Evolution of the thermostability of actin-depolymerizing factors enhances pollen germination at high temperature

In angiosperms, pollen germination leads to a period of extensive polarized growth of the pollen tube, which carries the sperm nuclei to the ovule. Studies of tip growth in both pollen tubes and root hairs have contributed to a descriptive model that involves polarized vesicle movement along the cytoskeleton to release materials needed for growth, such as cell wall precursors, at the growing tip. Key to this is the rapid disassembly and reassembly of the actin filaments due to the action of numerous actin-binding proteins, including actin-depolymerizing factors (ADFs). Here, Qian et al. investigated the ADF protein family in plants. In Arabidopsis, ADF proteins expressed highly in pollen display remarkable thermal stability, remaining undenatured even after boiling, whereas other ADFs denature at much lower temperatures. Genetic studies revealed that the thermostable ADFs are needed for pollen germination at the high temperatures that can occur in summertime, e.g., 37 degrees. The authors identified a few key residues as critical for thermostability. They then examined ADF proteins across the plant lineage and identified sequences of ancestral proteins; their results showed a trend of increasing and decreasing thermostability over evolutionary time. Interestingly, the trend maps to the changing global temperature over the past few hundred million years, and the emergence of the pollen-specific thermostable forms coincide with the rise of angiosperms. (Summary by Mary Williams @PlantTeaching) Plant Cell 10.1093/plcell/koad280

Subsidiary cells mediate stomatal closure in maize

Stomata are pores on leaves which enable gas exchange. In grasses, stomata are surrounded by dumbbell-shaped guard cells that are flanked by subsidiary cells. However, the role of the subsidiary cells in stomatal closure is not fully understood. Here, Liu et al. investigated this using two maize (Zea mays) mutants, called pangloss1 (pan1) and pangloss2 (pan2), which have altered subsidiary cell morphology. The mutations have incomplete penetrance, with 10-30% of stomata having at least one aberrantly shaped subsidiary cell. To test for stomatal closure defects, they moved plants into the dark or rapidly increased the carbon dioxide concentration and measured stomatal conductance. In both cases pan1 plants have normal stomatal closure, whereas it is greatly impaired in pan2 mutants. These experiments measure the entire population of stomata, even those with normal subsidiary cells, therefore, to study individual stomatal complexes confocal microscopy was used. In both pan1 and pan2 mutants, stomatal complexes with at least one aberrant subsidiary cell failed to close upon application of abscisic acid (ABA). Thus, subsidiary cells participate in stomatal closure. (Summary by Rose McNelly @rose_mcn) New Phytol. 10.1111/nph.19379

Ferns unleashed: Novel insecticidal proteins IPD113 challenge Bt resistance

Insect pests pose a significant threat to global crop production, with lepidopteran species like corn earworm and armyworms causing substantial losses. The widespread use of insecticidal proteins from Bacillus thuringiensis (Bt) in crops has been a key strategy to combat these pests. However, the emergence of field-evolved resistance in various pest species raises concerns about the sustainability of Bt crops. This study focuses on the discovery of a new family of insecticidal proteins, IPD113, isolated from fern (Pteris) species. Building on earlier observations that ferns are naturally resistant to insect infestation, the researchers identified IPD113 as having strong activity against lepidopteran pests in artificial diet-based assays and transgenic plants. Interestingly, IPD113s proved successful in eliminating insect colonies that were resistant to popular Bt proteins. The IPD113_Cow, member of the IPD113 family revealed a similar structure to Bt insecticidal proteins. Genomic analyses confirmed that these proteins are encoded by plant genes, challenging the initial hypothesis of horizontal gene transfer from bacteria. The discovery of IPD113 offers a viable path for developing insect-resistant crops with a new mode of action, potentially addressing challenges associated with Bt resistance in agricultural settings. (Summary by Arpita Yadav @arpita_yadav_). Proc. Natl. Acad. Sci. USA  10.1073/pnas.2306177120.

