Plant Life Cycle Research Articles

An Assessment of the Environmental Impact of Construction Materials of Monocrystalline and Perovskite Photovoltaic Power Plants Toward Their Sustainable Development

The interest in alternative energy sources, including the use of solar radiation energy, is growing year by year. Currently, the most frequently installed photovoltaic modules are made of single-crystalline silicon solar cells (sc-Si). However, one of the latest solutions are perovskite solar cells (PSC), which are considered the future of photovoltaics. Therefore, the main objective of this research was to assess the environmental impact of the construction materials of monocrystalline and perovskite photovoltaic power plants toward their sustainable development. The research object was the construction materials and components of two 1 MW photovoltaic power plants: one based on monocrystalline modules and the other on perovskite modules. The life cycle assessment (LCA) method was used for the analyses. The IMPACT World+, IPCC and CED models were used in it. The analyses were performed separately for five sets of elements: support structures, photovoltaic panels, inverter stations, electrical installations and transformers. Two post-consumer management scenarios were adopted: storage and recycling. The life cycle of a photovoltaic power plant based on photovoltaic modules made of perovskite cells is characterized by a smaller negative impact on the environment compared to traditional power plants with monocrystalline silicon modules. Perovskites, as a construction material of photovoltaic modules, fit better into the main assumptions of sustainable development compared to cells made of monocrystalline silicon. However, it is necessary to conduct further work which aims at reducing energy and material consumption in the life cycles of photovoltaic power plants.

Pemanfaatan Kebun Sayuran Sekolah sebagai Media Pembelajaran dalam Meningkatkan Produktivitas Peserta Didik di SDN Bulusidokare Sidoarjo

Vegetable garden does not only serve as a source of food but also as an educational tool that supports experiential learning. This research was conducted at Bulusidokare Sidoarjo Public Elementary School. Through gardening activities, students can develop practical skills, an understanding of the plant life cycle, and environmental values. This research aims to explore the utilization of vegetable gardens as an innovative learning medium to enhance student productivity. This research method uses a qualitative approach by conducting interviews with teachers and students, field observations, and collecting supporting documentation. The author describes the findings as data analysis. The research results show that student involvement in gardening activities can enhance motivation, teamwork skills, and a sense of disciplined responsibility. In addition, this experience strengthens the understanding of scientific and environmental concepts, as well as enhances the love for nature. Thus, vegetable gardens can be used as an effective learning medium to enhance the productivity and quality of students' education.

Mechanistic Insights into Leaf Senescence Regulation in Woody Plants: A Molecular Perspective.

Leaves, the primary carbon fixers in autotrophic plants, undergo a complex senescence process, which is critical for the redistribution of nutrients and supports ongoing growth and development. This natural aging phenomenon, often triggered at the end of a plant's life cycle or during the autumn season in perennial species, is finely regulated at multiple levels. Premature senescence can disrupt normal plant development, while the timing and pace of senescence significantly impact crop yield and quality. Notably, accelerated senescence under stress conditions may enhance the survival of future generations, suggesting an evolutionary strategy for plants to cope with harsh environments. Our review focuses on the molecular regulation of leaf senescence in woody plants, emphasizing the intricate determinants and regulatory mechanisms involved, including the role of phytohormones, environmental cues, and genetic factors. We highlight recent advances in understanding the regulation of leaf senescence in woody plants, integrating insights from multidisciplinary approaches and cutting-edge technologies.

Functions and Regulation of HAM Family Genes in Meristems During Gametophyte and Sporophyte Generations.

A fascinating feature of land plants is their ability to continually initiate new tissues and organs throughout their lifespan, driven by a pool of pluripotent stem cells located in meristems. In seed plants, various types of meristems are initiated and maintained during the sporophyte generation, while their gametophytes lack meristems and rely on sporophyte tissues for growth. In contrast, seed-free vascular plants, such as ferns, develop meristems during both the sporophyte and gametophyte generations, allowing for the independent growth of both generations. Recent findings have highlighted both conserved and lineage-specific roles of the HAIRY MERISTEM (HAM) family of GRAS-domain transcriptional regulators in various meristems throughout the land plant lifecycle. Here, we review and discuss how HAM genes maintain meristem indeterminacy in both sporophytes and gametophytes, with a focus on studies performed in two model species: the flowering plant Arabidopsis thaliana and the fern Ceratopteris richardii. Additionally, we summarize the crucial and tightly regulated functions of the microRNA171 (miR171)-HAM regulatory modules, which define HAM spatial patterns and activities during meristem development across various meristem identities in land plants.

