Structural insights into rice KAI2 receptor provide functional implications for perception and signal transduction.

KAI2 receptors, classified as plant α/ß hydrolase enzymes, are capable of perceiving smoke-derived butenolide signals and endogenous yet unidentified KAI2-ligands (KLs). While the number of functional KAI2 receptors varies among land plant species, rice has only one KAI2 gene. Rice, a significant crop and representative of grasses, relies on KAI2-mediated Arbuscular mycorrhiza (AM) symbioses to flourish in traditionally arid and nutrient-poor environments. This study presents the first crystal structure of an active rice (Oryza sativa, Os) KAI2 hydrolase receptor. Our structural and biochemical analyses uncover grass-unique pocket residues influencing ligand sensitivity and hydrolytic activity. Through structure-guided analysis, we identify a specific residue whose mutation enables the increase or decrease of ligand perception, catalytic activity, and signal transduction. Furthermore, we investigate OsKAI2-mediated signaling by examining its ability to form a complex with its binding partner, the F-box protein DWARF3 (D3) ubiquitin ligase and subsequent degradation of the target substrate OsSMAX1, demonstrating the significant role of hydrophobic interactions in the OsKAI2-D3 interface. This study provides new insights into the diverse and pivotal roles of the OsKAI2 signaling pathway in the plant kingdom, particularly in grasses.

KAR1-induced dormancy release in Avena fatua caryopses involves reduction of caryopsis sensitivity to ABA and ABA/GAs ratio in coleorhiza and radicle.

MAIN CONCLUSION: The dormancy release by KAR1 is associated with a reduction of coleorhiza and radicle sensitivity to ABA as well as with reduction the ABA/GAs ratio in the coleorhiza, by a decrease content of ABA, and in the radicle, by a decrease the ABA and an increase of the GAs contents. Both, karrikin 1 (KAR1) and gibberellin A3 (GA3), release dormancy in Avena fatua caryopses, resulting in the emergence of coleorhiza (CE) and radicle (RE). Moreover, KAR1 and GA3 stimulate CE and RE in the presence of abscisic acid (ABA), the stimulation being more effective in CE. The stimulatory effects of KAR1 and GA3 involve also the CE and RE rates. A similar effect was observed at KAR1 concentrations much lower than those of GA3. KAR1 increased the levels of bioactive GA5 and GA6 in embryos and the levels of GA1, GA5, GA3, GA6 and GA4 in radicles. The stimulatory effect of KAR1 on germination, associated with increased levels of gibberellins (GAs) and reduced levels of ABA in embryos, was counteracted by paclobutrazol (PAC), commonly regarded as a GAs biosynthesis inhibitor. Consequently, KAR1 decreased the ABA/GAs ratio, whereas PAC, used alone or in combination with KAR1, increased it. The ABA/GAs ratio was reduced by KAR1 in both coleorhiza and radicle, the effect being stronger in the latter. We present the first evidence that KAR1-induced dormancy release requires a decreased ABA/GAs ratio in coleorhiza and radicle. It is concluded that the dormancy-releasing effect of KAR1 in A. fatua caryopses includes (i) a reduction of the coleorhiza and radicle sensitivity to ABA, and (2) a reduction of the ABA/GAs ratio (i) in the coleorhiza, by decreasing the ABA content, and (ii) in the radicle, by decreasing the ABA and increasing the content GAs, particularly GA1. The results may suggest different mechanisms of dormancy release by KAR1 in monocot and dicot seeds.

Karrikin signalling: impacts on plant development and abiotic stress tolerance.

Plants rely upon a diverse range of metabolites to control growth and development, and to overcome stress that results from suboptimal conditions. Karrikins (KARs) are a class of butenolide compounds found in smoke that stimulate seed germination and regulate various developmental processes in plants. KARs are perceived via a plant α/ß-hydrolase called KARRIKIN INSENSITIVE2 (KAI2), which also functions as a receptor for a postulated phytohormone, provisionally termed KAI2 ligand (KL). Considered natural analogues of KL, KARs have been extensively studied for their effects on plant growth and their crosstalk with plant hormones. The perception and response pathway for KAR-KL signalling is closely related to that of strigolactones, another class of butenolides with numerous functions in regulating plant growth. KAR-KL signalling influences seed germination, seedling photomorphogenesis, root system architecture, abiotic stress responses, and arbuscular mycorrhizal symbiosis. Here, we summarize current knowledge of KAR-KL signalling, focusing on its role in plant development, its effects on stress tolerance, and its interaction with other signalling mechanisms.