Graduate student mentorship as a target for diversifying biology

The quality of student-advisor relationships is one of the top predictors of research progress and sense of belonging in science, yet in most cases the mentoring relationship is simply left to chance. In this preprint, Debray et al. surveyed a cohort of PhD students to understand what mentoring practices really support the students. The authors start with some interesting but not surprising statistics: 69% of faculty received no formal mentoring training, yet they rarely feel they mentor poorly. Conversely, nearly half of graduate students report frequent poor mentoring, and these students are disproportionally from minority groups. The study identified several strategies to improve how students are mentored. Good mentoring involves mentors who demonstrate empathy and welcome honest feedback, meet frequently with their mentees, and provide both positive encouragement and critical feedback. In cases where the mentor does not provide good support, the student can fill the gap with strong support from other students and faculty, as well as benefit from a supportive department. Note: The ROOT & SHOOT project is recruiting plant scientists at all career stages to participate in a Working Group on Culturally Responsive Mentoring – see https://rootandshoot.org/working-group-on-culturally-responsive-mentoring/. (Summary by Mary Williams @PlantTeaching) bioRxiv https://doi.org/10.1101/2023.08.18.553806

Plant Science Research Weekly: December 1, 2023

Review: Paternal imprinting in Marchantia polymorpha

Humans and flowering plants spend most of their lives in a diploid state with two copies of each chromosome in most cells, but to reproduce they produce haploid gametes through meiosis. By contrast, bryophytes (liverworts, hornworts, and mosses), spend most of their lives in the haploid state. They produce gametes through mitosis and differentiation, and then very briefly experience a diploid state before meiosis leads again to the dominant, haploid state. The process of toggling between diploid and haploids is actually quite complicated, as some genes should only be active in one state or the other, or when inherited from one parent or the other. Shutting off of specific genes by parent-of-origin is known as imprinting, and usually involves the polycomb repressive complex and histone methylation (H3K27me3). Recently, Montgomery and Berger and colleagues identified a new type of selective gene silencing in the liverwort Marchantia, which involves the complete silencing of the paternal genome for the duration of the diploid stage – effectively rendering it haploid. This review discusses this novel finding in the context of other forms of imprinting and the evolution of land plants. Check it out! (Summary by Mary Williams @PlantTeaching) New Phytol. doi.org/10.1111/nph.19377

Review: The exocyst complex is targeted by pathogen effectors

The exocyst complex is a conserved octameric protein complex in eukaryotic cells. Its primary function is to tether secretory vesicles to the plasma membrane during the exocytosis process, and it is also involved in autophagy and host-pathogen interactions. Intriguingly, EXO70, one of the subunits of the exocyst complex, is present in a single copy in animals and fungi, but has expanded into a large protein family in plants. In a recent review, De la Concepcion brings together current research about the exocyst complex, including its 3D structure and investigations into the functional diversification of EXO70 in plants. Most notably, the review focuses on the different ways that the exocyst complex helps plants combat pathogens, for example through direct or indirect interactions with immune proteins. Not surprisingly given its role in immunity, the exocyst complex is targeted by several pathogen effectors, which are summarized in a useful table listing pathogen effectors and their targeted exocyst subunits. The review ends by raising open important questions about the diversification of exocyst subunits and pathogen effector–exocyst interactions. (Summary by Xiaohui Li @Xiao_hui_Li) Curr. Opin. Plant Biol. 10.1016/j.pbi.2023.102482. (For further insights into the exocyst complex’s role in autophagy, refer to Viktor Žárský’s detailed review in FEBS Letters 10.1002/1873-3468.14430.)

Review: CLAVATA signaling in plant-environment interactions

CLAVATA 3/EMBRYO SURROUNDING REGION-related (CLE) peptides and CLAVATA type receptors have been well charecterized for their role in root and shoot apical meristem maintenance in Arabidopsis. CLE peptides are also referred to as “peptide hormones” for their role in contolling physiological and developmental changes in plants. In this review by Bashyal et al., the roles of these peptides in regulating plant responses to various environmental stimuli is comprehensively summarized. CLAVATA signaling has diverse roles ranging from plant developmental adaptations to nutrient availability, responses to abiotic and biotic stimuli, and symbiotic relationships with rhizobia and arbuscular mycorrhiza fungi. The review also describes interesting studies on CLE-like signaling peptides from parasitic nematodes that hijack the plant-CLAVATA signaling pathway for their own benefit. The authors anticipate that future functional characterization through advanced bioinformatic tools will help in understanding how CLE-specific plant responses integrate plant physiology and the environment. (Summary by Indrani Kakati @Indranik333) Plant Physiol. 10.1093/plphys/kiad591

