المؤلفون: Li, Chunlong, Krishnan, Srinivasan, Zhang, Mengxia, Hu, Dagang, Meng, Dong, Riedelsberger, Janin, Dougherty, Laura, Xu, Kenong, Piñeros, Miguel A., Cheng, Lailiang

المصدر: Advanced Science ; volume 11, issue 22 ; ISSN 2198-3844 2198-3844

الوصف: Vacuolar malic acid accumulation largely determines fruit acidity, a key trait for the taste and flavor of apple and other fleshy fruits. Aluminum‐activated malate transporter 9 ( ALMT9 / Ma1 ) underlies a major genetic locus, Ma , for fruit acidity in apple, but how the protein transports malate across the tonoplast is unclear. Here, it is shown that overexpression of the coding sequence of Ma1 ( Ma1α ) drastically decreases fruit acidity in “Royal Gala” apple, leading to uncovering alternative splicing underpins Ma1's function. Alternative splicing generates two isoforms: Ma1β is 68 amino acids shorter with much lower expression than the full‐length protein Ma1α. Ma1β does not transport malate itself but interacts with the functional Ma1α to form heterodimers, creating synergy with Ma1α for malate transport in a threshold manner (When Ma1β/Ma1α ≥ 1/8). Overexpression of Ma1α triggers feedback inhibition on the native Ma1 expression via transcription factor MYB73, decreasing the Ma1β level well below the threshold that leads to significant reductions in Ma1 function and malic acid accumulation in fruit. Overexpression of Ma1α and Ma1β or genomic Ma1 increases both isoforms proportionally and enhances fruit malic acid accumulation. These findings reveal an essential role of alternative splicing in ALMT9‐mediated malate transport underlying apple fruit acidity.

الإتاحة: https://doi.org/10.1002/advs.202310159Test

المؤلفون: Maity, Koustav, Heumann, John M, McGrath, Aaron P, Kopcho, Noah J, Hsu, Po-Kai, Lee, Chang-Wook, Mapes, James H, Garza, Denisse, Krishnan, Srinivasan, Morgan, Garry P, Hendargo, Kevin J, Klose, Thomas, Rees, Steven D, Medrano-Soto, Arturo, Saier, Milton H, Piñeros, Miguel, Komives, Elizabeth A, Schroeder, Julian I, Chang, Geoffrey, Stowell, Michael HB

المصدر: Proceedings of the National Academy of Sciences of the United States of America. 116(28)

مصطلحات موضوعية: Biochemistry and Cell Biology, Biological Sciences, 1.1 Normal biological development and functioning, Underpinning research, Generic health relevance, Amino Acid Sequence, Anoctamins, Arabidopsis, Arabidopsis Proteins, Calcium Channels, Cryoelectron Microscopy, Cytoplasm, Mass Spectrometry, Membrane Potentials, Oryza, Osmotic Pressure, Protein Conformation, Water, osmotic stress, channel, structure, cryo-EM, rice

الوصف: Sensing and responding to environmental water deficiency and osmotic stresses are essential for the growth, development, and survival of plants. Recently, an osmolality-sensing ion channel called OSCA1 was discovered that functions in sensing hyperosmolality in Arabidopsis Here, we report the cryo-electron microscopy (cryo-EM) structure and function of an OSCA1 homolog from rice (Oryza sativa; OsOSCA1.2), leading to a model of how it could mediate hyperosmolality sensing and transport pathway gating. The structure reveals a dimer; the molecular architecture of each subunit consists of 11 transmembrane (TM) helices and a cytosolic soluble domain that has homology to RNA recognition proteins. The TM domain is structurally related to the TMEM16 family of calcium-dependent ion channels and lipid scramblases. The cytosolic soluble domain possesses a distinct structural feature in the form of extended intracellular helical arms that are parallel to the plasma membrane. These helical arms are well positioned to potentially sense lateral tension on the inner leaflet of the lipid bilayer caused by changes in turgor pressure. Computational dynamic analysis suggests how this domain couples to the TM portion of the molecule to open a transport pathway. Hydrogen/deuterium exchange mass spectrometry (HDXMS) experimentally confirms the conformational dynamics of these coupled domains. These studies provide a framework to understand the structural basis of proposed hyperosmolality sensing in a staple crop plant, extend our knowledge of the anoctamin superfamily important for plants and fungi, and provide a structural mechanism for potentially translating membrane stress to transport regulation.

وصف الملف: application/pdf

الوصول الحر: https://escholarship.org/uc/item/50v441jmTest

المؤلفون: Scinto-Madonich, Nathan J., Baruah, Shivranjani, Young, Sameya, Vignona, Katherine, Read, Andrew C., Carpenter, Sara C.D., Wang, Li, Shi, Xinying, Chang, Geoffrey, Piñeros, Miguel A., Bogdanove, Adam J.

