Cytokinin Levels Research Articles

Transcriptome Analysis Revealed Hormone Pathways and bZIP Genes Responsive to Decapitation in Sunflower.

Decapitation is an essential agricultural practice and is a typical method for analyzing shoot branching. However, it is unclear exactly how decapitation controls branching. In this study, the decapitation of sunflower plants led to the development of lateral buds, accompanied by a decrease in indole-3-acetic acid (IAA) and abscisic acid (ABA) levels and an increase in cytokinin (CK) levels. Additionally, 82 members of the HabZIP family were discovered and categorized into 9 groups, using phylogenetic and conservative domain analysis. The intron/exon structure and motif compositions of HabZIP members were also investigated. Based on tissue-specific expression and expression analysis following decapitation derived from the transcriptome, several HabZIP members may be involved in controlling decapitation-induced bud outgrowth. Therefore, it is hypothesized that the dynamic variations in hormone levels, in conjunction with particular HabZIP genes, led to the development of axillary buds in sunflowers following decapitation.

Crosstalk of Cytokinin with Ethylene and Auxin for Cell Elongation Inhibition and Boron Transport in Arabidopsis Primary Root under Boron Deficiency

Several studies have shown the role of phytohormones in the regulation of root growth of Arabidopsis plants under boron (B) deficiency. Ethylene and auxin play an important role in the control of Arabidopsis primary root cell elongation under short-term B deprivation, whereas cytokinins regulate root growth inhibition under B deficiency by controlling meristem cell proliferation. In this work, we study the possible interaction among cytokinin, ethylene, and auxin in the primary root response to B-deprivation treatment, as well as their possible role in B uptake and transport. Wild type (WT) and two mutants related to auxin and ethylene (aux1 and acs11) Arabidopsis plants were grown in control (10 µM B) or B starvation (0 µM B) treatment, in the absence or presence of trans-zeatin, and their primary root growth was analyzed. The possible interaction between these hormones was also studied by analyzing AUX1 gene expression in the acs11 mutant and ACS11 gene expression in the aux1 mutant. The GUS reporter lines ARR5::GUS, IAA2::GUS, and EBS::GUS were used to observe changes in cytokinin, auxin, and ethylene levels in the root, respectively. The results of this work suggest that cytokinin inhibits root cell elongation under B deficiency through two different mechanisms: (i) an ethylene-dependent mechanism through increased expression of the ACS11 gene, which would lead to increased ethylene in the root, and (ii) an ethylene-independent mechanism through decreased expression of the AUX1 gene, which alters auxin signaling in the meristematic and elongation zones and stele. We also report that changes in the expression of several B transporters occur in response to auxin, ethylene, and cytokinin that may affect the plant B content.

Auxin and cytokinin control fate determination of cotyledons in the one-leaf plant Monophyllaea glabra.

One-leaf plants in the Gesneriaceae family initially have two cotyledons of identical size; one cotyledon stops growing shortly after germination, whereas the other continues indeterminate growth. Factors involved in the unequal growth have been investigated, and a competitive relationship between the two cotyledons was previously proposed. However, questions regarding the fate determination of the two cotyledons remain: Why does only one cotyledon grow indeterminately while the other stops; is the fate of the cotyledons reversible; and what role does light quality play in the fate determination of the cotyledons? In this study, physiological experiments using the one-leaf plant species Monophyllaea glabra suggest that a biased auxin concentration between the two cotyledons and subsequent cytokinin levels may determine the fate of the cotyledons. In addition, observation of relatively mature individuals without hormone treatment and younger individuals with cytokinin treatment under laboratory growth conditions revealed that the fate determination of the microcotyledon is reversible. Although light quality has been suggested to be important for the determination of cotyledon fate in Streptocarpus rexii, an anisocotylous species, we conclude that light quality is not important in M. glabra.

Cellular differentiation, hormonal gradient, and molecular alternation between the division zone and the elongation zone of bamboo internodes.