Identification of cell-type specificity, trans- and cis-acting functions of plant lincRNAs from single-cell transcriptomes.

Long noncoding RNAs, including intergenic lncRNAs (lincRNAs), play a key role in various biological processes throughout the plant life cycle, and the advent of single-cell RNA sequencing (scRNA-seq) technology has opened up a valuable avenue for scrutinizing the intricate roles of lincRNAs in cellular processes. Here, we identified a new batch of lincRNAs using scRNA-seq data from diverse tissues of plants (rice, Arabidopsis, tomato, and maize). Based on well-annotated single-cell transcriptome atlases, plant lincRNAs were found to possess the same level of cell-type specificity as mRNAs and to be involved in the differentiation of certain cell types based on pseudo-time analysis. Many lincRNAs were predicted to play a hub role in the cell-type-specific co-expression networks of lincRNAs and mRNAs, suggesting their trans-acting abilities. Besides, plant lincRNAs were revealed to have potential cis-acting properties based on their genomic distances and expression correlations with the neighboring mRNAs. Furthermore, an online platform, PscLncRNA (http://ibi.zju.edu.cn/psclncrna/), was constructed for searching and visualizing all identified plant lincRNAs with annotated potential functions. Our work provides new insights into plant lincRNAs at single-cell resolution and an important resource for understanding and further investigation of plant lincRNAs.

Warm temperature perceived at the vegetative stage affects progeny seed germination in natural accessions of Arabidopsis thaliana.

Temperatures perceived early in the life cycle of mother plants can affect the germination of the offspring seeds. In Arabidopsis thaliana, vernalisation-insensitive mutants showed altered germination response to elevated maternal temperature, hence revealing a strong genetic determinism. However, the genetic control of this maternal effect and its prevalence across natural populations remain unclear. Here, we exposed a collection of European accessions of A. thaliana to increased temperature during the vegetative phase and assessed germination in their progeny to identify the genetic basis of transgenerational germination response. We found that genotypes with rapidly germinating progeny after early maternal exposure to elevated temperature originated from regions with low-light radiation. Combining genome-wide association, expression analysis and functional assays across multiple genetic backgrounds, we show a central role for PHYB in mediating the response to maternally perceived temperature at the vegetative stage. Differential gene expression analysis in leaves identified a similar genetic network as previously found in seed endosperm under elevated temperature, supporting the pleiotropic involvement of PHYB signalling across different tissues and stages. This provides evidence that complex environmental responses modulated by the maternal genotype can rely on a consistent set of genes yet produce different effects at the different stages of exposure.

DESEMPENHO DA PRODUTIVIDADE DE MILHO A PARTIR DE APLICAÇÃO DE FUNGICIDA EM DIFERENTES ESTÁGIOS FENOLÓGICOS

Corn (Zea mays L.) is a crucial crop in terms of global production and economic value, but it is susceptible to fungal diseases such as common rust (Puccinia sorghi) and cercosporiosis (Cercospora zeae-maydis), which can reduce grain production and quality. Effective control of these diseases is vital for food security and agricultural economy. Proper management of corn fungal diseases depends on the plant's phenological stages, from planting to grain maturity. The application of fungicides at strategic moments during the plant's life cycle is essential to prevent infections and reduce disease impact. A study conducted in Patos de Minas, Minas Gerais, during 2023, evaluated the effect of fungicide application at different phenological stages of corn on final productivity. The results showed that fungicide application at multiple phenological stages (V4, V8, and VT) resulted in greater above-ground height, lower severity of fungal diseases, higher productivity, and uniform stalk diameter compared to the control treatment. This integrated disease management method proved effective in protecting plants against fungal diseases, promoting healthier and more vigorous growth, and increasing final corn productivity. These findings highlight the importance of fungicide application at strategic moments in the corn development cycle to maximize productivity and promote more efficient and sustainable agriculture.

MID1-COMPLEMENTING ACTIVITY regulates cell proliferation and development via Ca2+ signaling in Marchantia polymorpha.

