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In this study, agronomic and functional characteristics of potato (Solanum tuberosum ) plants constitutively overexpressing the protein phosphatase 2A (PP2A) catalytic subunit StPP2Ac2b (StPP2Ac2b-OE) were evaluated. StPP2Ac2b-OE plants display reduced vegetative growth, tuber yield and tuber weight under well-watered and drought conditions. Leaves of StPP2Ac2b-OE plants show an increased rate of water loss, associated with an impaired ability to close stomata in response to abscisic acid. StPP2Ac2b-OE lines exhibit larger stomatal size and reduced stomatal density. These altered stomatal characteristics might be responsible for the impaired stomatal closure and the elevated transpiration rates, ultimately leading to increased sensitivity to water-deficit stress and greater yield loss under drought conditions. Overexpression of StPP2Ac2b accelerates senescence in response to water-deficit stress, which could also contribute to the increased sensitivity to drought. Actively photosynthesising leaves of StPP2Ac2b-OE plants exhibit elevated levels of carbohydrates and a down-regulation of the sucrose transporter StSWEET11 , suggesting a reduced sucrose export from leaves to developing tubers. This effect, combined with the hindered vegetative development, may contribute to the reduced tuber weight and yield in StPP2Ac2b-OE plants. These findings offer novel insights into the physiological functions of PP2A in potato plants and provide valuable information for enhancing potato productivity by modulating the expression of StPP2Ac2b .

Sprouting negatively affects the quality of stored potato tubers. Understanding the molecular mechanisms that control this process is important for the development of potato varieties with desired sprouting characteristics. Serine/threonine protein phosphatase type 2A (PP2A) has been implicated in several developmental programs and stress responses in plants. PP2A comprises a catalytic (PP2Ac), a scaffolding (A), and a regulatory (B) subunit. In cultivated potato, six PP2Ac isoforms were identified, named StPP2Ac1, 2a, 2b, 3, 4, and 5. In this study we evaluated the sprouting behavior of potato tubers overexpressing the catalytic subunit 2b (StPP2Ac2b-OE). The onset of sprouting and initial sprout elongation is significantly delayed in StPP2Ac2b-OE tubers; however, sprout growth is accelerated during the late stages of development, due to a high degree of branching. StPP2Ac2b-OE tubers also exhibit a pronounced loss of apical dominance. These developmental characteristics are accompanied by changes in carbohydrate metabolism and response to gibberellic acid, and a differential balance between abscisic acid, gibberellic acid, cytokinins, and auxin. Overexpression of StPP2Ac2b alters the source-sink balance, increasing the source capacity of the tuber, and the sink strength of the sprout to support its accelerated growth., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

The serine/threonine protein phosphatases type 2A (PP2A) are involved in several physiological responses in plants, playing important roles in developmental programs, stress responses and hormone signaling. Six PP2A catalytic subunits (StPP2Ac) were identified in cultivated potato. Transgenic potato plants constitutively overexpressing the catalytic subunit StPP2Ac2b (StPP2Ac2b-OE) were developed to determine its physiological roles. The response of StPP2Ac2b-OE plants to the oomycete Phytophthora infestans, the causal agent of late blight, was evaluated. We found that overexpression of StPP2Ac2b enhances susceptibility to the pathogen. Further bioinformatics, biochemical, and molecular analyses revealed that StPP2Ac2b positively regulates developmental and pathogen-induced senescence, and that P. infestans infection promotes senescence, most likely through induction of StPP2Ac2b expression., Competing Interests: The authors have declared that no competing interests exist.

Key Message: In this study we show that expression of the Arabidopsis ABF4 gene in potato increases tuber yield under normal and abiotic stress conditions, improves storage capability and processing quality of the tubers, and enhances salt and drought tolerance. Potato is the third most important food crop in the world. Potato plants are susceptible to salinity and drought, which negatively affect crop yield, tuber quality and market value. The development of new varieties with higher yields and increased tolerance to adverse environmental conditions is a main objective in potato breeding. In addition, tubers suffer from undesirable sprouting during storage that leads to major quality losses; therefore, the control of tuber sprouting is of considerable economic importance. ABF (ABRE-binding factor) proteins are bZIP transcription factors that regulate abscisic acid signaling during abiotic stress. ABF proteins also play an important role in the tuberization induction. We developed transgenic potato plants constitutively expressing the Arabidopsis ABF4 gene (35S::ABF4). In this study, we evaluated the performance of 35S::ABF4 plants grown in soil, determining different parameters related to tuber yield, tuber quality (carbohydrates content and sprouting behavior) and tolerance to salt and drought stress. Besides enhancing salt stress and drought tolerance, constitutive expression of ABF4 increases tuber yield under normal and stress conditions, enhances storage capability and improves the processing quality of the tubers.