KARRIKIN INSENSITIVE2 regulates leaf development, root system architecture and arbuscular-mycorrhizal symbiosis in Brachypodium distachyon.

KARRIKIN INSENSITIVE2 (KAI2) is an α/ß-hydrolase required for plant responses to karrikins, which are abiotic butenolides that can influence seed germination and seedling growth. Although represented by four angiosperm species, loss-of-function kai2 mutants are phenotypically inconsistent and incompletely characterised, resulting in uncertainties about the core functions of KAI2 in plant development. Here we characterised the developmental functions of KAI2 in the grass Brachypodium distachyon using molecular, physiological and biochemical approaches. Bdkai2 mutants exhibit increased internode elongation and reduced leaf chlorophyll levels, but only a modest increase in water loss from detached leaves. Bdkai2 shows increased numbers of lateral roots and reduced root hair growth, and fails to support normal root colonisation by arbuscular-mycorrhizal (AM) fungi. The karrikins KAR1 and KAR2 , and the strigolactone (SL) analogue rac-GR24, each elicit overlapping but distinct changes to the shoot transcriptome via BdKAI2. Finally, we show that BdKAI2 exhibits a clear ligand preference for desmethyl butenolides and weak responses to methyl-substituted SL analogues such as GR24. Our findings suggest that KAI2 has multiple roles in shoot development, root system development and transcriptional regulation in grasses. Although KAI2-dependent AM symbiosis is likely conserved within monocots, the magnitude of the effect of KAI2 on water relations may vary across angiosperms.

The Karrikin Receptor Karrikin Insensitive2 Positively Regulates Heat Stress Tolerance in Arabidopsis thaliana.

In this study, we investigated the potential role of the karrikin receptor KARRIKIN INSENSITIVE2 (KAI2) in the response of Arabidopsis seedlings to high-temperature stress. We performed phenotypic, physiological and transcriptome analyses of Arabidopsis kai2 mutants and wild-type (WT) plants under control (kai2_C and WT_C, respectively) and 6- and 24-h heat stress conditions (kai2_H6, kai2_H24, WT_H6 and WT_H24, respectively) to understand the basis for KAI2-regulated heat stress tolerance. We discovered that the kai2 mutants exhibited hypersensitivity to high-temperature stress relative to WT plants, which might be associated with a more highly increased leaf surface temperature and cell membrane damage in kai2 mutant plants. Next, we performed comparative transcriptome analysis of kai2_C, kai2_H6, kai2_H24, WT_C, WT_H6 and WT_H24 to identify transcriptome differences between WT and kai2 mutants in response to heat stress. K-mean clustering of normalized gene expression separated the investigated genotypes into three clusters based on heat-treated and non-treated control conditions. Within each cluster, the kai2 mutants were separated from WT plants, implying that kai2 mutants exhibited distinct transcriptome profiles relative to WT plants. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses showed a repression in 'misfolded protein binding', 'heat shock protein binding', 'unfolded protein binding' and 'protein processing in endoplasmic reticulum' pathways, which was consistent with the downregulation of several genes encoding heat shock proteins and heat shock transcription factors in the kai2 mutant versus WT plants under control and heat stress conditions. Our findings suggest that chemical or genetic manipulation of KAI2 signaling may provide a novel way to improve heat tolerance in plants.

KAI2 Can Do: Karrikin Receptor Function in Plant Development and Response to Abiotic and Biotic Factors.

The α/ß hydrolase KARRIKIN INSENSITIVE 2 (KAI2) functions as a receptor for a yet undiscovered phytohormone, provisionally termed KAI2 ligand (KL). In addition, it perceives karrikin, a butenolide compound found in the smoke of burnt plant material. KAI2-mediated signaling is involved in regulating seed germination and in shaping seedling and adult plant morphology, both above and below ground. It also governs responses to various abiotic stimuli and stresses and shapes biotic interactions. KAI2-mediated signaling is being linked to an elaborate cross-talk with other phytohormone pathways such as auxin, gibberellin, abscisic acid, ethylene and salicylic acid signaling, in addition to light and nutrient starvation signaling. Further connections will likely be revealed in the future. This article summarizes recent advances in unraveling the function of KAI2-mediated signaling and its interaction with other signaling pathways.