MID1-COMPLEMENTING ACTIVITY (MCA) is a land plant-specific, plasma membrane protein and Ca2+ signaling component that responds to exogenous mechanical stimuli, such as touch, gravity, and hypotonic-osmotic stress, in various plant species. MCA is essential for cell proliferation and differentiation during growth and development in rice (Oryza sativa) and maize (Zea mays). However, the mechanism by which MCA mediates cell proliferation and differentiation via Ca2+ signaling remains unknown. Here, we address this question using the liverwort Marchantia polymorpha. We show that the M. polymorpha MCA ortholog, MpMCA, is highly expressed in actively dividing regions, such as apical notches in the thalli and developing gametangiophores, and that MpMCA is a plasma membrane protein. In vivo Ca2+ imaging using a Ca2+ sensor (yellow cameleon) revealed that MpMCA is required for maintaining proper [Ca2+]cyt levels in the apical notch region, egg cells, and antheridium cells. Mpmca mutant plants showed severe cell proliferation and differentiation defects in the thalli, gametangiophores, and gametangia, resulting in abnormal development and unsuccessful fertilization. Furthermore, expression of the Arabidopsis MCA1 gene complemented most of the defects in the growth and development of the Mpmca mutant plants. Our findings indicate that MpMCA is an evolutionarily conserved Ca2+-signaling component that regulates cell proliferation and development across the life cycle of land plants.

Study on carbon emissions of a small hydropower plant in Southwest China

Hydropower plants with a small installed capacity, which are widely distributed in mountainous areas with abundant rainfall and steep rivers, play an important role in resolving energy problems in remote rural areas. These plants are a crucial source of clean electricity generated from water power. Harnessing local water resources not only helps alleviate energy shortages, but also reduces reliance on fossil fuels, contributing significantly to China’s national goals of achieving peak carbon emissions and carbon neutrality. This study investigates the carbon footprint of the Huangshadong Reservoir Project in Chongqing, China. The entire life cycle of the hydropower plant is assessed, including the preparation, construction, operation and maintenance, and demolition phases. The uncertainty was evaluated using the error propagation method. Following analysis, suggestions for carbon footprint reduction measures were proposed. Results showed that the total carbon footprint and the carbon intensity of the Huangshadong Reservoir Project over its entire life cycle are 33,148.29 t CO2e and 417.75 g CO2e/kWh, respectively. Of the total carbon footprint, the preparation phase, construction phase, operation and maintenance phase, and demolition phase account for 0.04%, 67.06%, 26.2%, and 6.7%, respectively. It means that the requirement for cement during the construction phase represents an important contribution to the entire life cycle carbon footprint of a small hydropower plant. As an integrated water conservancy project, the carbon intensity of the Huangshadong Reservoir Project is higher than that of medium-sized and large hydropower plants. However, its carbon intensity is lower than the emission factor of fossil power plants. The research results provide reference for both planning and construction of small hydropower plants and low-carbon development of rural hydraulic engineering.

REGULATOR OF FATTY ACID SYNTHESIS proteins regulate de novo fatty acid synthesis by modulating hetACCase distribution.

In plants, heteromeric acetyl-CoA carboxylase (hetACCase) initiates de novo fatty acid synthesis (FAS) by generating malonyl-CoA in the first committed step of this process. hetACCase activity is precisely regulated to meet the cellular demand for acyl chains during the plant life cycle. In this study, we performed a systematic co-expression analysis of hetACCase and its regulators in Arabidopsis (Arabidopsis thaliana) to better understand the regulatory mechanism of hetACCase. Our analysis uncovered REGULATOR OF FATTY ACID SYNTHESIS 1 (RFS1), whose expression is positively correlated with that of other regulators of hetACCase. The RFS gene family encodes two plastid inner envelope membrane proteins with undiscovered roles. Further analysis revealed that RFS1 co-localizes and directly interacts with CARBOXYLTRANSFERASE INTERACTOR 1 (CTI1). CRISPR/Cas9-mediated knockouts of RFSs exhibit enhanced hetACCase activity, higher FAS rates, and increased fatty acid contents, with particularly marked accumulation of absolute triacylglycerol levels in leaves, similar to cti mutants. The mutations of rfs and cti alter the plastid membrane distribution pattern of α-carboxyltransferase, leading to reduced hetACCase activity on the membrane, which could potentially be the original mechanism through which RFSs restrain hetACCase activity. Thus, we reveal a unique regulatory module that regulates de novo FAS and a genetic locus that may contribute to breeding of improved oil crops.

Changes in SWEET-mediated sugar partitioning affect photosynthesis performance and plant response to drought.