Bamboo differentiates a cell division zone (DZ) and a cell elongation zone (EZ) to promote internode elongation during rapid growth. However, the biological mechanisms underlying this sectioned growth behavior are still unknown. Using histological, physiological, and genomic data, we found that the cell wall and other subcellular organelles such as chloroplasts are more developed in the EZ. Abundant hydrogen peroxide accumulated in the pith cells of the EZ, and stomata formed completely in the EZ. In contrast, most cells in the DZ were in an undifferentiated state with wrinkled cell walls and dense cytoplasm. Hormone detection revealed that the levels of gibberellin, auxin, cytokinin, and brassinosteroid were higher in the DZ than in the EZ. However, the levels of salicylic acid and jasmonic acid were higher in the EZ than in the DZ. Transcriptome analysis with qRT-PCR quantification revealed that the transcripts for cell division and primary metabolism had higher expression in the DZ, whereas the genes for photosynthesis, cell wall growth, and secondary metabolism were dramatically upregulated in the EZ. Overexpression of a MYB transcription factor, BmMYB83, promotes cell wall lignification in transgenic plants. BmMYB83 is specifically expressed in cells that may have lignin deposits, such as protoxylem vessels and fiber cells. Our results indicate that hormone gradient and transcriptome reprogramming, as well as specific expression of key genes such as BmMYB83, may lead to differentiation of cell growth in the bamboo internode.

Root-associated endophytic fungi modulate endogenous auxin and cytokinin levels to improve plant biomass and root morphology of trifoliate orange

Root-associated endophytic fungi like Serendipita indica and arbuscular mycorrhizal (AM) fungi can improve plant growth and root construction, but the potential mechanism is unclear. In this study, Funneliformis mosseae (an AM fungus) and S. indica, singly or in combination were inoculated into trifoliate orange (Poncirus trifoliata) seedlings, to assess changes in biomass and root morphological traits, coupled with auxins and cytokinins concentrations in leaves and roots and the expression of auxin synthesis and transporter protein genes. After 20 weeks of inoculation with these fungi, shoot and root biomass, root total length, taproot length, average diameter, surface area, volume, and the number of lateral roots in 1st-, 2nd-, and 3rd-order were improved, and S. indica showed a relatively greater effect than F. mosseae and dual inoculation. Endophytic fungal inoculation also significantly increased the concentration of indoleacetic acid, indole butyric acid, trans-zeatin, dihydrozeatin, and isopentenyl adenine in leaves and roots, whilst F. mosseae and S. indica exhibited relatively greater effects on leaves and roots, respectively. Correlation analysis revealed that both biomass and root morphological traits (except root projected area) were significantly positively associated with endogenous auxins and cytokinins. In addition, the inoculated plants recorded comparatively higher expression levels of indoleacetic acid synthesis genes (PtTAA1, PtTAR2, PtYUC3, PtYUC4, PtYUC6, and PtYUC8) and indoleacetic acid transporter protein genes (PtAUX1, PtLAX1, PtLAX2, PtLAX3, PtPIN1, PtPIN3, PtPIN4, PtABCB1, and PtABCB19) than the non-inoculated plants, among which F. mosseae and S. indica showed better effects in leaves and roots, respectively. These results suggest that root-associated endophytic fungi improved plant growth and root architecture, which were associated with changes in endogenous auxins and cytokinins.

Integrated transcriptome and hormone analyses provide insights into silver thiosulfate-induced "maleness" responses in the floral sex differentiation of pumpkin (Cucurbita moschata D.).

