Regulation Of Shoot Branching Research Articles

Characterization of MORE AXILLARY GROWTH Genes in Populus

BackgroundStrigolactones are a new class of plant hormones that play a key role in regulating shoot branching. Studies of branching mutants in Arabidopsis, pea, rice and petunia have identified several key genes involved in strigolactone biosynthesis or signaling pathway. In the model plant Arabidopsis, MORE AXILLARY GROWTH1 (MAX1), MAX2, MAX3 and MAX4 are four founding members of strigolactone pathway genes. However, little is known about the strigolactone pathway genes in the woody perennial plants.Methodology/Principal FindingHere we report the identification of MAX homologues in the woody model plant Populus trichocarpa. We identified the sequence homologues for each MAX protein in P. trichocarpa. Gene expression analysis revealed that Populus MAX paralogous genes are differentially expressed across various tissues and organs. Furthermore, we showed that Populus MAX genes could complement or partially complement the shoot branching phenotypes of the corresponding Arabidopsis max mutants.Conclusion/SignificanceThis study provides genetic evidence that strigolactone pathway genes are likely conserved in the woody perennial plants and lays a foundation for further characterization of strigolactone pathway and its functions in the woody perennial plants.

Conditional Auxin Response and Differential Cytokinin Profiles in Shoot Branching Mutants.

Strigolactone (SL), auxin, and cytokinin (CK) are hormones that interact to regulate shoot branching. For example, several ramosus (rms) branching mutants in pea (Pisum sativum) have SL defects, perturbed xylem CK levels, and diminished responses to auxin in shoot decapitation assays. In contrast with the last of these characteristics, we discovered that buds on isolated nodes (explants) of rms plants instead respond normally to auxin. We hypothesized that the presence or absence of attached roots would result in transcriptional and hormonal differences in buds and subtending stem tissues, and might underlie the differential auxin response. However, decapitated plants and explants both showed similar up-regulation of CK biosynthesis genes, increased CK levels, and down-regulation of auxin transport genes. Moreover, auxin application counteracted these trends, regardless of the effectiveness of auxin at inhibiting bud growth. Multivariate analysis revealed that stem transcript and CK changes were largely associated with decapitation and/or root removal and auxin response, whereas bud transcript profiles related more to SL defects. CK clustering profiles were indicative of additional zeatin-type CKs in decapitated stems being supplied by roots and thus promoting bud growth in SL-deficient genotypes even in the presence of added auxin. This difference in CK content may explain why rms buds on explants respond better to auxin than those on decapitated plants. We further conclude that rapid changes in CK status in stems are auxin dependent but largely SL independent, suggesting a model in which auxin and CK are dominant regulators of decapitation-induced branching, whereas SLs are more important in intact plants.

Strigolactones and the control of plant development: lessons from shoot branching

Strigolactones (SLs) were originally identified through their activities as root exudates in the rhizosphere; however, it is now clear that they have many endogenous signalling roles in plants. In this review we discuss recent progress in understanding SL action in planta, particularly in the context of the regulation of shoot branching, one of the best-characterized endogenous roles for SLs. Rapid progress has been made in understanding SL biosynthesis, but many questions remain unanswered. There are hints of as yet unidentified sources of SL, as well as unknown SL-like molecules with important signalling functions. SL signalling is even more enigmatic. Although a likely receptor has been identified, along with some candidate immediate downstream targets, our understanding of how these targets mediate SL signalling is limited. There is still considerable uncertainty about whether the targets of SL signalling are primarily transcriptional or not. There is at least one non-transcriptional target, because a rapid primary response to SL is the removal of PIN1 auxin exporter proteins from the plasma membrane in vascular-associated cells of the stem. We discuss how the various early events in SL signalling could result in the observed changes in shoot branching.

