Ruth R. Finkelstein

Abscisic Acid Signaling in Seeds and Seedlings

Abscisic acid (ABA) regulates many agronomically important aspects of plant development, including the synthesis of seed storage proteins and lipids, the promotion of seed desiccation tolerance and dormancy, and the inhibition of the phase transitions from embryonic to germinative growth and from

Abscisic Acid Inhibits Type 2C Protein Phosphatases via the PYR/PYL Family of START Proteins

ABA Receptor Rumbled? The plant hormone abscisic acid (ABA) is critical for normal development and for mediating plant responses to stressful environmental conditions. Now, two papers present analyses of candidate ABA receptors (see the news story by Pennisi). Ma et al. (p. 1064; published online 30 April) and Park et al. (p. 1068, published online 30 April) used independent strategies to search for proteins that physically interact with ABI family phosphatase components of the ABA response signaling pathway. Both groups identified different members of the same family of proteins, which appear to interact with ABI proteins to form a heterocomplex that can act as the ABA receptor. The variety of both families suggests that the ABA receptor may not be one entity, but rather a class of closely related complexes, which may explain previous difficulties in establishing its identity. Links between two ancient multimember protein families signal responses to the plant hormone abscisic acid. Type 2C protein phosphatases (PP2Cs) are vitally involved in abscisic acid (ABA) signaling. Here, we show that a synthetic growth inhibitor called pyrabactin functions as a selective ABA agonist. Pyrabactin acts through PYRABACTIN RESISTANCE 1 (PYR1), the founding member of a family of START proteins called PYR/PYLs, which are necessary for both pyrabactin and ABA signaling in vivo. We show that ABA binds to PYR1, which in turn binds to and inhibits PP2Cs. We conclude that PYR/PYLs are ABA receptors functioning at the apex of a negative regulatory pathway that controls ABA signaling by inhibiting PP2Cs. Our results illustrate the power of the chemical genetic approach for sidestepping genetic redundancy.

Abscisic Acid: Emergence of a Core Signaling Network

Abscisic acid (ABA) regulates numerous developmental processes and adaptive stress responses in plants. Many ABA signaling components have been identified, but their interconnections and a consensus on the structure of the ABA signaling network have eluded researchers. Recently, several advances have led to the identification of ABA receptors and their three-dimensional structures, and an understanding of how key regulatory phosphatase and kinase activities are controlled by ABA. A new model for ABA action has been proposed and validated, in which the soluble PYR/PYL/RCAR receptors function at the apex of a negative regulatory pathway to directly regulate PP2C phosphatases, which in turn directly regulate SnRK2 kinases. This model unifies many previously defined signaling components and highlights the importance of future work focused on defining the direct targets of SnRK2s and PP2Cs, dissecting the mechanisms of hormone interactions (i.e., cross talk) and defining connections between this new negative regulatory pathway and other factors implicated in ABA signaling.

The Arabidopsis Abscisic Acid Response Gene ABI5 Encodes a Basic Leucine Zipper Transcription Factor

The Arabidopsis abscisic acid (ABA)–insensitive abi5 mutants have pleiotropic defects in ABA response, including decreased sensitivity to ABA inhibition of germination and altered expression of some ABA-regulated genes. We isolated the ABI5 gene by using a positional cloning approach and found that it encodes a member of the basic leucine zipper transcription factor family. The previously characterized abi5-1 allele encodes a protein that lacks the DNA binding and dimerization domains required for ABI5 function. Analyses of ABI5 expression provide evidence for ABA regulation, cross-regulation by other ABI genes, and possibly autoregulation. Comparison of seed and ABA-inducible vegetative gene expression in wild-type and abi5-1 plants indicates that ABI5 regulates a subset of late embryogenesis–abundant genes during both developmental stages.

Molecular Aspects of Seed Dormancy

Seed dormancy provides a mechanism for plants to delay germination until conditions are optimal for survival of the next generation. Dormancy release is regulated by a combination of environmental and endogenous signals with both synergistic and competing effects. Molecular studies of dormancy have correlated changes in transcriptomes, proteomes, and hormone levels with dormancy states ranging from deep primary or secondary dormancy to varying degrees of release. The balance of abscisic acid (ABA):gibberellin (GA) levels and sensitivity is a major, but not the sole, regulator of dormancy status. ABA promotes dormancy induction and maintenance, whereas GA promotes progression from release through germination; environmental signals regulate this balance by modifying the expression of biosynthetic and catabolic enzymes. Mediators of environmental and hormonal response include both positive and negative regulators, many of which are feedback-regulated to enhance or attenuate the response. The net result is a slightly heterogeneous response, thereby providing more temporal options for successful germination.

The Arabidopsis Abscisic Acid Response Locus ABI4 Encodes an APETALA2 Domain Protein

Arabidopsis abscisic acid (ABA)–insensitive abi4 mutants have pleiotropic defects in seed development, including decreased sensitivity to ABA inhibition of germination and altered seed-specific gene expression. This phenotype is consistent with a role for ABI4 in regulating seed responses to ABA and/or seed-specific signals. We isolated the ABI4 gene by positional cloning and confirmed its identity by complementation analysis. The predicted protein product shows homology to a plant-specific family of transcriptional regulators characterized by a conserved DNA binding domain, the APETALA2 domain. The single mutant allele identified has a single base pair deletion, resulting in a frameshift that should disrupt the C-terminal half of the protein but leave the presumed DNA binding domain intact. Expression analyses showed that despite the seed-specific nature of the mutant phenotype, ABI4 expression is not seed specific.

Abscisic Acid synthesis and response.