Karrikin Receptor KAI2 Coordinates Salt Tolerance Mechanisms in Arabidopsis thaliana.

Plants activate a myriad of signaling cascades to tailor adaptive responses under environmental stresses, such as salinity. While the roles of exogenous karrikins (KARs) in salt stress mitigation are well comprehended, genetic evidence of KAR signaling during salinity responses in plants remains unresolved. Here, we explore the functions of the possible KAR receptor KARRIKIN-INSENSITIVE2 (KAI2) in Arabidopsis thaliana tolerance to salt stress by investigating comparative responses of wild-type (WT) and kai2-mutant plants under a gradient of NaCl. Defects in KAI2 functions resulted in delayed and inhibited cotyledon opening in kai2 seeds compared with WT seeds, suggesting that KAI2 played an important role in enhancing seed germination under salinity. Salt-stressed kai2 plants displayed more phenotypic aberrations, biomass reduction, water loss and oxidative damage than WT plants. kai2 shoots accumulated significantly more Na+ and thus had a lower K+/Na+ ratio, than WT, indicating severe ion toxicity in salt-stressed kai2 plants. Accordingly, kai2 plants displayed a lower expression of genes associated with Na+ homeostasis, such as SALT OVERLY SENSITIVE (SOS) 1, SOS2, HIGH-AFFINITY POTASSIUM TRANSPORTER 1;1 (HKT1;1) and CATION-HYDROGEN EXCHANGER 1 (NHX1) than WT plants. WT plants maintained a better glutathione level, glutathione-related redox status and antioxidant enzyme activities relative to kai2 plants, implying KAI2's function in oxidative stress mitigation in response to salinity. kai2 shoots had lower expression levels of genes involved in the biosynthesis of strigolactones (SLs), salicylic acid and jasmonic acid and the signaling of abscisic acid and SLs than those of WT plants, indicating interactive functions of KAI2 signaling with other hormone signaling in modulating plant responses to salinity. Collectively, these results underpin the likely roles of KAI2 in the alleviation of salinity effects in plants by regulating several physiological and biochemical mechanisms involved in ionic and osmotic balance, oxidative stress tolerance and hormonal crosstalk.

SUPPRESSOR of MAX2 1 (SMAX1) and SMAX1-LIKE2 (SMXL2) Negatively Regulate Drought Resistance in Arabidopsis thaliana.

Recent investigations in Arabidopsis thaliana suggest that SUPPRESSOR of MORE AXILLARY GROWTH 2 1 (SMAX1) and SMAX1-LIKE2 (SMXL2) are negative regulators of karrikin (KAR) and strigolactone (SL) signaling during plant growth and development, but their functions in drought resistance and related mechanisms of action remain unclear. To understand the roles and mechanisms of SMAX1 and SMXL2 in drought resistance, we investigated the drought-resistance phenotypes and transcriptome profiles of smax1 smxl2 (s1,2) double-mutant plants in response to drought stress. The s1,2 mutant plants showed enhanced drought-resistance and lower leaf water loss when compared with wild-type (WT) plants. Transcriptome comparison of rosette leaves from the s1,2 mutant and the WT under normal and dehydration conditions suggested that the mechanism related to cuticle formation was involved in drought resistance. This possibility was supported by enhanced cuticle formation in the rosette leaves of the s1,2 mutant. We also found that the s1,2 mutant plants were more sensitive to abscisic acid in assays of stomatal closure, cotyledon opening, chlorophyll degradation and growth inhibition, and they showed a higher reactive oxygen species detoxification capacity than WT plants. In addition, the s1,2 mutant plants had longer root hairs and a higher root-to-shoot ratio than the WT plants, suggesting that the mutant had a greater capacity for water absorption than the WT. Taken together, our results indicate that SMAX1 and SMXL2 negatively regulate drought resistance, and disruption of these KAR- and SL-signaling-related genes may therefore provide a novel means for improving crop drought resistance.

The karrikin signaling regulator SMAX1 controls Lotus japonicus root and root hair development by suppressing ethylene biosynthesis.

