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Over the last decade, remarkable progress has been made in our understanding the phytohormones, cytokinin's (CKs) biosynthesis, perception, and signalling pathways. Additionally, it became apparent that interfering with any of these steps has a significant effect on all stages of plant growth and development. As a result of their complex regulatory and cross-talk interactions with other hormones and signalling networks, they influence and control a wide range of biological activities, from cellular to organismal levels. In agriculture, CKs are extensively used for yield improvement and management because of their wide-ranging effects on plant growth, development and physiology. One of the primary targets in this regard is cytokinin oxidase/dehydrogenase (CKO/CKX), which is encoded by CKX gene, which catalyses the irreversible degradation of cytokinin. The previous studies on various agronomically important crops indicated that plant breeders have targeted CKX directly. In recent years, prokaryotic clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system has been increasingly used in editing the CKO/CKX gene and phenomenal results have been achieved. This review provides an updated information on the applications of CRISPR-based gene-editing tools in manipulating cytokinin metabolism at the genetic level for yield improvement. Furthermore, we summarized the current developments of RNPmediated DNA/transgene-free genomic editing of plants which would broaden the application of this technology. The current review will advance our understanding of cytokinins and their role in sustainably increase crop production through CRISPR/Cas genome editing tool. [ABSTRACT FROM AUTHOR]

Summary: Sugar transporter proteins (STPs) play critical roles in regulating plant stress tolerance, growth, and development. However, the role of STPs in regulating crop yield is poorly understood.This study elucidates the mechanism by which knockout of the sugar transporter OsSTP15 enhances grain yield via increasing the tiller number in rice.We found that OsSTP15 is specifically expressed in the shoot base and vascular bundle sheath of seedlings and encodes a plasma membrane‐localized high‐affinity glucose efflux transporter. OsSTP15 knockout enhanced sucrose and trehalose‐6‐phosphate (Tre6P) synthesis in leaves and improved sucrose transport to the shoot base by inducing the expression of sucrose transporters. Higher glucose, sucrose, and Tre6P contents were observed at the shoot base of stp15 plants. Transcriptome and metabolome analyses of the shoot base demonstrated that OsSTP15 knockout upregulated the expression of cytokinin (CK) synthesis‐ and signaling pathway‐related genes and increased CK levels.These findings suggest that OsSTP15 knockout represses glucose export from the cytoplasm and simultaneously enhances sugar transport from source leaves to the shoot base by promoting the synthesis of sucrose and Tre6P in leaves. Subsequent accumulation of glucose, sucrose, and Tre6P in the shoot base promotes tillering by stimulating the CK signaling pathway. [ABSTRACT FROM AUTHOR]

Cucumber (Cucumis sativus L.) lateral branch elongation is influenced by a variety of environmental signals, including light [e.g., far-red (FR) light] and hormones. In this experiment, the effect of FR light on the lateral branch elongation of cucumber ('Zhongnong No. 26') seedlings was investigated. The results showed that FR light significantly inhibited the lateral branch elongation of cucumber seedlings. In addition, FR light significantly increased the auxin (indole-3-acetic acid, IAA) content, decreased the cytokinin (CTK; Zeatin) content, and suppressed the expression of most CTK synthetic-related genes, such as IPTs, in cucumber seedlings. The lateral branch elongation of cucumber seedlings was assessed in response to decapitation and exogenous 6-BA treatment to further investigate the relationship between IAA and CTK on the lateral branch elongation of cucumber seedlings under FR light. Both decapitation and exogenous 6-BA treatment eliminated the inhibitory effect of FR light on the lateral branch elongation of cucumber seedlings. In conclusion, these results indicated that IAA and CTK were involved in the regulatory effects of FR light on cucumber seedling lateral branch elongation. [ABSTRACT FROM AUTHOR]

Plants constantly perceive and process environmental signals and balance between the energetic demands of growth and defense. Growth arrest upon pathogen attack was previously suggested to result from a redirection of the plants' metabolic resources towards the activation of plant defense. The energy sensor Target of Rapamycin (TOR) kinase is a conserved master coordinator of growth and development in all eukaryotes. Although TOR is positioned at the interface between development and defense, little is known about the mechanisms by which TOR may potentially regulate the relationship between these two modalities. The plant hormones cytokinin (CK) and gibberellin (GA) execute various aspects of plant development and defense. The ratio between CK and GA was reported to determine the outcome of developmental programmes. Here, investigating the interplay between TOR‐mediated development and TOR‐mediated defense in tomato, we found that TOR silencing resulted in rescue of several different aberrant developmental phenotypes, demonstrating that TOR is required for the execution of developmental cues. In parallel, TOR inhibition enhanced immunity in genotypes with a low CK/GA ratio but not in genotypes with a high CK/GA ratio. TOR‐inhibition mediated disease resistance was found to depend on developmental status, and was abolished in strongly morphogenetic leaves, while being strongest in mature, differentiated leaves. CK repressed TOR activity, suggesting that CK‐mediated immunity may rely on TOR downregulation. At the same time, TOR activity was promoted by GA, and TOR silencing reduced GA sensitivity, indicating that GA signalling requires normal TOR activity. Our results demonstrate that TOR likely acts in concert with CK and GA signalling, executing signalling cues in both defense and development. Thus, differential regulation of TOR or TOR‐mediated processes could regulate the required outcome of development‐defense prioritisation. Summary Statement: Target of Rapamycin (TOR) is positioned at the interface between development and defense, but little is known about the mechanisms by which TOR may regulate shifts between them. Here, we demonstrate that TOR likely acts in concert with cytokinin and gibberellin to execute signalling cues in both defense and development. [ABSTRACT FROM AUTHOR]

Phytohormones play a key role in regulating tiller number. Ascorbic acid (Asc)-phytohormone interaction plays a pivotal role in the regulation of senescence. We analysed the relationship between Asc and the enzyme concentrations and gene transcript abundances related to the signal perception of strigolactones (SLs), the contents of four phytohormones (abscisic acid, ABA; jasmonic acid, JA; indole acetic acid, IAA; cytokinin, CTK), the enzyme concentrations and gene transcript abundances related to the synthesis or transportation of these four phytohormones. Our results showed that Asc deficiency leads to the upregulation of enzyme concentrations, gene transcript abundances related to the SL signal perception, ABA synthesis and IAA transport. The altered level of Asc also leads to a change in the contents of ABA, JA, IAA and CTK. These findings support the conclusion that Asc or Asc/DHA play an important role in the signal perception and transduction of SLs, and Asc may affect the coordinated regulation of SL, IAA and CTK on rice (Oryza sativa) tillering. [ABSTRACT FROM AUTHOR]

