Plant Growth Responses Research Articles

Imaging the spatial distribution of structurally diverse plant hormones: A comprehensive review.

Plant hormones are essential and structurally diverse molecules that regulate various aspects of plant growth, development, and stress responses. However, the precise analysis of plant hormones in complex biological samples poses a challenge due to their low concentrations, dynamic levels, and intricate spatial distribution. Moreover, the complexity and interconnectedness of hormone signaling networks make it difficult to simultaneously trace multiple hormone distributions. In this review, we provide an overview of the currently recognized small-molecule plant hormones, signal peptide hormones, and plant growth regulators, along with the analytical methods employed for their analysis. We delve into the latest advancements in mass spectrometry imaging and in situ fluorescence techniques, which enable the examination of the spatial distribution of plant hormones. The advantages and disadvantages of these imaging techniques are further discussed. Finally, we propose potential avenues for future research in this field to further enhance our understanding of plant hormone biology.

The sucrose signalling route controls Flavescence dorée phytoplasma load in grapevine leaves.

Flavescence dorée (FD) is a phytoplasma disease transmitted by insects, causing severe damage to vineyards across Europe. Infected plants cannot be cured and must be removed to prevent further spread. Different grapevine cultivars show varying susceptibility to FD, and some exhibit symptom remission, known as recovery, although the mechanisms behind this are unclear. Diseased plants accumulate soluble sugars, including sucrose, which influences the concentration of trehalose-6P (T6P), a signalling molecule affecting plant growth and stress responses. It is hypothesized that sucrose-mediated signalling via T6P could trigger defence mechanisms, reducing FD pathogen load and increasing plant recovery. Testing this, two grapevine genotypes with different susceptibility to FD were compared, revealing increased sucrose level and TPS activity in the more tolerant cultivar. However, FD-infected plants showed inhibited sucrose-cleaving enzymes and no activation of TPS expression. Attempts to enhance sucrose levels through trunk infusion and girdling promoted sucrose metabolism, T6P biosynthesis, and defence gene expression, facilitating symptom recovery. Girdling particularly enhanced T6P biosynthesis and defence genes above the treatment point, reducing FD pathogen presence and promoting recovery. These findings suggest that elevated sucrose levels, possibly signalling through T6P, may limit FD pathogen spread, aiding in plant recovery.

Effect of nitrate, ammonium, and urea ratios and molybdenum level in the nutrient solution on vegetative growth and biochemical responses of basil plants

In hydroponics, preferring a higher nitrate to ammonia ratio can lead to nitrate accumulation in plant tissues. Reducing nitrate application and substituting nitrate-based fertilizers with ammonium or urea can help solve this issue. Additionally, adding molybdenum (Mo) can decrease nitrate accumulation by stimulating nitrate reductase activity. This study was carried out in a factorial design within a completely randomized experimental setup to investigate the effects of varying nitrate, ammonium, and urea ratios, as well as Mo concentration in the nutrient solution, on vegetative growth and biochemical parameters in hydroponically grown Jenovise basil. The treatments included different ratios of nitrate: ammonium: urea (75:25:0, 50:25:25, 25:25:50, 0:25:75, 50:0:50, 25:0:75, 0:0:100, or 100:0:0) and three Mo levels (0.001, 0.01, 0.1 mg. L−1) in a modified Hoagland solution. The results indicated that growth-associated characteristics improved when nitrate was partially substituted with ammonium and urea in the nutrient solution. The highest total phenolics (2.61 mg. g−1 DW) and flavonoids (2500.36 mg. g−1 DW) were observed at a nitrate: ammonium: urea ratio of 75:25:0 with 0.1 mg. L−1 Mo. Plants treated with a nutrient solution containing a nitrate: ammonium: urea ratio of 0:25:75 and 0.01 mg L−1 Mo showed a significant increase in chlorophyll b concentration, reaching 0.22 mg.g−1 FW, which was higher than all other treatments. In conclusion, our findings demonstrated that optimizing the combined nitrate, ammonium, and urea ratios along with Mo in the nutrient solution regulated nitrate accumulation and growth in hydroponically grown basil.

A Bamboo HD-Zip Transcription Factor PeHDZ72 Conferred Drought Tolerance by Promoting Sugar and Water Transport.