Strigolactone Promotes Degradation of DWARF14, an α/β Hydrolase Essential for Strigolactone Signaling inArabidopsis

Strigolactones (SLs) are phytohormones that play a central role in regulating shoot branching. SL perception and signaling involves the F-box protein MAX2 and the hydrolase DWARF14 (D14), proposed to act as an SL receptor. We used strong loss-of-function alleles of the Arabidopsis thaliana D14 gene to characterize D14 function from early axillary bud development through to lateral shoot outgrowth and demonstrated a role of this gene in the control of flowering time. Our data show that D14 distribution in vivo overlaps with that reported for MAX2 at both the tissue and subcellular levels, allowing physical interactions between these proteins. Our grafting studies indicate that neither D14 mRNA nor the protein move over a long range upwards in the plant. Like MAX2, D14 is required locally in the aerial part of the plant to suppress shoot branching. We also identified a mechanism of SL-induced, MAX2-dependent proteasome-mediated degradation of D14. This negative feedback loop would cause a substantial drop in SL perception, which would effectively limit SL signaling duration and intensity.

Carlactone is an endogenous biosynthetic precursor for strigolactones

Strigolactones (SLs) are a class of terpenoid plant hormones that regulate shoot branching as well as being known as root-derived signals for symbiosis and parasitism. SL has tricyclic-lactone (ABC-ring) and methyl butenolide (D-ring), and they are connected through an enol ether bridge. Recently, a putative biosynthetic intermediate called carlactone (CL), of which carbon skeleton is in part similar to those of SLs, was identified by biochemical analysis of three biosynthetic enzymes, DWARF27, CAROTENOID CLEAVAGE DIOXYGENASE 7 (CCD7), and CCD8 in vitro. However, CL has never been identified from plant tissues, and the conversion of CL to SLs has not been proven in vivo. To address these questions, we chemically synthesized (13)C-labeled CL. We show that (13)C-labeled CL is converted to (-)-[(13)C]-2'-epi-5-deoxystrigol ((-)-2'-epi-5DS) and [(13)C]-orobanchol, endogenous SLs in rice, in the dwarf10 mutant, which is defective in CCD8. In addition, we successfully identified endogenous CL by using liquid chromatography-quadrupole/time-of-flight tandem mass spectrometry in rice and Arabidopsis. Furthermore, we determined the absolute stereochemistry of endogenous CL to be (11R)-configuration, which is the same as that of (-)-2'-epi-5DS at the corresponding position. Feeding experiments showed that only the (11R)-isomer of CL, but not the (11S)-isomer, was converted to (-)-2'-epi-5DS in vivo. Taken together, our data provide conclusive evidence that CL is an endogenous SL precursor that is stereospecifically recognized in the biosynthesis pathway.

Strigolactone/MAX2-Induced Degradation of Brassinosteroid Transcriptional Effector BES1 Regulates Shoot Branching

Strigolactones (SLs), a class of the most recently identified terpenoid phytohormones, play essential roles in plant development, specifically in suppressing shoot branching. MAX2, a subunit of an SCF E3 ligase and a positive regulator that inhibits shoot branching, is likely a key SL signaling component. Here, we provide genetic and biochemical evidence to demonstrate that BES1 interacts with MAX2 and acts as its substrate to regulate SL-responsive gene expression. Additional AtD14, a putative receptor of SLs, can promote BES1 degradation. Knockdown of BES1 and its homologs dramatically suppressed the branching phenotype of max2-1 mutant. These results portray an SL signaling cascade from the putative receptor to downstream transcription factors. In addition, we demonstrate that the SL and brassinosteroid (BR) signaling pathways distinctly regulate the same transcription factor, BES1, to control specific developmental processes.

