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We studied the D3-type cyclin function during gynoecium development in Arabidopsis and how they are related to the hormone cytokinin and the transcription factor SPATULA. Growth throughout the life of plants is sustained by cell division and differentiation processes in meristematic tissues. In Arabidopsis, gynoecium development implies a multiphasic process where the tissues required for pollination, fertilization, and seed development form. The Carpel Margin Meristem (CMM) is a mass of undifferentiated cells that gives rise to the gynoecium internal tissues, such as septum, ovules, placenta, funiculus, transmitting tract, style, and stigma. Different genetic and hormonal factors, including cytokinin, control the CMM function. Cytokinin regulates the cell cycle transitions through the activation of cell cycle regulators as cyclin genes. D3-type cyclins are expressed in proliferative tissues, favoring the mitotic cell cycle over the endoreduplication. Though the role of cytokinin in CMM and gynoecium development is highly studied, its specific role in regulating the cell cycle in this tissue remains unclear. Additionally, despite extensive research on the relationship between CYCD3 genes and cytokinin, the regulatory mechanism that connects them remains elusive. Here, we found that D3-type cyclins are expressed in proliferative medial and lateral tissues. Conversely, the depletion of the three CYCD3 genes showed that they are not essential for gynoecium development. However, the addition of exogenous cytokinin showed that they could control the division/differentiation balance in gynoecium internal tissues and outgrowths. Finally, we found that SPATULA can be a mechanistic link between cytokinin and the D3-type cyclins. The data suggest that the role of D3-type cyclins in gynoecium development is related to the cytokinin response, and they might be activated by the transcription factor SPATULA.

Phytohormones are organic compounds produced in trace amounts within plants that regulate their physiological processes. Their physiological effects are highly complex and diverse. They influence processes ranging from cell division, elongation, and differentiation to plant germination and rooting. Therefore, phytohormones play a crucial regulatory role in plant growth and development. Recently, various studies have highlighted the role of PHs, such as auxin, cytokinin (CK), and abscisic acid (ABA), and newer classes of PHs, such as brassinosteroid (BR) and peptide hormone, in the plant responses toward environmental stresses. These hormones not only have distinct roles at different stages of plant growth but also interact to promote or inhibit each other, thus effectively regulating plant development. Roots are the primary organs for water and mineral absorption in plants. During seed germination, the radicle breaks through the seed coat and grows downward to form the primary root. This occurs because the root needs to quickly penetrate the soil to absorb water and nutrients, providing essential support for the plant's subsequent growth. Root development is a highly complex and precisely regulated process influenced by various signals. Changes in root architecture can affect the plant's ability to absorb nutrients and water, which in turn impacts crop yield. Thus, studying the regulation of root development is of great significance. Numerous studies have reported on the role of phytohormones, particularly auxins, in root regulation. This paper reviews recent studies on the regulation of root development by various phytohormones, both individually and in combination, providing a reference for researchers in this field and offering perspectives on future research directions for improving crop yields. [ABSTRACT FROM AUTHOR]

Developmental reprogramming allows for flexibility in growth and adaptation to changing environmental conditions. In plants, wounding events can result in new stem cell niches and lateral organs. Adventitious roots develop from aerial parts of the plant and are regulated by multiple stimuli, including wounding. Here, we find that Arabidopsis thaliana seedlings wounded at the hypocotyl-root junction reprogram certain pericycle cells to produce adventitious roots proximal to the wound site. We have determined that competence for this reprogramming is controlled; basal cells close to the wound site can produce adventitious roots, whereas cells distal from the wound site mostly cannot. We found that altering cytokinin response or indole-3-butyric acid (IBA)-to-(indole-3-acetic acid) IAA conversion resulted in an expanded adventitious root competence zone and delineated the connection between these pathways. Our work highlights the importance of endogenous IBA-derived auxin and its interaction with cytokinin in adventitious root formation and the regenerative properties of plants. [Display omitted] • Excision-induced adventitious roots (ARs) form proximal to the wound site • Endogenous IBA-derived auxin promotes AR formation • IBA-derived auxin regulates the zone of AR formation • Cytokinin regulates IBA-derived auxin to control the AR competence zone Damodaran and Strader determine that, in Arabidopsis, adventitious roots created in response to wounding occur proximal to the wound site, suggesting that the wound site sets a zone of competence for cellular reprogramming. Cytokinin and IBA-derived auxin dictate this competence zone. [ABSTRACT FROM AUTHOR]

Background: Plant breeding played a very important role in transforming strawberries from being a niche crop with a small geographical footprint into an economically important crop grown across the planet. But even modern marker assisted breeding takes a considerable amount of time, over multiple plant generations, to produce a plant with desirable traits. As a quicker alternative, plants with desirable traits can be raised through tissue culture by doing precise genetic manipulations. Overexpression of morphogenic regulators previously known for meristem development, the transcription factors Growth-Regulating Factors (GRFs) and the GRF-Interacting Factors (GIFs), provided an efficient strategy for easier regeneration and transformation in multiple crops. Results: We present here a comprehensive protocol for the diploid strawberry Fragaria vesca Hawaii 4 (strawberry) regeneration and transformation under control condition as compared to ectopic expression of different GRF4-GIF1 chimeras from different plant species. We report that ectopic expression of Vitis vinifera VvGRF4-GIF1 provides significantly higher regeneration efficiency during re-transformation over wild-type plants. On the other hand, deregulated expression of miRNA resistant version of VvGRF4-GIF1 or Triticum aestivum (wheat) TaGRF4-GIF1 resulted in abnormalities. Transcriptomic analysis between the different chimeric GRF4-GIF1 lines indicate that differential expression of FvExpansin might be responsible for the observed pleiotropic effects. Similarly, cytokinin dehydrogenase/oxygenase and cytokinin responsive response regulators also showed differential expression indicating GRF4-GIF1 pathway playing important role in controlling cytokinin homeostasis. Conclusion: Our data indicate that ectopic expression of Vitis vinifera VvGRF4-GIF1 chimera can provide significant advantage over wild-type plants during strawberry regeneration without producing any pleiotropic effects seen for the miRNA resistant VvGRF4-GIF1 or TaGRF4-GIF1. [ABSTRACT FROM AUTHOR]

In vitro cultivation of Melaleuca could contribute to the cloning of superior genotypes. Studies of factors affecting micropropagation are needed, such as the interaction with light-emitting diodes (LEDs) and plant growth regulators added to the culture media. This study aimed at better understanding the effects of spectra on the development and physiology of melaleuca cultivated in vitro, as well as the interaction of LEDs with the main cytokinin used in micropropagation, N6-Benzylaminopurine (6-BAP). 6-BAP, spectra, and their interaction had a significant effect on most of the variables analyzed, altering the in vitro development and chlorophyll concentrations in the plants, as well as changing different variables in the culture medium, such as pH, EC, and levels of Ca2+, Mg2+, and P, and nutrient accumulation in the shoots. The results demonstrate that the main effects of adding BAP to the in vitro cultivation of melaleuca are an increase in the number of shoots, which resulted in greater fresh and dry masses; a reduction in height and chlorophyll content; complete inhibition of adventitious rooting; higher consumption of Mg, and lower consumption of Ca and P from the culture medium; higher content of Fe, and lower content of P, S, Mn, Cu and B in the in vitro shoot tissues. [ABSTRACT FROM AUTHOR]

Summary Plant root system is significantly influenced by high soil levels of ammonium nitrogen, leading to reduced root elongation and enhanced lateral root branching. In Arabidopsis, these processes have been reported to be mediated by phytohormones and their downstream signaling pathways, while the controlling mechanisms remain elusive in crops. Through a transcriptome analysis of roots subjected to high/low ammonium treatments, we identified a cytokinin oxidase/dehydrogenase encoding gene, CKX3, whose expression is induced by high ammonium. Knocking out CKX3 and its homologue CKX8 results in shorter seminal roots, fewer lateral roots, and reduced sensitivity to high ammonium. Endogenous cytokinin levels are elevated by high ammonium or in ckx3 mutants. Cytokinin application results in shorter seminal roots and fewer lateral roots in wild‐type, mimicking the root responses of ckx3 mutants to high ammonium. Furthermore, CKX3 is transcriptionally activated by type‐B RR25 and RR26, and ckx3 mutants have reduced auxin content and signaling in roots under low ammonium. This study identified RR25/26‐CKX3‐cytokinin as a signal module that mediates root responses to external ammonium by modulating of auxin signaling in the root meristem and lateral root primordium. This highlights the critical role of cytokinin metabolism in regulating rice root development in response to ammonium. [ABSTRACT FROM AUTHOR]