Drought drastically affects plant growth, development and productivity. Plants respond to drought stress by enhancing sugar accumulation and water transport. Homeodomain-leucine zipper (HD-Zip) transcription factors (TFs) participate in various aspects of plant growth and stress response. However, the internal regulatory mechanism of HD-Zips in moso bamboo (Phyllostachys edulis) remains largely unknown. In this study, we identified an HD-Zip member, PeHDZ72, which was highly expressed in bamboo shoots and roots and was induced by drought. Furthermore, PeSTP_46019, PeSWEET_23178 and PeTIP4-3 were identified as downstream genes of PeHDZ72 in moso bamboo by DAP-seq. The expressions of these three genes were all induced by drought stress. Y1H, DLR and GUS activity assays demonstrated that PeHDZ72 could bind to three types of HD-motifs in the promoters of these three genes. Overexpression of PeHDZ72 led to a remarkable enhancement in drought tolerance in transgenic rice, with significantly improved soluble sugar and sucrose contents. Meanwhile, the expressions of OsSTPs, OsSWEETs and OsTIP were all upregulated in transgenic rice under drought stress. Overall, our results indicate that drought stress might induce the expression of PeHDZ72, which in turn activated downstream genes PeSTP_46019, PeSWEET_23178 and PeTIP4-3, contributing to the improvement of cellular osmotic potential in moso bamboo in response to drought stress.

Earthworm mucus contributes significantly to the accumulation of soil cadmium in tomato seedlings

Whether earthworm mucus affects Cd transport behavior in soil-plant systems remains uncertain. Consequently, this study thoroughly assessed the impacts of earthworm mucus on plant growth and physiological responses, plant Cd accumulation, translocation, and distribution, as well as soil characteristics and Cd fractionation in a soil-plant (tomato seedling) system. Results demonstrated that the earthworm inoculation considerably enhanced plant Cd uptake and decreased plant Cd translocation, the effects of which were appreciably less significant than those of the earthworm mucus. This suggested that earthworm mucus may play a crucial role in the way earthworms influence plant Cd uptake and translocation. Moreover, the artificial mucus, which contained identical inorganic nitrogen contents to those in earthworm mucus, had no significant effect on plant Cd accumulation or translocation, implying that components other than inorganic nitrogen in the earthworm mucus may have contributed significantly to the overall effects of the mucus. Compared with the control, the earthworm mucus most substantially increased the root Cd content, the Cd accumulation amount of root and whole plant, and root Cd BCF by 93.7 %, 221.3 %, 72.2 %, and 93.7 %, respectively, while notably reducing the Cd TF by 48.2 %, which may be ascribed to the earthworm mucus's significant impacts on tomato seedling growth and physiological indicators, its considerable influences on the subcellular components and chemical species of root Cd, and its substantial effects on the soil characteristics and soil Cd fractionation, as revealed by correlation analysis. Redundancy analysis further suggested that the most prominent impacts of earthworm mucus may have been due to its considerable reduction of soil pH, improvement of soil DOC content, and enhancement of the exchangeable Cd fraction in soil. This work may help better understand how earthworm mucus influences the transport behavior of metals in soil-plant systems.

Plant growth and physiological responses of the invasive plant Acmella radicans to contrasting light and soil water conditions

Acmella radicans (Jacquin) R.K. Jansen is a new invasive species record for Yunnan Province, China, as of 2017 and little is known about its invasion mechanisms. To better understand its invasive strategies, we investigated the growth, physiological and soil nutrient use parameters of the invader under combined conditions of light (25%, 50%, 75%, and 100% of light availability) and soil water content (full, high, medium, and low soil water content) in the glasshouse. The results showed that light level, soil water content and their interaction had a significant effect on all plant morphological, physiological and soil nutrient parameters for A. radicans (P < 0.05). For the most part, plant height, total branch length, leafstalk length, leaf area, inflorescence number, seed number, leaf biomass, stem biomass, aboveground biomass, and total biomass of A. radicans were significantly increased with increased shading rate and soil water content and were generally greater at intermediate light levels and intermediate to high soil water levels. Under high-irradiance conditions, the chlorophyll content was greatly reduced compared to other treatments. We also observed that leafstalk length, leaf area, Pn, chlorophyll a, and chlorophyll b of A. radicans were markedly increased with increased shading rate and soil water content, and were generally highest under intermediate irradiance and soil water conditions, but many plant physiological parameters also exhibited relatively high values under waterlogging conditions. The concentrations of organic matter, available N and available K of A. radicans soils at high-irradiance and full-high soil water content treatments and medium-irradiance and high-medium soil water content treatments were often less than those of other treatments indicated that hypothetically soil absorption was increased by moderate shading and high soil water. This was the first study demonstrating that A. radicans thrives best under moderate light conditions in combination with high soil water. Furthermore, we surmised that its higher phenotypic and physiological plasticity contributes to its invasion success.