The perfect mating of male and female flowers is the key to successful pollination. The regulation of ethylene with chemicals is a good option for inducing staminate or female flowers. Silver thiosulfate is often used to induce the formation of male flowers in subgynoecious and gynoecious crops, which is important to maintain their progenies. However, its effects on flower sex differentiation in pumpkin (Cucurbita moschata Duch.) and the underlying mechanism remain unclear. In this study, the application of silver thiosulfate to pumpkin seedlings significantly delayed the occurrence of the first female flower and increased the number of male flowers. We next investigated the underlying mechanism by employing transcriptome and endogenous hormone analyses of the treated plants. In total, 1,304 annotated differentially expressed genes (DEGs)were identified by comparing silver thiosulfate-treated and control plants. Among these genes, 835 were upregulated and 469 were downregulated. The DEGs were mainly enriched in the phenylpropanoid biosynthesis and metabolism pathways (phenylalanine ammonia-lyase, peroxidase) and plant hormone signal transduction pathways (auxin signaling, indole-3-acetic acid-amido synthetase, ethylene response factor). Silver thiosulfate significantly reduced the levels of 2-oxindole-3-acetic acid, para-topolin riboside, dihydrozeatin-O-glucoside riboside, and jasmonoyl-l-isoleucine but increased the levels of trans-zeatin-O-glucoside, cis-zeatin riboside, and salicylic acid 2-O-β-glucoside. The levels of auxin and jasmonic acid were decreased, whereas those of salicylic acid were increased. Different trends were observed for different types of cytokinins. We concluded that silver thiosulfate treatment not only affects the expression of auxin synthesis and signaling genes but also that of ethylene response factor genes and regulates the levels of auxin, salicylic acid, jasmonic acid, and cytokinins, which together might contribute to the maleness of pumpkin. This study provides useful information for understanding the mechanism underlying the effect of silver thiosulfate on floral sex differentiation in pumpkin, a widely cultivated vegetable crop worldwide, and gives a production guidance for the induction of maleness using STS for the reproduction of gynoecious lines of Cucurbitaceae crops.

Cytokinins regulate rice lamina joint development and leaf angle.

Leaf angle is determined by lamina joint inclination and is an important agronomic trait that determines plant architecture, photosynthetic efficiency, and crop yield. Cytokinins (CKs) are phytohormones involved in shaping rice (Oryza sativa L.) architecture, but their role in leaf angle remains unknown. Here, we report that CK accumulation mediated by rice CK OXIDASE/DEHYDROGENASE3 (OsCKX3) controls lamina joint development and negatively regulates leaf angle. Phenotypic analysis showed that rice osckx3 mutants had smaller leaf angles, while the overexpression lines (OsCKX3-OE) had larger leaf angles. Histological sections indicated that the leaf inclination changes in the osckx3 and OsCKX3-OE lines resulted from asymmetric proliferation of the cells and vascular bundles in the lamina joint. Reverse transcription quantitative PCR, promoter-fused β-glucuronidase expression, and subcellular localization assays indicated that OsCKX3 was highly expressed in the lamina joint, and OsCKX3-GFP fusion protein localized to the endoplasmic reticulum. The enzyme assays using recombinant protein OsCKX3 revealed that OsCKX3 prefers trans-zeatin (tZ) and isopentenyladenine (iP). Consistently, tZ and iP levels increased in the osckx3 mutants but decreased in the OsCKX3 overexpression lines. Interestingly, agronomic trait analysis of the rice grown in the paddy field indicated that osckx3 displayed a smaller leaf angle and enhanced primary branch number, grain size, 1,000-grain weight, and flag leaf size. Collectively, our results revealed that enhancing CK levels in the lamina joint by disrupting OsCKX3 negatively regulates leaf angle, highlighting that the CK pathway can be engineered to reduce leaf angle in rice and possibly in other cereals.

Cytokinin regulates female gametophyte development by cell cycle modulation in Arabidopsis thaliana