ABSTRACT Abscisic acid (ABA) is one of the “classical” plant hormones, i.e. discovered at least 50 years ago, that regulates many aspects of plant growth and development. This chapter reviews our current understanding of ABA synthesis, metabolism, transport, and signal transduction, emphasizing knowledge gained from studies of Arabidopsis. A combination of genetic, molecular and biochemical studies has identified nearly all of the enzymes involved in ABA metabolism, almost 200 loci regulating ABA response, and thousands of genes regulated by ABA in various contexts. Some of these regulators are implicated in cross-talk with other developmental, environmental or hormonal signals. Specific details of the ABA signaling mechanisms vary among tissues or developmental stages; these are discussed in the context of ABA effects on seed maturation, germination, seedling growth, vegetative stress responses, stomatal regulation, pathogen response, flowering, and senescence.

ABA and sugar interactions regulating development: cross-talk or voices in a crowd?

Plant growth and development are controlled by the concerted action of many signaling pathways that integrate information from environmental signals with that from developmental and metabolic cues. Physiological studies have demonstrated that abscisic acid and sugars have both similar and antagonistic effects on diverse processes, including seed development, germination, and seedling growth. Recent genetic studies have identified several loci that are involved in both sugar and hormonal responses. It is rarely clear whether these apparent linkages reflect direct or indirect interactions between sugar and hormone signaling pathways, but the identification of gene products that are encoded at these loci is allowing these possibilities to be tested.

Abscisic Acid Biosynthesis and Response

Abscisic acid (ABA) is an optically active 15-C weak acid that was first identified in the early 1960s as a growth inhibitor accumulating in abscising cotton fruit (“abscisin II”) and leaves of sycamore trees photoperiodically induced to become dormant (“dormin”) (reviewed in Addicott, 1983). It has since been shown to regulate many aspects of plant growth and development including embryo maturation, seed dormancy, germination, cell division and elongation, and responses to environmental stresses such as drought, salinity, cold, pathogen attack and UV radiation (reviewed in Leung and Giraudat, 1998; Rock, 2000). However, despite the name, it does not appear to control abscission directly; the presence of ABA in abscising organs reflects its role in promoting senescence and/or stress responses, the processes preceding abscission. Although ABA has historically been thought of as a growth inhibitor, young tissues have high ABA levels, and ABA-deficient mutant plants are severely stunted (Figure 1) because their ability to reduce transpiration and establish turgor is impaired. Exogenous ABA treatment of mutants restores normal cell expansion and growth. Figure 1. Exogenous ABA suppresses growth inhibition of ABA-deficient mutants. Plants with one of three mutant alleles of aba1 were grown with (bottom) or without (top) ABA treatment (spraying twice weekly with 10 µM ABA for 8 weeks). (Photograph courtesy ... ABA is ubiquitous in lower and higher plants. It is also produced by some phytopathogenic fungi (Assante et al., 1977; Neill et al., 1982; Kitagawa et al., 1995) and has even been found in mammalian brain tissue (Le Page-Degivry et al., 1986). As a sesquiterpenoid, it was long thought to be synthesized directly from farnesyl pyrophosphate, as in fungi (reviewed in Zeevaart and Creelman, 1988). However, it is actually synthesized indirectly from carotenoids. As a weak acid (pKa=4.8), ABA is mostly uncharged when present in the relatively acidic apoplastic compartment of plants and can easily enter cells across the plasma membrane. The major control of ABA distribution among plant cell compartments follows the “anion trap” concept: the dissociated (anion) form of this weak acid accumulates in alkaline compartments (e.g. illuminated chloroplasts) and may redistribute according to the steepness of the pH gradients across membranes. In addition to partitioning according to the relative pH of compartments, specific uptake carriers contribute to maintaining a low apoplastic ABA concentration in unstressed plants. Despite the ease with which ABA can enter cells, there is evidence for extracellular as well as intracellular perception of ABA (reviewed in Leung and Giraudat, 1998; Rock, 2000). Multiple receptor types are also implicated by the variation in stereospecificity among ABA responses. Genetic studies, especially in Arabidopsis, have identified many loci involved in ABA synthesis and response and analyzed their functional roles in ABA physiology (reviewed in Leung and Giraudat, 1998; Rock, 2000). Many likely signaling intermediates correlated with ABA response (e.g. ABA-activated or -induced kinases and DNA-binding proteins that specifically bind ABA-responsive promoter elements) have also been identified by molecular and biochemical studies, but the relationships among these proteins are unclear. Cell biological studies have identified secondary messengers involved in ABA response. Ongoing studies combine these approaches in efforts to determine coherent models of ABA signaling mechanism(s).

Redundant and distinct functions of the ABA response loci ABA- INSENSITIVE(ABI)5 and ABRE-BINDING FACTOR (ABF)3

Abscisic acid-responsive gene expression is regulated by numerous transcription factors, including a subgroup of basic leucine zipper factors that bind to the conserved cis-acting sequences known as ABA-responsive elements. Although one of these factors, ABA-insensitive 5 (ABI5), was identified genetically, the paucity of genetic data for the other family members has left it unclear whether they perform unique functions or act redundantly to ABI5 or each other. To test for potential redundancy with ABI5, we identified the family members with most similar effects and interactions in transient expression systems (ABF3 and ABF1), then characterized loss-of-function lines for those loci. The abf1 and abf3 monogenic mutant lines had at most minimal effects on germination or seed-specific gene expression, but the enhanced ABA- and stress-resistance of abf3 abi5 double mutants revealed redundant action of these genes in multiple stress responses of seeds and seedlings. Although ABI5, ABF3, and ABF1 have some overlapping effects, they appear to antagonistically regulate each other’s expression at specific stages. Consequently, loss of any one factor may be partially compensated by increased expression of other family members.