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Kirschner, G. K.

Publications and source records attributed to Kirschner, G. K..

7 recordsLinked to original sources

Cytokinin regulates the gravitropic setpoint angle of seminal roots in barley and wheat

Plant root growth angle determines the spatial distribution of roots in soil and thereby influences access to water and nutrients when these resources are heterogeneously distributed. In barley, multiple seminal roots emerge at distinct gravitropic setpoint angles (GSAs) and they reliably re-establish their characteristic growth angles following reorientation, indicating robust intrinsic control. However, the molecular basis underlying GSA variation among roots within a single root system remains unknown. Here, we combined individual root type-specific transcriptomics with laser-capture microdissection to profile gravity-responsive zones (root cap, meristem, and elongation zone). We identified two CYTOKININ OXIDASE/DEHYDROGENASE (CKX) genes whose expression in the root cap was associated with root-specific GSA. Functional analysis showed that ckx mutants, which show higher cytokinin levels, exhibit significantly steeper seminal root GSAs, implicating cytokinin catabolism in root angle regulation. Consistent with this, exogenous cytokinin treatment induced a dose-dependent steepening in root angle in barley. Analyses of gravitropic curvature after gravistimulation further showed that ckx mutants exhibit altered gravitropic responses, indicating that CKX-mediated regulation of the root angle depends on gravity-responsive pathways. Notably, GSA-associated CKX expression patterns and cytokinin-dependent root angle responses were also observed in wheat, suggesting a conservation of this mechanism across cereals. Together, these findings identify cytokinin as a key regulator of root specific setpoint angles within a root system architecture, with potential implications for optimizing crop resource acquisition and yield.

plant biology↗

Root phenotypic plasticity improves yield stability when directed toward an adaptive integrated phenotype

Root phenotypic plasticity is often proposed to improve crop performance under stress, yet it remains unclear how much plasticity is beneficial and whether adaptive responses require changes across many traits or adjustments in few specific traits. Using public data of 6,500 field-grown maize and barley plants, this study examined the extent and distribution of root plasticity, and when it is associated with yield stability. We quantified root plasticity across nine anatomical and architectural traits using complementary statistical models and applied a feature-discovery framework to identify the drought-associated optimal integrated phenotypes and determine whether plasticity toward these phenotypes improved yield stability. More plasticity did not mean greater yield stability. Neither the number of plastic traits nor the magnitude of plastic responses predicted yield stability. Rather, we identified species-specific high-yielding, stable integrated phenotypes defined by distinct trait configurations. Critically, genotypes whose plastic responses moved their root phenotype toward these targets achieved greater yield stability, whereas movement away from them was associated with lower stability. Root plasticity is adaptive when it shifts root phenotypes towards an optimal integrated phenotype. These findings show that the value of plasticity depends on the trajectory of phenotypic change rather than its magnitude alone.

plant biology↗

CLAVATA signalling shapes barley inflorescence architecture by controlling activity and determinacy of shoot apical and rachilla meristems

Grasses exhibit a large variety of diverse inflorescence architectures, from complex branched inflorescences in Oryzeae (rice) to simple spike-type inflorescences in Triticeae (e.g. barley, wheat). Inflorescence architecture depends on shape, longevity and determinacy of meristems that direct growth of the main rachis and lateral branches, but how individual meristem activities are determined and integrated within complex inflorescences is not yet understood. We found that activity of distinct meristems in the barley inflorescence is coordinated by a signalling pathway comprising the receptor like kinase Hordeum vulgare CLAVATA1 (HvCLV1) and the secreted CLAVATA3/ENDOSPERM SURROUNDING REGION (CLE)-family peptide FON2- LIKE CLE PROTEIN1 (HvFCP1). HvFCP1 interacts with HvCLV1 to promote spikelet formation but restricts inflorescence meristem and rachilla meristem proliferation. Hvfcp1 or Hvclv1 mutants generate branched inflorescences with additional rows of spikelets and supernumerary florets. Transcriptome analysis reveals that HvFCP1/HvCLV1 signalling controls inflorescence branching through the regulation of trehalose-6-phosphate synthesis and sugar transport. Our discoveries reveal the potential to engineer barley inflorescence architecture by manipulating regulation of distinct meristem activities.

plant biology↗

The PLETHORA/PIN-FORMED/AUXIN network mediates terminal prehaustorium formation in the parasitic plant Striga hermonthica

The parasitic plant Striga hermonthica invades the host root through the formation of a haustorium and has detrimental impacts on cereal crops. The haustorium is derived directly from the differentiation of the Striga radicle. Currently, how Striga root cell lineages are patterned and the molecular mechanisms leading to radicle differentiation shortly after germination remain unclear. In this study, we determined the developmental-morphodynamic programs that regulate terminal haustorium formation in S. hermonthica at spatiotemporal and cellular resolutions. We showed that in S. hermonthica roots, meristematic cells first undergo multiplanar divisions, which decrease during growth and correlate with reduced expression of the stem cell regulator PLETHORA1. We also found that PIN-FORMED (PIN) proteins undergo a shift in polarity. Using the layout of the root structure and the polarity of outer-membrane PIN proteins, we constructed a mathematical model of auxin transport that explains the auxin distribution patterns observed during S. hermonthica root growth. Our results reveal a fundamental molecular and cellular framework governing the switch of S. hermonthica roots from the vegetative to the invasive state by inducing meristem differentiation through auxin excretion to the environment and explain how asymmetric PIN polarity controls auxin distribution to maintain meristem activity and sustain root growth.