Male and female gametophyte development, double fertilization, and embryogenesis are key to alternating generations in angiosperms. The female gametophyte of Arabidopsis is an eight-nucleate haploid structure developed from functional megaspores (FMs) through three flawless mitoses regulated by a series of cell cycle genes. Cytokinin, an important phytohormone, plays a critical role in the regulation of plant growth and development. However, the mechanisms by which cytokinins regulate female gametophyte development remain largely unknown. In this study, we constructed transgenic plants (pES1::CKX1) with low cytokinin levels in the embryo sac. Phenotypic analysis showed that pES1::CKX1 inhibits female gametophyte development. Microscopic observation revealed that female gametophyte development of pES1::CKX1 was delayed. The promoters of all cell cycle genes were cloned and transformed into wild-type (WT). We crossed these transgenic plants of cell cycle genes expressed in ovules with pES1::CKX1 and compared the expression level of β-glucuronidase (GUS) in pES1::CKX1 and WT. Many cell cycle-regulated genes were up or downregulated in pES1::CKX1 compared with WT, and the embryo sac development cell cycle in cycd2;1/+ cycd3;3 was defective. Our results demonstrated that cytokinin affects cell division in the female gametophyte by affecting the expression of cell cycle genes.

Genome-Wide Analysis and Evolutionary Perspective of the Cytokinin Dehydrogenase Gene Family in Wheat (Triticum aestivum L.).

Cytokinin dehydrogenase (CKX; EC.1.5.99.12) regulates the level of cytokinin (CK) in plants and is involved in CK regulatory activities. In different plants, a small gene family encodes CKX proteins with varied numbers of members. These genes are expanded in the genome mainly due to segmental duplication events. Despite their biological importance, CKX genes in Triticum aestivum have yet to be studied in depth. A total of 11 CKX subfamilies were identified with similar gene structures, motifs, domains, cis-acting elements, and an average signal peptide of 25 amino acid length was found. Introns, ranging from one to four, were present in the coding regions at a similar interval in major CKX genes. Putative cis-elements such as abscisic acid, auxin, salicylic acid, and low-temperature-, drought-, and light-responsive cis-regulatory elements were found in the promoter region of majority CKX genes. Variation in the expression pattern of CKX genes were identified across different tissues in Triticum. Phylogenetic analysis shows that the same subfamily of CKX clustered into a similar clade that reflects their evolutionary relationship. We performed a genome-wide identification of CKX family members in the Triticum aestivum genome to get their chromosomal location, gene structure, cis-element, phylogeny, synteny, and tissue- and stage-specific expression along with gene ontology. This study has also elaborately described the tissue- and stage-specific expression and is the resource for further analysis of CKX in the regulation of biotic and abiotic stress resistance, growth, and development in Triticum and other cereals to endeavor for higher production and proper management.

Morpho-histology, endogenous hormone dynamics, and transcriptome profiling in Dacrydium pectinatum during female cone development.

Dacrydium pectinatum de Laubenfels is a perennial dioeciously gymnosperm species dominant in tropical montane rain forests. Due to deforestation, natural disasters, long infancy, and poor natural regeneration ability, the population of this species has been significantly reduced and listed as an endangered protected plant. To better understand the female cone development in D. pectinatum, we examined the morphological and anatomical changes, analyzed the endogenous hormone dynamics, and profiled gene expression. The female reproductive structures were first observed in January. The morpho-histological observations suggest that the development of the D. pectinatum megaspore can be largely divided into six stages: early flower bud differentiation, bract primordium differentiation, ovule primordium differentiation, dormancy, ovule maturity, and seed maturity. The levels of gibberellins (GA), auxin (IAA), abscisic acid (ABA), and cytokinin (CTK) fluctuate during the process of female cone development. The female cones of D. pectinatum need to maintain a low level of GA3-IAA-ABA steady state to promote seed germination. The first transcriptome database for female D. pectinatum was generated, revealing 310,621 unigenes. Differential expression analyses revealed several floral (MADS2, AGL62, and LFY) and hormone biosynthesis and signal transduction (CKX, KO, KAO, ABA4, ACO, etc.) genes that could be critical for female cone development. Our study provides new insights into the cone development in D. pectinatum and the foundation for female cone induction with hormones.

Association of jasmonic acid priming with multiple defense mechanisms in wheat plants under high salt stress.