Summary Hydrogen sulfide (H2S) is an endogenous gaseous signaling molecule, which has been shown to play an important role in plant growth and development by coupling with various phytohormones. However, the relationship between H2S and cytokinin (CTK) and the mechanisms by which H2S and CTK affect root growth remain poorly understood. Endogenous CTK was analyzed by UHPLC‐ESI‐MS/MS. Persulfidation of cytokinin oxidase/dehydrogenases (CKXs) was analyzed by mass spectrometry (MS). ckx2/CKX2wild‐type (WT), OE CKX2 and ckx2/CKX2Cys(C)62alanine(A) transgenic lines were isolated with the ckx2 background. H2S is linked to CTK content by CKX2, which regulates root system architecture (RSA). Persulfidation at cysteine (Cys)62 residue of CKX2 enhances CKX2 activity, resulting in reduced CTK content. We utilized 35S‐LCD/oasa1 transgenic lines to investigate the effect of endogenous H2S on RSA, indicating that H2S reduces the gravitropic set‐point angle (GSA), shortens root hairs, and increases the number of lateral roots (LRs). The persulfidation of CKX2Cys62 changes the elongation of cells on the upper and lower flanks of LR elongation zone, confirming that Cys62 of CKX2 is the specificity target of H2S to regulate RSA in vivo. In conclusion, this study demonstrated that H2S negatively regulates CTK content and affects RSA by persulfidation of CKX2Cys62 in Arabidopsis thaliana. [ABSTRACT FROM AUTHOR]

Cytokinins, which play crucial roles in shoot development, substantially affect grain yield. In rice, the OsRopGEF10-OsRAC3 module is associated with cytokinin signaling and crown root development. However, the effects of RopGEF-mediated cytokinin signaling on rice shoot development and grain yield remain unclear. In this study, we investigated the role of OsRopGEF10 in SAM development and the underlying mechanism. We showed that overexpression of OsRopGEF10 inhibited SAM and panicle development, leading to decreased grain yield. Intriguingly, the overexpression of a specific amino acid mutant of OsRopGEF10, designated gef10-W260S, was found to promote panicle development and grain yield. Further analysis using the BiFC assay revealed that the gef10-W260S mutation disrupted the recruitment of rice histidine phosphotransfer proteins (OsAHP1/2) to the plasma membrane (PM), thereby promoting cytokinin signaling. This effect was corroborated by a dark-induced leaf senescence assay, which revealed an increased cytokinin response in the gef10-W260S ectopic expression lines, whereas the overexpression lines presented a suppressed cytokinin response. Moreover, we revealed that the enhanced panicle development in the gef10-W260S lines was attributable to the upregulated expression of several type-B response regulators (RRs) that are crucial for panicle development. Collectively, these findings revealed the negative regulatory function of OsRopGEF10 in the development of the shoot apical meristem (SAM) via interference with cytokinin signaling. Our study highlights the promising role of OsRopGEF10 as a potential target for regulating SAM and panicle development in rice, revealing a valuable breeding strategy for increasing crop yield. [ABSTRACT FROM AUTHOR]

Micropropagation of explants in vitro has potency to address propagule demands for promoting large-scale vanilla production. Plant growth regulators (i.e., cytokinin, auxin) are necessary for the plant micropropagation success. Objective of this study is to determine the shoot multiplication and root development of Vanilla planifolia under the influence of different concentrations and combinations of BA and IBA for micropropagation. Sixty stem nodal segments of Vanilla planifolia were cultured on MS medium supplemented with IBA (0, 0.5, or 2.0 mg/L) and combined with BA (0, 0.5, 1.0, or 2.0 mg/L). Shoot multiplication and root induction were measured after 60 days of culture. The results show that the MS medium with 1 mg/L IBA hindered shoot growth, while the medium containing 1 mg/L BA yielded the highest number or weight of shoots per explant. For the root development, supplementing the medium with 0.5 mg/L or 1 mg/L IBA improved root length or number of roots per explant, respectively. This research establishes a valuable approach for vanilla micropropagation by utilising low concentrations of plant growth regulators and a rapid protocol. This paves the way for significant advancements in large-scale commercial production. [ABSTRACT FROM AUTHOR]

Potato tubers are reproductive and storage organs, enabling their survival. Unraveling the molecular mechanisms that regulate tuberization is crucial for understanding how potatorespond to environmental stress situations and for potato breeding. Previously, we did a transcriptomic analysis of potato microtuberization without light. This showed that important cellular processes like ribosomal proteins, cell cycle, carbon metabolism, oxidative stress, fatty acids, and phytosterols (PS) biosynthesis were closely connected in a protein–protein interaction (PPI) network. Research on PS function during potato tuberization has been scarce. PS plays a critical role in regulating membrane permeability and fluidity, and they are biosynthetic precursors of brassinosteroids (BRs) in plants, which are critical in regulating gene expression, cell division, differentiation, and reproductive biology. Within a PPI network, we found a module of 15 genes involved in the PS biosynthetic process. Darkness, as expected, activated the mevalonate (MVA) pathway. There was a tight interaction between three coding gene products for HMGR3, MVD2, and FPS1, and the gene products that synthetize PS, including CAS1, SMO1, BETAHSD, CPI1, CYP51, FACKEL, HYDRA1, SMT2, SMO2, STE1, and SSR1. Quantitative real-time polymerase chain reaction (qRT-PCR) confirmed the expression analysis of ten specific genes involved in the biosynthesis of PS. This manuscript discusses the potential role of genes involved in PS biosynthesis during microtuber development. [ABSTRACT FROM AUTHOR]

Abstract Background Plant breeding played a very important role in transforming strawberries from being a niche crop with a small geographical footprint into an economically important crop grown across the planet. But even modern marker assisted breeding takes a considerable amount of time, over multiple plant generations, to produce a plant with desirable traits. As a quicker alternative, plants with desirable traits can be raised through tissue culture by doing precise genetic manipulations. Overexpression of morphogenic regulators previously known for meristem development, the transcription factors Growth-Regulating Factors (GRFs) and the GRF-Interacting Factors (GIFs), provided an efficient strategy for easier regeneration and transformation in multiple crops. Results We present here a comprehensive protocol for the diploid strawberry Fragaria vesca Hawaii 4 (strawberry) regeneration and transformation under control condition as compared to ectopic expression of different GRF4-GIF1 chimeras from different plant species. We report that ectopic expression of Vitis vinifera VvGRF4-GIF1 provides significantly higher regeneration efficiency during re-transformation over wild-type plants. On the other hand, deregulated expression of miRNA resistant version of VvGRF4-GIF1 or Triticum aestivum (wheat) TaGRF4-GIF1 resulted in abnormalities. Transcriptomic analysis between the different chimeric GRF4-GIF1 lines indicate that differential expression of FvExpansin might be responsible for the observed pleiotropic effects. Similarly, cytokinin dehydrogenase/oxygenase and cytokinin responsive response regulators also showed differential expression indicating GRF4-GIF1 pathway playing important role in controlling cytokinin homeostasis. Conclusion Our data indicate that ectopic expression of Vitis vinifera VvGRF4-GIF1 chimera can provide significant advantage over wild-type plants during strawberry regeneration without producing any pleiotropic effects seen for the miRNA resistant VvGRF4-GIF1 or TaGRF4-GIF1.

Abstract Cytokinins, which play crucial roles in shoot development, substantially affect grain yield. In rice, the OsRopGEF10-OsRAC3 module is associated with cytokinin signaling and crown root development. However, the effects of RopGEF-mediated cytokinin signaling on rice shoot development and grain yield remain unclear. In this study, we investigated the role of OsRopGEF10 in SAM development and the underlying mechanism. We showed that overexpression of OsRopGEF10 inhibited SAM and panicle development, leading to decreased grain yield. Intriguingly, the overexpression of a specific amino acid mutant of OsRopGEF10, designated gef10-W260S, was found to promote panicle development and grain yield. Further analysis using the BiFC assay revealed that the gef10-W260S mutation disrupted the recruitment of rice histidine phosphotransfer proteins (OsAHP1/2) to the plasma membrane (PM), thereby promoting cytokinin signaling. This effect was corroborated by a dark-induced leaf senescence assay, which revealed an increased cytokinin response in the gef10-W260S ectopic expression lines, whereas the overexpression lines presented a suppressed cytokinin response. Moreover, we revealed that the enhanced panicle development in the gef10-W260S lines was attributable to the upregulated expression of several type-B response regulators (RRs) that are crucial for panicle development. Collectively, these findings revealed the negative regulatory function of OsRopGEF10 in the development of the shoot apical meristem (SAM) via interference with cytokinin signaling. Our study highlights the promising role of OsRopGEF10 as a potential target for regulating SAM and panicle development in rice, revealing a valuable breeding strategy for increasing crop yield.