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Cassava breeding faces obstacles due to late flowering and poor flower and seed set. The acceleration of breeding processes and the reduction in each cycle's duration hinge upon efficiently conducting crosses to yield ample progeny for subsequent cycles. Our primary objective was to identify methods that provide tools for cassava breeding programs, enabling them to consistently and rapidly generate offspring from a wide array of genotypes. In greenhouse trials, we examined the effects of the anti-ethylene silver thiosulfate (STS) and the cytokinin benzyladenine (BA). STS, administered via petiole infusion, and BA, applied as an apical spray, combined with the pruning of young branches, significantly augmented the number of flowers. Controls produced no flowers, whereas treatments with pruning plus either BA or STS alone produced an average maximum of 86 flowers per plant, and the combination of pruning, BA and STS yielded 168 flowers per plant. While STS had its primary effect on flower numbers, BA increased the fraction of female flowers from less than 20% to ≥87%, thus increasing the number of progeny from desired parents. Through field studies, we devised an optimal protocol that maintained acceptable levels of phytodamage ratings while substantially increasing seed production per plant compared to untreated plants. This protocol involves adjusting the dosage and timing of treatments to accommodate genotypic variations. As a result, cassava breeding programs can effectively leverage a diverse range of germplasm to develop cultivars with the desired traits. [ABSTRACT FROM AUTHOR]

Cytokinins (CKs) are widespread in a variety of organisms from bacteria to humans, and are particularly abundant in insects and hexapods. However, how organisms other than bacteria and plants obtain CKs has not been thoroughly studied. We examined the transcriptomes of 670 species of Hexapoda (predominantly Insecta) to determine if transcripts that encode proteins homologous to any of the known enzymes involved in CK biosynthesis and metabolism are widespread in these groups (occur in > 80% of species). We found that transcripts encoding proteins homologous to the enzymes tRNA-dimethylallyltransferase (EC: 2.5.1.75) and tRNA-2-methylthio-N6-dimethylallyladenosine synthase (EC: 2.8.4.3) are widespread in insects and hexapods. These enzymes could allow insects and hexapods to synthesize iP-based CKs and methylthiolated iP-based CKs via a tRNA-degradation pathway whereby tRNA is first prenylated and possibly methylthiolated prior to releasing CKs or methylthiolated CKs upon degradation. We also found widespread occurrence in insects and hexapods of transcripts encoding proteins that are homologous to five enzymes in the adenine salvage pathway: 5'- nucleotidase (EC: 3.1.3.5), adenosine kinase (EC:2.7.1.20), purine-nucleoside phosphorylase (EC: 2.4.2.1), purine nucleosidase (EC: 3.2.2.1), and adenine phosphoribosyltransferase (EC: 2.4.2.7). These enzymes could allow insects and hexapods to convert CK nucleotides to nucleosides and free base CKs. We found few transcripts encoding proteins homologous to enzymes that would convert CKs to storage forms such as their O-glucosides and no transcripts encoding proteins homologous to enzymes that would degrade CKs such as CK oxidases. We suggest that insects and hexapods have the enzymatic pathways necessary to synthesize and metabolize CKs, in contrast to the presumption that CKs are merely obtained via consumption and sequestration from plants or via microbial symbiosis. [ABSTRACT FROM AUTHOR]

The phytohormone cytokinin has various roles in plant development, including meristem maintenance, vascular differentiation, leaf senescence, and regeneration. Prior investigations have revealed that cytokinin acts via a phosphorelay similar to the two-component system by which bacteria sense and respond to external stimuli. The eventual targets of this phosphorelay are type-B ARABIDOPSIS RESPONSE REGULATORS (B-ARRs), containing the conserved N-terminal receiver domain (RD), middle DNA binding domain (DBD), and C-terminal transactivation domain. While it has been established for two decades that the phosphoryl transfer from a specific histidyl residue in ARABIDOPSIS HIS PHOSPHOTRANSFER PROTEINS (AHPs) to an aspartyl residue in the RD of B-ARRs results in a rapid transcriptional response to cytokinin, the underlying molecular basis remains unclear. In this work, we determine the crystal structures of the RD-DBD of ARR1 (ARR1RD-DBD) as well as the ARR1DBD-DNA complex from Arabidopsis. Analyses of the ARR1DBD-DNA complex have revealed the structural basis for sequence-specific recognition of the GAT trinucleotide by ARR1. In particular, comparing the ARR1RD-DBD and ARR1DBD-DNA structures reveals that unphosphorylated ARR1RD-DBD exists in a closed conformation with extensive contacts between the RD and DBD. In vitro and vivo functional assays have further suggested that phosphorylation of the RD weakens its interaction with DBD, subsequently permits the DNA binding capacity of DBD, and promotes the transcriptional activity of ARR1. Our findings thus provide mechanistic insights into phosphorelay activation of gene transcription in response to cytokinin. [ABSTRACT FROM AUTHOR]

Root system architecture (RSA) influences the acquisition of heterogeneously dispersed soil nutrients. Cytokinin and C-TERMINALLY ENCODED PEPTIDE (CEP) hormones affect RSA, in part by controlling the angle of lateral root (LR) growth. Both hormone pathways converge on CEP DOWNSTREAM 1 (CEPD1) and CEPD2 to control primary root growth; however, a role for CEPDs in controlling the growth angle of LRs is unknown. Using phenotyping combined with genetic and grafting approaches, we show that CEP hormone-mediated shallower LR growth requires cytokinin biosynthesis and perception in roots via ARABIDOPSIS HISTIDINE KINASE 2 (AHK2) and AHK3. Consistently, cytokinin biosynthesis and ahk2,3 mutants phenocopied the steeper root phenotype of cep receptor 1 (cepr1) mutants on agar plates, and CEPR1 was required for trans -Zeatin (t Z)-type cytokinin-mediated shallower LR growth. In addition, the cepd1,2 mutant was less sensitive to CEP and t Z, and showed basally steeper LRs on agar plates. Cytokinin and CEP pathway mutants were grown in rhizoboxes to define the role of these pathways in controlling RSA. Only cytokinin receptor mutants and cepd1,2 partially phenocopied the steeper-rooted phenotype of cepr1 mutants. These results show that CEP and cytokinin signaling intersect to promote shallower LR growth, but additional components contribute to the cepr1 phenotype in soil. [ABSTRACT FROM AUTHOR]

