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1.
Commun Biol ; 7(1): 1085, 2024 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-39232040

RESUMO

The developmental plasticity of the root system plays an essential role in the adaptation of plants to the environment. Among many other signals, auxin and its directional, intercellular transport are critical in regulating root growth and development. In particular, the PIN-FORMED2 (PIN2) auxin exporter acts as a key regulator of root gravitropic growth. Multiple regulators have been reported to be involved in PIN2-mediated root growth; however, our information remains incomplete. Here, we identified ROWY Bro1-domain proteins as important regulators of PIN2 sorting control. Genetic analysis revealed that Arabidopsis rowy1 single mutants and higher-order rowy1 rowy2 rowy3 triple mutants presented a wavy root growth phenotype. Cell biological experiments revealed that ROWY1 and PIN2 colocalized to the apical side of the plasma membrane in the root epidermis and that ROWYs are required for correct PM targeting of PIN2. In addition, ROWYs also affected PIN3 protein abundance in the stele, suggesting the potential involvement of additional PIN transporters as well as other proteins. A global transcriptome analysis revealed that ROWY genes are involved in the Fe2+ availability perception pathway. This work establishes ROWYs as important novel regulators of root gravitropic growth by connecting micronutrient availability to the proper subcellular targeting of PIN auxin transporters.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Regulação da Expressão Gênica de Plantas , Gravitropismo , Raízes de Plantas , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Gravitropismo/genética , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Ácidos Indolacéticos/metabolismo , Mutação
2.
J Agric Food Chem ; 72(33): 18507-18519, 2024 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-39113497

RESUMO

Small secreted peptides (SSPs) are essential for defense mechanisms in plant-microbe interactions, acting as danger-associated molecular patterns (DAMPs). Despite the first discovery of SSPs over three decades ago, only a limited number of SSP families, particularly within Solanaceae plants, have been identified due to inefficient approaches. This study employed comparative genomics screens with Solanaceae proteomes (tomato, tobacco, and pepper) to discover a novel SSP family, SolP. Bioinformatics analysis suggests that SolP may serve as an endogenous signal initiating the plant PTI response. Interestingly, SolP family members from tomato, tobacco, and pepper share an identical sequence (VTSNALALVNRFAD), named SlSolP12 (also referred to as NtSolP15 or CaSolP1). Biochemical and phenotypic analyses revealed that synthetic SlSolP12 peptide triggers multiple defense responses: ROS burst, MAPK activation, callose deposition, stomatal closure, and expression of immune defense genes. Furthermore, SlSolP12 enhances systemic resistance against Botrytis cinerea infection in tomato plants and interferes with classical peptides, flg22 and Systemin, which modulate the immune response. Remarkably, SolP12 activates ROS in diverse plant species, such as Arabidopsis thaliana, soybean, and rice, showing a broad spectrum of biological activities. This study provides valuable approaches for identifying endogenous SSPs and highlights SlSolP12 as a novel DAMP that could serve as a useful target for crop protection.


Assuntos
Botrytis , Genômica , Doenças das Plantas , Imunidade Vegetal , Proteínas de Plantas , Solanum lycopersicum , Solanum lycopersicum/imunologia , Solanum lycopersicum/genética , Solanum lycopersicum/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/imunologia , Proteínas de Plantas/metabolismo , Doenças das Plantas/microbiologia , Doenças das Plantas/imunologia , Doenças das Plantas/genética , Imunidade Vegetal/genética , Peptídeos/imunologia , Peptídeos/química , Resistência à Doença/genética , Resistência à Doença/imunologia , Regulação da Expressão Gênica de Plantas , Nicotiana/imunologia , Nicotiana/genética , Nicotiana/microbiologia , Nicotiana/metabolismo , Capsicum/imunologia , Capsicum/genética , Capsicum/microbiologia , Capsicum/química
3.
Plant Sci ; 348: 112227, 2024 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-39173887