Abstract Background Kentucky bluegrass (Poa pratensis L.) panicle development is a coordinated process of cell proliferation and differentiation with distinctive phases and architectural changes that are pivotal to determine seed yield. Cytokinin (CK) is a key factor in determining seed yield that might underpin the second “Green Revolution”. However, whether there is a difference between endogenous CK content and seed yields of Kentucky bluegrass, and how CK-related genes are expressed to affect enzyme regulation and downstream seed yield in Kentucky bluegrass remains enigmatic. Results In order to establish a potential link between CK regulation and seed yield, we dissected and characterized the Kentucky bluegrass young panicle, and determined the changes in nutrients, 6 types of endogenous CKs, and 16 genes involved in biosynthesis, activation, inactivation, re-activation and degradation of CKs during young panicle differentiation of Kentucky bluegrass. We found that high seed yield material had more meristems compared to low seed yield material. Additionally, it was found that seed-setting rate (SSR) and lipase activity at the stage of spikelet and floret primordium differentiation (S3), as well as 1000-grain weight (TGW) and zeatin-riboside (ZR) content at the stages of first bract primordium differentiation (S1) and branch primordium differentiation (S2) showed a significantly positive correlation in the two materials. And zeatin, ZR, dihydrozeatin riboside, isopentenyl adenosine and isopentenyl adenosine riboside contents were higher in seed high yield material than those in seed low yield material at S3 stage. Furthermore, the expressions of PpITP3, PpITP5, PpITP8 and PpLOG1 were positively correlated with seed yield, while the expressions of PpCKX2, PpCKX5 and PpCKX7 were negatively correlated with seed yield in Kentucky bluegrass. Conclusions Overall, our study established a relationship between CK and seed yield in Kentucky bluegrass. Perhaps we can increase SSR and TGW by increasing lipase activity and ZR content. Of course, using modern gene editing techniques to manipulate CK related genes such as PpITP3/5/8, PpLOG1 and PpCKX2/5/7, will be a more direct and effective method in Kentucky bluegrass, which requires further trial validation.

Abstract Background In plants, the leaf functions as a solar panel, where photosynthesis converts carbon dioxide and water into carbohydrates and oxygen. In soybean, leaf type traits, including leaf shape, leaf area, leaf width, and leaf width so on, are considered to be associated with yield. In this study, we performed morphological characterization, transcriptome analysis, and endogenous hormone analysis of a rolled and narrow leaf mutant line (rl) in soybean. Results Compared with wild type HX3, mutant line rl showed rolled and narrower leaflet, and smaller leaf, meanwhile rl also performed narrower pod and narrower seed. Anatomical analysis of leaflet demonstrated that cell area of upper epidermis was bigger than the cell area of lower epidermis in rl, which may lead rolled and narrow leaf. Transcriptome analysis revealed that several cytokinin oxidase/dehydrogenase (CKX) genes (Glyma.06G028900, Glyma.09G225400, Glyma.13G104700, Glyma.14G099000, and Glyma.17G054500) were up-regulation dramatically, which may cause lower cytokinin level in rl. Endogenous hormone analysis verified that cytokinin content of rl was lower. Hormone treatment results indicated that 6-BA rescued rolled leaf enough, rescued partly narrow leaf. And after 6-BA treatment, the cell area was similar between upper epidermis and lower epidermis in rl. Although IAA content and ABA content were reduced in rl, but exogenous IAA and ABA didn’t affect leaf type of HX3 and rl. Conclusions Our results suggest abnormal cytokinin metabolism caused rolled and narrow leaf in rl, and provide valuable clues for further understanding the mechanisms underlying leaf development in soybean.

Abstract Background Hulless barley (Hordeum vulgare L. var. nudum Hook. f.) is a significant cereal crop and a substantial source of forage for livestock. Long non-coding RNAs (lncRNAs) and metabolites play crucial roles in the nutrient accumulation and regeneration of hulless barley plants following mowing. The study aimed to identify differentially expressed lncRNAs and metabolites in hulless barley plants by analyzing transcriptomic and metabolomic datasets at 2 h, 24 h, and 72 h following mowing. Results The study revealed that 190, 90, and 438 lncRNA genes were differentially expressed at the 2 h, 24 h, and 72 h time points compared to the non-mowing control. We identified 14 lncRNA genes—11 downregulated and 3 upregulated—showing consistently significant differential expression across all time points after mowing. These differentially expressed lncRNAs target genes involved in critical processes such as cytokinin signaling, cell wall degradation, storage protein accumulation, and biomass increase. In addition, we identified ten differentially expressed metabolites targeting diverse metabolic pathways, including plant hormones, alkaloids, and flavonoids, before and after mowing at various time points. Endogenous hormone analysis revealed that cytokinin most likely played a crucial role in the regeneration of hulless barley after mowing. Conclusions This study created a comprehensive dataset of lncRNAs, metabolites, and hormones in hulless barley after mowing, revealing valuable insights into the functional characteristics of lncRNAs, metabolites, and hormones in regulating plant regeneration. The results indicated that cytokinin plays a significant role in facilitating the regeneration process of hulless barley after mowing. This comprehensive dataset is an invaluable resource for better understanding the complex mechanisms that underlie plant regeneration, with significant implications for crop improvement.

Abstract:The research was conducted in a greenhouse at the Department of Horticulture and Garden Engineering, College of Agriculture, University of Anbar, Ramadi City, to study the impact of NPK supplementation and CPPU cytokinin spraying on Ficus nitida L.'s vegetative and root growth properties. The study encompassed two primary factors. The first factor comprises the NPK chemical fertilizer in four distinct concentrations: F0 (no addition), F1 (1g urea, 3g DAB, 2g potassium sulfate), F2 (2g urea, 6g DAB, 4g potassium sulfate), and F3 (3g urea, 9g DAB, 4g potassium sulfate). The second factor involves the cytokine CPPU at three concentrations: C0 (no addition), C1 (3 mg/ liters), and C2 (6 mg/ liter). The study was structured using a randomized complete block design (RCBD) comprising three blocks, with three pots allocated per experimental unit. Each block consisted of 36 coefficients, resulting in a total of 108 experimental units across all blocks. Leaf area ,the number of leaves per plant, The increase in the number of branches, the percentage of nitrogen ,phosphorus ,and potassium in the leaves, the percentage of carbohydrates ,And the surface area of the roots 30.837cm2 , 185.083 leaf plant-1 , 40.500 Plant Branch-1 , 2.66% , 0.291% , 1.585% , 6.005%,43.041 cm-1.The increase in vegetative, root, and chemical growth characteristics is attributed to the fact that chemical fertilization with nitrogen plays a major role in the plant's life by participating in the formation of RNA and DNA acids, as well as the enzymatic co-factors NAD and NADP, and also participates in the photosynthetic process (El-Badawy and Sultan, 2020).

A two-year field experiment conducted under dryland conditions in semi-humid and drought-prone regions of China aimed to assess the effect of ammonia-oxidizing bacterial on maize water use efficiency and yield. A heterotrophic ammonia-oxidizing bacteria (HAOB) strain S2_8_1 was used. Six treatments were applied: (1) no irrigation + HAOB strain (DI), (2) no irrigation + blank culture medium (DM), (3) no irrigation control (DCK), (4) irrigation + HAOB (WI), (5) irrigation + blank culture medium (WM), and (6) irrigation control (WCK). Results revealed that HAOB treatment increased maize growth, yield, and water use efficiency over controls, regardless of whether the year was wet or dry. This improvement was attributed to the accelerated nitrification in the rhizosphere soil due to HAOB inoculation, which subsequently led to increased levels of leaf cytokinins. Overall, these findings suggest that HAOB inoculation holds promise as a strategy to boost water use efficiency and maize productivity in dryland agriculture.