Sugars, produced through photosynthesis, are at the core of all organic compounds synthesized and used for plant growth and their response to environmental changes. Their production, transport, and utilization are highly regulated and integrated throughout the plant life cycle. The maintenance of sugar partitioning between the different subcellular compartments and between cells is important in adjusting the photosynthesis performance and response to abiotic constraints. We investigated the consequences of the disruption of four genes coding for SWEET sugar transporters in Arabidopsis (SWEET11, SWEET12, SWEET16, and SWEET17) on plant photosynthesis and the response to drought. Our results show that mutations in both SWEET11 and SWEET12 genes lead to an increase of cytosolic sugars in mesophyll cells and phloem parenchyma cells, which impacts several photosynthesis-related parameters. Further, our results suggest that in the swt11swt12 double mutant, the sucrose-induced feedback mechanism on stomatal closure is poorly efficient. On the other hand, changes in fructose partitioning in mesophyll and vascular cells, measured in the swt16swt17 double mutant, positively impact gas exchanges, probably through an increased starch synthesis together with higher vacuolar sugar storage. Finally, we propose that the impaired sugar partitioning, rather than the total amount of sugars observed in the quadruple mutant, is responsible for the enhanced sensitivity upon drought. This work highlights the importance of considering SWEET-mediated sugar partitioning rather than global sugar content in photosynthesis performance and plant response to drought. Such knowledge will pave the way to design new strategies to maintain plant productivity in a challenging environment.

Towards a Standardized LCOE Calculation for Informed Decision-Making in Energy Policy and Investment: Application to the Colombian Context

The Levelized cost of electricity - LCOE represents a constant cost per unit of generated electrical energy, resulting from the analysis of the total costs throughout the life cycle of a generation plant. Its widespread use, thanks to its simplicity of calculation and potential of standardization, has made it one of the primary criteria for comparing electricity generation technologies. However, in the pursuit of simplification and generalization, certain aspects associated with the socioeconomic context of the region, as well as accounting and financial concepts, are excluded due to externalities or incentives related to regulatory and fiscal policies. These exclusions affect the accuracy of LCOE and introduce uncertainty when using it as a criterion for investment decision-making or allocation of public resources for the development of renewable sources. This article proposes a clear and standardized methodology for calculating LCOE, which allows for a fair and precise comparison of renewable energy generation options in rural areas. It contributes to informed and transparent decision-making in the fields of energy policy and public investment for expanding electricity service coverage. The proposed methodology is based on the financial cash flow matrix to address variability sources in the calculation, considering the local context of the generation project.

Comparative proteomic analysis provides insights into wood formation in immature xylem at different ages in Eucalyptus urophylla × Eucalyptus grandis.

Wood formation is a crucial developmental stage in the life cycle of a woody plant; this process has substantial scientific research implications and practical applications. However, the mechanisms underlying woody plant development, especially the process of wood formation, remain poorly understood. As eucalyptus is one of the fastest growing tree species in the world, understanding the mechanism of wood formation in eucalyptus will greatly promote the development of molecular breeding technology for forest trees. In this study, we investigated the proteomic profile of immature xylem at four different ages of Eucalyptus urophylla × Eucalyptus grandis (E. urograndis) using iTARQ technology. We identified 5236 proteins and 492 differentially abundant proteins (DAPs). The expression profiles of the DAPs corresponding to coding genes associated with wood formation were assessed using qRT-PCR. From the different expression profiles, it is inferred that the genes encoding kinesin, CDKD3, EXPA13, EXPA2, XTH27, EGases, UGT76E2, LAC, CCoAMT, CesA3, PAL, and CAD may undergo posttranscriptional regulation (PTR). Additionally, the genes encoding EIN2, ETR, MC4-like, and XCP may undergo posttranslational modifications (PTMs). We investigated changes in wood formation-related proteins at the protein abundance level in the immature xylem of E. urograndis, thereby elucidating potential regulatory mechanisms of key proteins involved in eucalyptus wood formation. This study may provide theoretical guidance for further research on molecular breeding techniques and genetic improvement related to the cultivation of rapidly growing and high-quality trees.

Advances in seed hypoxia research - an updated review.