Genome-wide analysis of the WOX gene family and function exploration of RhWOX331 in rose (R. 'The Fairy').

WOXs are a class of plant-specific transcription factors that play key roles in plant growth and stress responses. However, the mechanism by which WOXs influence adventitious root development in Rosa hybrida remains unclear. In this study, RcWOX gene family in rose was identified and phylogenetically analyzed using bioinformatics analysis. A total of 381 RcWOX gene members were localized on seven chromosomes except of nine members. The main cis-acting elements involved in hormonal, light, developmental, and abiotic stress responses were identified in the promoters of RcWOX genes, suggesting their regulation by these signals. Nine RhWOX genes had significant different expression during rooting process of rose. RhWOX331, RhWOX308, RhWOX318 were positive with the formation of rose roots. RhWOX331 was positively involved in the formation of adventitious root primordia, which gene coding a transcription factor localized in the nucleus. The HOX conserved domain in the protein contributed to the self-activating activity of RhWOX331. We obtained genetically modified Arabidopsis to validate the function of RhWOX331. Overexpression of RhWOX331 gene alleviated the inhibition of root length of A. thaliana primary roots by high concentration of IBA and NPA, and significantly increased the number of lateral roots on the primary roots, as well as the height of A. thaliana plants. Additionally, RhWOX331 promoted adventitious root formation in A. thaliana and mitigated hormonal inhibition by exogenous 6-BA, NPA, and GA3. The RhWOX331 promoter contained cis-acting elements such as ABRE, Box 4 and CGTCA-motif et.al. GUS activity analysis showed that the gene acted at the cotyledon attachment site. Taken together, these studies identified a significant expansion of the RcWOX gene family, inferred roles of certain branch members in adventitious root formation, elucidated the function of RhWOX331 in adventitious root initiation, and laid the foundation for further research on the function of WOX gene family in roses.

The knockout of SlMTC impacts tomato seed size and reduces resistance to salt stress in tomato

Members of the MT-A70 family are key catalytic proteins involved in m6A methylation modifications in plants. They play diverse roles at the posttranscriptional level by regulating RNA secondary structure, selective splicing, stability, and translational efficiency, which collectively affect plant growth, development, and stress responses. In this study, we explored the function of the gene SlMTC, a Class C member of the MT-A70 family, in tomatoes by using CRISPR/Cas9 technology. Compared with the wild-type (WT), the CR-slmtc mutants exhibited decreased seed size and slower growth rates during the seedling stage, along with weaker salt tolerance and significant downregulation of stress-related genes, such as PR1, PR5, and P5CS. The qRT-PCR results revealed that the expression levels of genes involved in auxin biosynthesis (FZY1, FZY3, and FZY4) and polar transport (PIN1, PIN4, and PIN8) were lower in CR-slmtc plants than in the WT plants. In addition, yeast two-hybrid assays showed that SlMTC could interact with SlMTA, a Class A member of the MT-A70 family, providing insights into the potential mode of action of SlMTC in tomatoes. Overall, our findings indicate the critical role of SlMTC in plant growth and development as well as in response to salt stress.