Abstract Non-hydraulic root source signaling (nHRS) is a unique positive response to soil drying in the regulation of plant growth and development. However, it is unclear how the nHRS mediates the tradeoff between source and sink at the late growth stages and its adaptive mechanisms in primitive wheat. To address this issue, a root-splitting design was made by inserting solid partition in the middle of the pot culture to induce the occurrence of nHRS using four wheat cultivars (MO1 and MO4, diploid; DM22 and DM31, tetraploid) as materials. Three water treatments were designed as 1) both halves watered (CK), 2) holistic root system watered then droughted (FS), 3) one-half of the root system watered and half droughted (PS). FS and PS were designed to compare the role of the full root system and split root system to induce nHRS. Leaves samples were collected during booting and anthesis to compare the role of nHRS at both growth stages. The data indicated that under PS treatment, ABA concentration was significantly higher than FS and CK, demonstrating the induction of nHRS in split root design and nHRS decreased cytokinin (ZR) levels, particularly in the PS treatment. Soluble sugar and proline accumulation were higher in the anthesis stage as compared to the booting stage. POD activity was higher at anthesis, while CAT was higher at the booting stage. Increased ABA (nHRS) correlated with source-sink relationships and metabolic rate (i.e., leaf) connecting other stress signals. Biomass density showed superior resource acquisition and utilization capabilities in both FS and PS treatment as compared to CK in all plants. Our findings indicate that nHRS-induced alterations in phytohormones and their effect on source-sink relations were allied with the growth stages in primitive wheat.

Plant growth regulators (PGRs) are chemical substances that imitate the functions of phytohormones to enhance the crop yield and the harvest process. Phenylurea-derived plant growth regulators are known for their excellent efficacy in promoting fruit growth, particularly in kiwifruit, grapes, and melons. Phenylurea derivatives represent one class of the highly efficient and versatile PGRs. Specifically, forchlorfenuron (CPPU, N-(2-chloro-4-pyridinyl)-N′-phenylurea) exhibits similar growth-regulating efficacy to cytokinins and has a significant impact on the plant growth and the crop yield. As a result, there is growing interest in exploring the incorporation of various phenylurea moieties into agrochemicals to enhance their regulatory properties on crops. This review aims to provide a comprehensive overview on representative synthetic approaches for phenylurea derived PGRs. Additionally, we provide our perspective on the future development in this active research field.

Summary: Wood is resulted from the radial growth paced by the division and differentiation of vascular cambium cells in woody plants, and phytohormones play important roles in cambium activity.Here, we identified that PagJAZ5, a key negative regulator of jasmonate (JA) signaling, plays important roles in enhancing cambium cell division and differentiation by mediating cytokinin signaling in poplar 84K (Populus alba × Populus glandulosa).PagJAZ5 is preferentially expressed in developing phloem and cambium, weakly in developing xylem cells. Overexpression (OE) of PagJAZ5m (insensitive to JA) increased cambium activity and xylem differentiation, while jaz mutants showed opposite results. Transcriptome analyses revealed that cytokinin oxidase/dehydrogenase (CKXs) and type‐A response regulators (RRs) were downregulated in PagJAZ5m OE plants. The bioactive cytokinins were significantly increased in PagJAZ5m overexpressing plants and decreased in jaz5 mutants, compared with that in 84K plants. The PagJAZ5 directly interact with PagMYC2a/b and PagWOX4b. Further, we found that the PagRR5 is regulated by PagMYC2a and PagWOX4b and involved in the regulation of xylem development.Our results showed that PagJAZ5 can increase cambium activity and promote xylem differentiation through modulating cytokinin level and type‐A RR during wood formation in poplar. [ABSTRACT FROM AUTHOR]

Blossom end enlargement (BEE) is a postharvest deformation that may be related to the influx of photosynthetic assimilates before harvest. To elucidate the mechanism by which BEE occurs, expression marker genes that indicate the physiological condition of BEE-symptomatic fruit are necessary. First, we discovered that preharvest treatment with a synthetic cytokinin, N-(2-Chloro-4-pyridyl)-N'-phenylurea (CPPU), promoted fruit growth and suppressed BEE occurrence. This suggests that excessive assimilate influx is not a main cause of BEE occurrence. Subsequently, the expression levels of seven sugar-starvation marker genes, CsSEF1, AS, CsFDI1, CsPID, CsFUL1, CsETR1, and CsERF1B, were compared among symptomatic and asymptomatic fruits, combined with and without CPPU treatment. Only CsSEF1 showed a higher expression level in asymptomatic fruits than in symptomatic fruits, regardless of CPPU treatment. This was then tested using fruits stored via the modified-atmosphere packaging technique, which resulted in a lower occurrence of BEE, and the asymptomatic fruits showed a higher CsSEF1 expression level than symptomatic fruits, regardless of the packaging method. CsSEF1 codes a CCCH-type zinc finger protein, and an increase in the expression of CsSEF1 was correlated with a decrease in the fruit respiration rate. Thus, CsSEF1 may be usable as a BEE expression marker gene. [ABSTRACT FROM AUTHOR]

Background and Objective: This research was undertaken to assess the impact of kinetin (Kin) and salicylic acid (SA) on yield and its components in soybean plants by reducing ionic, osmotic and oxidative stresses at different levels of salinity. Materials and Methods: The experiment with factorial arrangements based on randomized complete block design with three replications were conducted at the research greenhouse of Urmia University. The investigated factors included salinity (0, 4, 7 and 10 dS/m) and growth regulators (50 μM kinetin and 1 mM salicylic acid). 20 soybean seeds (cultivar M7) were planted at a depth of 3 cm in each pot containing 1 kg of perlite. Growth regulators were sprayed in the vegetative and reproductive stages. One week after the final treatment, tests were performed. Results: Salinity increased the contents of roots and leaves sodium, the amount of H2O2 and malondialdehyde, the activities of catalase, ascorbate peroxidase, superoxide dismutase, α-tocopherol, the contents of soluble sugars and proline of soybean leaves, but decreased potassium, calcium and magnesium ions, chlorophylls content, leaf area index, membrane stability index, leaf relative water content, number of pods per plant, number of seeds per plant, 100-seed weight, seed yield, biological yield and harvest index. Foliar spray of growth regulators, especially Kin + SA, reduced sodium content, H2O2, malondialdehyde and proline content, while enhancing potassium, calcium and magnesium ions, chlorophylls content, leaf area index, antioxidants activities, membrane stability index, soluble sugars and leaf relative water content. Kin + SA treatment also led to considerable improvements in seed yield (39%), biological yield (18.51%) and harvest index (18.16%) of soybean plants by increasing the number of pods per plant, the number of seeds per plant and the 100-seed weight. Conclusion: Foliar application of growth regulators under salt stress, especially Kin+ SA, increased the growth, seed yield and its components in soybean plants through reducing ionic toxicity and improving antioxidants activities. [ABSTRACT FROM AUTHOR]

Background: Kentucky bluegrass (Poa pratensis L.) panicle development is a coordinated process of cell proliferation and differentiation with distinctive phases and architectural changes that are pivotal to determine seed yield. Cytokinin (CK) is a key factor in determining seed yield that might underpin the second "Green Revolution". However, whether there is a difference between endogenous CK content and seed yields of Kentucky bluegrass, and how CK-related genes are expressed to affect enzyme regulation and downstream seed yield in Kentucky bluegrass remains enigmatic. Results: In order to establish a potential link between CK regulation and seed yield, we dissected and characterized the Kentucky bluegrass young panicle, and determined the changes in nutrients, 6 types of endogenous CKs, and 16 genes involved in biosynthesis, activation, inactivation, re-activation and degradation of CKs during young panicle differentiation of Kentucky bluegrass. We found that high seed yield material had more meristems compared to low seed yield material. Additionally, it was found that seed-setting rate (SSR) and lipase activity at the stage of spikelet and floret primordium differentiation (S3), as well as 1000-grain weight (TGW) and zeatin-riboside (ZR) content at the stages of first bract primordium differentiation (S1) and branch primordium differentiation (S2) showed a significantly positive correlation in the two materials. And zeatin, ZR, dihydrozeatin riboside, isopentenyl adenosine and isopentenyl adenosine riboside contents were higher in seed high yield material than those in seed low yield material at S3 stage. Furthermore, the expressions of PpITP3, PpITP5, PpITP8 and PpLOG1 were positively correlated with seed yield, while the expressions of PpCKX2, PpCKX5 and PpCKX7 were negatively correlated with seed yield in Kentucky bluegrass. Conclusions: Overall, our study established a relationship between CK and seed yield in Kentucky bluegrass. Perhaps we can increase SSR and TGW by increasing lipase activity and ZR content. Of course, using modern gene editing techniques to manipulate CK related genes such as PpITP3/5/8, PpLOG1 and PpCKX2/5/7, will be a more direct and effective method in Kentucky bluegrass, which requires further trial validation. [ABSTRACT FROM AUTHOR]