Seeds represent essential stages of the plant life cycle: embryogenesis, the intermittent quiescence phase and germination. Each stage has its own physiological requirements, genetic program and environmental challenges. Consequently, the effects of developmental and environmental hypoxia can vary from detrimental to beneficial. Past and recent evidence shows how low-oxygen signalling and metabolic adaptations to hypoxia affect seed development and germination. Here, we review the recent literature on seed biology in relation to hypoxia research, and present our perspective on key challenges and opportunities for future investigations.

Influence of vegetation phenological carryover effects on plant autumn phenology under climate change

Vegetation phenological carryover effects refer to the influence of previous phenological events on the current or subsequent ones. These effects can modulate the responses of autumn phenology to climate change, but the magnitude and duration of this effect remain poorly understood. Therefore, we employed multiple remote sensing datasets from 1982 to 2018 in the Northern Hemisphere (> 30°N) and partial correlation to investigate the influence of the carryover effect and environmental factors on the end of the growing season (EOS) over time. The importance of the variables in the projection score was used to quantify their relative importance. Our results showed that the previous year's EOS had a robust positive impact on the EOS for 40.02 % of the study area during 1982–2015, which was mainly located in the northern 50°N region. In contrast, the start of the growing season (SOS) was the main contributor to the EOS during 2001–2018, mainly at 40°N-50°N and in northern Russia. The carryover effect persisted into the subsequent year, but its strength and positive impacts varied dramatically in the next year. Concurrently, the mean correlation coefficient between climate factors and EOS rose from 0.20 to 0.22. Notably, the correlation coefficient for frost day frequency increased significantly (P < 0.05) from 0.15 to 0.36, with its influence area expanding from 10.29 % to 11.85 % of the study area. Compared with climate factors, the phenological carryover effect was the dominant driver of the EOS for each vegetation type, accounting for 72.8 % and 44.23 % of the total pixels during 1982–2015 and 2001–2018, respectively. Our study reveals a cascade of ecological consequences that extend beyond the initial occurrence and emphasizes the interconnectedness of growth stages in the plant life cycle, providing valuable insights into the adaptability and vulnerability of plant communities.

Application of high-throughput sequencing in research on plant endophytes

Plant endophytes spend at least part of their life cycle in plants without causing diseases in the hosts, being the microbial resources with rich species and diverse functions. With the advancement in sequencing technology, the microbiological study of endophytes has become increasingly intensive. Being praised for the targeted validation and low cost, Sanger sequencing has been preferred by researchers. However, Sanger sequencing is no longer suitable for deeper genomic study of endophytes due to the low throughput. In this paper, we briefly summarize the research history of endophytes, review the applications of next-generation sequencing characterized by high throughput and third-generation sequencing (single-molecule real-time sequencing) in the research on endophytes, and summarize the research results of different sequencing technologies. Furthermore, we summarize the advantages and limitations of different sequencing technologies and discuss how to choose the appropriate sequencing technology according to the research needs. This review provides a reference for researchers to further explore the potential value of plant endophytes.

Arabidopsis B-BOX DOMAIN PROTEIN14/15/16 form a feedback loop with ELONGATED HYPOCOTYL 5 and PHYTOCHROME-INTERACTING FACTORs to regulate hypocotyl elongation

Light-regulated developmental processes such as photomorphogenesis and flowering play important roles in the plant life cycle, from seedling emergence to reproduction. Three members of the Arabidopsis thaliana B-BOX DOMAIN PROTEIN (BBX) family, BBX14, BBX15, and BBX16 (hereafter BBX14/15/16), redundantly regulate flowering time, but whether this genetic redundancy also affects the regulation of photomorphogenesis remains unclear. Here, we show that light induces BBX14/15/16 expression primarily in the hypocotyl, where BBX14/15/16 redundantly repress hypocotyl elongation. PHYTOCHROME-INTERACTING FACTORs (PIFs) negatively regulate BBX14/15/16 expression mainly through GOLDEN-LIKE proteins (GLKs); however, analyses of ChIP-seq data showed that PIFs are recruited to the BBX14/15/16 loci and can also regulate these genes independently of GLKs. ELONGATED HYPOCOTYL 5 (HY5), a major regulator of photomorphogenesis, also directly binds to the BBX14/15/16 loci and regulates their expression. Simultaneous knockdown of BBX14/15/16 resulted in significant downregulation of HY5 and upregulation of PIFs, suggesting that these factors participate in a feedback regulatory loop. Indeed, BBX14/15/16 induced HY5 promoter activity by binding to the HY5 promoter. The brassinosteroid-responsive gene TOUCH4 (TCH4) and several auxin-responsive SMALL AUXIN UPREGULATED RNA (SAUR) genes were upregulated in the BBX14/15/16 knockdown plants, suggesting that auxin and brassinosteroids might participate in BBX14/15/16-mediated hypocotyl regulation. Mutating the predicted BBX-binding sites in SAUR4 and TCH4 impaired their regulation by BBX14/15/16. We propose that BBX14/15/16, together with HY5 and PIFs, form a feedback loop that regulates the expression of auxin- and brassinosteroid-related genes to modulate hypocotyl elongation.

