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Biology subjects

Mueller, L. M.

Publications and source records attributed to Mueller, L. M..

6 recordsLinked to original sources

Plant-parasitic nematodes produce functional mimics of plant PSK peptides to facilitate parasitism

Root-knot nematodes are obligate plant parasites that cause substantial agricultural losses worldwide. They induce highly specialized, metabolically hyperactive feeding sites within host roots, which serve as their sole source of nutrients throughout their life cycle. The formation and maintenance of these feeding sites depend on the manipulation of host developmental pathways by nematode-derived secretions. Phytosulfokines (PSKs) are small plant peptide hormones that regulate cell division, tissue expansion, and growth responses, processes essential for feeding site development. Here, we identify root-knot nematode genes predicted to encode peptides with a conserved PSK functional motif. These genes are predominantly expressed during the early stages of infection and localize to secretory glands, suggesting a role in early parasitism. Moreover, silencing PSK-like gene expression reduces root gall formation and nematode reproduction. Together, these findings reveal that root-knot nematodes deploy PSK-like peptides as virulence factors to promote successful parasitism, providing the first report of PSK peptide mimicry in any plant pathogen.

plant biology↗

Yeast-2-Hybrid-Seq and Bifluorescence Complementation Resources for assessing Protein:Protein Interactions in Arbuscular Mycorrhizal Roots: CKL2 as a Case Study

Reverse genetics, facilitated by CRISPR technologies and comprehensive sequence-indexed insertion mutant collections, has advanced the identification of plants genes essential for arbuscular mycorrhizal (AM) symbiosis. However, a mutant phenotype alone is generally insufficient to reveal the specific role of the protein in AM symbiosis and in many cases, identifying interacting partner proteins is useful. To enable identification of protein:protein interactions during AM symbiosis, we established a Medicago truncatula -Diversispora epigaea yeast-two-hybrid (Y2H) library which, through Y2H-seq screening, can provide a rank-ordered list of candidate interactors of a protein of interest. We also developed a vector system to facilitate bimolecular fluorescence complementation assays (BIFC) in mycorrhizal roots so that protein interactions can be assessed in their native cell types and sub-cellular locations. We demonstrate the utility of a Y2H-seq screen coupled with BIFC in mycorrhizal roots, with a search for proteins that interact with CYCLIN DEPENDENT LIKE KINASE 2 (CKL2), a kinase essential for AM symbiosis. The Y2H-seq screen identified three 14-3-3 proteins as the highest ranked CKL2 interacting proteins. BIFC assays in mycorrhizal roots provided evidence for a CKL2:14-3-3 interaction at the periarbuscular membrane (PAM) in colonized root cells. Down-regulation of 14-3-3 by RNA interference provides initial evidence for a function in AM symbiosis. Thus, CKL2 may utilize 14-3-3 proteins to direct signaling from the PAM. The Y2H and BIFC resources will accelerate understanding of protein functions during AM symbiosis.

plant biology↗

Medicago truncatula CORYNE regulates inflorescence meristem branching, nutrient signaling, and arbuscular mycorrhizal symbiosis

The CLAVATA signaling pathway regulates plant development and plant-environment interactions. CLAVATA signaling consists of mobile, cell-type or environment-specific CLAVATA3/ESR-related (CLE) peptides, which are perceived by a receptor complex consisting of leucine-rich repeat receptor-like kinases such as CLAVATA1 and receptor-like proteins such as CLAVATA2, which often functions with the pseudokinase CORYNE (CRN). CLAVATA signaling has been extensively studied in various plant species for its role in meristem maintenance and in legumes for modulating root interactions with nitrogen-fixing rhizobia. Some signaling proteins involved in development and nodulation, including CLAVATA1, also regulate plant interactions with mutualistic arbuscular mycorrhizal (AM) fungi. However, our knowledge on AM symbiosis regulation by CLAVATA signaling remains limited and only a handful of genetic regulators have been identified. Here we report that Medicago truncatula CRN controls inflorescence meristem branching and negatively regulates root interactions with AM fungi. MtCRN functions partially independently of the AM autoregulation signal MtCLE53. Transcriptomic data revealed that crn roots display signs of perturbed nutrient, symbiosis, and stress signaling, suggesting that MtCRN plays various roles in plant development and interactions with the environment.

