RESUMO
DNA methylation is an important epigenetic mark implicated in selective rRNA gene expression, but the DNA methylation readers and effectors remain largely unknown. Here, we report a protein complex that reads DNA methylation to regulate variant-specific 45S ribosomal RNA (rRNA) gene expression in Arabidopsis (Arabidopsis thaliana). The complex, consisting of METHYL-CpG-BINDING DOMAIN PROTEIN5 (MBD5), MBD6, ALPHA-CRYSTALLIN DOMAIN PROTEIN15.5 (ACD15.5), and ACD21.4, directly binds to 45S rDNA. While MBD5 and MBD6 function redundantly, ACD15.5 and ACD21.4 are indispensable for variant-specific rRNA gene expression. These 4 proteins undergo phase separation in vitro and in vivo and are interdependent for their phase separation. The α-crystallin domain of ACD15.5 and ACD21.4, which is essential for their function, enables phase separation of the complex, likely by mediating multivalent protein interactions. The effector MICRORCHIDIA6 directly interacts with ACD15.5 and ACD21.4, but not with MBD5 and MBD6, and is recruited to 45S rDNA by the MBD-ACD complex to regulate variant-specific 45S rRNA expression. Our study reveals a pathway in Arabidopsis through which certain 45S rRNA gene variants are silenced, while others are activated.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , alfa-Cristalinas , Arabidopsis/genética , Arabidopsis/metabolismo , Genes de RNAr , Metilação de DNA/genética , RNA Ribossômico/genética , RNA Ribossômico/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , DNA Ribossômico/genética , DNA Ribossômico/metabolismo , alfa-Cristalinas/genética , alfa-Cristalinas/metabolismoRESUMO
Apurinic/apyrimidinic (AP) sites are one of the most abundant DNA lesions and are mainly repaired by AP endonucleases (APEs). While most eukaryotic genomes encode two APEs, plants usually possess three APEs, namely APE1L, APE2, and ARP. To date, the biological relevance and functional divergence of plant APEs are unclear. Here, we show that the three plant APEs have ancient origins, with the APE1L clade being plant-specific. In Arabidopsis thaliana, simultaneously mutating APE1L and APE2, but not ARP alone or in combination with either APE1L or APE2, results in clear developmental defects linked to genotoxic stress. Genetic analyses indicated that the three plant APEs have different substrate preferences in vivo. ARP is mainly responsible for AP site repair, while APE1L and APE2 prefer to repair 3'-blocked single-stranded DNA breaks. We further determined that APEs play an important role in DNA repair and the maintenance of genomic integrity in meiotic cells. The ape1l ape2 double mutant exhibited a greatly enhanced frequency of sporulation 1 (SPO11-1)-dependent and SPO11-1-independent double-stranded DNA breaks. The DNA damage response (DDR) was activated in ape1l ape2 to trigger pollen abortion. Our findings suggest functional divergence of plant APEs and reveal important roles of plant APEs during vegetative and reproductive development.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Hominidae , Animais , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/genética , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/metabolismo , Reparo do DNA/genética , Dano ao DNA/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Endonucleases/genética , Hominidae/metabolismo , Proteínas de Arabidopsis/genéticaRESUMO
Base excision repair and active DNA demethylation produce repair intermediates with DNA molecules blocked at the 3'-OH end by an aldehyde or phosphate group. However, both the physiological consequences of these accumulated single-strand DNAs break with 3'-blocked ends (DNA 3'-blocks) and the signaling pathways responding to unrepaired DNA 3'-blocks remain unclear in plants. Here, we investigated the effects of DNA 3'-blocks on plant development using the zinc finger DNA 3'-phosphoesterase (zdp) AP endonuclease2 (ape2) double mutant, in which 3'-blocking residues are poorly repaired. The accumulation of DNA 3'-blocked triggered diverse developmental defects that were dependent on the ATM and RAD3-related (ATR)-suppressor of gamma response 1 (SOG1) signaling module. SOG1 mutation rescued the developmental defects of zdp ape2 leaves by preventing cell endoreplication and promoting cell proliferation. However, SOG1 mutation caused intensive meristematic cell death in the radicle of zdp ape2 following germination, resulting in rapid termination of radicle growth. Notably, mutating FORMAMIDOPYRIMIDINE DNA GLYCOSYLASE (FPG) in zdp ape2 sog1 partially recovered its radicle growth, demonstrating that DNA 3'-blocks generated by FPG caused the meristematic defects. Surprisingly, despite lacking a functional radicle, zdp ape2 sog1 mutants compensated the lack of root growth by generating anchor roots having low levels of DNA damage response. Our results reveal dual roles of SOG1 in regulating root establishment when seeds germinate with excess DNA 3'-blocks.