Strigolactone Can Promote or Inhibit Shoot Branching by Triggering Rapid Depletion of the Auxin Efflux Protein PIN1 from the Plasma Membrane

Author SummaryPlants can adapt their form to suit the environment in which they are growing. For example, genetically identical plants can develop as a single unbranched stem or as a highly ramified bush. This broad developmental potential is possible because the shoot system is produced continuously by growing tips, known as shoot meristems. Meristems produce the stem and leaves of a shoot, and at the base of each leaf, a new meristem is formed. This meristem can remain dormant as a small bud or activate to produce a branch. Thus, the shoot system is a community of shoot meristems, the combined activity and inactivity of which shape shoot form. Here we provide evidence that growth is balanced across the Arabidopsis shoot system by competition between the shoot meristems. This competition is likely mediated by the requirement of meristems to export the plant hormone auxin in order to activate bud outgrowth. In our model, auxin in the main stem, exported from active branches, can prevent auxin export by dormant buds, thus preventing their activation. Our findings show that a second hormone, strigolactone, increases the level of competition between branches by making auxin export harder to establish. Together, these hormones balance growth across the shoot system, adjusting it according to the environmental conditions in which a plant is growing.

Recent Advances in Strigolactone Research: Chemical and Biological Aspects

Strigolactones (SLs) are a group of terpenoid lactones that were discovered in the 1960s. They were initially characterized as allelochemicals secreted from roots to the rhizosphere, and have functions in parasitic and symbiotic interactions with root parasitic plants and arbuscular mycorrhizal (AM) fungi, respectively. In 2008, SLs were shown to act as endogenous hormones that regulate shoot branching. The discovery of a hormonal function for SLs has provided a link between genetically studied shoot branching mutants and chemically characterized SLs in earlier studies. This has offered new strategies and experimental tools to address a number of intriguing questions as to the biological function and molecular action of SLs. In this review, we will provide an overview of recent topics on SLs, and highlight new discoveries regarding its biosynthetic pathway and multiple hormonal roles in plant development and adaptive responses.

Grasses provide new insights into regulation of shoot branching

Tillering (branching) is a major determinant of crop yield that is controlled by complex interactions between hormonal, developmental, and environmental factors. Historically, research on shoot branching has focused on eudicots, mainly due to the ease of manipulating branching by shoot decapitation and grafting in these species. These studies demonstrated hormonal control of branching. Recent studies in monocots have contributed to our knowledge of tillering/branching by identifying novel branching genes and regulatory mechanisms. A comparison of branching controls in eudicots and monocots reveals that the regulatory signals and genes are broadly conserved, but that there are differences in the detail.

The Arabidopsis Ortholog of Rice DWARF27 Acts Upstream of MAX1 in the Control of Plant Development by Strigolactones

Strigolactones (SLs) are carotenoid-derived plant hormones that regulate shoot branching, secondary growth, root development, and responses to soil phosphate. In Arabidopsis (Arabidopsis thaliana), SL biosynthesis requires the sequential action of two carotenoid cleavage dioxygenases, MORE AXILLARY GROWTH3 (MAX3) and MAX4, followed by a cytochrome P450, MAX1. In rice (Oryza sativa), the plastid-localized protein DWARF27 (OsD27) is also necessary for SL biosynthesis, but the equivalent gene in Arabidopsis has not been identified. Here, we use phylogenetic analysis of D27-like sequences from photosynthetic organisms to identify AtD27, the likely Arabidopsis ortholog of OsD27. Using reverse genetics, we show that AtD27 is required for the inhibition of secondary bud outgrowth and that exogenous application of the synthetic SL GR24 can rescue the increased branching phenotype of an Atd27 mutant. Furthermore, we use grafting to demonstrate that AtD27 operates on a nonmobile precursor upstream of MAX1 in the SL biosynthesis pathway. Consistent with the plastid localization of OsD27, we also show that AtD27 possesses a functional plastid transit peptide. We demonstrate that AtD27 transcripts are subject to both local feedback and auxin-dependent signals, albeit to a lesser extent than MAX3 and MAX4, suggesting that early steps in SL biosynthesis are coregulated at the transcriptional level. By identifying an additional component of the canonical SL biosynthesis pathway in Arabidopsis, we provide a new tool to investigate the regulation of shoot branching and other SL-dependent developmental processes.