المصدر: Physiological and Molecular Plant Pathology ; volume 126, page 102031 ; ISSN 0885-5765

مصطلحات موضوعية: Plant Science, Genetics

الإتاحة: https://doi.org/10.1016/j.pmpp.2023.102031Test
https://api.elsevier.com/content/article/PII:S0885576523000863?httpAccept=text/xmlTest
https://api.elsevier.com/content/article/PII:S0885576523000863?httpAccept=text/plainTest

المؤلفون: Doshi, Rupak, McGrath, Aaron P, Piñeros, Miguel, Szewczyk, Paul, Garza, Denisse M, Kochian, Leon V, Chang, Geoffrey

المصدر: Scientific reports. 7(1)

مصطلحات موضوعية: Sorghum, Plant Roots, Aluminum, Membrane Transport Proteins, Plant Proteins, Phylogeny, Substrate Specificity

الوصف: About 50% of the world's arable land is strongly acidic (pH ≤ 5). The low pH solubilizes root-toxic ionic aluminium (Al3+) species from clay minerals, driving the evolution of counteractive adaptations in cultivated crops. The food crop Sorghum bicolor upregulates the membrane-embedded transporter protein SbMATE in its roots. SbMATE mediates efflux of the anionic form of the organic acid, citrate, into the soil rhizosphere, chelating Al3+ ions and thereby imparting Al-resistance based on excluding Al+3 from the growing root tip. Here, we use electrophysiological, radiolabeled, and fluorescence-based transport assays in two heterologous expression systems to establish a broad substrate recognition profile of SbMATE, showing the proton and/or sodium-driven transport of 14C-citrate anion, as well as the organic monovalent cation, ethidium, but not its divalent analog, propidium. We further complement our transport assays by measuring substrate binding to detergent-purified SbMATE protein. Finally, we use the purified membrane protein as an antigen to discover native conformation-binding and transport function-altering nanobodies using an animal-free, mRNA/cDNA display technology. Our results demonstrate the utility of using Pichia pastoris as an efficient eukaryotic host to express large quantities of functional plant transporter proteins. The nanobody discovery approach is applicable to other non-immunogenic plant proteins.

وصف الملف: application/pdf

الوصول الحر: https://escholarship.org/uc/item/69d5x7k3Test

المؤلفون: Huber, Annika E., Melcher, Peter J., Piñeros, Miguel A., Setter, Tim L., Bauerle, Taryn L.

المصدر: The New Phytologist, 2019 Oct 01. 224(2), 675-688.

الوصول الحر: https://www.jstor.org/stable/26800776Test

المؤلفون: Stuebler, Martin, Manzer, Zachary A., Liu, Han-Yuan, Miller, Julia, Richter, Annett, Krishnan, Srinivasan, Selivanovitch, Ekaterina, Banuna, Barituziga, Jander, Georg, Reimhult, Erik, Zipfel, Warren R., Roeder, Adrienne H. K., Piñeros, Miguel A., Daniel, Susan

المساهمون: United States - Israel Binational Agricultural Research and Development Fund, Deutsche Forschungsgemeinschaft, Cornell University, National Science Foundation

المصدر: ACS Applied Materials & Interfaces ; ISSN 1944-8244 1944-8252

الإتاحة: https://doi.org/10.1021/acsami.3c18562Test

المؤلفون: Koyama, Hiroyuki, Huang, Chao-Feng, Piñeros, Miguel A., Yamamoto, Yoko

المصدر: Frontiers in Plant Science ; volume 13 ; ISSN 1664-462X

مصطلحات موضوعية: Plant Science

الإتاحة: https://doi.org/10.3389/fpls.2022.925541Test

المؤلفون: Ishka, Maryam Rahmati, Sussman, Hayley, Hu, Yunfei, Alqahtani, Mashael Daghash Saeed, Craft, Eric, Sicat, Ronell Barrera, Wang, Minmin, Yu, Li'ang, Ait-Haddou, Rachid, Li, Bo, Drakakaki, Georgia, Nelson, Andrew, Pineros, Miguel, Korte, Arthur, Jaremko, Lukasz, Testerink, Christa, Tester, Mark A., Julkowska, Magdalena

المساهمون: Bioscience, Bioscience Program, Biological, Environmental Sciences and Engineering, Biological and Environmental Science and Engineering (BESE) Division, Visualization, Visual Computing Center (VCC), Computer, Electrical and Mathematical Sciences and Engineering, Computer, Electrical and Mathematical Science and Engineering (CEMSE) Division, Plant Science, Center for Desert Agriculture, Water Desalination & Reuse Center, Water Desalination and Reuse Research Center (WDRC), Visualization Laboratory, KAUST Visualization Laboratory (KVL), Boyce Thompson Institute, Ithaca, NY, USA, Lanzhou University, Lanzhou, China, USDA-ARS, Ithaca, NY, USA, UC Davis, Davis, CA, USA, Julius-von-Sachs-Institute & Center for Computational and Theoretical Biology, JMU, Wuerzburg, Germany, Wageningen Research and University, Wageningen, The Netherlands