Review: Increasing yields of barley and wheat through inflorescence architecture

The Triticeae is an important tribe of crops that contains both barley and wheat. As the global population increases, the yield of Triticeae crops must increase to meet global food demands. An important factor influencing yield is grain number per spike, which is affected by inflorescence architecture. Here, Zhang et al. suggest that altering the inflorescence architecture in Triticeae crops by increasing the amount of branching and the number of spikelets per inflorescence could lead to higher yields. They highlight many key regulators that could be targeted to achieve this, for instance the AP2-ERF (APETALA2/ETHYLENE RESPONSIVE FACTOR) transcription factor COMPOSITUM 2 in barley which when mutated results in a highly branched spike. Another potential target is the miRNA156-SPL (SQUAMOSA promoter binding protein-like) module; SPL knockout wheat lines have decreased spikelet number, so increasing SPL expression might increase spikelet number. Such regulators could be targeted using CRISPR/Cas9 genome editing or by searching for beneficial alleles in seed stores or wild relatives, leading to increased yields of these important crops. (Summary by Rose McNelly @rose_mcn) J. Exp. Bot. 10.1093/jxb/erad386

RALF4 prevents pollen tubes from growing crazy

Pollen tubes are characterized by extremely fast elongation growth. Previously, small peptides known as RALFs (RAPID ALKALINIZATION FACTORs) were identified as signaling peptides, some with a role in pollen tube growth; for instance, RALF4/19 are important in maintaining the cell wall integrity of pollen tubes. RALF peptides bind to a membrane-integral protein complex consisting of LLGs [LORELEI-like GLYCOLPHOSPHATIDYLINOSITOL (GPI)-ANCHORED PROTEINS] and CrRLK1Ls (Catharanthus roseus RLK1-like receptor kinases). In addition, RALF peptides also bind to cell wall-anchored LRX (LEUCINE-RICH REPEAT EXTENSIN) proteins. When RALF4 binds to LRX8, it exposes an alkaline surface patch hypothesized to interact with negatively charged cell wall polysaccharides like pectin. In a recent study, Moussu et al. used the Arabidopsis pollen tube to explore the LRX8-RALF4 complex’s role in cell wall patterning. Using biochemical assays, they discovered that the LRX8-RALF4 complex directly binds to oligogalacturonans (OGs, a type of pectin). Notably, neither RALF4 nor LRX8 alone could bind to OGs independently. This interaction is specific to demethylated pectins, found predominantly in the pollen tube’s shank. Super-resolution imaging with immunolabeling revealed that LRX8 and RALF4 not only colocalize but also associate with pectin, creating a reticulated pattern along the pollen tube cell wall. Surprisingly, RALF4 is present along the pollen tube cell wall as an integral component colocalizing with LRX8. In addition, they found that mutations of the exposed alkaline surface of RALF4 lead to abnormal cell wall patterning, increased pollen tube growth rates, and premature bursting. Collectively, these discoveries highlight the critical role of the LRX8-RALF4-pectin association in the assembly and patterning of the pollen tube cell wall. (Summary by Xiaohui Li @Xiao_hui_Li) Science 10.1126/science.adi4720. For an extended analysis of this study’s background and future directions, and to see a wonderful image of pectin network in an Arabidopsis pollen tube, I recommend to read the excellent perspective article from Debra Mohnen in Science (10.1126/science.adl1198).

Lateral root branching promoted by ammonia borane-dependent H2

Molecular hydrogen (H2) has been shown to have several cellular benefits, most notably serving as an antioxidant. Its effects in humans and plants have been primarily assayed using hydrogen-rich water (HRW, produced by bubbling H2 into water), but slower, more persistent release of H2 has been demonstrated from solid ammonia borane (NH3BH3). Here, Wang et al. examined how ammonia borane affects root growth, using hydroponically grown tomatoes treated with ammonia borane (AB). Upon treatment with AB, roots showed an increase in expression level of several genes involved in the synthesis of phytomelatonin and an accompanying increase in phytomelatonin levels. The AB-promoted increase in root branching is suppressed by an inhibitor of phytomelatonin synthesis, and root branching is enhanced by exogenous phytomelatonin treatment, suggesting that the effects of AB are mediated through phytomelatonin. AB or phytomelatonin further led to an increase in expression on several auxin-signaling genes, suggesting that auxin has a role in this response. (Summary by Mary Williams @PlantTeaching) Plant Physiol. 10.1093/plphys/kiad595