An evolutionarily ancient plant hormone receptor complex comprising the α/ß-fold hydrolase receptor KARRIKIN INSENSITIVE 2 (KAI2) and the F-box protein MORE AXILLARY GROWTH 2 (MAX2) mediates a range of developmental responses to smoke-derived butenolides called karrikins (KARs) and to yet elusive endogenous KAI2 ligands (KLs). Degradation of SUPPRESSOR OF MAX2 1 (SMAX1) after ligand perception is considered to be a key step in KAR/KL signaling. However, molecular events which regulate plant development downstream of SMAX1 removal have not been identified. Here we show that Lotus japonicus SMAX1 is specifically degraded in the presence of KAI2 and MAX2 and plays an important role in regulating root and root hair development. smax1 mutants display very short primary roots and elongated root hairs. Their root transcriptome reveals elevated ethylene responses and expression of ACC Synthase 7 (ACS7), which encodes a rate-limiting enzyme in ethylene biosynthesis. smax1 mutants release increased amounts of ethylene and their root phenotype is rescued by treatment with ethylene biosynthesis and signaling inhibitors. KAR treatment induces ACS7 expression in a KAI2-dependent manner and root developmental responses to KAR treatment depend on ethylene signaling. Furthermore, in Arabidopsis, KAR-induced root hair elongation depends on ACS7 Thus, we reveal a connection between KAR/KL and ethylene signaling in which the KAR/KL signaling module (KAI2-MAX2-SMAX1) regulates the biosynthesis of ethylene to fine-tune root and root hair development, which are important for seedling establishment at the beginning of the plant life cycle.

The MAX2-KAI2 module promotes salicylic acid-mediated immune responses in Arabidopsis.

Arabidopsis MORE AXILLARY GROWTH2 (MAX2) is a key component in the strigolactone (SL) and karrikin (KAR) signaling pathways and regulates the degradation of SUPPRESSOR OF MAX2 1/SMAX1-like (SMAX1/SMXL) proteins, which are transcriptional co-repressors that regulate plant architecture, as well as abiotic and biotic stress responses. The max2 mutation reduces resistance against Pseudomonas syringae pv. tomato (Pst). To uncover the mechanism of MAX2-mediated resistance, we evaluated the resistance of various SL and KAR signaling pathway mutants. The resistance of SL-deficient mutants and of dwarf 14 (d14) was similar to that of the wild-type, whereas the resistance of the karrikin insensitive 2 (kai2) mutant was compromised, demonstrating that the KAR signaling pathway, not the SL signaling pathway, positively regulates the immune response. We measured the resistance of smax1 and smxl mutants, as well as the double, triple, and quadruple mutants with max2, which revealed that both the smax1 mutant and smxl6/7/8 triple mutant rescue the low resistance phenotype of max2 and that SMAX1 accumulation diminishes resistance. The susceptibility of smax1D, containing a degradation-insensitive form of SMAX1, further confirmed the SMAX1 function in the resistance. The relationship between the accumulation of SMAX1/SMXLs and disease resistance suggested that the inhibitory activity of SMAX1 to resistance requires SMXL6/7/8. Moreover, the exogenous application of KAR2 enhanced resistance against Pst, but KAR-induced resistance depended on salicylic acid (SA) signaling. Inhibition of karrikin signaling delayed SA-mediated defense responses and inhibited pathogen-induced protein biosynthesis. Together, we propose that the MAX2-KAI2-SMAX1 complex regulates resistance with the assistance of SMXL6/7/8 and SA signaling and that SMAX1/SMXLs possibly form a multimeric complex with their target transcription factors to fine tune immune responses.

Karrikins control seedling photomorphogenesis and anthocyanin biosynthesis through a HY5-BBX transcriptional module.