Background: In plants, the leaf functions as a solar panel, where photosynthesis converts carbon dioxide and water into carbohydrates and oxygen. In soybean, leaf type traits, including leaf shape, leaf area, leaf width, and leaf width so on, are considered to be associated with yield. In this study, we performed morphological characterization, transcriptome analysis, and endogenous hormone analysis of a rolled and narrow leaf mutant line (rl) in soybean. Results: Compared with wild type HX3, mutant line rl showed rolled and narrower leaflet, and smaller leaf, meanwhile rl also performed narrower pod and narrower seed. Anatomical analysis of leaflet demonstrated that cell area of upper epidermis was bigger than the cell area of lower epidermis in rl, which may lead rolled and narrow leaf. Transcriptome analysis revealed that several cytokinin oxidase/dehydrogenase (CKX) genes (Glyma.06G028900, Glyma.09G225400, Glyma.13G104700, Glyma.14G099000, and Glyma.17G054500) were up-regulation dramatically, which may cause lower cytokinin level in rl. Endogenous hormone analysis verified that cytokinin content of rl was lower. Hormone treatment results indicated that 6-BA rescued rolled leaf enough, rescued partly narrow leaf. And after 6-BA treatment, the cell area was similar between upper epidermis and lower epidermis in rl. Although IAA content and ABA content were reduced in rl, but exogenous IAA and ABA didn't affect leaf type of HX3 and rl. Conclusions: Our results suggest abnormal cytokinin metabolism caused rolled and narrow leaf in rl, and provide valuable clues for further understanding the mechanisms underlying leaf development in soybean. [ABSTRACT FROM AUTHOR]

Some citrus orchards in China often experience nitrogen (N) deficiency. For the first time, targeted metabolomics was used to examine N-deficient effects on hormones in sweet orange (Citrus sinensis (L.) Osbeck cv. Xuegan) leaves and roots. The purpose was to validate the hypothesis that hormones play a role in N deficiency tolerance by regulating root/shoot dry weight ratio (R/S), root system architecture (RSA), and leaf and root senescence. N deficiency-induced decreases in gibberellins and indole-3-acetic acid (IAA) levels and increases in cis(+)-12-oxophytodienoic acid (OPDA) levels, ethylene production, and salicylic acid (SA) biosynthesis might contribute to reduced growth and accelerated senescence in leaves. The increased ethylene formation in N-deficient leaves might be caused by increased 1-aminocyclopropanecarboxylic acid and OPDA and decreased abscisic acid (ABA). N deficiency increased R/S, altered RSA, and delayed root senescence by lowering cytokinins, jasmonic acid, OPDA, and ABA levels and ethylene and SA biosynthesis, increasing 5-deoxystrigol levels, and maintaining IAA and gibberellin homeostasis. The unchanged IAA concentration in N-deficient roots involved increased leaf-to-root IAA transport. The different responses of leaf and root hormones to N deficiency might be involved in the regulation of R/S, RSA, and leaf and root senescence, thus improving N use efficiency, N remobilization efficiency, and the ability to acquire N, and hence conferring N deficiency tolerance. [ABSTRACT FROM AUTHOR]

Background: Hulless barley (Hordeum vulgare L. var. nudum Hook. f.) is a significant cereal crop and a substantial source of forage for livestock. Long non-coding RNAs (lncRNAs) and metabolites play crucial roles in the nutrient accumulation and regeneration of hulless barley plants following mowing. The study aimed to identify differentially expressed lncRNAs and metabolites in hulless barley plants by analyzing transcriptomic and metabolomic datasets at 2 h, 24 h, and 72 h following mowing. Results: The study revealed that 190, 90, and 438 lncRNA genes were differentially expressed at the 2 h, 24 h, and 72 h time points compared to the non-mowing control. We identified 14 lncRNA genes—11 downregulated and 3 upregulated—showing consistently significant differential expression across all time points after mowing. These differentially expressed lncRNAs target genes involved in critical processes such as cytokinin signaling, cell wall degradation, storage protein accumulation, and biomass increase. In addition, we identified ten differentially expressed metabolites targeting diverse metabolic pathways, including plant hormones, alkaloids, and flavonoids, before and after mowing at various time points. Endogenous hormone analysis revealed that cytokinin most likely played a crucial role in the regeneration of hulless barley after mowing. Conclusions: This study created a comprehensive dataset of lncRNAs, metabolites, and hormones in hulless barley after mowing, revealing valuable insights into the functional characteristics of lncRNAs, metabolites, and hormones in regulating plant regeneration. The results indicated that cytokinin plays a significant role in facilitating the regeneration process of hulless barley after mowing. This comprehensive dataset is an invaluable resource for better understanding the complex mechanisms that underlie plant regeneration, with significant implications for crop improvement. [ABSTRACT FROM AUTHOR]

A protocol for optimization of callus culture establishment from leaf explants of Portulaca oleracea L. was developed. The effect of ethyl methanesulphonate (EMS) on callus induction and synthesis of fatty acids was evaluated. Callus culture was initiated from leaf explants on Murashige and Skoog’s medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D), a combination of 2,4-D + 6-benzyl adenine (BA) or 2,4-D + kinetin. The maximum callus biomass was obtained at 2.5 µM BA + 2.5 µM 2,4-D (20.22 g leaf explant-1 fresh weight). The leaves treated with EMS (0.1 to 0.4% h-1) differentiated callus on this optimized medium combination. The gas chromatography-mass spectrometry analysis of fatty acids indicated that the leaves had a high linolenic acid content (17.72%). The callus cultures synthesized heptadecanoic, oleic, and tricosanoic acids, which were otherwise absent in the leaves. Eicosanoic and docosanoic acids in callus cultures were 2.78 and 3.18 times higher than their content in the leaves, respectively. The diversity of fatty acids in treated callus cultures decreased with increased EMS concentration, but the content of a few individual fatty acids was enhanced. Callus at the dose of 0.1% EMS synthesized linoleic acid, which was two times higher than in the untreated callus, while at 0.2% EMS stearic acid was synthesized, which was absent in the untreated callus. It was concluded that the callus of P. oleracea accumulated different fatty acids, and EMS treatment enhanced the content of a few fatty acids in the cultures. [ABSTRACT FROM AUTHOR]

Here, cytosine methylation in the whole genome of pear flower buds was mapped at a single-base resolution. There was 19.4% methylation across all sequenced C sites in the Pyrus pyrifolia cultivar 'Sucui 1' flower bud genome. Meantime, the CG, CHG, and CHH sequence contexts (where H = A, T or C) exhibited 47.4%, 33.3%, and 11.9% methylation, respectively. Methylation in different gene regions was revealed through combining methylome and transcriptome analysis, which presented various transcription trends. Genes with methylated promoters exhibited lower expression levels than genes with non-methylated promoters, while body-methylated genes displayed an obvious negative correlation with their transcription levels. The methylation profiles of auxin- and cytokinin-related genes were estimated. And some of them proved to be hypomethylated, with increased transcription levels, in wizened buds. More specifically, the expression of the genes PRXP73, CYP749A22, and CYP82A3 was upregulated as a result of methylation changes in their promoters. Finally, auxin and cytokinin concentrations were higher in wizened flower buds than in normal buds. The exogenous application of paclobutrazol (PP333) in the field influenced the DNA methylation status of some genes and changed their expression level, reducing the proportion of wizened flower buds in a concentration-dependent manner. Overall, our results demonstrated the relationship between DNA methylation and gene expression in wizened flower buds of P. pyrifolia cultivar 'Sucui 1', which was associated with changes in auxin and cytokinin concentrations. [ABSTRACT FROM AUTHOR]

Branching/tillering is a critical process for plant architecture and grain yield. However, Branching is intricately controlled by both endogenous and environmental factors. The underlying mechanisms of tillering in wheat remain poorly understood. In this study, we identified Less Tiller 1 (LT1) as a novel regulator of wheat tillering using an enhanced bulked segregant analysis (BSA) method, uni-BSA. This method effectively reduces alignment noise caused by the high repetitive sequence content in the wheat genome. Loss-of-function of LT1 results in fewer tillers due to defects in axillary meristem initiation and bud outgrowth. We mapped LT1 to a 6 Mb region on the chromosome 2D short arm and validated a nucleotide-binding (NB) domain encoding gene as LT1 using CRISPR/Cas9. Furthermore, the lower sucrose concentration in the shoot bases of lt1 might result in inadequate bud outgrowth due to disturbances in the sucrose biosynthesis pathways. Co-expression analysis suggests that LT1 controls tillering by regulating TaROX/TaLAX1, the ortholog of the Arabidopsis tiller regulator REGULATOR OF AXILLARY MERISTEM FORMATION (ROX) or the rice axillary meristem regulator LAX PANICLE1 (LAX1). This study not only offers a novel genetic resource for cultivating optimal plant architecture but also underscores the importance of our innovative BSA method. This uni-BSA method enables the swift and precise identification of pivotal genes associated with significant agronomic traits, thereby hastening gene cloning and crop breeding processes in wheat. [ABSTRACT FROM AUTHOR]