Salinity is a global conundrum that negatively affects various biometrics of agricultural crops. Jasmonic acid (JA) is a phytohormone that reinforces multilayered defense strategies against abiotic stress, including salinity. This study investigated the effect of JA (60 μM) on two wheat cultivars, namely ZM9 and YM25, exposed to NaCl (14.50 dSm−1) during two consecutive growing seasons. Morphologically, plants primed with JA enhanced the vegetative growth and yield components. The improvement of growth by JA priming is associated with increased photosynthetic pigments, stomatal conductance, intercellular CO2, maximal photosystem II efficiency, and transpiration rate of the stressed plants. Furthermore, wheat cultivars primed with JA showed a reduction in the swelling of the chloroplast, recovery of the disintegrated thylakoids grana, and increased plastoglobuli numbers compared to saline-treated plants. JA prevented dehydration of leaves by increasing relative water content and water use efficiency via reducing water and osmotic potential using proline as an osmoticum. There was a reduction in sodium (Na+) and increased potassium (K+) contents, indicating a significant role of JA priming in ionic homeostasis, which was associated with induction of the transporters, viz., SOS1, NHX2, and HVP1. Exogenously applied JA mitigated the inhibitory effect of salt stress in plants by increasing the endogenous levels of cytokinins and indole acetic acid, and reducing the abscisic acid (ABA) contents. In addition, the oxidative stress caused by increasing hydrogen peroxide in salt-stressed plants was restrained by JA, which was associated with increased α-tocopherol, phenolics, and flavonoids levels and triggered the activities of superoxide dismutase and ascorbate peroxidase activity. This increase in phenolics and flavonoids could be explained by the induction of phenylalanine ammonia-lyase activity. The results suggest that JA plays a key role at the morphological, biochemical, and genetic levels of stressed and non-stressed wheat plants which is reflected in yield attributes. Hierarchical cluster analysis and principal component analyses showed that salt sensitivity was associated with the increments of Na+, hydrogen peroxide, and ABA contents. The regulatory role of JA under salinity stress was interlinked with increased JA level which consequentially improved ion transporting, osmoregulation, and antioxidant defense.

Effects of temperature and light conditions during the late growth stage on delayed stem senescence and cytokinin levels in the xylem exudate of soybean

AbstractDelayed stem senescence (DSS) in soybean plants is defined as retarded leaf and stem maturity, even if pods mature normally. This phenomenon is a serious problem for the combine harvesting of soybean in Japan. Our previous research showed that DSS occurred more frequently in June sowing than in July sowing, and the plants sown in June were exposed to higher temperature and higher solar radiation intensity during reproductive growth stage as compared to those sown in July. We additionally observed higher levels of cytokinins, plant hormones with senescence inhibition activity, in xylem exudate after pod‐setting stage (R4) under the DSS‐induced condition. Thus, this study investigated the effects of the respective environmental factors, temperature and light intensity, after R4 on cytokinin levels in the xylem exudate and the occurrence of DSS. Pot experiments were conducted in the experimental field of Nihon University (Fujisawa‐city, Kanagawa, Japan). In temperature treatments, soybean plants grown in the field environment were moved to an air‐conditioned room at R4 stage, whereas control of light intensity was achieved by shading using cheese clothes under the field condition. The temperature control experiments revealed that high temperature after R4 promoted the DSS symptom, while low temperature after R4 inhibited the occurrence of DSS. On the other hand, decreased light intensity after R4 did not affect the severity of DSS. The cytokinin levels in the xylem exudate at R5 correlated with the severity of DSS in the temperature treatments, but the change in light intensity did not influence the cytokinin levels. These results indicated that high temperature after pod‐setting stage have a profound effect on the occurrence of DSS, and this effect is mediated by the increased cytokinin levels in the xylem exudate.

