Stress‐adaptive gene discovery by exploiting collective decision‐making of decentralised plant response systems

Despite having a network of cytoplasmic interconnections (plasmodesmata) facilitating rapid exchange of metabolites and signal molecules, plant cells use the extracellular matrix as an alternative route for cell‐cell communication. The need for extracellular signalling in plasmodesmata‐networked tissues is baffling. We propose the hypothesis that this phenomenon defines the plant extracellular matrix as a “democratic space” for collective decision‐making in a decentralised system, similar to quorum‐sensing in bacteria. Extracellular communication enables signal integration and coordination across several cell layers through ligand‐activated plasma membrane receptors. Recent results from drought stress‐adaptive responses and light‐mediated signalling in cell death activation show operational utility of this decision‐making process. We discuss opportunities for new innovations in drought gene discovery using platforms targeting the extracellular matrix

Heat Stress Triggers Differential Protein Accumulation in the Extracellular Matrix of Sorghum Cell Suspension Cultures

Plants reprogram gene expression as an adaptive response to survive high temperatures. While the identity and functions of intracellular heat stress-responsive proteins have been extensively studied, the heat response of proteins secreted to the extracellular matrix is unknown. Here, we used Sorghum bicolor, a species adapted for growth in hot climates, to investigate the extracellular heat-induced responses. When exposed to 40 C for 72 h, heat-sensitive Arabidopsis cell suspension cultures died, while ICSB338 sorghum cell cultures survived by activation of a transcriptional response characterized by the induction of HSP70 and HSP90 genes. Quantitative proteomic analysis of proteins recovered from cell culture medium revealed specific heat stress-induced protein accumulation within the sorghum secretome. Of the 265 secreted proteins identified, 31 responded to heat (2-fold change), with 84% possessing a predicted signal peptide for targeting to the classical secretory pathway. The differentially accumulated proteins have putative functions in metabolism, detoxification, and protein modifications. A germin (SORBI_3003G427700) was highly heat-inducible at both protein and gene level. Overall, our study reveals new insights into sorghum responses to heat and provides a useful resource of extracellular proteins that could serve as targets for developing thermotolerant crops. Data are available via ProteomeXchange with identifier PXD021536

Exogenous abscisic acid treatment regulates protein secretion in sorghum cell suspension cultures.

Drought stress adversely affects plant growth, often leading to total crop failure. Upon sensing soil water deficits, plants switch on biosynthesis of abscisic acid (ABA), a stress hormone for drought adaptation. Here, we used exogenous ABA application to dark-grown sorghum cell suspension cultures as an experimental system to understand how a drought-tolerant crop responds to ABA. We evaluated intracellular and secreted proteins using isobaric tags for relative and absolute quantification. While the abundance of only ~ 7% (46 proteins) intracellular proteins changed in response to ABA, ~32% (82 proteins) of secreted proteins identified in this study were ABA responsive. This shows that the extracellular matrix is disproportionately targeted and suggests it plays a vital role in sorghum adaptation to drought. Extracellular proteins responsive to ABA were predominantly defense/detoxification and cell wall-modifying enzymes. We confirmed that sorghum plants exposed to drought stress activate genes encoding the same proteins identified in the cell culture system with ABA. Our results suggest that ABA activates defense and cell wall remodeling systems during stress response. This could underpin the success of sorghum adaptation to drought stress

Carbon Monoxide Alleviates Salt-Induced Oxidative Damage in Sorghum bicolor by Inducing the Expression of Proline Biosynthesis and Antioxidant Genes

Crop growth and yield are affected by salinity, which causes oxidative damage to plant cells. Plants respond to salinity by maintaining cellular osmotic balance, regulating ion transport, and enhancing the expression of stress-responsive genes, thereby inducing tolerance. As a byproduct of heme oxygenase (HO)-mediated degradation of heme, carbon monoxide (CO) regulates plant responses to salinity. This study investigated a CO-mediated salt stress tolerance mechanism in sorghum seedlings during germination. Sorghum seeds were germinated in the presence of 250 mM NaCl only, or in combination with a CO donor (1 and 1.5 μM hematin), HO inhibitor (5 and 10 μM zinc protoporphyrin IX; ZnPPIX), and hemoglobin (0.1 g/L Hb). Salt stress decreased the germination index (47.73%) and root length (74.31%), while hydrogen peroxide (H2O2) (193.5%), and proline (475%) contents increased. This increase correlated with induced HO (137.68%) activity and transcripts of ion-exchanger and antioxidant genes. Salt stress modified vascular bundle structure, increased metaxylem pit size (42.2%) and the Na+/K+ ratio (2.06) and altered primary and secondary metabolites. However, exogenous CO (1 μM hematin) increased the germination index (63.01%) and root length (150.59%), while H2O2 (21.94%) content decreased under salt stress. Carbon monoxide further increased proline (147.62%), restored the vascular bundle structure, decreased the metaxylem pit size (31.2%) and Na+/K+ ratio (1.46), and attenuated changes observed on primary and secondary metabolites under salt stress. Carbon monoxide increased HO activity (30.49%), protein content, and antioxidant gene transcripts. The alleviatory role of CO was abolished by Hb, whereas HO activity was slightly inhibited by ZnPPIX under salt stress. These results suggest that CO elicited salt stress tolerance by reducing oxidative damage through osmotic adjustment and by regulating the expression of HO1 and the ion exchanger and antioxidant transcripts