Non-hydraulic root source signaling (nHRS) is a unique positive response to soil drying in the regulation of plant growth and development. However, it is unclear how the nHRS mediates the tradeoff between source and sink at the late growth stages and its adaptive mechanisms in primitive wheat. To address this issue, a root-splitting design was made by inserting solid partition in the middle of the pot culture to induce the occurrence of nHRS using four wheat cultivars (MO1 and MO4, diploid; DM22 and DM31, tetraploid) as materials. Three water treatments were designed as 1) both halves watered (CK), 2) holistic root system watered then droughted (FS), 3) one-half of the root system watered and half droughted (PS). FS and PS were designed to compare the role of the full root system and split root system to induce nHRS. Leaves samples were collected during booting and anthesis to compare the role of nHRS at both growth stages. The data indicated that under PS treatment, ABA concentration was significantly higher than FS and CK, demonstrating the induction of nHRS in split root design and nHRS decreased cytokinin (ZR) levels, particularly in the PS treatment. Soluble sugar and proline accumulation were higher in the anthesis stage as compared to the booting stage. POD activity was higher at anthesis, while CAT was higher at the booting stage. Increased ABA (nHRS) correlated with source-sink relationships and metabolic rate (i.e., leaf) connecting other stress signals. Biomass density showed superior resource acquisition and utilization capabilities in both FS and PS treatment as compared to CK in all plants. Our findings indicate that nHRS-induced alterations in phytohormones and their effect on source-sink relations were allied with the growth stages in primitive wheat. [ABSTRACT FROM AUTHOR]

In vitro mass propagation of apple plants plays an important role in the rapid multiplication of genetically uniform, disease-free scions and rootstocks with desired traits. Successful micropropagation of apple using axillary shoot cultures is influenced by several factors, the most critical of which is the cytokinin included in the culture medium. The impact of medium composition from single added cytokinins on shoot proliferation of apple scion Húsvéti rozmaring cultured on agar-agar gelled Murashige and Skoog medium fortified with indole butyric acid and gibberellic acid was investigated. The optimum concentration for efficient shoot multiplication differs according to the type of cytokinin. The highest significant multiplication rate (5.40 shoots/explant) was achieved using 2.0 μM thidiazuron while the longest shoots (1.80 cm) were observed on the medium containing benzyladenine at a concentration of 2.0 μM. However, application of either thidiazuron or benzyladenine as cytokinin source in the medium resulted in shoots of low quality, such as stunted and thickened shoots with small leaves. In the case of benzyladenine riboside, the 8 μM concentration was the most effective in increasing the multiplication rate (4.76 shoots/explant) but caused thickened stem development with tiny leaves. In the present study, meta-topolin was shown to be the most effective cytokinin that could be applied to induce sufficient multiplication (3.28 shoots/explant) and high-quality shoots along with shoot lengths of 1.46 cm when it was applied at concentrations of 4 μM. However, kinetin was the least active cytokinin; it practically did not induce the development of new shoots. The superior cytokinin for in vitro axillary shoot development of apple scion Húsvéti rozmaring with high-quality shoots was the meta-topolin, but it may be different depending on the variety/genotype under study. [ABSTRACT FROM AUTHOR]

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This study aimed to investigate the effect of salinity and water stress on the physiological and functional characteristics of winter wheat (Triticum aestivum L.) under the foliar application of plant growth regulators (PGRs). The experiment was carried out as a split plot based on a randomized complete block design with three replications in two environments. In each environment, water stress at two irrigation levels (after 90 and 120 mm of pan evaporation) and with two EC of 1.5 and 10 dS/m in the main plots and spraying of PGRs including salicylic acid (SA), gibberellic acid (GA3), and cytokinins (CK) (purine) content with a concentration of 100 ppm and the control treatment (spraying solution with normal water) were placed in subplots. Results indicated that all treatments caused significant increases in functional and qualitative characteristics and yield of Triticum aestivum L. The saline environment and irrigation level after 120 mm of pan evaporation caused a reduction in grain yield in all traits except for seed proline, seed nitrogen content, and seed protein content. Also, the combined foliar application of GA3 + CK + SA increased yield in most traits. The highest RWC of flag leaves was observed in the foliar application of GA3 + SA (3.36 kg/ha) and then in the foliar application of GA3 + SA + CK (57.87 kg/ha). GA3 interacts with PGR spraying to balance another development under saline and non-saline conditions. [ABSTRACT FROM AUTHOR]

In plants, the robust maintenance of tissue structure is crucial to supporting its functionality. The multi-layered shoot apical meristem (SAM) of Arabidopsis, containing stem cells, is an approximately radially symmetric tissue whose shape and structure is maintained throughout the life of the plant. In this paper, a new biologically calibrated pseudo-three-dimensional (P3D) computational model of a longitudinal section of the SAM is developed. It includes anisotropic expansion and division of cells out of the cross-section plane, as well as representation of tension experienced by the SAM epidermis. Results from the experimentally calibrated P3D model provide new insights into maintenance of the structure of the SAM epidermal cell monolayer under tension and quantify dependence of epidermal and subepidermal cell anisotropy on the amount of tension. Moreover, the model simulations revealed that out-of-plane cell growth is important in offsetting cell crowding and regulating mechanical stresses experienced by tunica cells. Predictive model simulations show that tension-determined cell division plane orientation in the apical corpus may be regulating cell and tissue shape distributions needed for maintaining structure of the wild-type SAM. This suggests that cells' responses to local mechanical cues may serve as a mechanism to regulate cell- and tissue-scale patterning.

Plant somatic embryogenesis encompasses somatic cells switch into embryogenic cells that can later produce somatic embryos with the ability to produce plantlets. Previously, we defined in vitro culture settings for the somatic embryogenesis process of Coffea canephora that comprise adequate plantlets with auxin plus cytokinin followed by cut-leaf explant cultivation with cytokinin, producing embryos with the ability to regenerate plantlets. Here, we confirmed that cultivating cut-leaf explants with cytokinin is sufficient to promote somatic embryos proliferation and the high yield of somatic embryos in the protocol requires adequate plantlets with auxin plus cytokinin. Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels reveal auxin-plus cytokinin-dependent regulated proteins in plantlets with up and down abundance. Chitinase A class III, proteins involved in the metabolism and folding of proteins, photosynthesis, antioxidant activity, and chromatin organization were identified. The RPN12 protein, which is a subunit of the proteasome 26S, has an abundance that is not associated with transcript changes, suggesting post-translational regulation.

Common centaury (Centaurium eryhtraea Rafn) is a medicinal plant species with vigorous morphogenic potential in vitro. The process of spontaneous shoot regeneration in a solid root culture is characteristic for this plant species. In this context, the aim of this work was to investigate the dynamic changes of endogenous phytohormones and carbohydrates content in root explants at different time points (0, 2, 4, 7, 14, 21, 28, and 60 days) during spontaneous centaury morphogenesis in vitro. Detailed analysis of cytokinins (CKs) showed that trans-zeatin (tZ) was the major bioactive CK at all time points. The corresponding riboside, tZ9R, was also determined in the majority of the identified transport forms, at all time-points. Further analysis of endogenous auxin revealed a significant increase in endogenous indole-3-acetic acid (IAA) after 21 days, when a huge jump in the ratio of IAA/bioactive CKs was also observed. The maximum total soluble sugar content was measured after 14 days, while a significant decrease was determined after 21 days, when the first regenerated adventitious shoots appeared. This undoubtedly indicates an increased energy requirement prior to the actual regeneration of the shoots. The obtained results indicate that the period from day 14 to day 21 involves the most dramatic disturbances in endogenous bioactive CKs, IAA and carbohydrate balance, which are very important and valuable factors for the onset of shoot regeneration.