Effects of soil microplastic heterogeneity on plant growth vary with species and microplastic types

Microplastics are heterogeneously distributed in soils. However, it is unknown whether soil microplastic heterogeneity affects plant growth and root foraging responses and whether such effects vary with plant species and microplastic types. We grew each of seven herbaceous species (Platycodon grandiflorus, Trifolium repens, Portulaca oleracea, Medicago sativa, Taraxacum mongolicum, Perilla frutescenst, and Paspalum notatum) in heterogeneous soil (patches without microplastics and patches with 0.2 % microplastics) and homogeneous soil (patches with 0.1 % microplastics). Three microplastic types were tested: polypropylene (PP), polyacrylonitrile (PAN), and polyester (PET). P. frutescens showed no response to soil microplastic heterogeneity. For P. grandiflora, microplastic heterogeneity tended to decrease its biomass (total, shoot and root) when the microplastic was PAN and also shoot biomass when it was PET, but had no effect when it was PP. For T. repens, microplastic heterogeneity promoted biomass when PAN was used, decreased total and root biomass when PET was used, but showed no effect when PP was used. Microplastic heterogeneity increased biomass of P. oleracea and decreased that of M. sativa when PET was used, but had no effect when PP or PAN was used. For T. mongolicum, microplastic heterogeneity reduced biomass when the microplastic was PAN, tended to increase total and root biomass when it was PP, but showed no effect when it was PET. For P. notatum, microplastic heterogeneity increased biomass when the microplastic was PP, decreased it when PET was used, but had no effect when PAN was used. However, biomass of none of the seven species showed root foraging responses at the patch level. Therefore, soil microplastic heterogeneity can influence plant growth, but such effects depend on species and microplastic types and are not associated with root foraging. Our findings highlight the roles of soil microplastic heterogeneity, which may influence species interactions and community structure and productivity.

Addition of polyester microplastic fibers to soil alters the diversity and abundance of arbuscular mycorrhizal fungi and affects plant growth and nutrition

Microplastics (MPs) represent an emerging factor in global environmental change and are increasingly found in soils. However, the extent to which they affect plants and their interactions with the soil microbiome is poorly understood. Here, we test the hypothesis that increasing levels of polyester MP fibers in soil alter plant growth and nutrient acquisition responses to arbuscular mycorrhizal (AM) fungi via changes in AM fungal colonization and community composition. We used Sorghum drummondii as a model species in a fully factorial greenhouse experiment. Plants were exposed to soil treatments with 0, 0.2, 1, and 3 % MP polyester fibers either in the presence or absence of an assembled AM fungal community comprising 13 species across three families with contrasting life-history strategies. We found that the 1 % MP treatment promoted plant biomass irrespective of the presence of AM fungi. While no changes in macronutrient concentrations in plant tissues were seen, there was a significant increase in B and Mn when relatively low amounts of MPs were added, and this effect was modulated by AM fungi. Furthermore, there were shifts in AM fungal community composition in response to MP, favoring taxa such as Gigaspora sp. while negatively affecting ruderal taxa like Glomus sp. Overall, our data indicate that MP polyester fibers present in soil can in some cases be beneficial to plants and AM fungal interactions. However, the implications of these findings over the long-term and in the context of ecological repercussions of MP pollution in the environment remain to be seen.

Overexpression of VaSS4 negatively regulates cold tolerance by disturbing ROS balance and decreasing soluble sugar content

Cold stress is one of the major limiting factors for grapevine growth and yield. Sucrose synthetase (SS) is a glycosyl transferase enzyme involved in plant sugar metabolism, which is essential for plant growth and development and response to abiotic stress. However, the function of SS genes in grape cold resistance has not been systematically understood. In the present study, a SS gene from Vitis amurensis named VaSS4 was identified, which was significantly suppressed in response to cold stress. The physicochemical properties, evolutionary relationships, synteny, and selection pressure of VaSS4 were analyzed. Furthermore, VaSS4-overexpressed Arabidopsis and grape calli were obtained to characterize their tolerance to cold stress. Amino acid sequence analysis demonstrated that VaSS4 protein encoded 806 amino acids and contained conserved sucrose synthase domain and glycosyl transferase domain. Overexpression of VaSS4 decreased SS decomposition activity and soluble sugar accumulation under cold stress. Meanwhile, VaSS4 overexpression decreased the reactive oxygen scavenging ability and cold tolerance of Arabidopsis and grape calli. Importantly, overexpression of VaSS4 negatively regulated the expression of cold-related genes in the CBF-dependent pathway compared with WT plants. Overall, these findings not only provide extensive systematic information for Vitis amurensis VaSS4 gene research, but also provide valuable germplasm resources for grape cold resistance breeding.