Strigolactone, a key regulator of nutrient allocation in plants

Strigolactones (SLs) are a group of terpenoid lactones that are derived from carotenoids. SLs have been found in a number of plant species and appear to serve several diverse physiological functions. SLs were first identified by their ability to stimulate seed germination of root-parasitic plants. Later, SLs were isolated as hyphal-branching inducers of arbuscular mycorrhizal fungi, which facilitate the uptake of soil nutrients by plants. Most recently, SLs (or their derivatives) were found to be a new class of plant hormones that inhibit shoot branching. Considering these three roles of SLs, it was unclear at first why communication signals in the rhizosphere would regulate shoot branching in the host plant. Recent reports, however, suggest that plants produce SLs in response to nitrogen and phosphorus deficiency, stimulating changes in plant shoot and root architecture that enable them to adapt to environmental conditions. Excess SLs produced in roots are released into the soil, where they stimulate the growth of arbuscular mycorrhizal fungi. These symbiotic fungi supply inorganic nutrients that can be used by the plant. This review paper focuses on the physiological roles of SLs as a key regulator of nutrient allocation in plants.

The role of strigolactones in host specificity ofOrobancheandPhelipancheseed germination

AbstractStrigolactones are apocarotenoids regulating shoot branching. They are also known to be exuded by plant roots at very low concentrations, stimulating hyphal branching of arbuscular mycorrhizal fungi and germination of root parasitic weed seeds. We show that strigolactones play a major role in host specificity ofOrobancheandPhelipanche(the broomrapes) seed germination. This observation confirms that host-derived germination stimulants are an important component determining the host specificity of these parasitic plants. Weedy broomrape species were less specialized in germination requirements than the non-weedy species except forO. cumanaandO. foetidavar.broteri. Similar results were obtained with the root exudates. Some species, such asP. aegyptiacaandO. minor, showed a broad spectrum of host specificity in terms of seed germination, which was stimulated by exudates from the majority of species tested, whereas others, such asO. cumana,O. hederaeandO. densiflora, were highly specific. Some species, such asO. minor,P. aegyptiacaandP. nana, were responsive to the three strigolactones studied, whereas others were induced by only one of them, or did not respond to them at all. The synthetic strigolactone analogue GR24, generally used as a standard for germination tests, was not effective on someOrobancheandPhelipanchespecies. Seeds of some species that did not respond to GR24 were induced to germinate in the presence of fabacyl acetate or strigol, confirming the role of strigolactones in host specificity.

MAX4 gene is involved in the regulation of low inorganic phosphate stress responses in Arabidopsis thaliana

MAX4 gene has been shown to be involved in the regulation of shoot branching in Arabidopsis (Arabidopsis thaliana). However, little is known about the role of MAX4 gene in low inorganic phosphate (Pi) stress response in Arabidopsis. Here we showed that MAX4 gene is involved in the regulation of low Pi stress response in Arabidopsis. MAX4 gene was repressed by low Pi stress, and the max4 mutants showed lower anthocyanin content and longer primary root length. In addition, max4 mutant plants also displayed altered root architecture such as increased root-to-shoot ratio, lower lateral root number and root hair density compared with wild-type plants under low Pi stress. Higher total Pi contents were detected in shoots and roots of max4 plants than those of wild-type plants when subjected to low Pi stress, which was associated, at least in part, with increase in expression of WRKY75 as well as AtPT1 and AtPT2 genes encoding high-affinity Pi transporters. Taken together, all these results suggest that MAX4 gene mediates low Pi stress response, at least in part, by regulating the expression of WRKY75 as well as AtPT1 and AtPT2 genes.