Vegetative growth of Cattleya orchids is influenced by environmental factors. In addition, growth and development are also determined by the availability of nutrients and hormones. One of the methods to accelerate the vegetative growth of orchids is by the application of NPK fertilizers and plant growth regulators such as benzyl amino purine (BAP). The objective of this study was to obtain the best concentration of NPK and BAP concentration on the vegetative growth of Cattleya orchids. The experiment was conducted at the screenhouse of the Experimental Garden of Bale Tatanen, Faculty of Agriculture, Padjadjaran University with a Factorial Randomized Block Design consisting of two factors, namely the concentration of NPK fertilizer (0, 1, 2, 3, and 4 g L-1 of NPK-28:10:10 and 2 g L-1 of NPK-32:10:10 as positive control), and the second factor was BAP concentration (0, 0.5 and 1 mg L-1) with three replications. The results showed that there was an interaction effect between the application of 2 g L-1 of NPK 28:10:10 and 1 mg L-1 BAP on the number of pseudobulbs per clump. Independently, the treatment of 2 g L-1 of NPK 28:10:10 showed the best results in the increase of plant height and the number of new shoots. While 0.5 mg L-1 of BAP showed the best results in the increase of plant height, the number of new shoots and the SPAD value.

ABSTRACT The cultivation of beans (Phaseolus vulgaris L.) is of great importance to the national economy since they are grown in almost all regions of Brazil and the use of biostimulants based on phytohormones and plant vitamins can improve the yield of the crop. This work aimed to evaluate the effects of the application of the vitamin nicotinamide and phytohormones as biostimulants on bean plants. The experimental design was carried out in randomized blocks in a 2 x 5 factorial scheme with 4 replicates. The treatments consisted of the application of five doses of nicotinamide (0, 200, 400, 600 and 800 mg L-1) in the presence or absence of phytohormones containing auxin, gibberellin and cytokinin. The growth characteristics, production components and productivity of the bean grains were evaluated. The use of phytohormones and nicotinamide alone or together favored the growth, production components and grain productivity of the bean plant. The range of 366 to 469 mL L-1 nicotinamide achieved the highest values for the growth variables, production components and grain productivity of the bean plant, with 414 mL L-1 being the most suitable to achieve higher grain productivity.

Abstract This study aimed to investigate the effect of salinity and water stress on the physiological and functional characteristics of winter wheat (Triticum aestivum L.) under the foliar application of plant growth regulators (PGRs). The experiment was carried out as a split plot based on a randomized complete block design with three replications in two environments. In each environment, water stress at two irrigation levels (after 90 and 120 mm of pan evaporation) and with two EC of 1.5 and 10 dS/m in the main plots and spraying of PGRs including salicylic acid (SA), gibberellic acid (GA3), and cytokinins (CK) (purine) content with a concentration of 100 ppm and the control treatment (spraying solution with normal water) were placed in subplots. Results indicated that all treatments caused significant increases in functional and qualitative characteristics and yield of Triticum aestivum L. The saline environment and irrigation level after 120 mm of pan evaporation caused a reduction in grain yield in all traits except for seed proline, seed nitrogen content, and seed protein content. Also, the combined foliar application of GA3 + CK + SA increased yield in most traits. The highest RWC of flag leaves was observed in the foliar application of GA3 + SA (3.36 kg/ha) and then in the foliar application of GA3 + SA + CK (57.87 kg/ha). GA3 interacts with PGR spraying to balance another development under saline and non-saline conditions.

Abstract Background Gaseous phytohormone ethylene levels are directly influenced by the production of its immediate non-volatile precursor 1-aminocyclopropane-1-carboxylic acid (ACC). Owing to the strongly acidic character of the ACC molecule, its quantification has been difficult to perform. Here, we present a simple and straightforward validated method for accurate quantification of not only ACC levels, but also major members of other important phytohormonal classes – auxins, cytokinins, jasmonic acid, abscisic acid and salicylic acid from the same biological sample. Results The presented technique facilitates the analysis of 15 compounds by liquid chromatography coupled with tandem mass spectrometry. It was optimized and validated for 10 mg of fresh weight plant material. The extraction procedure is composed of a minimal amount of necessary steps. Accuracy and precision were the basis for evaluating the method, together with process efficiency, recovery and matrix effects as validation parameters. The examined compounds comprise important groups of phytohormones, their active forms and some of their metabolites, including six cytokinins, four auxins, two jasmonates, abscisic acid, salicylic acid and 1-aminocyclopropane-1-carboxylic acid. The resulting method was used to examine their contents in selected Arabidopsis thaliana mutant lines. Conclusion This profiling method enables a very straightforward approach for indirect ethylene study and explores how it interacts, based on content levels, with other phytohormonal groups in plants.

This study involves the evaluation of in vitro shoot regeneration during cotyledon culture in black sesame (Sesamum indicum L. cv. TMV3) under various concentrations of sucrose and cytokinin (BAP) in Murashige and Skoog (MS) nutrient medium. Cotyledon explants were cultured on MS basal medium supplemented with various concentrations of sucrose (2%, 4%, 6%, and 8%) and combinations of BAP (1.0mg/L, 2.5mg/L, 3.5mg/L, 4.5mg/L, 5.5mg/L, 6.5mg/L, and 7.5mg/L) and IAA (1.0mg/L). The multiple-shoot regeneration was achieved after 5-weeks of culture initiation. Significantly, the maximum mean percentage of shoot regeneration (75±4.3%) and number of shoots/explants (1.3±0.8) was recorded with explants that were treated with MS medium supplemented with sucrose (4%) including BAP (3.5mg/L) and IAA (1.0mg/L) while the high concentration (8%) of sucrose was proved to be completely ineffective to induce shoot regeneration on same nutrient medium (BAP+IAA). Furthermore, results indicate that sucrose (8%) could be observed slightly effective for shoot regeneration with higher concentrations of BAP (6.5mg/L and 7.5mg/L) in presence of IAA (1.0mg/L). Significantly, cotyledon explants that were growing with low concentration of sucrose (2%) were failed to show regeneration if BAP concentration exceeds to (5.5mg/L). Moreover, optimal frequency of shoot regeneration (64.4±1.4%) could be recorded with 2% of sucrose in presence of lower concentration of BAP (2.5mg/L) and IAA (1.0mg/L). The regenerated shoots were further transferred to half strength of MS medium supplemented with various concentrations of IBA (0.5, 1.0, and 1.5mg/L) for induction of roots and complete plantlets formation. Significantly, optimal mean percentage of shoots showing root formation (90±2.8%) was obtained in shoots regenerated with 4% of sucrose in presence of IBA (1.0mg/L) and shoot-root length ratio was recorded to be the maximum (4.2±0.11cm/2.81±0.12cm). However, in contrast, shoots that were regenerated on medium containing high sucrose (8%) with high BAP (7.5mg/L) + IAA (1.0mg/L) were found to exhibit lack of root formation irrespective of IBA concentrations. Present study reports that sucrose and BAP contents of the medium influences the sesame cotyledon explants regeneration. In comparison to control (2%) of sucrose, increase in sucrose concentration (4%) with BAP (3.5mg/L) and IAA (1.0mg/L) proved to be the optimal combination for shoot regeneration and moreover, regenerated shoots could induce the maximum root length in MS1/2 medium supplemented with IBA (1.0mg/L). In contrast, the IBA medium containing very high concentration of sucrose (8%) was turned out to be completely inhibitory for root regeneration. Study indicates that concentration of sucrose and BAP cytokinin in nutrient medium influences considerably the regeneration potential of explants tissues in black sesame oil crop. Moreover, the regenerated plantlets were further gradually acclimatized and transferred to plastic cup soil under the greenhouse conditions.

Background: Panax ginseng, one of the valuable perennial medicinal plants, stores numerous pharmacological substrates in its storage roots. Given its perennial growth habit, organ regeneration occurs each year, and cambium stem cell activity is necessary for secondary growth and storage root formation. Cytokinin (CK) is a phytohormone involved in the maintenance of meristematic cells for the development of storage organs; however, its physiological role in storage-root secondary growth remains unknown. Methods: Exogenous CK was repeatedly applied to P. ginseng, and morphological and histological changes were observed. RNA-seq analysis was used to elucidate the transcriptional network of CK that regulates P. ginseng growth and development. The HISTIDINE KINASE 3 (PgHK3) and RESPONSE REGULATOR 2 (PgRR2) genes were cloned in P. ginseng and functionally analyzed in Arabidopsis as a two-component system involved in CK signaling. Results: Phenotypic and histological analyses showed that CK increased cambium activity and dormant axillary bud formation in P. ginseng, thus promoting storage-root secondary growth and bud formation. The evolutionarily conserved two-component signaling pathways in P. ginseng were sufficient to restore CK signaling in the Arabidopsis ahk2/3 double mutant and rescue its growth defects. Finally, RNA-seq analysis of CK-treated P. ginseng roots revealed that plant-type cell wall biogenesis-related genes are tightly connected with mitotic cell division, cytokinesis, and auxin signaling to regulate CK-mediated P. ginseng development. Conclusion: Overall, we identified the CK signaling-related two-component systems and their physiological role in P. ginseng. This scientific information has the potential to significantly improve the field-cultivation and biotechnology-based breeding of ginseng.