This study presents the characterization of the plasma membrane (PM) H+-ATPases in potato, focusing on their role in stolon and tuber development. Seven PM H+-ATPase genes were identified in the Solanum tuberosum genome, designated PHA1-PHA7. PHA genes show distinct expression patterns in different plant tissues and under different stress treatments. Application of PM H+-ATPase inhibitors arrests stolon growth, promotes tuber induction, and reduces tuber size, indicating that PM H+-ATPases are involved in tuberization, acting at different stages of the process. Transgenic potato plants overexpressing PHA1 were generated (PHA1-OE). At early developmental stages, PHA1-OE stolons elongate faster and show longer epidermal cells than wild-type stolons; this accelerated growth is accompanied by higher cell wall invertase activity, lower starch content, and higher expression of the sucrose-H+ symporter gene StSUT1. PHA1-OE stolons display an increased branching phenotype and develop larger tubers. PHA1-OE plants are taller and also present a highly branched phenotype. These results reveal a prominent role for PHA1 in plant growth and development. Regarding tuberization, PHA1 promotes stolon elongation at early stages, and tuber growth later on. PHA1 is involved in the sucrose-starch metabolism in stolons, possibly providing the driving force for sugar transporters to maintain the apoplastic sucrose transport during elongation., (© The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

Key Message: This study provides the first genetic evidence for the role of PP2A in tuberization, demonstrating that the catalytic subunit StPP2Ac2b positively modulates tuber induction, and that its function is related to the regulation of gibberellic acid metabolism. The results contribute to a better understanding of the molecular mechanism controlling tuberization induction, which remains largely unknown. The serine/threonine protein phosphatases type 2A (PP2A) are implicated in several physiological processes in plants, playing important roles in hormone responses. In cultivated potato (Solanum tuberosum), six PP2A catalytic subunits (StPP2Ac) were identified. The PP2Ac of the subfamily I (StPP2Ac1, 2a and 2b) were suggested to be involved in the tuberization signaling in leaves, where the environmental and hormonal signals are perceived and integrated. The aim of this study was to investigate the role of PP2A in the tuberization induction in stolons. We selected one of the catalytic subunits of the subfamily I, StPP2Ac2b, to develop transgenic plants overexpressing this gene (StPP2Ac2b-OE). Stolons from StPP2Ac2b-OE plants show higher tuber induction rates in vitro, as compared to wild type stolons, with no differences in the number of tubers obtained at the end of the process. This effect is accompanied by higher expression levels of the gibberellic acid (GA) catabolic enzyme StGA2ox1. GA up-regulates StPP2Ac2b expression in stolons, possibly as part of the feedback system by which the hormone regulates its own level. Sucrose, a tuber-promoting factor in vitro, increases StPP2Ac2b expression. We conclude that StPP2Ac2b acts in stolons as a positive regulator tuber induction, integrating different tuberization-related signals mainly though the modulation of GA metabolism.

Calcium-dependent protein kinases, CDPKs, decode calcium (Ca2+) transients and initiate downstream responses in plants. In order to understand how CDPKs affect plant physiology, their specific target proteins must be identified. In tobacco, the bZIP transcription factor Repression of Shoot Growth (NtRSG) that modulates gibberellin (GA) content is a specific target of NtCDPK1. StCDPK3 from potato is homologous (88% identical) to NtCDPK1 even in its N-terminal variable domain. In this work, we observe that NtRSG is also phosphorylated by StCDPK3. The potato RSG family of transcription factors is composed of three members that share similar features. The closest homologue to NtRSG, which was named StRSG1, was amplified and sequenced. qRT-PCR data indicate that StRSG1 is mainly expressed in petioles, stems, lateral buds, and roots. In addition, GA treatment affected StRSG1 expression. StCDPK3 transcripts were detected in leaves, petioles, stolons, roots, and dormant tubers, and transcript levels were modified in response to GA. The recombinant StRSG1-GST protein was produced and tested as a substrate for StCDPK3 and StCDPK1. 6xHisStCDPK3 was able to phosphorylate the potato StRSG1 in a Ca2+-dependent way, while 6xHisStCDPK1 could not. StCDPK3 also interacts and phosphorylates the transcription factor StABF1 (ABRE binding factor 1) involved in ABA signaling, as shown by EMSA and phosphorylation assays. StABF1 transcripts were mainly detected in roots, stems, and stolons. Our data suggest that StCDPK3 could be involved in the cross-talk between ABA and GA signaling at the onset of tuber development., Competing Interests: The authors have declared that no competing interests exist.