Vanilla is a popular flavouring essence derived from the pods of vanilla orchid plants. Due to the high demand for vanilla flavour, high yielding vanilla plantlets are necessary for establishing vanilla plantations. Clonal micropropagation is a viable technique for the mass production of high yielding vanilla plantlets. This study reports an efficient regeneration protocol by using cytokinin as the sole plant growth regulator to regenerate plantlets from the root tips of a commercial vanilla orchid species, Vanilla planifolia. Most studies to date have reported using seeds and nodes as starting explants for in vitro micropropagation of vanilla orchids. So far, regeneration from roots has not been very successful. Previous studies favoured the use of auxins only or high auxin to cytokinin ratios to induce callus, and sole cytokinins were used for direct shoot regeneration. However, it was sporadically observed in plantlets regeneration of V. planifolia that multiple shoots were regenerated from the tips of intact aerial roots submerged in media. This study therefore investigated the regeneration of excised vanilla root tips through the application of most commonly used auxins (1-naphthaleneacetic acid and 2,4-dichlorophenoxyacetic acid) and cytokinins (6-benzylaminopurine and thidiazuron). High auxin presence is known to promote callusing in in vitro plants. However, in this study, auxin treatment inhibits callusing in root tips. While cytokinin treatments, even at low levels, has promoted high rate of callusing. These callus cells regenerate into protocorm-like-body (PLB) shoots when cytokinin levels are increased to 0.5 mg/mL 6-benzylaminopurine (BAP) under light conditions. The findings of the study have the potential of providing large quantity of high yielding vanilla plantlets through clonal micropropagation. [ABSTRACT FROM AUTHOR]

Cytokinins (CKs) are a class of growth‐promoting signaling molecules that affect multiple cellular and developmental processes. These phytohormones are well studied in plants, but their presence continues to be uncovered in organisms spanning all kingdoms, which poses new questions about their roles and functions outside of plant systems. Cytokinin production can be initiated by one of two different biosynthetic enzymes, adenylate isopentenyltransfases (IPTs) or tRNA isopentenyltransferases (tRNA‐IPTs). In this study, the social amoeba, Dictyostelium discoideum, was used to study the role of CKs by generating deletion and overexpression strains of its single adenylate‐IPT gene, iptA. The life cycle of D. discoideum is unique and possesses both single‐ and multicellular stages. Vegetative amoebae grow and divide while food resources are plentiful, and multicellular development is initiated upon starvation, which includes distinct life cycle stages. CKs are produced in D. discoideum throughout its life cycle and their functions have been well studied during the later stages of multicellular development of D. discoideum. To investigate potential expanded roles of CKs, this study focused on vegetative growth and early developmental stages. We found that iptA‐deficiency results in cytokinesis defects, and both iptA‐deficiency and overexpression results in dysregulated tricarboxylic acid (TCA) cycle and amino acid metabolism, as well as increased levels of adenosine monophosphate (AMP). Collectively, these findings extend our understanding of CK function in amoebae, indicating that iptA loss and overexpression alter biological processes during vegetative growth that are distinct from those reported during later development. [ABSTRACT FROM AUTHOR]

Summary: Grain size is one of the important traits in wheat breeding programs aimed at improving yield, and cytokinins, mainly involved in cell division, have a positive impact on grain size. Here, we identified a novel wheat gene TaMADS‐GS encoding type I MADS‐box transcription factor, which regulates the cytokinins signalling pathway during early stages of grain development to modulate grain size and weight in wheat. TaMADS‐GS is exclusively expressed in grains at early stage of seed development and its knockout leads to delayed endosperm cellularization, smaller grain size and lower grain weight. TaMADS‐GS protein interacts with the Polycomb Repressive Complex 2 (PRC2) and leads to repression of genes encoding cytokinin oxidase/dehydrogenases (CKXs) stimulating cytokinins inactivation by mediating accumulation of the histone H3 trimethylation at lysine 27 (H3K27me3). Through the screening of a large wheat germplasm collection, an elite allele of the TaMADS‐GS exhibits higher ability to repress expression of genes inactivating cytokinins and a positive correlation with grain size and weight, thus representing a novel marker for breeding programs in wheat. Overall, these findings support the relevance of TaMADS‐GS as a key regulator of wheat grain size and weight. [ABSTRACT FROM AUTHOR]

This article explores the role of brassinosteroids (BRs) in regulating bud outgrowth and crop architecture. Short stature crops developed during the green revolution disrupted the gibberellin (GA) pathway, leading to reduced height but increased tillering. BRs have emerged as an alternative dwarfing pathway that can decouple tillering from height. The article discusses how BRs integrate with the branching signaling network, particularly through the transcription factor TB1, and highlights the potential for genetic manipulation of BR biosynthesis or signaling components to modify shoot architecture. Further research is needed to understand the effects of BRs on other branching hormones and to uncover new targets for modifying crop architecture. [Extracted from the article]

SUMMARY: Root hairs are crucial in the uptake of essential nutrients and water in plants. This study showed that a zinc finger protein, GIS3 is involved in root hair growth in Arabidopsis. The loss‐of‐function gis3 and GIS3 RNAi transgenic line exhibited a significant reduction in root hairs compared to the wild type. The application of 1‐aminocyclopropane‐1‐carboxylic acid (ACC), an exogenous ethylene precursor, and 6‐benzyl amino purine (BA), a synthetic cytokinin, significantly restored the percentage of hair cells in the epidermis in gis3 and induced GIS3 expression in the wild type. More importantly, molecular and genetic studies revealed that GIS3 acts upstream of ROOT HAIR DEFECTIVE 2 (RHD2) and RHD4 by binding to their promoters. Furthermore, exogenous ACC and BA application significantly induced the expression of RHD2 and RHD4, while root hair phenotype of rhd2‐1, rhd4‐1, and rhd4‐3 was insensitive to ACC and BA treatment. We can therefore conclude that GIS3 modulates root hair development by directly regulating RHD2 and RHD4 expression through ethylene and cytokinin signals in Arabidopsis. Significance Statement: In this paper, we demonstrated that zinc finger protein GIS3 is the key regulator to control root hair development in Arabidopsis. Further experiments demonstrated that GIS3, RHD2, and RHD4 are required for root hair development in response to ethylene and cytokinin signaling. More importantly, the molecular and genetic experiments reveal that GIS3 directly binds to the promoter of RHD2 and RHD4 to regulate root hair elongation in Arabidopsis. These results will provide a novel function of zinc finger protein interacting with hormone signal to control root hair development and a better understanding of the network of zinc finger protein interact with root hair‐related genes to control root hair development in Arabidopsis. [ABSTRACT FROM AUTHOR]