RESUMO

Torreya grandis, a dioecious Taxaceae species of significant economic value in southeast China, presents challenges for natural pollination due to asynchronous maturation of its sex organs and low pollen vitality. In order to enhance fertilization success through artificial pollination of T. grandis, this study investigated the optimal conditions for in vitro pollen germination and pollen tube growth of T. grandis. The optimal in vitro growth medium was found to contain 29 mM sucrose, 0.8 mM H3BO3, 0.72 mM CaCl2, and 0.32 mM MgSO4, supplemented with 4 µM NAA, 2 µM GA3, and 5 µM 2,4-D at pH=5.6. Under these conditions, we achieved a maximum pollen germination ratio of 69.99 ± 5.17 % and a pollen tube length of 34.38 ± 6.04 µm after 6 days germination at 28°C. FM4-64 dye and Mitotracker Red staining revealed highly dynamics of vesicles and mitochondria during germination, which were accumulated at the tip of pollen tube and exhibited biphasic movement patterns. The total number, motion rate, and movement velocity of vesicles as well as mitochondria showed an initially increase followed by a gradual decrease pattern. The presence of sucrose in the medium significantly increased the dynamics and metabolic activity of both vesicles and mitochondria, which may relate with higher pollen germination ratio and faster pollen tube growth compared to sucrose-depleted conditions. Thus, these findings shed light on the physiological characteristics of Torreya pollen germination and provide scientific information for improving Torreya fruit yield through artificial pollination.


Assuntos
Germinação , Tubo Polínico , Tubo Polínico/crescimento & desenvolvimento , Tubo Polínico/fisiologia , Tubo Polínico/metabolismo , Germinação/fisiologia , Pólen/crescimento & desenvolvimento , Pólen/fisiologia , Polinização , Sacarose/metabolismo
4.
Methods Mol Biol ; 2841: 157-164, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39115774

RESUMO

Agrobacterium-mediated transient expression is a flexible and efficient technique for introducing genes into plants, allowing for rapid and temporary gene expression. Agroinfiltration of Arabidopsis seedlings is a newly developed Agrobacterium-based transient expression system. The expression of target genes and the localization of relevant proteins can be observed within 3 days using this method. In this chapter, we present the detailed protocol for transient transformation in Arabidopsis thaliana seedlings utilizing vacuum infiltration of Agrobacterium. This procedure enables rapid and temporary gene expression by introducing exogenous DNA into Arabidopsis seedlings, particularly in easily accessible tissues such as cotyledons. This protocol provides a detailed description of experimental procedures, including Arabidopsis seedlings cultivation, the preparation of Agrobacterium suspensions, and subsequent steps leading to confocal microscope observation. Through this protocol, researchers can efficiently investigate gene function and subcellular localization in Arabidopsis cotyledons within 8 days in total.


Assuntos
Arabidopsis , Plântula , Arabidopsis/genética , Arabidopsis/metabolismo , Plântula/genética , Plântula/metabolismo , Plântula/crescimento & desenvolvimento , Vácuo , Cotilédone/genética , Cotilédone/metabolismo , Transformação Genética , Expressão Gênica , Plantas Geneticamente Modificadas/genética , Agrobacterium/genética , Regulação da Expressão Gênica de Plantas , Microscopia Confocal
5.
Methods Mol Biol ; 2841: 207-214, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39115780

RESUMO

High-pressure freezing/freeze substitution has been used to preserve biological samples for ultrastructure study instead of chemical fixation. For most plant samples, the water content is too high and cannot be properly preserved during cryofixation. Additionally, the cell wall is a barrier that prevents the substitution of water with the resin. In this chapter, we will discuss modified high-pressure freezing and subsequent processing protocols based on our routinely used methodology for examining Arabidopsis seeds in transmission electron microscopy and electron tomography.