Analysis of cytokinin content and associated genes at different developmental stages in pak choi (Brassica rapa ssp. chinensis Makino)

Pak choi (Brassica rapa ssp. chinensis Makino) is a typical seed vernalization vegetable. Because premature bolting in spring causes significant economic losses, it is very important to clarify the flower formation mechanism to prevent this. Cytokinins are important plant hormones involved in regulating plant growth and development. To understand the relationship between cytokinin metabolism and flowering of pak choi, in this study, we determined the cytokinin trans-zeatin content in shoot apices of pak choi at different developmental stages by enzyme-linked immunosorbent assay. The results showed that cytokinin levels increased significantly after low temperature treatment at 4 °C, and continued to increase with vegetative growth after transplanting, reaching a peak at the critical period which is immediately prior to flower buds’ differentiation (S0), then decreased thereafter. The levels for low temperature treatment were consistently higher than controls. To explore the molecular mechanism underpinning the cytokinin changes, expression of homologous genes encoding cytokinin metabolic enzymes was analysed by transcriptome sequencing. The expression levels of nine genes (Bra004037, Bra023701, Bra002204, Bra014968, Bra028326, Bra028182, Bra034022, Bra009143 and Bra005869) were consistent with the changes in cytokinin content. The correlation between differentially expressed genes and cytokinin content in different developmental stages of apexes was analysed, and six closely related genes (Bra023701, Bra002204, Bra014968, Bra028182, Bra005869 and Bra009143) were identified. The results help to illuminate the molecular mechanisms controlling flowering of pak choi.

The Citrus sinensis TILLER ANGLE CONTROL 1 (CsTAC1) gene regulates tree architecture in sweet oranges by modulating the endogenous hormone content

Citrus is a major fruit crop cultivated on a global scale. Citrus trees are long lived perennials with a large canopy. Understanding the genetic control of tree architecture could provide tools for breeding and selection of citrus cultivars suitable for high density planting with improved light exposure. Tree architecture is modulated by the TILLER ANGLE CONTROL 1 (TAC1) gene which plays an important role in the regulation of the shoot angle. Herein, we used CRISPR/Cas9 technology to knockout the CsTAC1 gene for the biochemical and molecular analysis of its function. Nine transgenic lines were obtained, and five edited plants were confirmed based on T7EI mismatch detection assay and Sanger sequencing. The transgenic citrus lines exhibited pleiotropic phenotypes, including differences in branch angle and stem growth. Additionally, silencing CsTAC1 led to enhanced CsLAZY1 transcript levels in the tested lines. Analysis of the phytohormonal profile revealed that TAC1-edited plants exhibited lower auxin contents and increased cytokinin levels in the leaves compared to the wild-type plants. The GA7 gibberellin level was enhanced in most of the edited lines. Collectively, TAC1 affects branch angle in association with hormone signals in citrus.

Gene expression profiling before and after internode culture for adventitious shoot formation in ipecac

BackgroundIn ipecac (Carapichea ipecacuanha (Brot.) L. Andersson), adventitious shoots can be induced simply by placing internodal segments on phytohormone-free culture medium. The shoots form locally on the epidermis of the apical region of the segments, but not the basal region. Levels of endogenous auxin and cytokinin transiently increase in the segments after 1 week of culture.ResultsHere, we conducted RNA-seq analysis to compare gene expression patterns in apical and basal regions of segments before culture and after 1 week of culture for adventitious shoot formation. The results revealed 8987 differentially expressed genes in a de novo assembly of 76,684 genes. Among them, 276 genes were upregulated in the apical region after 1 week of culture relative to before culture and the basal region after 1 week of culture. These genes include 18 phytohormone-response genes and shoot-formation-related genes. Validation of the gene expression by quantitative real-time PCR assay confirmed that the expression patterns were similar to those of the RNA-seq data.ConclusionsThe transcriptome data show that expression of cytokinin biosynthesis genes is induced along with the acquisition of cellular pluripotency and the initiation of cell division by wounding in the apical region of internodal segments, that trigger adventitious shoot formation without callusing.

Killing two birds with a single stone-genetic manipulation of cytokinin oxidase/dehydrogenase (CKX) genes for enhancing crop productivity and amelioration of drought stress response.