The butenolide molecule, karrikin (KAR), emerging in smoke of burned plant material, enhances light responses such as germination, inhibition of hypocotyl elongation, and anthocyanin accumulation in Arabidopsis. The KAR signaling pathway consists of KARRIKIN INSENSITIVE 2 (KAI2) and MORE AXILLARY GROWTH 2 (MAX2), which, upon activation, act in an SCF E3 ubiquitin ligase complex to target the downstream signaling components SUPPRESSOR OF MAX2 1 (SMAX1) and SMAX1-LIKE 2 (SMXL2) for degradation. How degradation of SMAX1 and SMXL2 is translated into growth responses remains unknown. Although light clearly influences the activity of KAR, the molecular connection between the two pathways is still poorly understood. Here, we demonstrate that the KAR signaling pathway promotes the activity of a transcriptional module consisting of ELONGATED HYPOCOTYL 5 (HY5), B-BOX DOMAIN PROTEIN 20 (BBX20), and BBX21. The bbx20 bbx21 mutant is largely insensitive to treatment with KAR2 , similar to a hy5 mutant, with regards to inhibition of hypocotyl elongation and anthocyanin accumulation. Detailed analysis of higher order mutants in combination with RNA-sequencing analysis revealed that anthocyanin accumulation downstream of SMAX1 and SMXL2 is fully dependent on the HY5-BBX module. However, the promotion of hypocotyl elongation by SMAX1 and SMXL2 is, in contrast to KAR2 treatment, only partially dependent on BBX20, BBX21, and HY5. Taken together, these results suggest that light- and KAR-dependent signaling intersect at the HY5-BBX transcriptional module.

SMAX1 Integrates Karrikin and Light Signals into GA-Mediated Hypocotyl Growth during Seedling Establishment.

Morphogenic adaptation of young seedlings to light environments is a critical developmental process that ensures plant survival and propagation, as they emerge from the soil. Photomorphogenic responses are facilitated by a network of light and growth hormonal signals, such as auxin and gibberellic acid (GA). Karrikins (KARs), a group of butenolide compounds produced from burning plant materials in wildfires, are known to stimulate seed germination in fire-prone plant species. Notably, recent studies support that they also regulate seedling growth, while underlying molecular mechanisms have been unexplored yet. Here, we demonstrate that SUPPRESSOR OF MAX2 1 (SMAX1), a negative regulator of KAR signaling, integrates light and KAR signals into GA-DELLA pathways that regulate hypocotyl growth during seedling establishment. We found that SMAX1 facilitates degradation of DELLA proteins in the hypocotyls. Interestingly, light induces the accumulation of SMAX1 proteins, and SMAX1-mediated degradation of DELLA is elevated in seedling establishment during the dark-to-light transition. Our observations indicate that SMAX1-mediated integration of light and KAR signals into GA pathways elaborately modulates seedling establishment.

Karrikin signaling regulates hypocotyl shade avoidance response by modulating auxin homeostasis in Arabidopsis.

Shade affects all aspects of plant growth and development, including seed germination, hypocotyl elongation, petiole growth, leaf hyponasty, and flowering time. Here, we found that mutations in the key Arabidopsis karrikins signal perception-associated KARRIKIN INSENSITIVE 2 (KAI2) gene, encoding an α/ß-fold hydrolase, and the MORE AXILLARY GROWTH 2 (MAX2) gene, encoding an F-box protein, led to greater hypocotyl elongation under shade avoidance conditions. We further verified that these phenotypes were caused by perception of the endogenous KAI2-ligands (KLs), and that this phenotype is independent of strigolactone biosynthetic or signaling pathways. Upon perception of a KL, it is probable that the target protein forms a complex with the KAI2/MAX2 proteins, which are degraded through the action of the 26S proteasome. We demonstrated that SUPPRESSOR OF MAX2-1 (SMAX1) is the degradation target for the KAI2/MAX2 complex in the context of shade avoidance. KAI2 and MAX2 require SMAX1 to limit the hypocotyl growth associated with shade avoidance. Treatment with l-kynurenine, an inhibitor of auxin accumulation, partially restored elongation of kai2 mutant hypocotyls under simulated shade. Furthermore, KAI2 is involved in regulating auxin accumulation and polar auxin transport, which may contribute to the hypocotyl shade response. In addition, SMAX1 gene overexpression promoted the hypocotyl shade response. RNA-sequencing analysis revealed that SMAX1-overexpression affected the expression of many auxin homeostasis genes, especially under simulated shade. Altogether, our data support the conclusion that KL signaling regulates shade avoidance by modulating auxin homeostasis in the hypocotyl.

Synthesis of sulfur karrikin bioisosteres as potential neuroprotectives.