This investigation was aimed to study the effect of soil application of Bio health and foliar application with Tecamin max and Benzyl adenine on growth characteristic of sweet orange (Citrus sinensis L.) seedlings during two successive seasons (2020 and 2021). Bio health was added with three concentrations (0, 6 and 12 g.L-1 ) to the soil and foliar application of Tecamin max with three concentration (0, 5 and 10 ml.L-1 ), Benzyl adenine with three concentration (0, 100 and 200 mg. L-1 ) on the sweet orange seedlings which were brought from private nursery in Duhok city and have two- years old and nearly uniform in growth vigor. The results proved that all parameters such as increases in plant high, stem diameter, branch numbers, single leaf area, total chlorophyll as soon as leaves carbohydrate and leaves dry weight in both seasons were increased significantly as compared with control, accept Tecamin max had no significant effect on leaves carbohydrates and leaves dry weight, Benzyl adenine no significant effect on leaves dry weight on both seasons, Furthermore, combination among high concentration of Bio health, Tecamin max and Benzyl adenine improved all parameters in comparison with the control. [ABSTRACT FROM AUTHOR]

In Eastern Europe, the traditional marketing of cyclamen is in the period from the middle of February to the middle of March. The poor light of winter months and the higher number of plants (because of the reduction in heating costs) often result in elongated plants. To avoid this, it is recommended to use plant growth retardants. At the same time, another problem is that flowers do not rise from the level of rosette due to the unfavorable cultivation conditions. This can be solved with growth stimulants. In the experiment, we tested the effect of growth regulators on the growth of a frequently used variety of cyclamen. We used daminozide and paclobutrazol plant growth retardants for height control and gibberellic acid (GA) and benzyladenine (BAP) as growth promoters for increasing the number of flowers and the length of stems. The results show that daminozide and paclobutrazol are both effective for height control in cyclamen production. [ABSTRACT FROM AUTHOR]

Background: The number of grains per panicle is an important factor in determining rice yield. The DST-OsCKX2 module has been demonstrated to regulate panicle development in rice by controlling cytokinin content. However, to date, how the function of DST-OsCKX2 module is regulated during panicle development remains obscure. Result: In this study, the ABNORMAL PANICLE 1 (ABP1), a severely allele of FRIZZY PANICLE (FZP), exhibits abnormal spikelets morphology. We show that FZP can repress the expression of DST via directly binding to its promotor. Consistently, the expression level of OsCKX2 increased and the cytokinin content decreased in the fzp mutant, suggesting that the FZP acts upstream of the DST-OsCKX2 to maintain cytokinin homeostasis in the inflorescence meristem. Conclusions: Our results indicate that FZP plays an important role in regulating spikelet development and grain number through mediating cytokinin metabolism. [ABSTRACT FROM AUTHOR]

In potatoes, tuber secondary growth, especially sprouting, deforms the tubers and severely lowers their commercial value. Tuber sprouting is induced by signal substances, such as gibberellin (GA), which are transported to the tuber from the plant body. The molecular mechanism underlying GA-induced sprouting remains ambiguous. Here, we tried to recreate tuber secondary growth using in vitro stemmed microtubers (MTs) (with the nodal stem attached) and MT halves (with the nodal stem entirely removed). Our experiments showed that GA alone could initiate the sprouting of stemmed microtubers; however, GA failed to initiate MT halves unless 6-benzyladenine, a synthetic cytokinin CK, was co-applied. Here, we analyzed the transcriptional profiles of sprouting buds using these in vitro MTs. RNA-seq analysis revealed a downregulation of cytokinin-activated signaling but an upregulation of the "Zeatin biosynthesis" pathway, as shown by increased expression of CYP735A, CISZOG, and UGT85A1 in sprouting buds; additionally, the upregulation of genes, such as IAA15, IAA22, and SAUR50, associated with auxin-activated signaling and one abscisic acid (ABA) negative regulator, PLY4, plays a vital role during sprouting growth. Our findings indicate that the role of the nodal stem is synonymous with CK in sprouting growth, suggesting that CK signaling and homeostasis are critical to supporting GA-induced sprouting. To effectively control tuber sprouting, more effort is required to be devoted to these critical genes. [ABSTRACT FROM AUTHOR]

SUMMARY: Auxins and cytokinins are two major families of phytohormones that control most aspects of plant growth, development and plasticity. Their distribution in plants has been described, but the importance of cell‐ and subcellular‐type specific phytohormone homeostasis remains undefined. Herein, we revealed auxin and cytokinin distribution maps showing their different organelle‐specific allocations within the Arabidopsis plant cell. To do so, we have developed Fluorescence‐Activated multi‐Organelle Sorting (FAmOS), an innovative subcellular fractionation technique based on flow cytometric principles. FAmOS allows the simultaneous sorting of four differently labelled organelles based on their individual light scatter and fluorescence parameters while ensuring hormone metabolic stability. Our data showed different subcellular distribution of auxin and cytokinins, revealing the formation of phytohormone gradients that have been suggested by the subcellular localization of auxin and cytokinin transporters, receptors and metabolic enzymes. Both hormones showed enrichment in vacuoles, while cytokinins were also accumulated in the endoplasmic reticulum. [ABSTRACT FROM AUTHOR]

Somatic embryogenesis (SE) has many applications in grapevine biotechnology including micropropagation, eradicating viral infections from infected cultivars, mass production of hypocotyl explants for micrografting, as a continuous source for haploid and doubled haploid plants, and for germplasm conservation. It is so far the only pathway for the genetic modification of grapevines through transformation. The single-cell origin of somatic embryos makes them an ideal explant for mutation breeding as the resulting mutants will be chimera-free. In the present research, two combinations of plant growth regulators and different explants from flower buds at two stages of maturity were tested in regard to the efficiency of callusing and embryo formation from the callus produced in three white grape cultivars. Also, the treatment of somatic embryos with the chemical mutagen ethyl methanesulfonate (EMS) was optimised. Medium 2339 supplemented with β-naphthoxyacetic acid (5 μM) and 6-benzylaminopurine (BAP—9.0 μM) produced significantly more calluses than medium 2337 supplemented with 2,4-dichlorophenoxyacetic acid (4.5 µM) and BAP (8.9 µM) in all explants. The calluses produced on medium 2337 were harder and more granular and produced more SEs. Although the stage of the maturity of floral bud did not have a significant effect on the callusing of the explants, calluses produced from immature floral bud explants in the premeiotic stage produced significantly more SEs than those from more mature floral buds. Overall, immature ovaries and cut floral buds exposing the cut ends of filaments, style, etc., tested for the first time in grapevine SE, produced the highest percentage of embryogenic calluses. It is much more efficient to cut the floral bud and culture than previously reported explants such as anthers, ovaries, stigmas and styles during the short flowering period when the immature flower buds are available. When the somatic embryos of the three cultivars were incubated for one hour with 0.1% EMS, their germination was reduced by 50%; an ideal treatment considered to obtain a high frequency of mutations for screening. Our research findings will facilitate more efficient SE induction in grapevines and inducing mutations for improving individual traits without altering the genetic background of the cultivar. [ABSTRACT FROM AUTHOR]