The plant root absorbs water and nutrients, anchors the plant in the soil, and promotes plant development. Root is developed from root apical meristem (RAM), which is formed during embryo stage and is maintained by dividing stem cells. Plant hormones have a predominant role in RAM maintenance. This review evaluates the functional crosstalk among three major hormones (auxin, cytokinin, and brassinolide) in RAM development in Arabidopsis, integrating a variety of experimental data into a regulatory network and revealing multiple layers of complexity in the crosstalk among these three hormones. We also discuss possible directions for future research on the roles of hormones in regulating RAM development and maintenance.

Nitric oxide (NO) and cytokinins (CKs) are known for their crucial contributions to plant development, growth, senescence, and stress response. Despite the importance of both signals in stress responses, their interaction remains largely unexplored. The interplay between NO and CKs emerges as particularly significant not only regarding plant growth and development but also in addressing plant stress response, particularly in the context of extreme weather events leading to yield loss. In this review, we summarize NO and CKs metabolism and signaling. Additionally, we emphasize the crosstalk between NO and CKs, underscoring its potential impact on stress response, with a focus on hypoxia tolerance. Finally, we address the most urgent questions that demand answers and offer recommendations for future research endeavors.

Many previous studies have suggested that various plant hormones play essential roles in the grafting process. In this study, to understand the plant hormones that accumulate in the graft junctions, whether these are supplied from the scion or rootstock, and how these hormones play a role in the grafting process, we performed a hormonome analysis that accumulated in the incision site of the upper plants from the incision as “ungrafted scion” and lower plants from the incision as “ungrafted rootstock” in Nicotiana benthamiana. The results revealed that indole-3-acetic acid (IAA) and gibberellic acid (GA), which regulate cell division; abscisic acid (ABA) and jasmonic acid (JA), which regulate xylem formation; cytokinin (CK), which regulates callus formation, show different accumulation patterns in the incision sites of the ungrafted scion and rootstock. In addition, to try discussing the differences in the degree and speed of each event during the grafting process between intra- and inter-family grafting by determining the concentration and accumulation timing of plant hormones in the graft junctions, we performed hormonome analysis of graft junctions of intra-family grafted plants with N. benthamiana as scion and Solanum lycopersicum as rootstock (Nb/Sl) and inter-family grafted plants with N. benthamiana as scion and Arabidopsis thaliana as rootstock (Nb/At), using the ability of Nicotiana species to graft with many plant species. The results revealed that ABA and CK showed different accumulation timings; IAA, JA, and salicylic acid (SA) showed similar accumulation timings, while different accumulated concentrations in the graft junctions of Nb/Sl and Nb/At. This information is important for understanding the molecular mechanisms of plant hormones in the grafting process and the differences in molecular mechanisms between intra- and inter-family grafting.

Root hair, single-celled tubular structures originating from the epidermis, plays a vital role in the uptake of nutrients from the soil by increasing the root surface area. Therefore, optimizing root hair growth is crucial for plants to survive in fluctuating environments. Root hair length is determined by the action of various plant hormones, among which the roles of auxin and ethylene have been extensively studied. However, evidence for the involvement of cytokinins has remained elusive. We recently reported that the cytokinin-activated B-type response regulators, ARABIDOPSIS RESPONSE REGULATOR 1 (ARR1) and ARR12 directly upregulate the expression of ROOT HAIR DEFECTIVE 6-LIKE 4 (RSL4), which encodes a key transcription factor that controls root hair elongation. However, depending on the nutrient availability, it is unknown whether the ARR1/12–RSL4 pathway controls root hair elongation. This study shows that phosphate deficiency induced the expression of RSL4 and increased the root hair length through ARR1/12, though the transcript and protein levels of ARR1/12 did not change. These results indicate that cytokinins, together with other hormones, regulate root hair growth under phosphate starvation conditions.

In many plant species, petal abscission can be considered the final step of petal senescence. Cytokinins (CKs) are powerful suppressors of petal senescence; however, their role in petal abscission is ambiguous. Here, we observed that, in rose (Rosa hybrida), biologically active CK is accumulated during petal abscission and acts as an accelerator of the abscission process. Using a combination of reverse genetics, and molecular and biochemical techniques, we explored the roles of a LESION SIMULATING DISEASE1 (LSD1) family member RhLOL1 interacting with a bHLH transcription factor RhILR3 in CK-induced petal abscission. Silencing RhLOL1 delays rose petal abscission, while the overexpression of its ortholog SlLOL1 in tomato (Solanum lycopersicum) promotes pedicel abscission, indicating the conserved function of LOL1 in activating plant floral organ abscission. In addition, we identify a bHLH transcription factor, RhILR3, that interacts with RhLOL1. We show that RhILR3 binds to the promoters of the auxin signaling repressor auxin/indole-3-acetic acid (Aux/IAA) genes to inhibit their expression; however, the interaction of RhLOL1 with RhILR3 activates the expression of the Aux/IAA genes including RhIAA4-1. Silencing RhIAA4-1 delays rose petal abscission. Our results thus reveal a RhLOL1-RhILR3 regulatory module involved in CK-induced petal abscission via the regulation of the expression of the Aux/IAA genes.

Defoliation is an inevitable abiotic stress for forage and turf grasses because harvesting, grazing, and mowing are general processes for their production and management. Vegetative regrowth occurs upon defoliation, a crucial trait determining the productivity and persistence of these grasses. However, the information about the molecular regulation of this trait is limited because it is still challenging to perform molecular analyses in forage and turf grasses. Here, we used rice as a model to investigate vegetative regrowth upon defoliation at physiological and molecular levels. This study analyzed stubble and regrown leaves following periodic defoliation using two rice varieties with contrasting regrowth vigor. Vigorous regrowth was associated with maintained chlorophyll content and photosystem II performance; a restricted and promoted mRNA accumulation of sucrose synthase (SUS) I and III subfamilies, respectively; and reduced enzymatic activity of SUS. These results suggest that critical factors affecting vegetative regrowth upon defoliation are de novo carbohydrate synthesis by newly emerged leaves and proper carbohydrate management in leaves and stubble. Physiological and genetic analyses have demonstrated that the reduced sensitivity to and inhibited biosynthesis of cytokinin enhance regrowth vigor. Proper regulation of these metabolic and hormonal pathways identified in this study can lead to the development of new grass varieties with enhanced regrowth vigor following defoliation. [ABSTRACT FROM AUTHOR]

A key question in plant biology is how oriented cell divisions are integrated with patterning mechanisms to generate organs with adequate cell type allocation. In the root vasculature, a gradient of miRNA165/6 controls the abundance of HD-ZIP III transcription factors, which in turn control cell fate and spatially restrict vascular cell proliferation to specific cells. Here, we show that vascular development requires the presence of ARGONAUTE10, which is thought to sequester miRNA165/6 and protect HD-ZIP III transcripts from degradation. Our results suggest that the miR165/6-AGO10-HDZIP III module acts by buffering cytokinin responses and restricting xylem differentiation. Mutants of AGO10 show faster growth rates and strongly enhanced survival under severe drought conditions. However, this superior performance is offset by markedly increased variation and phenotypic plasticity in sub-optimal carbon supply conditions. Thus, AGO10 is required for the control of formative cell division and coordination of robust cell fate specification of the vasculature, while altering its expression provides a means to adjust phenotypic plasticity. Synopsis: In the Arabidopsis root vascular tissue, a gradient of miRNA165/6 controls the abundance of HD-ZIP III transcription factors, which in turn control cell fate and spatially restrict vascular cell proliferation to specific cells. This work shows that vascular development requires ARGONAUTE10 to sequester miRNA165/6, thus protecting HD-ZIP III transcripts from degradation. AGO10 controls formative cell divisions in the centre of the root stele, with its loss leading to ectopic xylem strands, increased procambial cell number, and enhanced root growth. Consistent with a role in protecting HD-ZIP III transcripts from miRNA165/6-mediated degradation, AGO10 is mostly expressed in the centre of the meristematic stele. AGO10 promotes HD-ZIP III activity to suppress cytokinin responses in the stele. AGO10 mutants outperform the wild type under water-limiting conditions. However, AGO10 is required for phenotypic robustness upon reduced carbon availability. AGO10 sequesters miRNA165/6 to suppress cytokinin-dependent responses in the root apical meristem. [ABSTRACT FROM AUTHOR]