Boosting soil literacy in schools can help improve understanding of soil/human health linkages in Generation Z

Soil health underpins ecosystem services like food security and therefore underpins human health. Poor soil health is a global problem which is hindering attempts to deliver the UN’s Sustainable Development Goals. We focus on goals 3 (human health), 13 (climate change) which are intimately linked to goal 15 (soil health). Soil health is arguably most fragile in regions such as sub-Saharan Africa (SSA) where aged soils are characterised by poor nutrient and water holding capacity, and are largely deficient in micronutrients such as Zinc. Poor soil health coupled with the largely cereal-based diets can mean that micronutrient malnutrition is high in the region. In sub-Saharan Africa, where much of the population is too poor to purchase mineral supplements, poor soil health (SDG15) can therefore negatively impact on human health (SDG3). We surveyed 3661 school children aged 13–15 in three African countries, Ghana, South Africa and Zimbabwe, for their ‘Attitudes, Behaviours and Competencies’ of soil, which we termed ‘ABC’. The ‘ABC’ survey results showed significant soil illiteracy. The survey showed that although students were generally equipped with a good attitude to (overall 52% positive) and behaviour towards soil (overall 60% engagement), they had little competency as to how to improve soil health (overall 23% knowledge). For example, less than 35% of respondents across all countries know that soil is living. Less than 13% of students are aware of the important role of soil in climate change mitigation. We believe that these two knowledge gaps must be addressed for Generation Z to understand the important linkages between climate change, soil and human health. We propose a hands-on ‘ethics of care’ approach to engage society with soil, piggybacking on existing climate change educational resources by building terrariums with living soil can empower children to learn about soil, plant, human and planetary health. The future of food security depends on Generation Z having soil literacy. Our survey clearly shows that students who think farming is a good way to make money have significantly higher levels of overall soil literacy. We propose that the future of human health depends on soil literacy

Isolation of Arabidopsis extracellular ATP‐binding proteins by affinity proteomics and identification of PHOSPHOLIPASE C‐LIKE 1 as an extracellular protein essential for fumonisin B1 toxicity

ATP is secreted to the extracellular matrix where it activates plasma membrane receptors for controlling plant growth and stress‐adaptive processes. DOES NOT RESPOND TO NUCLEOTIDES 1 (DORN1), the first plant ATP receptor was identified, but key downstream proteins are sought after. Here, we identified 120 proteins secreted by Arabidopsis cell cultures and screened them for putative stress‐responsive proteins using ATP‐affinity purification. We report three Arabidopsis proteins isolated by ATP‐affinity: PEROXIDASE 52, SUBTILASE‐LIKE SERINE PROTEASE 1.7, and PHOSPHOLIPASE C‐LIKE 1. In wildtype Arabidopsis, expression of genes encoding all three proteins responded to fumonisin B1, a cell death‐activating mycotoxin. Expression of PEROXIDASE 52 and PHOSPHOLIPASE C‐LIKE 1 genes was altered in fumonisin B1‐resistant salicylic acid induction‐deficient (sid2) mutants. Exposure to fumonisin B1 suppressed PHOSPHOLIPASE C‐LIKE 1 expression in sid2 mutants, suggesting that inactivation of this gene might provide mycotoxin tolerance. Accordingly, gene knockout mutants of PHOSPHOLIPASE C‐LIKE 1 were resistant to fumonisin B1‐induced death. Activation of PHOSPHOLIPASE C‐LIKE 1 gene expression by exogenous ATP was not blocked in dorn1 loss‐of‐function mutants, indicating that DORN1 is not required. Furthermore, exogenous ATP rescued both wildtype and dorn1 mutants from fumonisin B1 toxicity, suggesting that different ATP receptor(s) are operational in this process. Our results point to the existence of additional plant ATP receptor(s) and provide crucial downstream targets for use in designing screens to identify these receptors. Finally, PHOSPHOLIPASE C‐LIKE 1 serves as a convergence point for fumonisin B1 and extracellular ATP signalling, and functions in Arabidopsis stress response to fumonisin B1

Sorghum’s Whole-Plant Transcriptome and Proteome Responses to Drought Stress: A Review

Sorghum is a cereal crop with key agronomic traits of drought and heat stress tolerance, making it an ideal food and industrial commodity for hotter and more arid climates. These stress tolerances also present a useful scientific resource for studying the molecular basis for environmental resilience. Here we provide an extensive review of current transcriptome and proteome works conducted with laboratory, greenhouse, or field-grown sorghum plants exposed to drought, osmotic stress, or treated with the drought stress-regulatory phytohormone, abscisic acid. Large datasets from these studies reveal changes in gene/protein expression across diverse signaling and metabolic pathways. Together, the emerging patterns from these datasets reveal that the overall functional classes of stress-responsive genes/proteins within sorghum are similar to those observed in equivalent studies of other drought-sensitive model species. This highlights a monumental challenge of distinguishing key regulatory genes/proteins, with a primary role in sorghum adaptation to drought, from genes/proteins that change in expression because of stress. Finally, we discuss possible options for taking the research forward. Successful exploitation of sorghum research for implementation in other crops may be critical in establishing climate-resilient agriculture for future food security