Soybean (Glycine max) is one of the most important crop plants for seed protein and oil content, and for its capacity to fix atmospheric nitrogen through symbioses with soil-borne microorganisms. We sequenced the 1.1-gigabase genome by a whole-genome shotgun approach and integrated it with physical and high-density genetic maps to create a chromosome-scale draft sequence assembly. We predict 46,430 protein-coding genes, 70% more than Arabidopsis and similar to the poplar genome which, like soybean, is an ancient polyploid (palaeopolyploid). About 78% of the predicted genes occur in chromosome ends, which comprise less than one-half of the genome but account for nearly all of the genetic recombination. Genome duplications occurred at approximately 59 and 13 million years ago, resulting in a highly duplicated genome with nearly 75% of the genes present in multiple copies. The two duplication events were followed by gene diversification and loss, and numerous chromosome rearrangements. An accurate soybean genome sequence will facilitate the identification of the genetic basis of many soybean traits, and accelerate the creation of improved soybean varieties.

Abscisic acid (ABA) signaling is important for stress responses and developmental processes in plants. A subgroup of protein phosphatase 2C (group A PP2C) or SNF1-related protein kinase 2 (subclass III SnRK2) have been known as major negative or positive regulators of ABA signaling, respectively. Here, we demonstrate the physical and functional linkage between these two major signaling factors. Group A PP2Cs interacted physically with SnRK2s in various combinations, and efficiently inactivated ABA-activated SnRK2s via dephosphorylation of multiple Ser/Thr residues in the activation loop. This step was suppressed by the RCAR/PYR ABA receptors in response to ABA. However the abi1-1 mutated PP2C did not respond to the receptors and constitutively inactivated SnRK2. Our results demonstrate that group A PP2Cs act as 'gatekeepers' of subclass III SnRK2s, unraveling an important regulatory mechanism of ABA signaling.A s sessile organisms, plants have to rapidly recognize and adapt to environmental changes. The phytohormone abscisic acid (ABA) plays a central role in such responses (1, 2), and is also involved in many developmental processes (3) and defense systems (4). Thus, ABA functions as a key molecule that unifies and regulates biotic and abiotic stress responses and the developmental status of the plant. Hence, the biological and agricultural importance of ABA has led to extensive studies on its signaling mechanism, and many putative signal transducers have been reported (5). Although it has been difficult to integrate all of the current findings, significant progress was recently made by two independent research groups. They identified the RCAR/ PYR family proteins as ABA receptors that inhibit protein phosphatase 2C (PP2C) in an ABA-dependent manner (6, 7). Among plant PP2Cs, a group A subfamily (e.g., ABI1 and ABI2) is annotated as negative regulators of the ABA response in seeds through to adult plants (5). Such PP2C-dependent negative regulation can be canceled by RCAR/PYR in response to ABA, leading to activation of some positive regulatory pathways (6, 7). Previously, we demonstrated that ABI1 interacts with a protein kinase, SRK2E (OST1/SnRK2.6) (8). SRK2E belongs to the SNF1-related protein kinase 2 (SnRK2) family, which is activated by ABA or osmotic stress and positively regulates the ABA response in various tissues (9 -11). Furthermore, ABAdependent activation of SRK2E was repressed in an abi1-1 mutant, suggesting that SnRK2 functions downstream of PP2C (8, 9). Based on these findings, a model was hypothesized in which RCAR/PYR and PP2C negatively regulate SnRK2 (7). However, there is no direct evidence demonstrating how PP2C regulates SnRK2, and the molecular process between them remains a question in ABA signaling. Our presented data clearly demonstrated the biochemical relation between PP2C and SnRK2 and elucidated an important regulatory mechanism of ABA signaling. Results and DiscussionRequirement of SnRK2 Activity for ABA Responses. In Arabidopsis, subclass III of the SnRK2 family is composed of...