Evolution of the thermostability of actin-depolymerizing factors enhances the adaptation of pollen germination to high temperature

In angiosperms, pollen germination leads to a period of extensive polarized growth of the pollen tube, which carries the sperm nuclei to the ovule. Studies of tip growth in both pollen tubes and root hairs have contributed to a descriptive model that involves polarized vesicle movement along the cytoskeleton to release materials needed for growth, such as cell wall precursors, at the growing tip. Key to this is the rapid disassembly and reassembly of the actin filaments due to the action of numerous actin-binding proteins, including actin-depolymerizing factors (ADFs). Here, Qian et al. investigated the ADF protein family in plants. In Arabidopsis, ADF proteins expressed highly in pollen display remarkable thermal stability, remaining undenatured even after boiling, whereas other ADFs denature at much lower temperatures. Genetic studies revealed that the thermostable ADFs are needed for pollen germination at the high temperatures that can occur in summertime, e.g., 37 degrees. The authors identified a few key residues as critical for thermostability. They then examined ADF proteins across the plant lineage and identified sequences of ancestral proteins; their results showed a trend of increasing and decreasing thermostability over evolutionary time. Interestingly, the trend maps to the changing global temperature over the past few hundred million years, and the emergence of the pollen-specific thermostable forms coincide with the rise of angiosperms. (Summary by Mary Williams @PlantTeaching) Plant Cell 10.1093/plcell/koad280

Subsidiary cells mediate stomatal closure in maize

Stomata are pores on leaves which enable gas exchange. In grasses, stomata are surrounded by dumbbell-shaped guard cells that are flanked by subsidiary cells. However, the role of the subsidiary cells in stomatal closure is not fully understood. Here, Liu et al. investigated this using two maize (Zea mays) mutants, called pangloss1 (pan1) and pangloss2 (pan2), which have altered subsidiary cell morphology. The mutations have incomplete penetrance, with 10-30% of stomata having at least one aberrantly shaped subsidiary cell. To test for stomatal closure defects, they moved plants into the dark or rapidly increased the carbon dioxide concentration and measured stomatal conductance. In both cases pan1 plants have normal stomatal closure, whereas it is greatly impaired in pan2 mutants. These experiments measure the entire population of stomata, even those with normal subsidiary cells, therefore, to study individual stomatal complexes confocal microscopy was used. In both pan1 and pan2 mutants, stomatal complexes with at least one aberrant subsidiary cell failed to close upon application of abscisic acid (ABA). Thus, subsidiary cells participate in stomatal closure. (Summary by Rose McNelly @rose_mcn) New Phytol. 10.1111/nph.19379

Ferns unleashed: Novel insecticidal proteins IPD113 challenge Bt resistance

Insect pests pose a significant threat to global crop production, with lepidopteran species like corn earworm and armyworms causing substantial losses. The widespread use of insecticidal proteins from Bacillus thuringiensis (Bt) in crops has been a key strategy to combat these pests. However, the emergence of field-evolved resistance in various pest species raises concerns about the sustainability of Bt crops. This study focuses on the discovery of a new family of insecticidal proteins, IPD113, isolated from fern (Pteris) species. Building on earlier observations that ferns are naturally resistant to insect infestation, the researchers identified IPD113 as having strong activity against lepidopteran pests in artificial diet-based assays and transgenic plants. Interestingly, IPD113s proved successful in eliminating insect colonies that were resistant to popular Bt proteins. The IPD113_Cow, member of the IPD113 family revealed a similar structure to Bt insecticidal proteins. Genomic analyses confirmed that these proteins are encoded by plant genes, challenging the initial hypothesis of horizontal gene transfer from bacteria. The discovery of IPD113 offers a viable path for developing insect-resistant crops with a new mode of action, potentially addressing challenges associated with Bt resistance in agricultural settings. (Summary by Arpita Yadav @arpita_yadav_). Proc. Natl. Acad. Sci. USA  10.1073/pnas.2306177120.