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Reparo do DNA/fisiologia , Fatores de Transcrição/metabolismo , Arabidopsis/citologia , Proteínas de Arabidopsis/genética , Proteínas Mutadas de Ataxia Telangiectasia/genética , Morte Celular/genética , Proliferação de Células/genética , DNA de Plantas/genética , DNA de Plantas/metabolismo , DNA-Formamidopirimidina Glicosilase/metabolismo , Endonucleases/genética , Endonucleases/metabolismo , Regulação da Expressão Gênica de Plantas , Pleiotropia Genética , Germinação/genética , Meristema/citologia , Meristema/genética , Células Vegetais , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Sementes/fisiologia , Transdução de Sinais , Fatores de Transcrição/genéticaRESUMO
Cultivated sweet potato (Ipomoea batatas) is an important source of food for both humans and domesticated animals. Here, we show that the B-box (BBX) family transcription factor IbBBX24 regulates the jasmonic acid (JA) pathway in sweet potato. When IbBBX24 was overexpressed in sweet potato, JA accumulation increased, whereas silencing this gene decreased JA levels. RNA sequencing analysis revealed that IbBBX24 modulates the expression of genes involved in the JA pathway. IbBBX24 regulates JA responses by antagonizing the JA signaling repressor IbJAZ10, which relieves IbJAZ10's repression of IbMYC2, a JA signaling activator. IbBBX24 binds to the IbJAZ10 promoter and activates its transcription, whereas it represses the transcription of IbMYC2 The interaction between IbBBX24 and IbJAZ10 interferes with IbJAZ10's repression of IbMYC2, thereby promoting the transcriptional activity of IbMYC2. Overexpressing IbBBX24 significantly increased Fusarium wilt disease resistance, suggesting that JA responses play a crucial role in regulating Fusarium wilt resistance in sweet potato. Finally, overexpressing IbBBX24 led to increased yields in sweet potato. Together, our findings indicate that IbBBX24 plays a pivotal role in regulating JA biosynthesis and signaling and increasing Fusarium wilt resistance and yield in sweet potato, thus providing a candidate gene for developing elite crop varieties with enhanced pathogen resistance but without yield penalty.
Assuntos
Ciclopentanos/metabolismo , Resistência à Doença , Fusarium/fisiologia , Ipomoea batatas/imunologia , Ipomoea batatas/microbiologia , Oxilipinas/metabolismo , Doenças das Plantas/microbiologia , Proteínas de Plantas/metabolismo , Acetatos/farmacologia , Sequência de Bases , Ciclopentanos/farmacologia , DNA de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genoma de Planta , Ipomoea batatas/genética , Ipomoea batatas/crescimento & desenvolvimento , Modelos Biológicos , Oxilipinas/farmacologia , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas/genética , Ligação Proteica/efeitos dos fármacos , Nicotiana/genética , Nicotiana/microbiologia , Transcrição Gênica/efeitos dos fármacosRESUMO
Nitrogen (N) availability is a major limiting factor for plant growth and agricultural productivity. Although the gene regulation network in response to N starvation has been extensively studied, it remains unknown whether N starvation has an impact on the activity of transposable elements (TEs). Here, we report that TEs can be transcriptionally activated in Arabidopsis under N starvation conditions. Through genetic screening of idm1-14 suppressors, we cloned GLU1, which encodes a glutamate synthase that catalyzes the synthesis of glutamate in the primary N assimilation pathway. We found that glutamate synthase 1 (GLU1) and its functional homologs GLU2 and glutamate transport 1 (GLT1) are redundantly required for TE silencing, suggesting that N metabolism can regulate TE activity. Transcriptome and methylome analyses revealed that N starvation results in genome-wide TE activation without inducing obvious alteration of DNA methylation. Genetic analysis indicated that N starvation-induced TE activation is also independent of other well-established epigenetic mechanisms, including histone methylation and heterochromatin decondensation. Our results provide new insights into the regulation of TE activity under stressful environments in planta.