الوصف: Soil salinity is one of the major threats to agricultural productivity worldwide. Salt stress exposure alters root and shoot growth rates, thereby affecting overall plant performance. While past studies have extensively documented the effect of salt stress on root elongation and shoot development separately, here we take an innovative approach by examining the coordination of root and shoot growth under salt stress conditions. Utilizing a newly developed tool for quantifying the root:shoot ratio in agar-grown Arabidopsis seedlings, we found that salt stress results in a loss of coordination between root and shoot growth rates. We identify a specific gene cluster encoding domain-of-unknown-function 247 (DUF247), and characterize one of these genes as Salt Root:shoot Ratio Regulator Gene (SR3G). Further analysis elucidates the role of SR3G as a negative regulator of salt stress tolerance, revealing its function in regulating shoot growth, root suberization, and sodium accumulation. We further characterize that SR3G expression is modulated by WRKY75 transcription factor, known as a positive regulator of salt stress tolerance. Finally, we show that the salt stress sensitivity of wrky75 mutant is completely diminished when it is combined with sr3g mutation. Together, our results demonstrate that utilizing root:shoot ratio as an architectural feature leads to the discovery of new stress resilience gene. The study's innovative approach and findings not only contribute to our understanding of plant stress tolerance mechanisms but also open new avenues for genetic and agronomic strategies to enhance crop environmental resilience. ; The authors would like to thank BTI and KAUST Greenhouse Teams for their care of the plants. The authors would like to acknowledge support from the NSF-IOS #2023310 (ADLN) and NSF-IOS #2102120 (ADLN). The majority of funding for this work was generously provided from KAUST baseline funding awarded to Mark Tester, and BTI’s startup funds awarded to Magdalena Julkowska.

وصف الملف: application/pdf

العلاقة: http://hdl.handle.net/10754/698041Test

الإتاحة: https://doi.org/10.1101/2024.04.09.588564Test
http://hdl.handle.net/10754/698041Test

المؤلفون: Sharma, Tripti, Dreyer, Ingo, Kochian, Leon, Pineros, Miguel A

الوصف: About a decade ago, members of a new protein family of anion channels were discovered on the basis of their ability to confer on plants the tolerance toward toxic aluminum ions in the soil. The efflux of Al3+-chelating malate anions through these channels is stimulated by external Al3+ ions. This feature of a few proteins determined the name of the entire protein family as Aluminum-activated Malate Transporters (ALMT). Meanwhile, after several years of research, it is known that the physiological roles of ALMTs go far beyond Al-detoxification. In this review article we summarize the current knowledge on this transporter family and assess their involvement in diverse physiological processes. Keywords. Author Keywords:anion channel ; ALMT ; aluminum tolerance ; nutrient transport ; malate transport ; citrate transport ; review ; Regular 2015 ; FONDECYT ; FONDECYT

العلاقة: handle/10533/111557; handle/10533/111541; handle/10533/108045; https://doi.org/10.3389/fpls.2016.01488Test; 1150054; WOS:000384692200001; https://hdl.handle.net/10533/250538Test

الإتاحة: https://doi.org/10.3389/fpls.2016.01488Test
https://hdl.handle.net/10533/250538Test

المؤلفون: Riedelsberger, Janin, Vergara-Jaque, Ariela, Pineros, Miguel, Dreyer, Ingo, González, Wendy

الوصف: BackgroundHKT channels mediate sodium uniport or sodium and potassium symport in plants. Monocotyledons express a higher number of HKT proteins than dicotyledons, and it is only within this clade of HKT channels that cation symport mechanisms are found. The prevailing ion composition in the extracellular medium affects the transport abilities of various HKT channels by changing their selectivity or ion transport rates. How this mutual effect is achieved at the molecular level is still unknown. Here, we built a homology model of the monocotyledonous OsHKT2;2, which shows sodium and potassium symport activity. We performed molecular dynamics simulations in the presence of sodium and potassium ions to investigate the mutual effect of cation species.ResultsBy analyzing ion-protein interactions, we identified a cation coordination site on the extracellular protein surface, which is formed by residues P71, D75, D501 and K504. Proline and the two aspartate residues coordinate cations, while K504 forms salt bridges with D75 and D501 and may be involved in the forwarding of cations towards the pore entrance. Functional validation via electrophysiological experiments confirmed the biological relevance of the predicted ion coordination site and identified K504 as a central key residue. Mutation of the cation coordinating residues affected the functionality of HKT only slightly. Additional in silico mutants and simulations of K504 supported experimental results.ConclusionWe identified an extracellular cation coordination site, which is involved in ion coordination and influences the conduction of OsHKT2;2. This finding proposes a new viewpoint in the discussion of how the mutual effect of variable ion species may be achieved in HKT channels. Keywords Author Keywords:Ion channel ; HKT ; Sodium transport ; Potassium transport ; Ion coordination site ; Plant ; Structure-function ; Regular 2015 ; FONDECYT ; FONDECYT

العلاقة: handle/10533/111557; handle/10533/111541; handle/10533/108045; https://doi.org/10.1186/s12870-019-1909-5Test; 1150054; WOS:000475743400006; https://hdl.handle.net/10533/250427Test

الإتاحة: https://doi.org/10.1186/s12870-019-1909-5Test
https://hdl.handle.net/10533/250427Test