Assuntos
Arabidopsis , Tomografia com Microscopia Eletrônica , Sementes , Sementes/ultraestrutura , Tomografia com Microscopia Eletrônica/métodos , Arabidopsis/ultraestrutura , Congelamento , Temperatura Baixa , Pressão , Microscopia Eletrônica de Transmissão/métodos , Criopreservação/métodos , Substituição ao Congelamento/métodos
6.
Toxins (Basel) ; 16(8)2024 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-39195768

RESUMO

HxTx-Hv1h, a neurotoxic peptide derived from spider venom, has been developed for use in commercial biopesticide formulations. Cell Penetrating Peptides (CPPs) are short peptides that facilitate the translocation of various biomolecules across cellular membranes. Here, we evaluated the aphidicidal efficacy of a conjugated peptide, HxTx-Hv1h/CPP-1838, created by fusing HxTx-Hv1h with CPP-1838. Additionally, we aimed to establish a robust recombinant expression system for HxTx-Hv1h/CPP-1838. We successfully achieved the secretory production of HxTx-Hv1h, its fusion with Galanthus nivalis agglutinin (GNA) forming HxTx-Hv1h/GNA and HxTx-Hv1h/CPP-1838 in yeast. Purified HxTx-Hv1h exhibited contact toxicity against Megoura crassicauda, with a 48 h median lethal concentration (LC50) of 860.5 µg/mL. Fusion with GNA or CPP-1838 significantly enhanced its aphidicidal potency, reducing the LC50 to 683.5 µg/mL and 465.2 µg/mL, respectively. The aphidicidal efficacy was further improved with the addition of surfactant, decreasing the LC50 of HxTx-Hv1h/CPP-1838 to 66.7 µg/mL-over four times lower compared to HxTx-Hv1h alone. Furthermore, we engineered HxTx-Hv1h/CPP-1838 multi-copy expression vectors utilizing the BglBrick assembly method and achieved high-level recombinant production in laboratory-scale fermentation. This study is the first to document a CPP fusion strategy that enhances the transdermal aphidicidal activity of a natural toxin like HxTx-Hv1h and opens up the possibility of exploring the recombinant production of HxTx-Hv1h/CPP-1838 for potential applications.


Assuntos
Peptídeos Penetradores de Células , Neurotoxinas , Venenos de Aranha , Venenos de Aranha/química , Venenos de Aranha/genética , Venenos de Aranha/toxicidade , Animais , Peptídeos Penetradores de Células/farmacologia , Peptídeos Penetradores de Células/química , Neurotoxinas/toxicidade , Neurotoxinas/farmacologia , Neurotoxinas/genética , Lectinas de Plantas/farmacologia , Lectinas de Plantas/genética , Lectinas de Plantas/química , Inseticidas/farmacologia , Inseticidas/toxicidade
9.
J Agric Food Chem ; 2024 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-38836320

RESUMO

The CLE (CLAVATA3/Embryo Surrounding Region-related) family, a group of peptides with hormone-like features, plays a pivotal role in plant growth, development, and adaptation to stress. Through homology-based blast analysis of 32 Arabidopsis thaliana CLE peptide sequences, we have identified 5, 14, and 10 CLE family members in Nicotiana tabacum, Capsicum annuum, and Solanum melongena, respectively. Chemical synthesis and functional assays of the peptides led to the discovery that NtCLE3 substantially enhances the drought resistance of these three Solanaceae crops. Our transcriptome, RT-qPCR, and antioxidant enzyme activity data showed that NtCLE3 increased antioxidant capacity and ABA synthesis in tobacco. Moreover, the recombinant protein RPNtCLE3, composed of 6*NtCLE3, preserved the capacity to foster drought resilience and proved to be a promising drought resistance regulator, which presents a more favorable alternative for field applications compared to ABA which degrades rapidly under sunlight exposure. This research unveils the prospective utility of NtCLE3 in enhancing drought tolerance in Solanaceae crops and provides new ideas for the development of novel bioregulators aimed at mitigating drought stress.

