Cytokinin Levels Research Articles

Multifaceted activity of cytokinin in leaf development shapes its size and structure in Arabidopsis.

The phytohormone cytokinin has been shown to affect many aspects of plant development ranging from the regulation of the shoot apical meristem to leaf senescence. However, some studies have reported contradictory effects of cytokinin on leaf physiology. Therefore cytokinin treatments cause both chlorosis and increased greening and both lead to decrease or increase in cell size. To elucidate this multifaceted role of cytokinin in leaf development, we have employed a system of temporal controls over the cytokinin pool and investigated the consequences of modulated cytokinin levels in the third leaf of Arabidopsis. We show that, at the cell proliferation phase, cytokinin is needed to maintain cell proliferation by blocking the transition to cell expansion and the onset of photosynthesis. Transcriptome profiling revealed regulation by cytokinin of a gene suite previously shown to affect cell proliferation and expansion and thereby a molecular mechanism by which cytokinin modulates a molecular network underlying the cellular responses. During the cell expansion phase, cytokinin stimulates cell expansion and differentiation. Consequently, a cytokinin excess at the cell expansion phase results in an increased leaf and rosette size fueled by higher cell expansion rate, yielding higher shoot biomass. Proteome profiling revealed the stimulation of primary metabolism by cytokinin, in line with an increased sugar content that is expected to increase turgor pressure, representing the driving force of cell expansion. Therefore, the developmental timing of cytokinin content fluctuations, together with a tight control of primary metabolism, is a key factor mediating transitions from cell proliferation to cell expansion in leaves.

Transcriptome analysis of the irregular shape of shoot apical meristem in dt (dou tou)\xa0mutant of Brassica napus L.

Rapeseed (Brassica napus L.) is an important oil crop in the world. In order to fulfill the requirement of mechanized harvesting and raised production, it is highly desirable to gain a better understanding of the regulatory networks controlling the main agronomic traits of this crop. In this study, we obtained a natural mutant of rapeseed with more main stems named as dt (duo tou, meaning more main stems in Chinese). The dt mutant exhibits abnormal differentiation of stems, increased leaves, and decreased plant height. Phenotype and tissue section analysis showed that abnormal development of the shoot apical meristem (SAM) led to the dt phenotype. Genes that participated in SAM activity maintenance, cytokinin biosynthesis, and signal transduction displayed greatly variation at transcriptional level, which was associated with the high level of cytokinin in the dt mutant. These results provide desirable material for improving the breeding and production of Brassica napus.

Antagonistic activity of auxin and cytokinin in shoot and root organs.

The hormones auxin and cytokinin are essential for plant growth and development. Because of the central importance of root and shoot apical meristems in plant growth, auxin/cytokinin interactions have been predominantly analyzed in relation to apical meristem formation and function. In contrast, the auxin/cytokinin interactions during organ growth have remained largely unexplored. Here, we show that a specific interaction between auxin and cytokinin operates in both the root and the shoot where it serves as an additional determinant of plant development. We found that auxin at low concentrations limits the action of cytokinin. An increase in cytokinin level counteracts this inhibitory effect and leads to an inhibition of auxin signaling. At higher concentrations of both hormones, these antagonistic interactions between cytokinin and auxin are absent. Thus, our results reveal a bidirectional and asymmetrical interaction of auxin and cytokinin beyond the bounds of apical meristems. The relation is bidirectional in that both hormones exert inhibitory effects on each other's signaling mechanisms. However, this relation is also asymmetrical because under controlled growth conditions, auxin present in nontreated plants suppresses cytokinin signaling, whereas the reverse is not the case.

Production and Role of Hormones During Interaction of Fusarium Species With Maize (Zea mays L.) Seedlings.