MiR1432 negatively regulates cold tolerance by targeting OsACAs.

miRNAs function as negative regulators that significantly influence plant growth and stress responses. Within rice and other monocotyledonous plants, miR1432 plays a conserved role in seed development and disease resistance. However, its involvement in the response to abiotic stresses remains unclear. Our study aimed to elucidate this mechanism by predicting the targeting of the rice P-type IIB Ca2+ ATPase gene OsACAs by miR1432 and identifying its cleavage sites via 5'RACE. We observed induced expression of miR1432 and its target gene, OsACA6, under abiotic stresses. Overexpression (OX) of miR1432 and suppression of OsACA6 resulted in reduced cold, salt, and drought tolerance, while OsACA6 suppression/knockout and OX had opposite effects on cold tolerance. Additionally, miR1432 may target other OsACA6 homologs. RNA-sequencing data highlighted the differential expression of stress-related genes in miR1432-overexpressing rice. Furthermore, miR1432-overexpressing rice exhibited weakened vigor, dwarfism, yellowing leaves and reduced fertility. Collectively, our results strongly suggest that miR1432 not only negatively modulates abiotic stress tolerance by suppressing Ca2+ ATPase gene(s) but also influences plant growth and development.

Genome-Wide Identification of the Class III Peroxidase Gene Family in Ginger and Expression Analysis under High Temperature and Intense Light Stress

Ginger, valued for its medicinal properties and economic significance, is vulnerable to environmental stressors such as intense light and high temperatures, which can hinder its growth and development. Class III peroxidases (PRXs) are plant-specific oxidoreductases essential for plant development, growth, and stress responses. Despite their importance, there is limited information available on the function of the class III peroxidase gene family in ginger (ZoPRX). In this study, 103 ZoPRX members within the ginger genome were identified, unevenly distributed across 11 chromosomes. The identified ZoPRX members were categorized into five subfamilies based on gene structures, protein motifs, and phylogenetic analysis. Gene duplication analysis revealed that ZoPRX has primarily undergone segmental duplication. Interspecies homology analysis between ginger and Arabidopsis thaliana, Oryza sativa, and Musa acuminata suggested most ZoPRXs in ginger originated after the divergence of dicotyledon and monocotyledon. Analysis of promoter cis-acting elements identified defense and stress response elements in 39 genes and hormone response elements in 95 genes, indicating their potential roles in responding to environmental stresses. Quantitative Real-Time PCR (qRT-PCR) analysis confirmed that the majority of ZoPRX members are responsive to high temperature and intense light stress. This study provides a comprehensive understanding of the PRX family in ginger, thereby laying the groundwork for future investigations into the functional role of ZoPRX genes under high-temperature and intense light-stress conditions.

Genome wide identification of phenylalanine ammonia-lyase (PAL) gene family in Cucumis sativus (cucumber) against abiotic stress

Phenylalanine ammonia lyase (PAL) is a widely studied enzyme in plant biology due to its role in connecting primary metabolism to secondary phenylpropanoid metabolism, significantly influencing plant growth, development, and stress response. Although PAL genes have been extensively studied in various plant species but their exploration in cucumber has been limited. This study successfully identified 11 CsPAL genes in Cucumis sativus (cucumber). These CsPAL genes were categorized based on their conserved sequences revealing patterns through MEME analysis and multiple sequence alignment. Interestingly, cis-elements related to stress were found in the promoter regions of CsPAL genes, indicating their involvement in responding to abiotic stress. Furthermore, these gene’s promoters contained components associated with light, development and hormone responsiveness. This suggests that they may have roles in hormone developmental processes. MicroRNAs were identified as a key regulators for the CsPAL genes, playing a crucial role in modulating their expression. This discovery underscores the complex regulatory network involved in the plant’s response to various stress conditions. The influence of these microRNAs further highlights the complicated mechanisms that plants use to manage stress. Gene expression patterns were analyzed using RNA-seq data. The significant upregulation of CsPAL9 during HT3h (heat stress for 3 h) and the heightened upregulation of both CsPAL9 and CsPAL7 under HT6h (heat stress for 6 h) in the transcriptome study suggest a potential role for these genes in cucumber’s tolerance to heat stress. This comprehensive investigation aims to enhance our understanding of the PAL gene family’s versatility, offering valuable insights for advancements in cucumber genetics.