plant biology↗

ENHANCED GRAVITROPISM 2 encodes a STERILE ALPHA MOTIVE containing protein that controls root growth angle in barley and wheat

The root growth angle defines how roots grow towards the gravity vector and is among the most important determinants of root system architecture. It controls water uptake capacity, nutrient use efficiency, stress resilience and as a consequence yield of crop plants. We demonstrated that the egt2 (enhanced gravitropism 2) mutant of barley exhibits steeper root growth of seminal and lateral roots and an auxin independent higher responsiveness to gravity compared to wild type plants. We cloned the EGT2 gene by a combination of bulked segregant analysis and whole genome sequencing. Subsequent validation experiments by an independent CRISPR/Cas9 mutant allele demonstrated that egt2 encodes a STERILE ALPHA MOTIF domain containing protein. In situ hybridization experiments illustrated that EGT2 is expressed from the root cap to the elongation zone. Subcellular localization experiments revealed that EGT2 localizes to the nucleus and cytoplasm. We demonstrated the evolutionary conserved role of EGT2 in root growth angle control between barley and wheat by knocking out the EGT2 orthologs in the A and B genomes of tetraploid durum wheat. By combining laser capture microdissection with RNA-seq, we observed that seven expansin genes were transcriptionally downregulated in the elongation zone. This is consistent with a role of EGT2 in this region of the root where the effect of gravity sensing is executed by differential cell elongation. Our findings suggest that EGT2 is an evolutionary conserved regulator of root growth angle in barley and wheat that could be a valuable target for root-based crop improvement strategies in cereals. Significance StatementTo date the potential of utilizing root traits in plant breeding remains largely untapped. In this study we cloned and characterized the ENHANCED GRAVITROPISM2 (EGT2) gene of barley that encodes a STERILE ALPHA MOTIF domain containing protein. We demonstrated that EGT2 is a key gene of root growth angle regulation in response to gravity which is conserved in barley and wheat and could be a promising target for crop improvement in cereals.

plant biology↗

PLETHORA and WOX5 interaction and subnuclear localisation regulates Arabidopsis root stem cell maintenance

Maintenance and homeostasis of the stem cell niche (SCN) in the Arabidopsis root is essential for growth and development of all root cell types. The SCN is organized around a quiescent center (QC) maintaining the stemness of cells in direct contact. The key transcription factors (TFs) WUSCHEL-RELATED HOMEOBOX 5 (WOX5) and PLETHORAs (PLTs) are expressed in the SCN where they maintain the QC and regulate distal columella stem cell (CSC) fate. Here, we describe the concerted mutual regulation of the key TFs WOX5 and PLTs on a transcriptional and protein interaction level. Additionally, by applying a novel SCN staining method, we demonstrate that both WOX5 and PLTs regulate root SCN homeostasis as they control QC quiescence and CSC fate interdependently. Moreover, we uncover that some PLTs, especially PLT3, contain intrinsically disordered prion-like domains (PrDs) that are necessary for complex formation with WOX5 and its recruitment to subnuclear microdomains/nuclear bodies (NBs) in the CSCs. We propose that this partitioning of PLT-WOX5 complexes to NBs, possibly by phase separation, is important for CSC fate determination.

plant biology↗

Fluorescent reporter lines for auxin and cytokinin signalling in barley (Hordeum vulgare)

The phytohormones auxin and cytokinin influence the development and maintenance of plant stem cell niches. Although barley (Hordeum vulgare) is the fourth most abundant cereal crop plant, the knowledge about these important phytohormones in regard to the root and shoot stem cell niche in barley is still negligible. In this study, we analyse the influence of auxin and cytokinin on the barley root meristem and present reporter lines to describe the auxin and cytokinin signalling output. Application of high concentrations of auxin and cytokinin to barley seedlings had a negative influence on barley root and meristem growth. The expression of the cytokinin reporter TCSn revealed that cytokinin signalling mostly takes place in the stele cells proximal to the QC and in the differentiated root cap cells, but can additionally be activated in the root stem cell niche by cytokinin application. Analysing signalling targets of auxin showed that a homologue of AtPLT1, HvPLT1, is expressed in a similar way as AtPLT1 in Arabidopsis, in particular in the QC and the surrounding cells. Furthermore, a homologue of the auxin PIN transporters PIN1, HvPIN1, was expressed in the root and the shoot meristem and polarly localizes to the plasma membrane. Its expression is regulated by cytokinin and the intracellular localisation is affected by BFA. With this study, we provide a valuable tool set of fluorescent barley reporter lines for auxin and cytokinin.

plant biology↗