MicroRNA171c-Targeted SCL6-II, SCL6-III, and SCL6-IV Genes Regulate Shoot Branching in Arabidopsis

MicroRNAs (miRNAs) are ∼21-nucleotide noncoding RNAs that play critical roles in regulating plant growth and development through directing the degradation of target mRNAs. Axillary meristem activity, and hence shoot branching, is influenced by a complicated network that involves phytohormones such as auxin, cytokinin, and strigolactone. GAI, RGA, and SCR (GRAS) family members take part in a variety of developmental processes, including axillary bud growth. Here, we show that the Arabidopsis thaliana microRNA171c (miR171c) acts to negatively regulate shoot branching through targeting GRAS gene family members SCARECROW-LIKE6-II (SCL6-II), SCL6-III, and SCL6-IV for cleavage. Transgenic plants overexpressing MIR171c (35Spro-MIR171c) and scl6-II scl6-III scl6-IV triple mutant plants exhibit a similar reduced shoot branching phenotype. Expression of any one of the miR171c-resistant versions of SCL6-II, SCL6-III, and SCL6-IV in 35Spro-MIR171c plants rescues the reduced shoot branching phenotype. Scl6-II scl6-III scl6-IV mutant plants exhibit pleiotropic phenotypes such as increased chlorophyll accumulation, decreased primary root elongation, and abnormal leaf and flower patterning. SCL6-II, SCL6-III, and SCL6-IV are located to the nucleus, and show transcriptional activation activity. Our results suggest that miR171c-targeted SCL6-II, SCL6-III, and SCL6-IV play an important role in the regulation of shoot branch production.

Over-expression of the IGI1 leading to altered shoot-branching development related to MAX pathway in Arabidopsis

Shoot branching and growth are controlled by phytohormones such as auxin and other components in Arabidopsis. We identified a mutant (igi1) showing decreased height and bunchy branching patterns. The phenotypes reverted to the wild type in response to RNA interference with the IGI1 gene. Histochemical analysis by GUS assay revealed tissue-specific gene expression in the anther and showed that the expression levels of the IGI1 gene in apical parts, including flowers, were higher than in other parts of the plants. The auxin biosynthesis component gene, CYP79B2, was up-regulated in igi1 mutants and the IGI1 gene was down-regulated by IAA treatment. These results indicated that there is an interplay regulation between IGI1 and phytohormone auxin. Moreover, the expression of the auxin-related shoot branching regulation genes, MAX3 and MAX4, was down-regulated in igi1 mutants. Taken together, these results indicate that the overexpression of the IGI1 influenced MAX pathway in the shoot branching regulation.Electronic supplementary materialThe online version of this article (doi:10.1007/s11103-010-9645-0) contains supplementary material, which is available to authorized users.

Structural requirements of strigolactones for hyphal branching in AM fungi.

Strigolactones are a group of terpenoid lactones that act as a host-derived signal in the rhizosphere communication of plants with arbuscular mycorrhizal (AM) fungi and root parasitic weeds as well as an endogenous plant hormone regulating shoot branching in plants. Strigolactones induce hyphal branching in AM fungi at very low concentrations, suggesting a highly sensitive perception system for strigolactones present in AM fungi. However, little is known about the structural requirements of strigolactones for hyphal branching in AM fungi. Here, we tested a series of natural and synthetically modified strigolactones as well as non-strigolactone-type germination stimulants for hyphal branching-inducing activity in germinating spores of the AM fungus Gigaspora margarita. All tested compounds with a tricyclic lactone coupled to a methylbutenolide via an enol ether bond showed activity, but differed in the active concentration and in the branching pattern of hyphae. Truncation of the A- and AB-rings in the tricyclic ABC lactone of strigolactones resulted in a drastic reduction in hyphal branching activity. Although the connection of the C-ring in the tricyclic lactone to the methylbutenolide D-ring was shown to be essential for hyphal branching, the bridge structure in the C–D part was found not necessarily to be enol ether, being replaceable with either alkoxy or imino ethers. These structural requirements in AM fungi are very similar but not identical to those observed in root parasitic weeds, especially with respect to the enol ether bridge in the C–D part.

Auxin and strigolactones in shoot branching: intimately connected?