Potato (Solanum tuberosum L.) tuberization is regulated by many signals, such as abscisic acid (ABA), sucrose and gibberellic acid (GA). ABA and sucrose are positive modulators, while GA is an inhibitor of the process. ABF (ABRE-binding factor) proteins are transcription factors involved in ABA and stress signaling. Previously, we reported that S. tuberosum StABF1 could mediate the ABA effects on tuberization. The aim of the present study was to evaluate the potential use of ABF genes to enhance tuberization and to determine the molecular mechanism involved. For this purpose, transgenic potato plants expressing the Arabidopsis ABF4 or ABF2 genes were generated, and their tuberization capacity and response to tuberization-related signals were analyzed in vitro. The results indicate that both ABF4 and ABF2 proteins positively regulate potato tuber induction; however, only ABF4 expression significantly increases the number and weight of the tubers obtained, without stunting growth. ABF4 and ABF2 transgenic plants exhibit ABA hypersensitivity during tuberization, accompanied by a GA-deficient phenotype. ABF4 expression triggers a significant rise in ABA levels in stolons under tuber-inducing conditions as compared with wild-type plants and a transcriptional deregulation of GA metabolism genes. Our results demonstrate that Arabidopsis ABF4 functions in potato ABA-GA signaling crosstalk during tuberization by regulating the expression of ABA- and GA-metabolism genes. ABF4 gene might be a potential tool to increase tuber production, since its heterologous expression in potato enhances tuber induction without affecting plant growth.

Plasma membrane proton pumps (PM H(+)-ATPases) are involved in several physiological processes, such as growth and development, and abiotic stress responses. The major regulators of the PM H(+)-ATPases are proteins of the 14-3-3 family, which stimulate its activity. In addition, a novel interaction partner of the AHA1 PM H(+)-ATPase, named PPI1 (proton pump interactor, isoform 1), was identified in Arabidopsis thaliana. This protein stimulates the activity of the proton pump in vitro. In this work, we report the characterization of an A. thaliana PPI1 homolog in potato (Solanum tuberosum L.) named StPPI1. The full-length coding sequence of StPPI1 was obtained. The open reading frame (ORF) encodes a protein of 629 amino acids showing 50% identity with A. thaliana PPI1 protein. The StPPI1 ORF is divided into seven exons split by six introns. Southern blot analysis suggests that StPPI1 belongs to a family of related genes. Recombinant StPPI1 stimulates H(+)-ATPase activity in vitro. Basal levels of StPPI1 transcripts are observed in all tissues, however, StPPI1 expression is higher in proliferative regions (shoot apex and flower buds), flowers and leaves than in shoots and roots. StPPI1 mRNA levels significantly increase during tuber development. StPPI1 is induced by salt stress and cold. Drought and mechanical wounding slightly increase StPPI1 transcript levels. In addition, the expression of SlPPI1, the tomato homolog of StPPI1, was determined under adverse environmental conditions in tomato plants. SlPPI1 mRNA levels are increased by drought and cold, but are unaffected by salt stress. Mechanical wounding slightly increases SlPPI1 expression.

Tuber formation in potato (Solanum tuberosum L.) is regulated by hormonal and environmental signals that are thought to be integrated in the leaves. The molecular mechanisms that mediate the responses to tuberization-related signals in leaves remain largely unknown. In this study we analyzed the roles of protein phosphatase type 2A catalytic subunits (PP2Ac) in the leaf responses to conditions that affect tuberization. The responses were monitored by analyzing the expression of the "tuber-specific" genes Patatin and Pin2, which are induced in tubers and leaves during tuber induction. Experiments using PP2A inhibitors, together with PP2Ac expression profiles under conditions that affect tuberization indicate that high sucrose/nitrogen ratio, which promotes tuber formation, increases the transcript levels of Patatin and Pin2, by increasing the activity of PP2As without affecting PP2Ac mRNA or protein levels. Gibberellic acid (GA), a negative regulator of tuberization, down-regulates the transcription of catalytic subunits of PP2As from the subfamily I and decreases their enzyme levels. In addition, GA inhibits the expression of Patatin and Pin2 possibly by a PP2A-independent mechanism. PP2Ac down-regulation by GA may inhibit tuberization signaling downstream of the inductive effects of high sucrose/nitrogen ratio. These results are consistent with the hypothesis that PP2As of the subfamily I may positively modulate the signaling pathways that lead to the transcriptional activation of "tuber-specific" genes in leaves, and act as molecular switches regulated by both positive and negative modulators of tuberization.