10.
Front Public Health ; 12: 1352894, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38887255

RESUMO

Aim: In China, with the increase of life expectancy and the decrease of fertility rate, the aging problem has become increasingly prominent, and the physical problems of the older people over 70 years are the key and difficult problems. Method: Based on the interactive logic between the aging problem and the older people health, in the study, a questionnaire survey and a nationwide physical fitness test were carried out on the older people over 70, to divide into different age groups (70-74 years old, 75-79 years old, 80-84 years old, 85 years old and older) and different genders. There were 8,400 valid samples, and 1,050 persons in each group. One-way ANOVA was used to compare the differences among groups of different ages, and a broken line chart was drawn to discuss the aging characteristics of various physical indexes of the older people over 70 in China. Result: (1) Body morphology: male waist circumference, male waist-to-height ratio and female BMI showed a gradual downward trend with the increase of age; (2) Physiological function: male and female vital capacity showed a decreasing trend with the increase of age, while female pulse pressure showed a gradual upward trend. (3) Physical quality: the indicators of male and female muscle strength, flexibility quality, aerobic endurance and balance showed a downward trend with the increase of age. Conclusion: Vital capacity, flexibility quality, muscle strength, aerobic endurance, balance ability and so on, decreased significantly with the growth of age. 80 years old is the inflection point of the rapid decline of various indicators. Blood pressure, silent pulse, BMI, waist-to-hip ratio, waist-to-height ratio and other indicators did not change regularly with age. Indicators such as blood pressure, BMI, waist-to-hip ratio and waist-to-height ratio were in the high-risk range of metabolic diseases and cardiovascular and cerebrovascular diseases. The study conducted physical fitness test on the older people over 70 years old in 7 geographical regions of China, which is the first nationwide physical fitness test for the older people, which is an extension and expansion of the national physical fitness monitoring system, and also shows that the test indicators involved in the "Health fitness scale" are simple and feasible. And the study added a series of test data over 70 years old, which is the basis for scientific and reasonable formulation of physical fitness evaluation standards for the older people, and is of great significance for improving the national physical fitness database and grasping the dynamic changes of national physical health status, and providing data support for scientific guidance of physical exercise for the older people.


Assuntos
Envelhecimento , Aptidão Física , Humanos , Idoso , Masculino , China/epidemiologia , Feminino , Idoso de 80 Anos ou mais , Aptidão Física/fisiologia , Envelhecimento/fisiologia , Inquéritos e Questionários , Índice de Massa Corporal , Força Muscular/fisiologia
11.
Small Methods ; : e2400312, 2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38654560

RESUMO

Gate-controlled ionic intercalation in the van der Waals gap of 2D layered materials can induce novel phases and unlock new properties. However, this strategy is often unsuitable for densely packed 2D non-layered materials. The non-layered rhombohedral Cr2S3 is an intrinsic heterodimensional superlattice with alternating layers of 2D CrS2 and 0D Cr1/3. Here an innovative chemical vapor deposition method is reported, utilizing strategically modified metal precursors to initiate entirely new seed layers, yields ultrathin inclined-standing grown 2D Cr2S3 nanosheets with edge instead of face contact with substrate surfaces, enabling rapid all-dry transfer to other substrates while ensuring high crystal quality. The unconventional ordered vacancy channels within the 0D Cr1/3 layers, as revealed by cross-sectional scanning transmission electron microscope, permitting the insertion of Li+ ions. An unprecedented metal-insulator transition, with a resistance modulation of up to six orders of magnitude at 300 K, is observed in Cr2S3-based ionic field-effect transistors. Theoretical calculations corroborate the metallization induced by Li-ion intercalation. This work sheds light on the understanding of growth mechanism, structure-property correlation and highlights the diverse potential applications of 2D non-layered Cr2S3 superlattice.