Tillering is an important part in strawberry growth, and strawberries can reproduce nutritionally through stolons to generate genetically stable offspring. However, excessive tillering during the fruit-growing stage can negatively impact fruit yield and quality. In this study, different concentrations of exogenous rac-GR24 (GR24) are used to treat the strawberry plants. It was found that GR24 effectively inhibited the sprouting of strawberry stolons, while promoting the growth of the stems and leaves. Among the treatments, the most effective concentration was found to be 5 μmol/L GR24. This treatment resulted in a decrease in the glucose content in the strawberry crowns and also caused changes in the contents of two endogenous phytohormones, gibberellic acid (GA3) and trans-zeatin riboside (tZR). Transcriptome data further suggested that exogenous GR24 may inhibit strawberry plant tillering by affecting various phytohormone signaling pathways and the sugar metabolism pathway. In 5 μmol/L GR24-treated plants, the expression level of type-B response regulator (B-ARR) was down-regulated and the expression level of CYTOKININ RESPONSE 1 (CRE1), histidine-containing phosphotransfer protein (AHP), and type-A response regulator (A-ARR) were up-regulated, suggesting the inhibition of the cytokinin (CTK) signaling pathway. The down-regulation of auxin (AUX) and auxin response factor (ARF), as well as the up-regulation of auxin/indole-3-acetic acid (AUX/IAA), led to the inhibition of the indole-3-acetic acid (IAA) signaling pathway. Additionally, the up-regulation of pyrabactin resistance 1/ pyrabactin resistance 1-like (PYR/PYL), non-fermenting 1-related protein kinase 2 (SnRK2), and ABRE binding factors (ABF) and the down-regulation of protein phosphatase 2C (PP2C) were observed in the up-regulated abscisic acid (ABA) signaling pathways. In the sugar metabolism pathway, the up-regulation of invertase (INV), hexokinase (HK), and fructokinase (FRK) and the down-regulation of trehalase (TREH) and beta-amylase (BMY) led to a decreased glucose synthesis and an increased glucose consumption. Therefore, GR24 can effectively inhibit strawberry plant tillering through these pathways, making it an effective reagent for tillering inhibition. [ABSTRACT FROM AUTHOR]

A micropropagation protocol for growing Clematis 'Warszawska Nike' was developed. The MS medium supplemented with 1 mg∙dm−3 BAP showed good results in the case of microshoot initiation (80%). The addition of BAP to the medium at higher concentrations resulted in the formation of a large amount of callus tissue at the base of the explant. Of the explants growing on the medium with the lowest cytokinin concentration, 8% flowered. Very quickly, after just 14 days, the explants began to die: some of the leaves that developed in in vitro cultures began to turn yellow and wither. The propagation of shoots was performed in two steps. In the first step, cytokinin BAP and Kin in various concentrations (0.5–2 mg∙dm−3) were added to the MS medium. In the second step, MS medium with the combinations of BAP (0.5 and 1 mg∙dm−3) with IAA or GA3 (1 and 2 mg∙dm−3) was used. The MS medium with 0.5 mg∙dm−3 BAP and 2 mg∙dm−3 GA3 was the best medium for the multiplication stage of clematis. Plants growing on this medium had the largest number of leaves, shoots, and internodes, and were also heavier compared to plants propagated on other media. The proliferated clematis explants were rooted on MS medium with the addition of IAA or IBA in different concentrations (0.5 to 4 mg∙dm−3). Of the concentrations tested, 0.5 mg∙dm−3 IAA was the most effective one for in vitro root induction. The highest percentage of acclimatized plants (75%) was observed when the shoots were rooted on MS medium with 0.5 mg∙dm−3 IAA. [ABSTRACT FROM AUTHOR]

Poa pratensis is widely distributed in cold temperate regions and can be used as a species for stress restoration and as a forage for livestock. Studying the genetic characteristics of tillering occurrence in bluegrass provides a theoretical basis for studying plant yield formation, environmental adaptation, and improving survival competitiveness. The regulating effects of endogenous hormone IAA content and the expression of related genes ARF1, ARF12, ARF14, ZT content and the expression of related genes CKX2, CKX3, CKX4, SL content and the expression of related genes D14-like, D14.1-like and D14 in wild Kentucky bluegrass were investigated. Kentucky bluegrass from Sunan and Qingshui was used to evaluate the influence of hormone and gene expression on tillering behavior. Endogenous hormone contents and expression levels of related genes in stems and roots of both materials were measured at prophase, peak, and anaphase of tillering. The results showed that among the three materials, the Sunan material had a better tillering ability for Poa pratensis, while the Qingshui material had poorer tillering ability. The downregulation of CKX2, CKX3, and CKX4 gene expression levels promotes the synthesis of ZT, thereby improving the tillering ability of the germplasm. Upregulation of ARF14, D14, and D14.1-like gene expression levels enhances the synthesis of IAA and SL, thereby inhibiting tillering. More importantly, the interaction between hormones affects the tillering ability of bluegrass, and high levels of ZT/IAA, ZT/SL, and ZT/(IAA+SL) values promote tillering. In summary, this study reveals the mechanism by which hormones regulate the occurrence of tillering in Kentucky bluegrass, providing a theoretical basis for understanding the genetic characteristics of plant type, effectively regulating tillering, studying yield development, environmental adaptation, and improving survival rate. [ABSTRACT FROM AUTHOR]

Cytokinins (CTKs) are a diverse collection of evolutionarily conserved adenine‐derived signaling molecules classically studied as phytohormones; however, their roles and production have been less studied in mammalian systems. Skeletal muscles are sensitive to cellular cues such as inflammation and in response, alter their secretome to regulate the muscle stem cell and myofiber niche. Using cultured C2C12 muscle cells, we profiled CTK levels to understand (1) whether CTKs are part of the muscle secretome and (2) whether CTKs are responsive to cellular stress. To induce cellular stress, C2C12 myotubes were treated with lipopolysaccharides (LPS) for 24 h and then media and cell fractions were collected for ultra high‐performance liquid chromatography tandem mass spectrometry with electrospray ionization (UHPLC‐(ESI+)‐HRMS/MS) for metabolomics and CTK profiling. Across LPS‐treated and control cells, 11 CTKs were detected in the extracellular space while 6 were detected intracellularly. We found that muscle cells are enriched in isopentenyladenine (iP) species (from free base, riboside to nucleotide forms), and that extracellular levels are increased after LPS treatment. Our study establishes that muscle cells express various forms of CTKs, and that CTK levels are responsive to LPS‐induced cell stress, suggesting a role for CTKs in intra‐ and extracellular signaling of mammalian cells. [ABSTRACT FROM AUTHOR]