It has long been known that hormones affect the interaction of a phytopathogen with its host plant. The pathogen can cause changes in plant hormone homeostasis directly by affecting biosynthesis or metabolism in the plant or by synthesizing and secreting the hormone itself. We previously demonstrated that pathogenic fungi of the Fusarium species complex are able to produce three major types of hormones: auxins, cytokinins, and gibberellins. In this work, we explore changes in the levels of these hormones in maize and mango plant tissues infected with Fusarium. The ability to produce individual phytohormones varies significantly across Fusarium species and such differences likely impact host specificity inducing the unique responses noted in planta during infection. For example, the production of gibberellins by F. fujikuroi leads to elongated rice stalks and the suppression of gibberellin biosynthesis in plant tissue. Although all Fusarium species are able to synthesize auxin, sometimes by multiple pathways, the ratio of its free form and conjugates in infected tissue is affected more than the total amount produced. The recently characterized unique pathway for cytokinin de novo synthesis in Fusarium appears silenced or non-functional in all studied species during plant infection. Despite this, a large increase in cytokinin levels was detected in F. mangiferae infected plants, caused likely by the up-regulation of plant genes responsible for their biosynthesis. Thus, the accumulation of active cytokinins may contribute to mango malformation of the reproductive organs upon infection of mango trees. Together, our findings provide insight into the complex role fungal and plant derived hormones play in the fungal–plant interactions.

Low salinity promotes the growth of broccoli sprouts by regulating hormonal homeostasis and photosynthesis

Broccoli sprouts produce several bioactive compounds and are recognized as a health-promoting vegetable. In this study, the effect of salinity (NaCl) on the growth of broccoli sprouts was investigated. Broccoli seeds were germinated for 4 and 8 d with spraying 0–120 mM NaCl and then harvested to evaluate changes in endogenous hormones, photosynthetic indices, chlorophyll fluorescence parameters and chloroplast ultrastructure. The growth of sprouts was significantly promoted by low salinity (40 and 80 mM NaCl) and inhibited by high salinity treatment (120, 160 and 200 mM NaCl). In 8-day-old sprouts treated with 80 mM NaCl, levels of abscisic acid, cytokinin, brassinolide, indole-3-acetic acid and gibberellic acid were significantly enhanced, while net photosynthetic rate increased in low salinity conditions was due to the elevated chlorophyll content and increased photosystem II activity. Furthermore, low salinity increased the leaf area maximally in both 4- and 8-day-old sprouts. Enlarged chloroplast and an increased number of grana also contributed to improved photosynthesis. Low salinity conditions induced endogenous growth hormone synthesis and improved photosynthesis, thereby promoting the growth of broccoli sprouts. This study provides a theoretical basis for the improved production of broccoli sprouts in low salinity conditions.

Identification and Genetic Characterization of a Novel Tillering Dwarf Semi-Sterile Mutant tdr 1 in Rice

Tillering and plant height are important components of plant architecture and grain production in rice. We identified a novel high tillering, dwarf and semi-sterile mutant, as named tdr1 in a rice maintainer line E20 derived from the cross between between IR68888B and Luxiang 90. The investigation of tiller dynamic in the tdr1 line displayed 3 different phases: rapid increasing of tillers in the vegetative growth stage, producing no new tillers in the transition stage from the vegetative growth to reproductive growth, and regeneration of new tillers after heading. The assay of hormones showed the significant reduction of brassinolide level and no change of the levels of gibberellic acid, cytokinin and strigolactone in the tdr1 line. Genetic analysis indicated the phenotype of high tillering, dwarfism and semi-sterility is controlled by a recessive gene in several different segregation populations. The TDR1 gene was mapped in the 105 kb interval between RM3288 and RM6590 on chromosome 4. Cloning of TDR1 gene would provide a new opportunity to uncover the molecular mechanism of the development of plant height and tiller in rice.

Structural and functional insights into the modulation of the activity of a flax cytokinin oxidase by flax rust effector AvrL567-A.

SummaryDuring infection, plant pathogens secrete effector proteins to facilitate colonization. In comparison with our knowledge of bacterial effectors, the current understanding of how fungal effectors function is limited. In this study, we show that the effector AvrL567‐A from the flax rust fungus Melampsora lini interacts with a flax cytosolic cytokinin oxidase, LuCKX1.1, using both yeast two‐hybrid and in planta bimolecular fluorescence assays. Purified LuCKX1.1 protein shows catalytic activity against both N6‐(Δ2‐isopentenyl)‐adenine (2iP) and trans‐zeatin (tZ) substrates. Incubation of LuCKX1.1 with AvrL567‐A results in increased catalytic activity against both substrates. The crystal structure of LuCKX1.1 and docking studies with AvrL567‐A indicate that the AvrL567 binding site involves a flexible surface‐exposed region that surrounds the cytokinin substrate access site, which may explain its effect in modulating LuCKX1.1 activity. Expression of AvrL567‐A in transgenic flax plants gave rise to an epinastic leaf phenotype consistent with hormonal effects, although no difference in overall cytokinin levels was observed. We propose that, during infection, plant pathogens may differentially modify the levels of extracellular and intracellular cytokinins.