12.
Mol Plant Pathol ; 25(4): e13447, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38561315

RESUMO

Genetic engineering using negative regulators of plant immunity has the potential to provide a huge impetus in agricultural biotechnology to achieve a higher degree of disease resistance without reducing yield. Type 2C protein phosphatases (PP2Cs) represent the largest group of protein phosphatases in plants, with a high potential for negative regulatory functions by blocking the transmission of defence signals through dephosphorylation. Here, we established a PP2C functional protoplast screen using pFRK1::luciferase as a reporter and found that 14 of 56 PP2Cs significantly inhibited the immune response induced by flg22. To verify the reliability of the system, a previously reported MAPK3/4/6-interacting protein phosphatase, PP2C5, was used; it was confirmed to be a negative regulator of PAMP-triggered immunity (PTI). We further identified PP2C15 as an interacting partner of BRI1-associated receptor kinase 1 (BAK1), which is the most well-known co-receptor of plasma membrane-localized pattern recognition receptors (PRRs), and a central component of PTI. PP2C15 dephosphorylates BAK1 and negatively regulates BAK1-mediated PTI responses such as MAPK3/4/6 activation, defence gene expression, reactive oxygen species bursts, stomatal immunity, callose deposition, and pathogen resistance. Although plant growth and 1000-seed weight of pp2c15 mutants were reduced compared to those of wild-type plants, pp2c5 mutants did not show any adverse effects. Thus, our findings strengthen the understanding of the mechanism by which PP2C family members negatively regulate plant immunity at multiple levels and indicate a possible approach to enhance plant resistance by eliminating specific PP2Cs without affecting plant growth and yield.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Reprodutibilidade dos Testes , Fosfoproteínas Fosfatases/genética , Fosfoproteínas Fosfatases/metabolismo , Fosfoproteínas Fosfatases/farmacologia , Imunidade Vegetal/fisiologia , Regulação da Expressão Gênica de Plantas , Proteínas Quinases/genética , Proteínas Quinases/metabolismo
13.
Front Plant Sci ; 14: 1208549, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38078092

RESUMO

PAMP-induced secreted peptide (PIP), one of the small post-translationally modified peptides (PTMPs), plays a crucial role in plant development and stress tolerance. However, little is known about functional divergence among this peptide family. Here, we studied the evolution of the PIP family in 23 plant species (10 monocotyledons and 13 dicotyledons from 7 families) and their functional divergence in Arabidopsis. A total of 128 putative PIP precursors were identified and classified into two subfamilies through phylogenetic analysis. Functional studies on AtPIP1 which represents Clade I family and AtPIP2 which represents Clade II family have shown that AtPIP2 displayed stronger immunity induction activity but weaker root growth inhibition than AtPIP1 in Arabidopsis. Transcriptome analysis of Arabidopsis seedlings treated with AtPIP1 and AtPIP2 showed that differential genes for both polypeptides were significantly enriched in similar plant defense pathways. However, Co-expression and Protein-protein interaction (PPI) analysis showed that the functions of AtprePIP2 co-expressed genes were more enriched in plant defense pathways than AtprePIP1. Molecular docking results show that AtPIP1 binds to RLK7 receptor with a more stable free energy and less binding area than AtPIP2, while hydrogen bond transfer occurs at the SGP motif position. The above results suggest that the PIP family have undergone functional divergence during evolution. Collectively, this work illustrates the relationship between PIP structure and function using Arabidopsis PIP as an example, and provides new insights into the current understanding between growth inhibition and immune responses which may be correlated but not fully coupled.