bZIP-type transcription factors AREBs͞ABFs bind an abscisic acid (ABA)-responsive cis-acting element named ABRE and transactivate downstream gene expression in Arabidopsis. Because AREB1 overexpression could not induce downstream gene expression, activation of AREB1 requires ABA-dependent posttranscriptional modification. We confirmed that ABA activated 42-kDa kinase activity, which, in turn, phosphorylated Ser͞Thr residues of R-X-X-S͞T sites in the conserved regions of AREB1. Amino acid substitutions of R-X-X-S͞T sites to Ala suppressed transactivation activity, and multiple substitution of these sites resulted in almost complete suppression of transactivation activity in transient assays. In contrast, substitution of the Ser͞Thr residues to Asp resulted in high transactivation activity without exogenous ABA application. A phosphorylated, transcriptionally active form was achieved by substitution of Ser͞Thr in all conserved R-X-X-S͞T sites to Asp. Transgenic plants overexpressing the phosphorylated active form of AREB1 expressed many ABA-inducible genes, such as RD29B, without ABA treatment. These results indicate that the ABA-dependent multisite phosphorylation of AREB1 regulates its own activation in plants.transactivation activity ͉ transcription factor AREB1 ͉ transgenic Arabidopsis T he phytohormone abscisic acid (ABA) plays important roles in seed maturation and dormancy and is also involved in the adaptation of vegetative tissues to abiotic environmental stresses, such as drought and high salinity. ABA promotes stomatal closure in guard cells and regulates the expression of many genes, the products of which may function in dehydration tolerance in both vegetative tissues and seeds. Many ABA-inducible genes contain a conserved element named ABA-responsive element (ABRE) (Py-ACGTGG͞TC) in their promoter regions. The ABRE functions as a cis-acting element and is involved in ABA-responsive gene expression (for reviews, see refs. 1 and 2).The RD29B promoter region contains two ABREs, and analyses with the ABA-deficient and insensitive mutants aba1 and abi1 revealed that the drought-inducible expression of RD29B is controlled mainly by ABA (3-6). Yeast one-hybrid screening with the RD29B promoter including ABREs enabled us to clone three independent cDNAs encoding ABRE-binding proteins (AREB1, AREB2, and AREB3) in Arabidopsis (7). Each AREB protein contained a single bZIP-type DNA-binding domain, and expression of AREB1 and AREB2 was up-regulated by ABA, drought, and high-salinity stresses, shown to function as trans-acting activators by using transient expression in protoplasts (7). Choi et al. (8) also reported the cloning of four independent cDNAs for ABREbinding factors (ABF1, ABF2, ABF3, and ABF4) from Arabidopsis. ABF2 and ABF4 were identical to AREB1 and AREB2, respectively.In the Arabidopsis genome, 75 distinct bZIP transcription factors exist, and nine members are classified as a homologous subfamily of AREBs that contain three N-terminal (C1, C2, and C3) and one C-terminal (C4) conserved domains (9-...

ABA is a major phytohormone that regulates a broad range of plant traits and is especially important for adaptation to environmental conditions. Our understanding of the molecular basis of ABA responses in plants improved dramatically in 2009 and 2010, banner years for ABA research. There are three major components; PYR/PYL/ RCAR (an ABA receptor), type 2C protein phosphatase (PP2C; a negative regulator) and SNF1-related protein kinase 2 (SnRK2; a positive regulator), and they offer a double negative regulatory system, [PYR/PYL/RCAR—| PP2C—| SnRK2]. In the absence of ABA, PP2C inactivates SnRK2 by direct dephosphorylation. In response to environmental or developmental cues, ABA promotes the interaction of PYR/PYL/RCAR and PP2C, resulting in PP2C inhibition and SnRK2 activation. This signaling complex can work in both the nucleus and cytosol, as it has been shown that SnRK2 phosphorylates basic-domain leucine zipper (bZIP) transcription factors or membrane proteins. Several structural analyses of PYR/PYL/RCAR have provided the mechanistic basis for this ‘core signaling’ model, by elucidating the mechanism of ABA binding of receptors, or the ‘gate–latch–lock’ mechanism of interaction with PP2C in inhibiting activity. On the other hand, intercellular ABA transport had remained a major issue, as had intracellular ABA signaling. Recently, two plasma membrane-type ABC transporters were identified and shed light on the influx/efflux system of ABA, resolving how ABA is transported from cell to cell in plants. Our knowledge of ABA responses in plants has been greatly expanded from intracellular signaling to intercellular transport of ABA.

ABA is an important phytohormone regulating various plant processes, including stress tolerance, seed development and germination. SRK2D/SnRK2.2, SRK2E/ SnRK2.6/OST1 and SRK2I/SnRK2.3 are redundant ABAactivated SNF1-related protein kinases 2 (SnRK2s) in Arabidopsis thaliana. We examined the role of these protein kinases in seed development and germination. These SnRK2 proteins were mainly expressed in the nucleus during seed development and germination. The triple mutant (srk2d srk2e srk2i) was sensitive to desiccation and showed severe growth defects during seed development. It exhibited a loss of dormancy and elevated seed ABA content relative to wild-type plants. The severity of these phenotypes was far stronger than that of any single or double SRK2D , SRK2E and SRK2I mutants, including the srk2d srk2i mutant. The triple mutant had greatly reduced phosphorylation activity in in-gel kinase experiments using basic leucine zipper (bZIP) transcription factors including ABI5. Microarray experiments revealed that 48 and 30 % of the down-regulated genes in abi5 and abi3 seeds were suppressed in the triple mutant seeds, respectively. Moreover, disruption of the three protein kinases induced global changes in the up-regulation of ABA-repressive gene expression, as well as the downregulation of ABA-inducible gene expression. These alterations in gene expression result in a loss of dormancy and severe growth defects during seed development. Collectively, these results indicate that SRK2D, SRK2E and SRK2I protein kinases involved in ABA signaling are essential for the control of seed development and dormancy through the extensive control of gene expression.