Assuntos
Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Elementos de DNA Transponíveis/genética , Inativação Gênica , Glutamato Sintase/genética , Metilação de DNA/genética , Glutamatos/genética , Glutamatos/metabolismo , Regulação da Expressão Gênica de Plantas/genéticaRESUMO
Sucrose non-fermenting-1-related protein kinase-1 (SnRK1) is an essential energy-sensing regulator and plays a key role in the global control of carbohydrate metabolism. The SnRK1 gene has been found to increase starch accumulation in several plant species. However, its roles in improving starch quality have not been reported to date. In this study, we found that the IbSnRK1 gene was highly expressed in the storage roots of sweet potato and strongly induced by exogenous sucrose. Its expression followed the circandian rhythm. Its overexpression not only increased starch content, but also decreased proportion of amylose, enlarged granule size and improved degree of crystallinity and gelatinization in transgenic sweet potato, which revealed, for the first time, the important roles of SnRK1 in improving starch quality of plants. The genes involved in starch biosynthesis pathway were systematically up-regulated, and the content of ADP-glucose as an important precursor for starch biosynthesis and the activities of key enzymes were significantly increased in transgenic sweet potato. These findings indicate that IbSnRK1 improves starch content and quality through systematical up-regulation of the genes and the increase in key enzyme activities involved in starch biosynthesis pathway in transgenic sweet potato. This gene has the potential to improve starch content and quality in sweet potato and other plants.
Assuntos
Genes de Plantas/genética , Ipomoea batatas/genética , Plantas Geneticamente Modificadas/genética , Amido/metabolismo , Amilose/metabolismo , Metabolismo dos Carboidratos/genética , Grânulos Citoplasmáticos/metabolismo , Regulação da Expressão Gênica de Plantas , Ipomoea batatas/química , Ipomoea batatas/metabolismo , Tubérculos/química , Tubérculos/metabolismo , Plantas Geneticamente Modificadas/química , Plantas Geneticamente Modificadas/metabolismo , Amido/análise , Sacarose/metabolismoRESUMO
Phenolic acids are known to reduce root biomass and hinder plant development, but it is unclear how they affect potato root traits. Over a 10 year field experiment, we found a negative correlation between the potato yield and continuous cropping years. The substantial reduction in adventitious root (AR) numbers was found to be primarily inhibited by soil vanillin accumulation. The study also found that vanillin had a more pronounced inhibitory effect on the potato yield than commonly reported ferulic acid and p-hydroxybenzoic acid. The decrease in yield was attributed to the reduction of root indole-3-acetic acid (IAA) content, which impeded the formation of AR. Exogenous IAA was found to increase the root IAA content and stimulate AR formation under vanillin stress, ultimately leading to an increase in the potato yield. This study provides valuable insights into potential strategies for the degradation of autotoxic substances and breeding of potato cultivars with enhanced resistance to autotoxicity.
RESUMO
High temperature is one of the major environmental stresses affecting plant growth and fitness. Heat stress transcription factors (HSFs) play critical roles in regulating the expression of heat-responsive genes. However, how HSFs are regulated remains obscure. Here, we show that ALBA4, ALBA5 and ALBA6, which phase separate into stress granules (SGs) and processing bodies (PBs) under heat stress, directly bind selected messenger RNAs, including HSF mRNAs, and recruit them into SGs and PBs to protect them from degradation under heat stress in Arabidopsis. The alba456 triple mutants, but not single and double mutants, display pleiotropic developmental defects and hypersensitivity to heat stress. Mutations in XRN4, a cytoplasmic 5' to 3' exoribonuclease, can rescue the observed developmental and heat-sensitive phenotypes of alba456 seedlings. Our study reveals a new layer of regulation for HSFs whereby HSF mRNAs are stabilized by redundant action of ALBA proteins in SGs and PBs for plant thermotolerance.