Graduate student mentorship as a target for diversifying biology

The quality of student-advisor relationships is one of the top predictors of research progress and sense of belonging in science, yet in most cases the mentoring relationship is simply left to chance. In this preprint, Debray et al. surveyed a cohort of PhD students to understand what mentoring practices really support the students. The authors start with some interesting but not surprising statistics: 69% of faculty received no formal mentoring training, yet they rarely feel they mentor poorly. Conversely, nearly half of graduate students report frequent poor mentoring, and these students are disproportionally from minority groups. The study identified several strategies to improve how students are mentored. Good mentoring involves mentors who demonstrate empathy and welcome honest feedback, meet frequently with their mentees, and provide both positive encouragement and critical feedback. In cases where the mentor does not provide good support, the student can fill the gap with strong support from other students and faculty, as well as benefit from a supportive department. Note: The ROOT & SHOOT project is recruiting plant scientists at all career stages to participate in a Working Group on Culturally Responsive Mentoring – see https://rootandshoot.org/working-group-on-culturally-responsive-mentoring/. (Summary by Mary Williams @PlantTeaching) bioRxiv https://doi.org/10.1101/2023.08.18.553806

Understanding secondary cell wall biosynthesis to improve fiber quality in cotton

Wang et al. investigate cotton fiber secondary cell wall biosynthesis in Gossypium hirsutum. The Plant Cell (2023).

By Yao Wang and Xue-Bao Li

Background: Cotton (Gossypium hirsutum) produces natural fibers (unicellular trichomes) on the seed, making it a valuable crop for the worldwide textile industry. Cotton fibers possess a unique secondary cell wall (SCW) that contains more than 90% cellulose but almost no hemicellulose or lignin. The phytohormones ethylene and auxin play indispensable roles in plant growth and development, but the molecular mechanisms for ethylene/auxin–mediated regulation of fiber SCW formation in cotton remain largely underexplored.

Question: We wanted to know if and how ethylene and auxin are involved in regulating cotton fiber SCW development.

Findings: We tested the role of these phytohormones in cotton fiber development using an in vitro cotton ovule culture assay with ethylene and auxin treatments. We also silenced GhERF108, which encodes an AP2/ERF transcription factor from the ETHYLENE RESPONSE FACTOR (ERF) family, which showed that GhERF108 positively regulates fiber SCW development. Moreover, both ethylene and auxin responses were reduced in GhMYBL1 silenced plants. GhERF108 interacts with two auxin response factors (GhARF7-1 and GhARF7-2), which respond to auxin signals to promote fiber SCW thickening. GhERF108 and the GhARF7s enhance the activation of GhMYBL1 in fibers where the SCW is thickening. GhMYBL1 directly binds to the promoters of cellulose synthase genes to activate their expression, promoting cellulose biosynthesis and boosting fiber SCW formation. Collectively, our findings demonstrate that the collaboration between GhERF108 and GhARF7s establishes crosstalk in ethylene-auxin signaling to activate GhMYBL1, ultimately leading to the activation of fiber SCW biosynthesis.

Next steps: Scientists aim to improve cotton fiber quality by genetic manipulation. Our work demonstrates that ethylene-auxin signal crosstalk may play a crucial role in improving fiber quality through regulating expression of the response genes (such as ERF and ARF transcription factors). We hope our work provides the theoretical basis for cotton breeding.

Reference:

Yao Wang, Yang Li, Shao-Ping He, Shang-Wei Xu, Li Li, Yong Zheng, Xue-Bao Li (2023). The transcription factor ERF108 interacts with AUXIN RESPONSE FACTORs to mediate cotton fiber secondary cell wall biosynthesis. https://doi.org/10.1093/plcell/koad214

N6-methyladenosine mRNA modification dynamics in plant immunity

Prall et al. explore how post-transcriptional regulation of mRNA mediated by methylation at the N6 position of adenine affects plants preparedness and reactivity to pathogen stress.

 Wil Prall and Brian Gregory, University of Pennsylvania Department of Biology

 Background: To determine how plants respond to bacterial and fungal pathogens, researchers are interested in analyzing the transcriptome, the global collection of RNAs in eukaryotic cells to see how plants react. Not only changes in individual RNA copy number (abundance) are observed, but also significant changes in the covalent additions to specific nucleotides in messenger RNAs (mRNAs) (i.e., nucleotide modification). In this work we investigate the addition of a methyl group onto the mRNA nucleotide adenosine (N6-methyladenosine (m6A)). m6A addition to mRNA is of great interest because its presence in individual mRNAs is known to affect RNA stability, translation, and maturation.