The development of high-yielding, bio-fortified, stress-tolerant crop cultivars is the need of the hour in the wake of increasing global food insecurity, abrupt climate change, and continuous shrinking of resources and landmass suitable for agriculture. The cytokinin group of phytohormones positively regulates seed yield by simultaneous regulation of source capacity (leaf senescence) and sink strength (grain number and size). Cytokinins also regulate root-shoot architecture by promoting shoot growth and inhibiting root growth. Cytokinin oxidase/dehydrogenase (CKX) are the only enzymes that catalyze the irreversible degradation of active cytokinins and thus negatively regulate the endogenous cytokinin levels. Genetic manipulation of CKX genes is the key to improve seed yield and root-shoot architecture through direct manipulation of endogenous cytokinin levels. Downregulation of CKX genes expressed in sink tissues such as inflorescence meristem and developing seeds, through reverse genetics approaches such as RNAi and CRISPR/Cas9 resulted in increased yield marked by increased number and size of grains. On the other hand, root-specific expression of CKX genes resulted in decreased endogenous cytokinin levels in roots which in turn resulted in increased root growth indicated by increased root branching, root biomass, and root-shoot biomass ratio. Enhanced root growth provided enhanced tolerance to drought stress and improved micronutrient uptake efficiency. In this review, we have emphasized the role of CKX as a genetic factor determining yield, micronutrient uptake efficiency, and response to drought stress. We have summarised the efforts made to increase crop productivity and drought stress tolerance in different crop species through genetic manipulation of CKX family genes.

Carpel-specific down-regulation of GhCKXs in cotton significantly enhances seed and fiber yield.

Cytokinin is considered to be an important driver of seed yield. To increase the yield of cotton while avoiding the negative consequences caused by constitutive overproduction of cytokinin, we down-regulated specifically the carpel genes for cytokinin oxidase/dehydrogenase (CKX), a key negative regulator of cytokinin levels, in transgenic cotton. The carpel-specific down-regulation of CKXs significantly enhanced cytokinin levels in the carpels. The elevated cytokinin promoted the expression of carpel- and ovule-development-associated genes, GhSTK2, GhAG1, and GhSHP, boosting ovule formation and thus producing more seeds in the ovary. Field experiments showed that the carpel-specific increase of cytokinin significantly increased both seed yield and fiber yield of cotton, without resulting in detrimental phenotypes. Our study details the regulatory mechanism of cytokinin signaling for seed development, and provides an effective and feasible strategy for yield improvement of seed crops.

Genes Impacting Grain Weight and Number in Wheat (Triticum aestivum L. ssp. aestivum).

The primary goal of common wheat (T. aestivum) breeding is increasing yield without negatively impacting the agronomic traits or product quality. Genetic approaches to improve the yield increasingly target genes that impact the grain weight and number. An energetic trade-off exists between the grain weight and grain number, the result of which is that most genes that increase the grain weight also decrease the grain number. QTL associated with grain weight and number have been identified throughout the hexaploid wheat genome, leading to the discovery of numerous genes that impact these traits. Genes that have been shown to impact these traits will be discussed in this review, including TaGNI, TaGW2, TaCKX6, TaGS5, TaDA1, WAPO1, and TaRht1. As more genes impacting the grain weight and number are characterized, the opportunity is increasingly available to improve common wheat agronomic yield by stacking the beneficial alleles. This review provides a synopsis of the genes that impact grain weight and number, and the most beneficial alleles of those genes with respect to increasing the yield in dryland and irrigated conditions. It also provides insight into some of the genetic mechanisms underpinning the trade-off between grain weight and number and their relationship to the source-to-sink pathway. These mechanisms include the plant size, the water soluble carbohydrate levels in plant tissue, the size and number of pericarp cells, the cytokinin and expansin levels in developing reproductive tissue, floral architecture and floral fertility.