Parasitic plants invade their host through their invasive organ, the haustorium. This organ connects to the vasculature of the host roots and hijacks water and nutrients. Although parasitism has evolved independently in plants, haustoria formation follows a similar mechanism throughout different plant species, highlighting the developmental plasticity of plant tissues. Here, we compare three types of haustoria formed by the root and shoot in the plant parasites Striga and Cuscuta. We discuss mechanisms underlying the interactions with their hosts and how different approaches have contributed to major understanding of haustoria formation and host invasion. We also illustrate the role of auxin and cytokinin in controlling this process. [ABSTRACT FROM AUTHOR]

Vascular tissues serve a dual function in plants, both providing physical support and controlling the transport of nutrients, water, hormones, and other small signaling molecules. Xylem tissues transport water from root to shoot; phloem tissues transfer photosynthates from shoot to root; while divisions of the (pro)cambium increase the number of xylem and phloem cells. Although vascular development constitutes a continuous process from primary growth in the early embryo and meristem regions to secondary growth in the mature plant organs, it can be artificially separated into distinct processes including cell type specification, proliferation, patterning, and differentiation. In this review, we focus on how hormonal signals orchestrate the molecular regulation of vascular development in the Arabidopsis primary root meristem. Although auxin and cytokinin have taken center stage in this aspect since their discovery, other hormones including brassinosteroids, abscisic acid, and jasmonic acid also take leading roles during vascular development. All these hormonal cues synergistically or antagonistically participate in the development of vascular tissues, forming a complex hormonal control network. [ABSTRACT FROM AUTHOR]

Zelkova schneideriana is a fast-growing tree species endemic to China. Recent surveys and reports have highlighted a continued decline in its natural populations; therefore, it is included in the Red List of Threatened Species by The International Union for Conservation of Nature. A new variety "HenTianGao" (H) has been developed with smaller plant height, slow growth, and lower branching points. In this study, we attempted to understand the differences in plant height of Z. schneideriana (J) and its dwarf variety H. We determined the endogenous hormone content in the annual grafted branches of both J and H. J exhibited higher gibberellic acid (GA)-19 and trans-Zeatin (tZ) levels, whereas H had higher levels of indole-3-acetic acid (IAA) catabolite 2-oxindole-3-acetic acid (OxIAA), IAA-Glu conjugate, and jasmonic acid (JA) (and its conjugate JA-Ile). The transcriptome comparison showed differential regulation of 20,944 genes enriched in growth and development, signaling, and metabolism-related pathways. The results show that the differential phytohormone level (IAA, JA, tZ, and GA) was consistent with the expression of the genes associated with their biosynthesis. The differences in relative OxIAA, IAA-Glu, GA19, trans-Zeatin, JA, and JA-Ile levels were linked to changes in respective signaling-related genes. We also observed significant differences in the expression of cell size, number, proliferation, cell wall biosynthesis, and remodeling-related genes in J and H. The differences in relative endogenous hormone levels, expression of biosynthesis, and signaling genes provide a theoretical basis for understanding the plant height differences in Z. schneideriana. [ABSTRACT FROM AUTHOR]

Introduction: Cotton (Gossypium hirsutum L.) is the most widely used natural fiber and the most important dual-purpose industrial plant in the world. Delayed transplanting of cotton using early cultivars is one of the effective methods to increase the cultivated area and increase the production of this strategic crop. Plant growth regulators can be effective in the source-sink balance in different plants and are progressively used to increase yield in many plants. Cytokinins are one of the most important plant growth regulators, which increase cell division and differentiation, reduce the effect of terminal dominance, increase leaf surface development, increase the number of lateral branches, increase mobility of nutrients, prevent chlorophyll decomposition, and prevent aging. Growth regulators containing mepiquat chloride, such as pix, reduce vegetative growth in cotton by preventing the synthesis of gibberellic acid. Controlling the growth of cotton as a result of the use of pix, allocates the photosynthetic assimilates to the fruit organs and growing bolls, which increases the weight of the bolls. Materials and Methods: To investigate the effect of foliar application of 6-benzylaminopurine and pix on the growth and yield of cotton (cv. May 344) in delayed transplanting, a factorial experiment was conducted in a randomized complete block design with three replications in Sabzevar during 2020. The factors studied were foliar application of pix at two levels of no foliar application and foliar application at the rate of 1.5 liters per hectare and 6-benzylaminopurine (BAP) foliar application at five concentrations of 0, 5, 25, 50 and 100 mg l-1. May 344 cotton cultivar was used in this experiment. Results and Discussion: The results showed that foliar application of pix significantly increased the content of chlorophyll a compared to the no foliar application conditions. Foliar application with concentrations of 50 and 100 mg l-1 BAP also caused a significant increase in the content of chlorophyll a compared to the no foliar application conditions. The highest content of chlorophyll a was observed in the concentration of 50 mg l-1 BAP, which had an increase of 17.7% compared to the control treatment. The highest height of the plant (67.9 cm) was obtained by foliar application of 25 mg l-1 BAP alone, which showed an increase of 15.1% compared to the control. The lowest plant height (51.6 cm) was also observed in the treatment of foliar application of pix along with 100 mg l-1 BAP. Foliar application of pix along with 25 mg l-1 BAP produced the highest number of bolls per plant, which was 29.8% higher than the control. The highest average boll weight (5.4 g) and the highest percentage of open bolls (83.8%) were obtained in the foliar application treatment of pix with 100 mg l-1 BAP, which was not significantly different from the control treatment. Foliar application of 1.5 liters per hectare of pix alone produced the highest seed cotton yield (2754.3 kg ha-1), which had a 40.2% increase compared to the control. The highest biological yield was produced in the foliar application treatment of PIX along with 5 mg l-1 of BAP, which had an increase of 26.5% compared to the control treatment. The lowest biological yield was obtained in the treatment of foliar application with 100 mg l-1 of BAP, which was not significantly different from the control treatment. The highest percentage of lint (52.2%) was observed in the pix foliar application treatment along with 50 mg l-1 BAP, which showed an increase of 20.3% compared to the control. Conclusion: According to the results, foliar application of 1.5 liters per hectare of pix is recommended to produce high seed cotton yield and increase the percentage of lint in delayed transplanting cotton. [ABSTRACT FROM AUTHOR]