14.
Plant Cell ; 35(12): 4217-4237, 2023 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-37647529

RESUMO

Membrane protein homeostasis is fine-tuned by the cellular pathways for vacuolar degradation and recycling, which ultimately facilitate plant growth and cell-environment interactions. The endosomal sorting complex required for transport (ESCRT) machinery plays important roles in regulating intraluminal vesicle (ILV) formation and membrane protein sorting to vacuoles. We previously showed that the plant-specific ESCRT component FYVE DOMAIN PROTEIN REQUIRED FOR ENDOSOMAL SORTING1 (FREE1) performs multiple functions in plants, although the underlying mechanisms remain elusive. In this study, we performed a suppressor screen of the FREE1-RNAi mutant and identified and characterized 2 suppressor of free1 (sof) mutants in Arabidopsis (Arabidopsis thaliana). These mutants, sof10 and sof641, result in a premature stop codon or a missense mutation in AT5G10370, respectively. This gene was named DEAH and RING domain-containing protein as FREE1 suppressor 1 (DRIF1). DRIF1 has a homologous gene, DRIF2, in the Arabidopsis genome with 95% identity to DRIF1. The embryos of drif1 drif2 mutants arrested at the globular stage and formed enlarged multivesicular bodies (MVBs) with an increased number of ILVs. DRIF1 is a membrane-associated protein that coordinates with retromer component sorting nexin 1 to regulate PIN-FORMED2 recycling to the plasma membrane. Altogether, our data demonstrate that DRIF1 is a unique retromer interactor that orchestrates FREE1-mediated ILV formation of MVBs and vacuolar sorting of membrane proteins for degradation in plants.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Nexinas de Classificação/genética , Nexinas de Classificação/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Plantas/metabolismo , Complexos Endossomais de Distribuição Requeridos para Transporte/metabolismo , Proteostase , Transporte Proteico/genética , Plantas/metabolismo , Proteínas de Transporte Vesicular/genética , Proteínas de Transporte Vesicular/metabolismo
15.
New Phytol ; 240(1): 41-60, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37507353

RESUMO

The endomembrane system consists of various membrane-bound organelles including the endoplasmic reticulum (ER), Golgi apparatus, trans-Golgi network (TGN), endosomes, and the lysosome/vacuole. Membrane trafficking between distinct compartments is mainly achieved by vesicular transport. As the endomembrane compartments and the machineries regulating the membrane trafficking are largely conserved across all eukaryotes, our current knowledge on organelle biogenesis and endomembrane trafficking in plants has mainly been shaped by corresponding studies in mammals and yeast. However, unique perspectives have emerged from plant cell biology research through the characterization of plant-specific regulators as well as the development and application of the state-of-the-art microscopical techniques. In this review, we summarize our current knowledge on the plant endomembrane system, with a focus on several distinct pathways: ER-to-Golgi transport, protein sorting at the TGN, endosomal sorting on multivesicular bodies, vacuolar trafficking/vacuole biogenesis, and the autophagy pathway. We also give an update on advanced imaging techniques for the plant cell biology research.


Assuntos
Endossomos , Plantas , Plantas/metabolismo , Endossomos/metabolismo , Vacúolos/metabolismo , Corpos Multivesiculares/metabolismo , Transporte Proteico , Complexo de Golgi/metabolismo , Rede trans-Golgi/metabolismo
16.
New Phytol ; 239(5): 1740-1753, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37301984

RESUMO

Pollen tubes have dynamic tubular vacuoles. Functional loss of AP-3, a regulator of one vacuolar trafficking route, reduces pollen tube growth. However, the role of canonical Rab5 GTPases that are responsible for two other vacuolar trafficking routes in Arabidopsis pollen tubes is obscure. By using genomic editing, confocal microscopy, pollen tube growth assays, and transmission electron microscopy, we demonstrate that functional loss of canonical Rab5s in Arabidopsis, RHA1 and ARA7, causes the failure of pollen tubes to grow through style and thus impairs male transmission. Functional loss of canonical Rab5s compromises vacuolar trafficking of tonoplast proteins, vacuolar biogenesis, and turgor regulation. However, rha1;ara7 pollen tubes are comparable to those of wild-type in growing through narrow passages by microfluidic assays. We demonstrate that functional loss of canonical Rab5s compromises endocytic and secretory trafficking at the plasma membrane (PM), whereas the targeting of PM-associated ATPases is largely unaffected. Despite that, rha1;ara7 pollen tubes contain a reduced cytosolic pH and disrupted actin microfilaments, correlating with the mis-targeting of vacuolar ATPases (VHA). These results imply a key role of vacuoles in maintaining cytoplasmic proton homeostasis and in pollen tube penetrative growth through style.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Tubo Polínico , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , GTP Fosfo-Hidrolases/metabolismo , Adenosina Trifosfatases/metabolismo
17.
J Integr Plant Biol ; 65(9): 2157-2174, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37252889