Question: What is the m6A transcriptomic landscape before and after exposure to simulated and real pathogen infection in Arabidopsis? How global reduction in m6A across all mRNAs affects plants preparedness and reactivity to pathogen stress?

Findings: Global depletion of m6A in mutant and transgenic seedlings and adult Arabidopsis plants (mta mutant and 35S:ALKBH10B over-expressor), results in increased resistance to bacterial and fungal infections, altering fundamental plant immune response. m6A deposition is specifically coordinated on RNA transcripts directly involved in defense and immunity prior to and following pathogen signaling and stress. The modulation of m6A on specific transcripts is correlated with changes in abundance and internal mRNA cleavage demonstrating this modification as another layer of control before and during the immune response.

Next Steps: The m6A modification is added to transcripts by ‘writer’ proteins, removed by ‘erasers’ and bound by ‘readers’. Understanding how these proteins are regulated themselves, and how they regulate the selective addition and removal of these marks in the context of pathogen infection is of critical importance.

Reference:

Wil Prall, Arsheed H. Sheikh, Jeremie Bazin, Jean Bigeard, Marilia Almeida-Trapp, Martin Crespi, Heribert Hirt, and Brian D. Gregory (2023) Pathogen-induced m6A dynamics affect plant immunity. https://doi.org/10.1093/plcell/koad224

Regulation of a calcium-dependent protein kinase in response to osmotic stress

Fan et al. explore how ubiquitin-dependent proteolysis regulates the levels of active kinase in plant responses to osmotic stress.

https://doi.org/10.1093/plcell/koad173

Wei Fan and Zixing Li

School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China

Background: Plants experience osmotic stress during drought conditions and in soils with high salt levels. Osmotic stress triggers a rapid rise in intracellular calcium ion (Ca2+) concentrations in plants. Upon the binding of Ca2+, the calcium-dependent protein kinase (CPK) undergoes a conformational change, releasing the kinase active site from the repression of an autoinhibitory junction. CPKs become active to phosphorylate their downstream targets to transduce osmotic stress signals. CPK4 plays a positive role in plant osmotic stress responses.

Question: How are active CPK4 protein levels dynamically and precisely regulated in response to hyperosmotic stress?

Findings: We found that salt/mannitol-induced osmotic stress promotes CPK4 protein accumulation by disrupting 26S proteasome-mediated degradation of CPK4. The U-BOX E3 ligase PUB44 interacts with and mediates the ubiquitination of CPK4, which results in the proteasomal degradation of CPK4. PUB44 exhibits a CPK4-dependent negative role in plant osmotic stress responses. In addition, we found that Ca-binding and kinase activation decrease the ubiquitination of CPK4 and enhance CPK4 protein stabilization.

Next steps: We plan to investigate how the binding of Ca2+ to CPK determines the ubiquitination and degradation of CPK4.

Reference:

Wei Fan, Xiliang Liao, Yanqiu Tan, Xiruo Wang, Julian I. Schroeder, Zixing Li. (2023). Arabidopsis PLANT U-BOX44 down-regulates osmotic stress signaling by mediating Ca2+-DEPENDENT PROTEIN KINASE4 degradation. https://doi.org/10.1093/plcell/koad173

背景回顾:渗透胁迫条件下,植物胞质自由钙离子浓度会快速上升。钙依赖的蛋白激酶(calcium-dependent protein kinase,CPK)结合细胞内钙离子,蛋白构象发生改变,释放自抑制域对激酶活性的抑制,并通过自磷酸化或其他激酶的磷酸化而激活,激活的钙依赖的蛋白激酶磷酸化下游底物蛋白,激发植物渗透胁迫信号,使植物响应渗透胁迫。CPK4在植物响应干旱胁迫时作为正调因子发挥重要作用,促使植物细胞调节自身生理活动以响应环境渗透胁迫

科学问题:在渗透胁迫条件下,植物如何精确调控CPK4蛋白的机制尚不清楚?