The only known sulfur-containing karrikin, 3-methyl-2H-thiopyrano[3,4-b]furan-2-one, has been recently identified as an extremely efficient neuroprotective butenolide. Herein, we report the targeted synthesis of this compound as well as new synthetic protocols toward a class of compounds derived from 2H-furo[2,3-c]pyran-2-ones (karrikins) via bioisosteric exchange of oxygen with sulfur. In particular, we present synthetic procedures toward bioisosteres of karrikins with one or two sulfur heteroatoms incorporated into the core backbone together with evaluation of their biological activity in inhibition of acetylcholinesterase.

Comparative functional analyses of DWARF14 and KARRIKIN INSENSITIVE 2 in drought adaptation of Arabidopsis thaliana.

Functional analyses of various strigolactone-deficient mutants have demonstrated that strigolactones enhance drought resistance; however, the mechanistic involvement of the strigolactone receptor DWARF14 (D14) in this trait remains elusive. In this study, loss-of-function analysis of the D14 gene in Arabidopsis thaliana revealed that d14 mutant plants were more drought-susceptible than wild-type plants, which was associated with their larger stomatal aperture, slower abscisic acid (ABA)-mediated stomatal closure, lower anthocyanin content and delayed senescence under drought stress. Transcriptome analysis revealed a consistent alteration in the expression levels of many genes related to the observed physiological and biochemical changes in d14 plants when compared with the wild type under normal and dehydration conditions. A comparative drought resistance assay confirmed that D14 plays a less critical role in Arabidopsis drought resistance than its paralog karrikin receptor KARRIKIN INSENSITIVE 2 (KAI2). In-depth comparative analyses of the single mutants d14 and kai2 and the double mutant d14 kai2, in relation to various drought resistance-associated mechanisms, revealed that D14 and KAI2 exhibited a similar effect on stomatal closure. On the other hand, D14 had a lesser role in the maintenance of cell membrane integrity, leaf cuticle structure and ABA-induced leaf senescence, but a greater role in drought-induced anthocyanin biosynthesis, than KAI2. Interestingly, a possible additive relationship between D14 and KAI2 could be observed in regulating cell membrane integrity and leaf cuticle development. In addition, our findings also suggest the existence of a complex interaction between the D14 and ABA signaling pathways in the adaptation of Arabidopsis to drought.

Avena fatua caryopsis dormancy release is associated with changes in KAR1 and ABA sensitivity as well as with ABA reduction in coleorhiza and radicle.

MAIN CONCLUSION: The dormancy release in Avena fatua caryopses was associated with a reduction in the ABA content in embryos, coleorhiza and radicle. The coleorhiza proved more sensitive to KAR1 and less sensitive to ABA than the radicle. The inability of dormant caryopses and ABA-treated non-dormant caryopses to complete germination is related to inhibition and delayed of cell-cycle activation, respectively. As freshly harvested Avena fatua caryopses are dormant at 20 °C, they cannot complete germination; the radicle is not able to emerge. Both karrikin 1 (KAR1) and dry after-ripening release dormancy, enabling the emergence of, first, the coleorhiza and later the radicle. The after-ripening removes caryopse sensitivity to KAR1 and decreases the sensitivity to abscisic acid (ABA). The coleorhiza was found to be more sensitive to KAR1, and less sensitive to ABA, than radicles. Effects of KAR1 and after-ripening were associated with a reduction of the embryo's ABA content during caryopsis germination. KAR1 was found to decrease the ABA content in the coleorhiza and radicles. Germination of after-ripened caryopses was associated with the progress of cell-cycle activation before coleorhiza emergence. Inhibition of the germination completion due to dormancy or treating the non-dormant caryopses with ABA was associated with a total and partial inhibition of cell-cycle activation, respectively.

Desmethyl butenolides are optimal ligands for karrikin receptor proteins.