Integrated transcriptome and hormonal analysis of naphthalene acetic acid-induced adventitious root formation of tea cuttings (Camellia sinensis)

BackgroundTea plant breeding or cultivation mainly involves propagation via cuttings, which not only ensures the inheritance of the excellent characteristics of the mother plant but also facilitates mechanized management. The formation of adventitious root (AR) determines the success of cutting-based propagation, and auxin is an essential factor involved in this process. To understand the molecular mechanism underlying AR formation in nodal tea cuttings, transcriptome and endogenous hormone analysis was performed on the stem bases of red (mature)- and green (immature)-stem cuttings of ‘Echa 1 hao’ tea plant as affected by a pulse treatment with naphthalene acetic acid (NAA).ResultsIn this study, NAA significantly promoted AR formation in both red- and green-stem cuttings but slightly reduced callus formation. External application of NAA reduced the levels of endogenous indole-3-acetic acid (IAA) and cytokinin (TZR, trans-zeatin riboside). The number of DEGs (NAA vs. CK) identified in the green-stem cuttings was significantly higher than that in the red-stem cuttings, which corresponded to a higher rooting rate of green-stem cuttings under the NAA treatment. A total of 82 common DEGs were identified as being hormone-related and involved in the auxin, cytokinin, abscisic acid, ethylene, salicylic acid, brassinosteroid, and jasmonic acid pathways. The negative regulation of NAA-induced IAA and GH3 genes may explain the decrease of endogenous IAA. NAA reduced endogenous cytokinin levels and further downregulated the expression of cytokinin signalling-related genes. By the use of weighted gene co-expression network analysis (WGCNA), several hub genes, including three [cellulose synthase (CSLD2), SHAVEN3-like 1 (SVL1), SMALL AUXIN UP RNA (SAUR21)] that are highly related to root development in other crops, were identified that might play important roles in AR formation in tea cuttings.ConclusionsNAA promotes the formation of AR of tea cuttings in coordination with endogenous hormones. The most important endogenous AR inductor, IAA, was reduced in response to NAA. DEGs potentially involved in NAA-mediated AR formation of tea plant stem cuttings were identified via comparative transcriptome analysis. Several hub genes, such as CSLD2, SVL1 and SAUR21, were identified that might play important roles in AR formation in tea cuttings.

Excessive nitrogen in field-grown rice suppresses grain filling of inferior spikelets by reducing the accumulation of cytokinin and auxin

Excessive nitrogen fertilizer is often applied to achieve higher rice yield in many regions of the world. However, the effect of excessive nitrogen on grain filling of inferior spikelets and its underlying mechanisms are still poorly understood. A three-year field experiment was conducted to reveal the mechanisms by which excessive nitrogen application regulated grain filling. We found that excessive application of nitrogen reduced the starch biosynthesis and grain weight of inferior spikelets but not superior spikelets. Plant hormone detection revealed that both cytokinin (CTK) and 3-Indoleacetic acid (IAA) were reduced by excessive nitrogen at the early stage of grain filling in inferior spiekelts. Quantitative real-time PCR results showed that inhibition of biosynthesis genes led to a decrease in IAA content, while elevation of CTK oxidase genes resulted in a lower level of CTK under excessive nitrogen treatment. Application of exogenous IAA or CTK significantly impaired the inhibitory effect of excessive nitrogen on starch accumulation and grain weight of inferior spikelets. Further enzyme assays revealed that excessive nitrogen application significantly suppressed the enzyme activities of sucrose synthase (SuS), adenosine diphosphate glucose pyrophosphorylase (AGP) and starch synthase (StS) in inferior spikelets. Application of exogenous IAA or CTK to the panicle under excessive nitrogen treatment significantly increased the enzyme activities of SuS, AGP and StS. Taken together, our results revealed that excessive nitrogen application in field-grown rice suppressed grain filling of inferior spikelets by reducing the accumulation of cytokinin and auxin. • Grain filling of inferior spikelets was reduced by excessive nitrogen application. • HN application reduced the contents of CTK and auxin in inferior spikelets at 6 DAA. • Exogenous CTK and auxin both increased grain filling under HN conditions. • HN downregulated auxin biosynthesis genes while upregulated CTK catabolism genes. • Activities of key enzymes for starch biosynthesis were regulated by CTK and auxin.