RESUMO

Arabidopsis plastid antiporters KEA1 and KEA2 are critical for plastid development, photosynthetic efficiency, and plant development. Here, we show that KEA1 and KEA2 are involved in vacuolar protein trafficking. Genetic analyses found that the kea1 kea2 mutants had short siliques, small seeds, and short seedlings. Molecular and biochemical assays showed that seed storage proteins were missorted out of the cell and the precursor proteins were accumulated in kea1 kea2. Protein storage vacuoles (PSVs) were smaller in kea1 kea2. Further analyses showed that endosomal trafficking in kea1 kea2 was compromised. Vacuolar sorting receptor 1 (VSR1) subcellular localizations, VSR-cargo interactions, and p24 distribution on the endoplasmic reticulum (ER) and Golgi apparatus were affected in kea1 kea2. Moreover, plastid stromule growth was reduced and plastid association with the endomembrane compartments was disrupted in kea1 kea2. Stromule growth was regulated by the cellular pH and K+ homeostasis maintained by KEA1 and KEA2. The organellar pH along the trafficking pathway was altered in kea1 kea2. Overall, KEA1 and KEA2 regulate vacuolar trafficking by controlling the function of plastid stromules via adjusting pH and K+ homeostasis.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Antiporters/genética , Antiporters/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Vacúolos/metabolismo , Plastídeos/metabolismo , Cátions/metabolismo , Transporte Proteico
18.
Nat Commun ; 14(1): 1768, 2023 03 30.
Artigo em Inglês | MEDLINE | ID: mdl-36997511

RESUMO

The energy sensor AMP-activated protein kinase (AMPK) can activate autophagy when cellular energy production becomes compromised. However, the degree to which nutrient sensing impinges on the autophagosome closure remains unknown. Here, we provide the mechanism underlying a plant unique protein FREE1, upon autophagy-induced SnRK1α1-mediated phosphorylation, functions as a linkage between ATG conjugation system and ESCRT machinery to regulate the autophagosome closure upon nutrient deprivation. Using high-resolution microscopy, 3D-electron tomography, and protease protection assay, we showed that unclosed autophagosomes accumulated in free1 mutants. Proteomic, cellular and biochemical analysis revealed the mechanistic connection between FREE1 and the ATG conjugation system/ESCRT-III complex in regulating autophagosome closure. Mass spectrometry analysis showed that the evolutionary conserved plant energy sensor SnRK1α1 phosphorylates FREE1 and recruits it to the autophagosomes to promote closure. Mutagenesis of the phosphorylation site on FREE1 caused the autophagosome closure failure. Our findings unveil how cellular energy sensing pathways regulate autophagosome closure to maintain cellular homeostasis.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Autofagossomos , Proteínas de Transporte Vesicular , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Complexos Endossomais de Distribuição Requeridos para Transporte/metabolismo , Proteínas de Transporte Vesicular/química , Proteínas de Transporte Vesicular/metabolismo , Motivos de Aminoácidos , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo
19.
Eur J Cell Biol ; 102(2): 151309, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36933283

RESUMO

The study of endomembrane trafficking is crucial for understanding how cells and whole organisms function. Moreover, there is a special interest in investigating endomembrane trafficking in plants, given its role in transport and accumulation of seed storage proteins and in secretion of cell wall material, arguably the two most essential commodities obtained from crops. The mechanisms of anterograde transport in the biosynthetic and endocytic pathways of plants have been thoroughly discussed in recent reviews, but, comparatively, retrograde trafficking pathways have received less attention. Retrograde trafficking is essential to recover membranes, retrieve proteins that have escaped from their intended localization, maintain homeostasis in maturing compartments, and recycle trafficking machinery for its reuse in anterograde transport reactions. Here, we review the current understanding on retrograde trafficking pathways in the endomembrane system of plants, discussing their integration with anterograde transport routes, describing conserved and plant-specific retrieval mechanisms at play, highlighting contentious issues and identifying open questions for future research.


Assuntos
Plantas , Transporte Biológico , Plantas/metabolismo , Transporte Proteico
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