parasitologia , Reguladores de Crescimento de Plantas/biossíntese , Imunidade Vegetal/fisiologia , Solanum lycopersicum/imunologia , Tylenchoidea/fisiologia , Animais , Solanum lycopersicum/parasitologia , Raízes de Plantas/metabolismoRESUMO
Interplay of hormones with reactive oxygen species (ROS) fine-tunes the response of plants to stress; however, the crosstalk between brassinosteroids (BRs) and ROS in nematode resistance is unclear. In this study, we found that low BR biosynthesis or lack of BR receptor increased, whilst exogenous BR decreased the susceptibility of tomato plants to Meloidogyne incognita. Hormone quantification coupled with hormone mutant complementation experiments revealed that BR did not induce the defence response by triggering salicylic acid (SA), jasmonic acid/ethylene (JA/ET) or abscisic acid (ABA) signalling pathway. Notably, roots of BR-deficient plants had decreased apoplastic ROS accumulation, transcript of RESPIRATORY BURST OXIDASE HOMOLOG1 (RBOH1) and WHITEFLY INDUCED1 (WFI1), and reduced activation of mitogen-activated protein kinase 1/2 (MPK1/2) and MPK3. Silencing of RBOH1, WFI1, MPK1, MPK2 and MPK3 all increased the root susceptibility to nematode and attenuated BR-induced resistance against the nematode. Significantly, suppressed transcript of RBOH1 compromised BR-induced activation of MPK1/2 and MPK3. These results strongly suggest that RBOH-dependent MPK activation is involved in the BR-induced systemic resistance against the nematode.
Assuntos
Brassinosteroides/metabolismo , Resistência à Doença , Doenças das Plantas/imunologia , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Solanum lycopersicum/fisiologia , Tylenchoidea/fisiologia , Ácido Abscísico/metabolismo , Animais , Ciclopentanos/metabolismo , Etilenos/metabolismo , Regulação da Expressão Gênica de Plantas , Solanum lycopersicum/enzimologia , Solanum lycopersicum/genética , Solanum lycopersicum/imunologia , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NADPH Oxidases/genética , NADPH Oxidases/metabolismo , Oxilipinas/metabolismo , Doenças das Plantas/parasitologia , Proteínas de Plantas/genética , Espécies Reativas de Oxigênio/metabolismo , Ácido Salicílico/metabolismo , Transdução de SinaisRESUMO
L-lysine is an essential amino acid with broad applications in the animal feed, human food, and pharmaceutical industries. The fermentation production of L-lysine by Escherichia coli has limitations such as poor substrate utilization efficiency and low saccharide conversion rates. We deleted the global regulatory factor gene mlc and introduced heterologous genes, including the maltose phosphotransferase genes (malAP) from Bacillus subtilis, to enhance the use efficiency of disaccharides and trisaccharides. The engineered strain E. coli XC3 demonstrated improved L-lysine production, yield, and productivity, which reached 160.00 g/L, 63.78%, and 4.44 g/(Lâ§h), respectively. Furthermore, we overexpressed the glutamate dehydrogenase gene (gdhA) and assimilated nitrate reductase genes (BsnasBC) from B. subtilis, along with nitrite reductase genes (EcnirBD) from E. coli, in strain E. coli XC3. This allowed the construction of E. coli XC4 with a nitrate assimilation pathway. The L-lysine production, yield, and productivity of E. coli XC4 were elevated to 188.00 g/L, 69.44%, and 5.22 g/(Lâ§h), respectively. After optimization of the residual sugar concentration and carbon to nitrogen ratio, the L-lysine production, yield, and productivity were increased to 204.00 g/L, 72.32%, and 5.67 g/(Lâ§h), respectively, in a 5 L fermenter. These values represented the increases of 40.69%, 20.03%, and 40.69%, respectively, compared with those of the starting strain XC1. By engineering the substrate utilization pathway, we successfully constructed a high-yield L-lysine producing strain, laying a solid foundation for the industrial production of L-lysine.
Assuntos
Bacillus subtilis , Escherichia coli , Fermentação , Lisina , Engenharia Metabólica , Escherichia coli/genética , Escherichia coli/metabolismo , Lisina/biossíntese , Lisina/metabolismo , Bacillus subtilis/genética , Bacillus subtilis/metabolismo , Glutamato Desidrogenase/metabolismo , Glutamato Desidrogenase/genética , Nitrato Redutase/genética , Nitrato Redutase/metabolismoRESUMO
Strigolactones are carotenoid-derived phytohormones that impact plant growth and development in diverse ways. However, the roles of strigolactones in the responses to temperature stresses are largely unknown. Here, we demonstrated that strigolactone biosynthesis is induced in tomato (Solanum lycopersicum) by heat and cold stresses. Compromised strigolactone biosynthesis or signaling negatively affected heat and cold tolerance, while application of the synthetic strigolactone analog GR245DS enhanced heat and cold tolerance. Strigolactone-mediated heat and cold tolerance was associated with the induction of abscisic acid (ABA), heat shock protein 70 (HSP70) accumulation, C-REPEAT BINDING FACTOR 1 (CBF1) transcription, and antioxidant enzyme activity. Importantly, a deficiency in ABA biosynthesis compromised the GR245DS effects on heat and cold stresses and abolished the GR245DS-induced transcription of HSP70, CBF1, and antioxidant-related genes. These results support that strigolactones positively regulate tomato heat and cold tolerance and that they do so at least partially by the induction of CBFs and HSPs and the antioxidant response in an ABA-dependent manner.
RESUMO
This study describes the use of 14 nm nonionic fluorosurfactant-capped gold nanoparticles (FSN-capped AuNPs) for the simultaneous detection of cysteine (Cys) and homocysteine (Hcy) using colorimetric method, requiring no use of separation techniques. It was found that the kinetics of Cys/Hcy-induced aggregation of the 14 nm FSN-capped AuNPs strongly depends on the pH value of gold colloidal solution. At a pH of 6.5, the Cys-induced aggregation kinetics of the FSN-capped AuNPs was almost identical to that induced by Hcy, facilitating simultaneous detection of total Cys and Hcy up to a concentration as low as 0.15 µM; while at pH 12.0, the kinetics of Cys-induced aggregation was much faster than that inducted by Hcy, leading to selective detection of Cys at concentration as low as 1.0 µM in the presence of Hcy. The applicability of the method was validated by spiking known amount of Cys and Hcy in human urine and plasma samples, obtaining a recovery of 95.4-105.5%. The present approach is simple, high selective and provides high reproducibility, and has a great potentiality in disease diagnosis.