研究发现:我们鉴定到一个U-BOX类型的E3泛素连接酶PUB44,能够识别并泛素化修饰CPK4 促其通过26S蛋白酶体途径降解,调节细胞内CPK4的蛋白丰度。渗透胁迫信号下,CPK4结合钙离子,蛋白发生构象改变,减弱PUB44泛素连接酶对CPK4蛋白的泛素化修饰程度。同时,渗透胁迫信号还会抑制PUB44蛋白的泛素酶活性,进一步降低CPK4蛋白的泛素化修饰水平。通过这些方式,植物细胞在正常环境下维持一定CPK4蛋白的丰度,并在胁迫条件下快速增加CPK4蛋白的含量,激活细胞适应性生理活动来响应渗透胁迫

展望未来:我们将进一步探究CPK4结合钙离子,蛋白构象的变化是如何影响CPK4蛋白的泛素化修饰。为什么CPK4蛋白丰度特异受到盐、渗透胁迫,而非温度和营养胁迫的调节。

The NLR receptor ZAR1 has been guarding receptor-like cytoplasmic kinases since the Jurassic

Adachi et al. explore the conservation of a nucleotide-binding leucine-rich repeat immune receptor.

Hiroaki Adachi1,2,3, Toshiyuki Sakai1,2, Jiorgos Kourelis1, Hsuan Pai1, Jose L. Gonzalez Hernandez4, Yoshinori Utsumi5, Motoaki Seki5,6,7, Abbas Maqbool1 and Sophien Kamoun1*

 1The Sainsbury Laboratory, University of East Anglia, Norwich Research Park, NR4 7UH, Norwich, UK

2Laboratory of Crop Evolution, Graduate School of Agriculture, Kyoto University, Mozume, Muko, Kyoto 617-0001, Japan

3JST-PRESTO, Saitama, Japan

4Agronomy, Horticulture and Plant Sciences Department, South Dakota State University, Brookings, South Dakota, US

5Plant Genomic Network Research Team, RIKEN Center for Sustainable Resource Science, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan

6Plant Epigenome Regulation Laboratory, RIKEN Cluster for Pioneering Research, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan

7Kihara Institute for Biological Research, Yokohama City University, 641-12 Maioka-cho, Totsuka-ku, Yokohama, Kanagawa 244-0813, Japan

Background: In plants, nucleotide-binding leucine-rich repeat (NLR) immune receptors generally exhibit hallmarks of rapid evolution even at the intraspecific level. NLRs evolve primarily through a birth-and-death process: new NLRs emerge by recurrent cycles of gene duplication and loss—some genes are maintained in the genome and acquire new pathogen detection specificities, whereas others are deleted or become non-functional through the accumulation of deleterious mutations. Such dynamic patterns of evolution enable the NLR immune system to keep up with fast-evolving effector repertoires of pathogenic microbes. Unlike typical NLRs, HOPZ-ACTIVATED RESISTANCE1 (ZAR1) is conserved across angiosperms.

Question: Can we use a molecular evolution framework to determine the critical features of a conserved plant NLR?

Findings: We performed iterative sequence similarity searches coupled with phylogenetic analyses to reconstruct the evolutionary history of ZAR1. ZAR1 is an atypically conserved NLR that traces its origin to early flowering plant lineages ~220 to 150 million years ago (Jurassic period). Ortholog sequence analyses revealed highly conserved features of ZAR1, including regions for pathogen recognition and immune activation. We functionally reconstructed the immune activity of ZAR1 and its host partner receptor-like cytoplasmic kinases (RLCKs) from distantly related plant species, supporting the hypothesis that ZAR1 has evolved to partner with RLCKs early in its evolution. ZAR1 stands out among angiosperm NLRs for having experienced relatively limited gene duplication and expansion throughout its deep evolutionary history.

Next steps: Further comparative analyses, combining molecular evolution and structural biology, of plant and animal NLR systems will yield experimentally testable hypotheses for NLR research.

Reference:

Hiroaki Adachi, Toshiyuki Sakai, Jiorgos Kourelis, Hsuan Pai, Jose L. Gonzalez Hernandez, Yoshinori Utsumi, Motoaki Seki, Abbas Maqbool and Sophien Kamoun (2023) Jurassic NLR: conserved and dynamic evolutionary features of the atypically ancient immune receptor ZAR1. https://doi.org/10.1093/plcell/koad175