plant biology↗

Recurrent drought increases grassland community seasonal synchrony

1. Climate change increases the frequency and severity of drought events with strong repercussions on grassland ecosystems. While the effects of single drought events on ecosystem structure and functioning are well understood, it is largely unknown whether and how drought frequency modifies ecosystem responses to drought. 2. Here, we assessed how the increase in frequency of severe, annual summer drought impacted grassland communities. We examined these effects in a species-rich sub-alpine mountain meadow with a drought frequency of one, three, and 13 years, as well as ambient conditions. 3. We found that high drought frequency increased seasonal plant community synchrony through a reduction in species richness, a shift of plant functional groups, a loss of early-seasonal plant species, and the constrained establishment of seedlings throughout the growing season. These changes were associated with a decreased fraction of biomass as drought frequency increased. 4. Furthermore, we show that negative drought effects were enhanced with an increasing drought frequency, and that negative drought effects on plant communities outweighed the weak adaptive effects of species. 5. Synthesis. We conclude that single and low-frequency drought studies may not adequately predict longer-term changes in our rapidly shifting climate. With the ongoing increase in drought frequency due to climate change, we predict that grassland plant communities will increase in seasonal synchrony. We suggest that this increase in synchrony will leave ecosystems highly vulnerable to future disturbances, because asynchrony is a critical component of stability. Moreover, given the weak adaptive effects of plant species to long-term recurrent drought, we conclude that plant communities are unlikely to be able to adapt to the rapid increase in recurrent drought events.

ecology↗

Heterozygous missense variant in GLI2 impairs human endocrine pancreas development

Missense variants are the most common type of coding genetic variants. Their functional assessment is fundamental for defining any implication in human diseases and may also uncover genes that are essential for human organ development. Here, we applied CRISPR-Cas9 gene editing on human iPSCs to study a heterozygous missense variant in GLI2 identified in a paediatric hyperglycaemic patient and family members. GLI2 is a primary mediator of the Hedgehog pathway, which regulates pancreatic {beta}-cell development in mice. However, neither mutations in GLI2 nor Hedgehog dysregulation have been reported as cause or predisposition to diabetes. A set of isogenic iPSC lines harbouring the missense variant were studied for their ability to differentiate into pancreatic {beta}-like cells. Interestingly, iPSCs carrying the missense variant showed altered GLI2 transcriptional activity and impaired endocrine differentiation. Collectively, our findings underscore an essential role for GLI2 in human endocrine development and identify a novel variant that predisposes to diabetes.

developmental biology↗

Temperature but not the circadian clock determines nocturnal carbohydrate availability for growth in cereals

The circadian clock is considered a key target for crop improvement because it controls metabolism and growth in Arabidopsis. Here, we show that the clock gene EARLY FLOWERING 3 (ELF3) controls vegetative growth in Arabidopsis but not in the cereal crop barley. Growth in Arabidopsis is determined by the degradation of leaf starch reserves at night, which is controlled by ELF3. The vegetative growth of barley, however, is determined by the depletion of leaf sucrose stores through an exponential kinetics, presumably catalyzed by the vacuolar sucrose exporter SUCROSE TRANSPORTER 2 (SUT2). This process depends on the sucrose content and the nighttime temperature but not on ELF3. The regulation of starch degradation and sucrose depletion in barley ensures efficient growth at favorable temperature as stores become exhausted at dawn. On cool nights, however, only the starch degradation rate is compensated against low nighttime temperatures, whereas the sucrose depletion rate is reduced. This coincides with reduced biomass in barley but not in Arabidopsis after growth in consecutive cool nights. The sucrose depletion metabolism determines growth in the cereal crops barley, wheat, and rice but is not generally conserved in monocot species and is not a domestication-related trait. Therefore, the control of growth by endogenous (clock) versus external factors (temperature) is species-specific and depends on the predominant carbohydrate store. Our results give new insights into the physiology of growth in cereals and provide a basis for studying the genetics and evolution of different carbohydrate stores and their contribution to plant productivity and adaptation.\n\nSignificance StatementThe circadian clock controls growth in the model plant Arabidopsis thaliana by regulating the starch degradation rate so that reserves last until dawn. This prevents nocturnal starvation until photosynthesis resumes. The cereal crops barley, wheat and rice, however, predominantly consume sucrose instead of starch as carbohydrate source. We find that carbohydrate supply from sucrose at night is regulated by enzyme kinetics and night-time temperature, but not the circadian clock. We postulate that the regulation of growth depends on the predominant carbohydrate store, where starch degradation is controlled by endogenous cues (clock) and sucrose depletion by external factors (temperature). These differences in the regulation of carbohydrate availability at night may have important implications for adapting crops yields to climate change.

plant biology↗