Molecular characterization of PSEUDO RESPONSE REGULATOR family in Rosaceae and function of PbPRR59a and PbPRR59b in flowering regulation

BackgroundPSEUDO RESPONSE REGULATOR (PRR) genes are essential components of circadian clock, playing vital roles in multiple processes including plant growth, flowering and stress response. Nonetheless, little is known about the evolution and function of PRR family in Rosaceae species.ResultsIn this study, a total of 43 PRR genes in seven Rosaceae species were identified through comprehensive analysis. The evolutionary relationships were analyzed with phylogenetic tree, duplication events and synteny. PRR genes were classified into three groups (PRR1, PRR5/9, PRR3/7). The expansion of PRR family was mainly derived from dispersed and whole-genome duplication events. Purifying selection was the major force for PRR family evolution. Synteny analysis indicated the existence of multiple orthologous PRR gene pairs between pear and other Rosaceae species. Moreover, the conserved motifs of eight PbPRR proteins supported the phylogenetic relationship. PRR genes showed diverse expression pattern in various tissues of pear (Pyrus bretschneideri). Transcript analysis under 12-h light/ dark cycle and constant light conditions revealed that PRR genes exhibited distinct rhythmic oscillations in pear. PbPRR59a and PbPRR59b highly homologous to AtPRR5 and AtPRR9 were cloned for further functional verification. PbPRR59a and PbPRR59b proteins were localized in the nucleus. The ectopic overexpression of PbPRR59a and PbPRR59b significantly delayed flowering in Arabidopsis transgenic plants by repress the expression of AtGI, AtCO and AtFT under long-day conditions.ConclusionsThese results provide information for exploring the evolution of PRR genes in plants, and contribute to the subsequent functional studies of PRR genes in pear and other Rosaceae species.

Cotton HD-Zip I transcription factor GhHB4-like regulates the plant response to salt stress

Soil salinity is a major environmental constraint to plant production. The homeodomain-leucine zipper I (HD-Zip I) transcription factors play a crucial role in growth, development and defence responses of plants. However, the function and underlying mechanism of HD-Zip I in cotton remain unexplored. This study investigated the role of GhHB4-like, a cotton HD-Zip I gene, in plant tolerance to salt stress. Ectopic expression of GhHB4-like gene enhanced, while its silencing impaired the salt tolerance in Arabidopsis. Y1H and effector-reporter assays revealed that GhHB4-like activated the expression of GhNAC007, which is essential for salt resistance. Knock-down of GhNAC007 also impaired salt resistance of cotton plants. In addition, GhHB4-like-GhNAC007 might have positively regulated the expression of GhMYB96 and ABA signalling-related genes, thereby leading to enhanced salt resistance. Interestingly, deleting motifs 3 and 5 near the 3′-end of GhHB4-like significantly enhanced GhNAC007 activation, indicating that both motifs acted as transcriptional activation inhibitory domains. The results suggest that GhHB4-like-GhNAC007 regulated plant response to salt stress, potentially by modulating GhMYB96 and ABA signalling-related genes.

The Role of Brassinosteroids in Plant Cold Stress Response.

Temperature affects plant growth and geographical distribution. Cold stress occurs when temperatures fall below the physiologically optimal range for plants, causing permanent and irreversible damage to plant growth, development, and production. Brassinosteroids (BRs) are steroid hormones that play an important role in plant growth and various stress responses. Recent studies have shown that low temperatures affect BR biosynthesis in many plant species and that BR signaling is involved in the regulation of plant tolerance to low temperatures, both in the CBF-dependent and CBF-independent pathways. These two regulatory pathways correspond to transient and acclimation responses of low temperature, respectively. The crosstalk between BRs and other hormones is a significant factor in low-temperature tolerance. We provide an overview of recent developments in our knowledge of BRs' function in plant responses to cold stress and how they interact with other plant hormones in this review.

Transcriptome-Wide Identification of Dark- and Salt-Induced Senescence-Related NAC Gene Family Members in Alfalfa.