Developmental phytohormones regulate many physiological responses in plants, including growth and immunity in response to pathogens. Disease resistance mediated by developmental phytohormones such as cytokinin (CK), gibberellin (GA), and auxin (Aux), is the result of complex interactions between these phytohormones with the salicylic acid (SA)-jasmonate (JA)-ethylene (ET) backbone of immune signaling circuitry. Based on the available findings concerning phytohormone mediated plant resistance, this review attempts to develop the understanding of the defense roles of CKs, GAs, and Auxs, and their interplay with pathogen targeted defense hormones such as SA, JA and ET in plants, during biotic stress. Several phytohormone pathway genes and proteins have been reported to influence biotic stress resistance. In our analysis, we relate to specific roles uncovered for the Aux pathway TRANSPORT INHIBITOR RESISTANT 1 (TIR)/AUXIN SIGNALING F-BOX (AFB) and Gretchen Hagen 3 (GH3) proteins, the CK pathway isopentenyl transferase (IPT), CK oxidase (CKX), and Arabidopsis histidine kinase (AHK) proteins, and the GA pathway DELLA and GA20 oxidase proteins, in plant immunity and disease resistance. Additionally, we discuss microbial phytohormones that can enhance plant growth and resistance against pathogens, and the effect of phytohormones on modulation of the plant microbiome. Finally, we provide a concise overview of the possible approaches for modulating hormonal cross talk to improve plant health and resistance. [ABSTRACT FROM AUTHOR]

The present study was conducted to determine the effect of osmotic stress on the plant growth hormone production by six osmotolerant plant growth promoting bacterial strains. These strains originated from the phytomicrobiome of chilli cultivated in the drought prone areas of Andhra Pradesh. They possessed multiple plant growth promotion traits including the ability to produce a variety of plant growth hormones. The effect of osmotic stress on the plant growth hormone production was determined by High Performance Liquid Chromatography (HPLC) under normal and in vitro osmotic stress conditions using 25% Poly Ethylene Glycol (PEG) 8000. In general, it was observed that osmotic stress impacted the plant growth hormone production of the isolates, but nevertheless plant hormones were detected in all the bacterial strains. An exception to this was the cytokinin molecule zeatin riboside, which was produced at higher levels by five of the six bacterial isolates under osmotic stressed conditions. [ABSTRACT FROM AUTHOR]

Plant cell walls are dynamic structures that play crucial roles in growth, development, and stress responses. Despite our growing understanding of cell wall biology, the connections between cell wall integrity (CWI) and cell cycle progression in plants remain poorly understood. This review aims to explore the intricate relationship between CWI and cell cycle progression in plants, drawing insights from studies in yeast and mammals. We provide an overview of the plant cell cycle, highlight the role of endoreplication in cell wall composition, and discuss recent findings on the molecular mechanisms linking CWI perception to cell wall biosynthesis and gene expression regulation. Furthermore, we address future perspectives and unanswered questions in the field, such as the identification of specific CWI sensing mechanisms and the role of CWI maintenance in the growth-defense trade-off. Elucidating these connections could have significant implications for crop improvement and sustainable agriculture. Key message: In this review, we provide a comprehensive exploration of the intricate relationship between plant CWI and cell cycle progression. By drawing parallels from studies in yeast and mammals, we offer new insights into this critical facet of plant cell biology. We underscore the role of endoreplication in cell wall composition and the molecular mechanisms linking CWI perception to cell wall biosynthesis and gene expression regulation. The review also identifies key research avenues that remain to be explored, such as specific CWI sensing mechanisms and the role of CWI maintenance in the growth-defense trade-off. Elucidating these links holds significant potential for crop improvement and sustainable agriculture, thus rendering our work of broad interest to researchers in plant biology, agricultural science, and sustainable farming practices. [ABSTRACT FROM AUTHOR]

ABSTRACT In this study, a tissue culture method was developed for the propagation and conservation of Cattleya gaskelliana (N.E.Br.) B.S. Williams. Initially, protocorms with leaf primordium, obtained via asymbiotic seed germination, were used as explants. Half-strength Murashige and Skoog (MS) medium supplemented with 1.5 mg L−1 6-benzyl aminopurine (BA) and 2.0 mg L−1, indole butyric acid (IBA) was found to be optimal for the proliferation of protocorm-like bodies (PLB). Most explants (81%) proliferated, each producing 11.1 ± 1.87 units per explant. The optimum number of PLB differentiated to plantlets on a cultured medium with 0.5 mg L−1 Thidiazuron (TDZ) + 100 mL L−1 coconut water. Firmly rooted plantlets with vigorous growth were regenerated on ½ MS medium + 1.0 mg L−1 Indole- 3-acetic acid (IAA) and 50 g L−1 banana puree. Plants regenerated via in vitro processes were wrapped with sphagnum moss and acclimatized in a substrate containing equal proportions of bark, perlite, and peat with a 100% ex-vitro survival rate after acclimation for 70 days. The protocol developed in this study can be used to obtain several thousand plants within one year, and thus, is an effective method.

Cucumber (Cucumis sativus L.) belongs to the cucumber genus of the Cucurbitaceae family, and the selection of cultivars with minimal or no lateral branches can enhance the cultivation management efficiency. The growth of lateral branches is inhibited by strigolactone. To investigate the regulatory mechanism of strigolactone on the lateral branch development in cucumber, the cultivar LZ1 exhibiting multiple lateral branches was selected as the experimental material. The axillae of the plants were infiltrated with 1, 5, and 10 μmol·L−1 germination releaser 24 (GR24) at the four- to five-leaf stage. It was identified that 1 μmol·L−1 GR24 exhibited the most potent inhibitory effect on cucumber lateral branches. Additionally, exogenous strigolactone decreased the auxin content in the apical bud and axillae and increased the auxin content in the stem. This inhibited polar auxin transport in the axillary bud and promoted polar auxin transport in the apical bud. The content of strigolactone in the axilla region of cucumbers was elevated, whereas the synthesis and expression of cytokinin in the same area were reduced. A low concentration of GR24 induced the expression of cucumber branched 1 (csbrc1), whereas a high concentration of GR24 downregulated the expression of cucumber lateral suppressor (cscls) and blind (csblind), which inhibited the growth of cucumber lateral branches.