Axillary meristems form in the axils of leaves. After an initial phase of meristematic activity during which a small axillary bud is produced, they often enter a state of suspended growth from which they may be released to form a shoot branch. This post-embryonic growth plasticity is typical of plants and allows them to adapt to changing environmental conditions. The shoot architecture of genotypically identical plants may display completely contrasting phenotypes when grown in distinct environmental niches, with one having only a primary inflorescence and many arrested axillary meristems and the other displaying higher orders of branches. In order to cease and resume growth as required, the plant must co-ordinate its intrinsic developmental programme with the responses to environmental cues. It is thought that information from the environment is integrated throughout the plant using plant hormones as long-distance signals. In the present review, we focus primarily on how two of these hormones, auxin and strigolactones, may be acting to regulate shoot branching.

Emergence of tissue polarization from synergy of intracellular and extracellular auxin signaling

Plant development is exceptionally flexible as manifested by its potential for organogenesis and regeneration, which are processes involving rearrangements of tissue polarities. Fundamental questions concern how individual cells can polarize in a coordinated manner to integrate into the multicellular context. In canalization models, the signaling molecule auxin acts as a polarizing cue, and feedback on the intercellular auxin flow is key for synchronized polarity rearrangements. We provide a novel mechanistic framework for canalization, based on up-to-date experimental data and minimal, biologically plausible assumptions. Our model combines the intracellular auxin signaling for expression of PINFORMED (PIN) auxin transporters and the theoretical postulation of extracellular auxin signaling for modulation of PIN subcellular dynamics. Computer simulations faithfully and robustly recapitulated the experimentally observed patterns of tissue polarity and asymmetric auxin distribution during formation and regeneration of vascular systems and during the competitive regulation of shoot branching by apical dominance. Additionally, our model generated new predictions that could be experimentally validated, highlighting a mechanistically conceivable explanation for the PIN polarization and canalization of the auxin flow in plants.

Studies on the Host Recognition Mechanism of Root Parasitic Plants

Among parasitic angiosperms, witchweeds (Striga spp.) and broomrapes (Orobanche and Phelipanche spp.) are the two most devastating root parasitic plants, causing enormous crop losses around the world. These root parasites have evolved special strategies to ensure their survival; the seeds germinate only when they are within the host rhizosphere so that after germination the seedlings can reach and parasitize the host roots. To locate host roots, root parasites exploit strigolactones (SLs), host-derived chemical signals that are released by host plants for symbiotic arbuscular mycorrhizal (AM) fungi. In addition to these functions as rhizosphere signaling molecules, SLs act as a class of plant hormones regulating shoot branching. Using high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS), SLs in root exudates from various plants have been characterized. During studies, in addition to the known SLs, more than 10 novel SLs have been isolated and their structures determined. Plants produce and exude mixtures of SLs, and nutrient availability affects SL production and exudation. These results suggest that quantitative and qualitative differences in SL production and/or exudation are important in host recognition of root parasitic plants.

Computational modeling and molecular physiology experiments reveal new insights into shoot branching in pea.

Bud outgrowth is regulated by the interplay of multiple hormones, including auxin, cytokinin, strigolactones, and an unidentified long-distance feedback signal that moves from shoot to root. The model of bud outgrowth regulation in pea (Pisum sativum) includes these signals and a network of five RAMOSUS (RMS) genes that operate in a shoot-root-shoot loop to regulate the synthesis of, and response to, strigolactones. The number of components in this network renders the integration of new and existing hypotheses both complex and cumbersome. A hypothesis-driven computational model was therefore developed to help understand regulation of shoot branching. The model evolved in parallel with stepwise laboratory research, helping to define and test key hypotheses. The computational model was used to verify new mechanisms involved in the regulation of shoot branching by confirming that the new hypotheses captured all relevant biological data sets. Based on cytokinin and RMS1 expression analyses, this model is extended to include subtle but important differences in the function of RMS3 and RMS4 genes in the shoot and rootstock. Additionally, this research indicates that a branch-derived signal upregulates RMS1 expression independent of the other feedback signal. Furthermore, we propose xylem-sap cytokinin promotes sustained bud outgrowth, rather than acting at the earlier stage of bud release.