Strigolactones and karrikins are butenolide molecules that regulate plant growth. They are perceived by the α/ß-hydrolase DWARF14 (D14) and its homologue KARRIKIN INSENSITIVE2 (KAI2), respectively. Plant-derived strigolactones have a butenolide ring with a methyl group that is essential for bioactivity. By contrast, karrikins are abiotic in origin, and the butenolide methyl group is nonessential. KAI2 is probably a receptor for an endogenous butenolide, but the identity of this compound remains unknown. Here we characterise the specificity of KAI2 towards differing butenolide ligands using genetic and biochemical approaches. We find that KAI2 proteins from multiple species are most sensitive to desmethyl butenolides that lack a methyl group. Desmethyl-GR24 and desmethyl-CN-debranone are active by KAI2 but not D14. They are more potent KAI2 agonists compared with their methyl-substituted reference compounds both in vitro and in plants. The preference of KAI2 for desmethyl butenolides is conserved in Selaginella moellendorffii and Marchantia polymorpha, suggesting that it is an ancient trait in land plant evolution. Our findings provide insight into the mechanistic basis for differential ligand perception by KAI2 and D14, and support the view that the endogenous substrates for KAI2 and D14 have distinct chemical structures and biosynthetic origins.

Molecular basis of strigolactone perception in root-parasitic plants: aiming to control its germination with strigolactone agonists/antagonists.

The genus Striga, also called "witchweed", is a member of the family Orobanchaceae, which is a major family of root-parasitic plants. Striga can lead to the formation of seed stocks in the soil and to explosive expansion with enormous seed production and stability once the crops they parasitize are cultivated. Understanding the molecular mechanism underlying the communication between Striga and their host plants through natural seed germination stimulants, "strigolactones (SLs)", is required to develop the technology for Striga control. This review outlines recent findings on the SL perception mechanism, which have been accumulated in Striga hermonthica by the similarity of the protein components that regulate SL signaling in nonparasitic model plants, including Arabidopsis and rice. HTL/KAI2 homologs were identified as SL receptors in the process of Striga seed germination. Recently, this molecular basis has further promoted the development of various types of SL agonists/antagonists as seed germination stimulants or inhibitors. Such chemical compounds are also useful to elucidate the dynamic behavior of SL receptors and the regulation of SL signaling.

Impairment in karrikin but not strigolactone sensing enhances root skewing in Arabidopsis thaliana.

Roots form highly complex systems varying in growth direction and branching pattern to forage for nutrients efficiently. Here mutations in the KAI2 (KARRIKIN INSENSITIVE) α/ß-fold hydrolase and the MAX2 (MORE AXILLARY GROWTH 2) F-box leucine-rich protein, which together perceive karrikins (smoke-derived butenolides), caused alteration in root skewing in Arabidopsis thaliana. This phenotype was independent of endogenous strigolactones perception by the D14 α/ß-fold hydrolase and MAX2. Thus, KAI2/MAX2 effect on root growth may be through the perception of endogenous KAI2-ligands (KLs), which have yet to be identified. Upon perception of a ligand, a KAI2/MAX2 complex is formed together with additional target proteins before ubiquitination and degradation through the 26S proteasome. Using a genetic approach, we show that SMAX1 (SUPPRESSOR OF MAX2-1)/SMXL2 and SMXL6,7,8 (SUPPRESSOR OF MAX2-1-LIKE) are also likely degradation targets for the KAI2/MAX2 complex in the context of root skewing. In A. thaliana therefore, KAI2 and MAX2 act to limit root skewing, while kai2's gravitropic and mechano-sensing responses remained largely unaffected. Many proteins are involved in root skewing, and we investigated the link between MAX2 and two members of the SKS/SKU family. Though KLs are yet to be identified in plants, our data support the hypothesis that they are present and can affect root skewing.

Perception of karrikins by plants: a continuing enigma.

Karrikins are small butenolide molecules with the capacity to promote germination and enhance seedling establishment. Generated abiotically from partial combustion of vegetation, karrikins are comparatively rare in the environment, but studying their mode of action has been most informative in revealing a new regulatory pathway for plant development that uses the karrikin perception machinery. Recent studies suggest that the karrikin receptor protein KAI2 and downstream transcriptional co-repressors in the SMXL family influence seed germination, seedling photomorphogenesis, root morphology, and responses to abiotic stress such as drought. Based on taxonomic distribution, this pathway is ubiquitous and likely to be evolutionarily ancient, originating prior to land plants. However, we still do not have a good grasp on how karrikins actually activate the receptor protein, and we have yet to discover the assumed endogenous ligand for KAI2 that karrikins are thought to mimic. This review covers recent progress in this field, as well as current gaps in our knowledge.