In the realm of genetically transformed crops, the process of plant regeneration holds utmost significance. However, the low regeneration efficiency of several wheat varieties currently restricts the use of genetic transformation for gene functional analysis and improved crop production. This research explores overexpression of TaLAX PANICLE1 (TaLAX1), which markedly enhances regeneration efficiency, thereby boosting genetic transformation and genome editing in wheat. Particularly noteworthy is the substantial increase in regeneration efficiency of common wheat varieties previously regarded as recalcitrant to genetic transformation. Our study shows that increased expression of TaGROWTH-REGULATING FACTOR (TaGRF) genes, alongside that of their co-factor, TaGRF-INTERACTING FACTOR 1 (TaGIF1), enhances cytokinin accumulation and auxin response, which may play pivotal roles in the improved regeneration and transformation of TaLAX1-overexpressing wheat plants. Overexpression of TaLAX1 homologs also significantly increases the regeneration efficiency of maize and soybean, suggesting that both monocot and dicot crops can benefit from this enhancement. Our findings shed light on a gene that enhances wheat genetic transformation and elucidate molecular mechanisms that potentially underlie wheat regeneration.

Climate change-induced environmental stress significantly affects crop yield and quality. In response to environmental stressors, plants use defence mechanisms and growth suppression, creating a resource trade-off between the stress response and development. Although stress-responsive genes have been widely engineered to enhance crop stress tolerance, there is still limited understanding of the interplay between stress signalling and plant growth, a research topic that can provide promising targets for crop genetic improvement. This review focuses on Cytokinin Response Factors (CRFs) transcription factor’s role in the balance between abiotic stress adaptation and sustained growth. CRFs, known for their involvement in cytokinin signalling and abiotic stress responses, emerge as potential targets for delaying senescence and mitigating yield penalties under abiotic stress conditions. Understanding the molecular mechanisms regulated by CRFs paves the way for decoupling stress responses from growth inhibition, thus allowing the development of crops that can adapt to abiotic stress without compromising development. This review highlights the importance of unravelling CRF-mediated pathways to address the growing need for resilient crops in the face of evolving climatic conditions.

Signals are exchanged at all stages of the arbuscular mycorrhizal (AM) symbiosis between fungi and their host plants. Root-exuded strigolactones are well-known early symbiotic cues, but the role of other phytohormones as interkingdom signals has seldom been investigated. Here we focus on ethylene and cytokinins, for which candidate receptors have been identified in the genome of the AM fungus Rhizophagus irregularis. Ethylene is known from the literature to affect asymbiotic development of AM fungi, and in the present study, we found that three cytokinin forms could stimulate spore germination in R. irregularis. Heterologous complementation of a Saccharomyces cerevisiae mutant strain with the candidate ethylene receptor RiHHK6 suggested that this protein can sense and transduce an ethylene signal. Accordingly, its N-terminal domain expressed in Pichia pastoris displayed saturable binding to radiolabeled ethylene. Thus, RiHHK6 displays the expected characteristics of an ethylene receptor. In contrast, the candidate cytokinin receptor RiHHK7 did not complement the S. cerevisiae mutant strain or Medicago truncatula cytokinin receptor mutants and seemed unable to bind cytokinins, suggesting that another receptor is involved in the perception of these phytohormones. Taken together, our results support the hypothesis that AM fungi respond to a range of phytohormones and that these compounds bear multiple functions in the rhizosphere beyond their known roles as internal plant developmental regulators. Our analysis of two phytohormone receptor candidates also sheds new light on the possible perception mechanisms in AM fungi. [Graphic: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Abstract Cytokinin oxidase/dehydrogenase (CKX) is a key regulatory enzyme for the irreversible degradation of the plant hormone cytokinin (CK), which is important in growth and development and response to abiotic stresses in cotton. In this study, 27, 28, 14 and 14 CKXs were screened by FAD structural domain and cytokinin binding structural domain in Gossypium hirsutum, Gossypium barbadense, Gossypium arboreum and Gossypium raimondii, respectively. Their phylogenetic relationships and expression patterns were analyzed, and most GhCKXs were found to be tissue-specific and responsive to various abiotic stresses such as cold, heat, salt and PEG. GhCKX6b-Dt was selected for gene silencing in evolutionary branch II for salt stress, because its expression increased after salt stress in cotton plants. An increase in PRO and MDA content and a decrease in SOD activity due to this gene were found after inducing salt stress, contributing to oxidative damage and decreased salt tolerance. In this study, CKXs were analyzed to reveal the possible role of GhCKXs against abiotic stresses in cotton, which provides a basis for further understanding of the biological functions of CK in plants such as growth and development and stress resistance.

This study addresses the propagation challenges faced by ‘Shine Muscat’, a newly introduced premium grapevine cultivar in South Korea, where multiple viral infections pose considerable economic loss. The primary objective was to establish a robust in vitro propagation method for producing disease-free grapes and to identify effective plant growth regulators to facilitate large-scale mass cultivation. After experimentation, 2.0 µM 6-benzyladenine (BA) exhibited superior shoot formation in the Murashige and Skoog medium compared with kinetin and thidiazuron. Conversely, α-naphthaleneacetic acid (NAA) hindered shoot growth and induced callus formation, while indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA) demonstrated favorable root formation, with IBA showing better results overall. Furthermore, inter simple sequence repeat analysis confirmed the genetic stability of in vitro-cultivated seedlings using 2.0 μM BA and 1.0 μM IBA, validating the suitability of the developed propagation method for generating disease-free ‘Shine Muscat’ grapes. These findings offer promising prospects for commercial grape cultivation, ensuring a consistent supply of healthy grapes in the market.

BackgroundThe heterologous expression of isopentenyl transferase (IPT) under the transcriptional control of the senescence-associated receptor-like kinase (SARK) promoter delayed cellular senescence and, through it, increased drought tolerance in plants. To evaluate the effect of pSARK::IPT expression in bread wheat, six independent transgenic events were obtained through the biolistic method and evaluated transgene expression, phenology, grain yield and physiological biomass components in plants grown under both drought and well-irrigating conditions. Experiments were performed at different levels: (i) pots and (ii) microplots inside a biosafety greenhouse, as well as under (iii) field conditions.ResultsTwo transgenic events, called TR1 and TR4, outperformed the wild-type control under drought conditions. Transgenic plants showed higher yield under both greenhouse and field conditions, which was positively correlated to grain number (given by more spikes and grains per spike) than wild type. Interestingly, this yield advantage of the transgenic events was observed under both drought and well-watered conditions.ConclusionsThe results obtained allow us to conclude that the SARK promoter-regulated expression of the IPT gene in bread wheat not only reduced the yield penalty produced by water stress but also led to improved productivity under well-watered conditions.

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.

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.

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.

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.