Leaves are a key forage part for livestock, and the aging of leaves affects forage biomass and quality. Preventing or delaying premature leaf senescence leads to an increase in pasture biomass accumulation and an improvement in alfalfa quality. NAC transcription factors have been reported to affect plant growth and abiotic stress responses. In this study, 48 NAC genes potentially associated with leaf senescence were identified in alfalfa under dark or salt stress conditions. A phylogenetic analysis divided MsNACs into six subgroups based on similar gene structure and conserved motif. These MsNACs were unevenly distributed in 26 alfalfa chromosomes. The results of the collinearity analysis show that all of the MsNACs were involved in gene duplication. Some cis-acting elements related to hormones and stress were screened in the 2-kb promoter regions of MsNACs. Nine of the MsNAC genes were subjected to qRT-PCR to quantify their expression and Agrobacterium-mediated transient expression to verify their functions. The results indicate that Ms.gene031485, Ms.gene032313, Ms.gene08494, and Ms.gene77666 might be key NAC genes involved in alfalfa leaf senescence. Our findings extend the understanding of the regulatory function of MsNACs in leaf senescence.

Plant responses to nitrate and ammonium availability in Australian soils as measured by diffusive gradients in thin-films (DGT) and KCl extraction

Determining soil nitrogen (N) availability is essential in agriculture to minimise over-application, maximise growers’ returns and reduce potential environmental consequences. The present study assesses soil mineral N (nitrate-N and ammonium-N) using the diffusive gradients in thin-films (DGT) technique against the conventional potassium chloride (KCl) extraction. The DGT technique has demonstrated reliable predictability for plant-available P, Cu and Zn. However, the use of DGT to quantify soil N bioavailability is underreported and N measurements made with DGT have not been compared to plant growth responses or N uptake. A pot trial using wheat was performed to determine the suitability of the DGT technique to predict N plant uptake and plant biomass. Four contrasting soil types from South Australia were used, and four rates of N were applied to the soil. DGT devices and KCl extraction were used at sowing to measure soil mineral N. These data were then compared with plant relative yield (YR) and N uptake after harvesting the plants. Soil mineral N, as measured by both the DGT and KCl extraction techniques, demonstrated a significant positive correlation with YR, with an R2 value of 0.6; however, DGT-N extracted comparatively more nitrate (NO3–, >87 % of CN) than KCl-N (65 % of EN). Mineral N and NO3– extracted by both DGT and KCl significantly correlated with plant N uptake albeit this correlation was stronger for KCl (R2 = 0.8) than DGT (R2 = 0.6). The same parameters also positively and significantly correlated with YR, however in this case, both correlations were similar and only modest (R2 < 0.6 in all cases). These results are explained in terms of the differences between the pools of N accessed by these techniques and limitations related to soil N dynamics. In conclusion, KCl showed similar or better predictive ability for N uptake and yield response (YR) in wheat compared to DGT across four Australian soils. Given its low cost and ease of application, KCl presents a competitive advantage in this study.

Five unaddressed questions about cytokinin biosynthesis.

Cytokinins, a class of phytohormones, play crucial roles in regulating plant growth and stress responses through finely tuned feedback loops involving metabolic and signaling cascades. Cytokinin metabolism modulates the abundance of these biologically active molecules. Over the past 25 years, studies have identified key genes involved in cytokinin biosynthesis and inactivation pathways. Nevertheless, several gaps remain in our understanding, particularly regarding the movement of intermediate metabolites between subcellular compartments and the discrepancy between the product of adenosine phosphate-isopentenyltransferase (IPT) and the substrate preferences of subsequent reactions. In addition, recent gene discoveries related to lonely guy (LOG)-independent pathways suggest a spatial extension of cytokinin biosynthesis into the apoplast. Other intriguing issues remain to be addressed, i.e., elucidating the synthetic pathway for cis-zeatin and unraveling the molecular mechanisms governing selective substrate use by the cytokinin biosynthetic enzyme tumor morphology root (Tmr) derived from the phytopathogen Agrobacterium tumefaciens during crown gall formation. Further studies are needed to reveal a fully comprehensive picture of cytokinin metabolism.