Investigating and Analyzing the Influence of a Solar Power Plant’s Life Cycle on the Depletion of Natural Materials and Mineral Resources

The production process requires massive amounts of minerals, fossil fuels, and energy. The efficient use of energy and natural resources appears to be crucial to the state of affairs. It should be noted that the post-consumer management of solar power plant elements results in a certain amount of power and matter, as well as harmful effects on the natural world. The major goal of this study was to examine the environmental effect of the solar power plant throughout its life cycle, taking into consideration the depletion of natural materials and mineral resources, using the ReCiPe 2016 model. A life cycle study was performed on an actual 2 MW solar power facility located in northern Poland. This study was conducted using the ReCiPe 2016 model and the Life Cycle Assessment (LCA) methodology. The analyzed renewable energy system’s impact was assessed utilizing 22 impact categories, focusing mostly on the depletion of natural resources. A Life Cycle Assessment was conducted for two post-consumer development scenarios (landfill and recycling). This research focuses on the full solar power plant, not just the photovoltaic panels. Recycling, as a kind of post-consumer development, can provide major environmental benefits and minimize negative environmental consequences throughout the solar power plant’s life cycle. The exceedingly harmful effects can be evident in losses related to water and the aquatic environment. The obtained study findings enabled the development of sustainable-friendly recommendations towards the continuous advancement of the life cycle of solar power plants, thereby reducing the use of rare earth minerals.

Phosphorylation of ADF7-Mediated by AGC1.7 Regulates Pollen Germination in Arabidopsis thaliana.

Actin depolymerizing factors (ADFs), like other actin-binding proteins (ABPs), are modified by phosphorylation to regulate the dynamics of the actin filaments, thereby functioning in various processes throughout the plant lifecycle. In this study, we found that the Arabidopsis thaliana cytoplasmic kinase AGC1.7 interacts with ADF7 in vitro and in vivo. AGC1.7 phosphorylates ADF7 at its Ser-6, Ser-103 and Ser-104 residues in vitro, while replacing these residues with alanine promotes ADF7-mediated actin depolymerization in vitro. Expression of the phosphorylation-mimetic mutant protein ADF7S6/103/104D driven by the pollen-specific LAT52 promoter fully rescues the defects in germination rate, silique length and seeds per silique in both adf7-2 and agc1.5 agc1.7 (agcdm) mutants. Our data establish a model whereby AGC1.7-mediated ADF7 phosphorylation plays an important role in pollen germination and pollen tube growth.

Maize Abiotic Stress Treatments in Controlled Environments.

Maize (Zea mays) is one of the world's most important crops, providing food for humans and livestock and serving as a bioenergy source. Climate change and the resulting abiotic stressors in the field reduce crop yields, threatening food security and the global economy. Water deficit (i.e., drought), heat, and insufficient nutrients (e.g., nitrogen and phosphorus) are major environmental stressors that affect maize yields, and impact growth and development at all stages of the plant life cycle. Understanding the biological processes underlying these responses in maize has the potential to increase yields in the face of abiotic stress. Optimizing individual or combined abiotic stress treatments in controlled environments reduces potential noise in data collection that can be present under less controlled growth conditions. Here, we describe methods and conditions for controlled abiotic stress treatments and associated controls during early vegetative growth of maize, conducted in greenhouses or growth chambers. This includes the environmental conditions, equipment, soil preparation, and intensity and duration of heat, drought, nitrogen deficiency, and phosphorous deficiency. Controlled experiments at early growth stages are informative for future in-field studies that require greater labor and inputs, saving researchers time and growing space, and thus research funds, before testing plants across later stages of development. We suggest that stress treatments be severe enough to result in a measurable phenotype, but not so severe that all plants die prior to sample collection. This protocol is designed to set important standards for replicable research in maize.