A member of the TERMINAL FLOWER 1/CENTRORADIALIS gene family controls sprout growth in potato tubers.

Potato tuber bud dormancy break followed by premature sprouting is a major commercial problem which results in quality losses and decreased tuber marketability. An approach to controlling premature tuber sprouting is to develop potato cultivars with a longer dormancy period and/or reduced rate of sprout growth. Our recent studies using a potato diploid population have identified several quantitative trait loci (QTLs) that are associated with tuber sprout growth. In the current study, we aim to characterize a candidate gene associated with one of the largest effect QTLs for rapid tuber sprout growth on potato chromosome 3. Underlying this QTL is a gene encoding a TERMINAL FLOWER 1/CENTRORADIALIS homologue (PGSC0003DMG400014322). Here, we use a transgenic approach to manipulate the expression level of the CEN family member in a potato tetraploid genotype (cv. Désirée). We demonstrate a clear effect of manipulation of StCEN expression, with decreased expression levels associated with an increased rate of sprout growth, and overexpressing lines showing a lower rate of sprout growth than controls. Associated with different levels of StCEN expression were different levels of abscisic acid and cytokinins, implying a role in controlling the levels of plant growth regulators in the apical meristem.

English

Cytokinins (CK) are key determinants of seed size and their level can impact crop yield. The biological activities and chemistry of cytokinins are well defined but less is known about their molecular mode of action during maize kernel development, especially in late stage kernels. To understand the regulation of cytokinin levels at late stage kernel development, we examined the levels of active and inactive cytokinins in embryo and endosperm across kernel development. Active trans-zeatin is high during early stage of kernel growth, but decreases during later stages. We also undertook a genome-wide survey of cytokinin metabolic genes involved in biosynthesis, metabolism, conversion, transport, and signaling and examined their expression profiles during kernel development. Several major genes were identified which may contribute to changes in cytokinin levels during the late stage. Our systematic analysis of cytokinin pathways genes and metabolites provides an overall insight into the regulation of CK level across maize kernel development. Our data on late stage kernel add to the existing knowledge of CK biology in maize and could facilitate the modulation of CK levels using biotechnology. Key words: Maize, cytokinin, kernel development, metabolism, gene regulation.

Transcriptome Profiles Reveal the Crucial Roles of Auxin and Cytokinin in the "Shoot Branching" of Cremastra appendiculata.

Cremastra appendiculata has become endangered due to reproductive difficulties. Specifically, vegetative reproduction is almost its only way to reproduce, and, under natural conditions, it cannot grow branches, resulting in an extremely low reproductive coefficient (reproductive percentage). Here, we performed RNA-Seq and a differentially expressed gene (DEG) analysis of the three stages of lateral bud development in C. appendiculata after decapitation—dormancy (D2), transition (TD2), and emergence (TG2)—and the annual axillary bud natural break (G1) to gain insight into the molecular regulatory network of shoot branching in this plant. Additionally, we applied the auxin transport inhibitors N-1-naphthylphthalamic acid (NPA) and 2,3,5-triiodibenzoic acid (TIBA) to a treated pseudobulb string of C. appendiculata to verify the conclusions obtained by the transcriptome. RNA-Seq provided a wealth of valuable information. Successive pairwise comparative transcriptome analyses revealed 5988 genes as DEGs. GO (Gene Ontology) and KEGG (Kyoto encyclopedia of genes and genomes) analyses of DEGs showed significant enrichments in phytohormone biosynthesis and metabolism, regulation of hormone levels, and a hormone-mediated signaling pathway. qRT-PCR validation showed a highly significant correlation (p < 0.01) with the RNA-Seq generated data. High-performance liquid chromatography (HPLC) and qRT-PCR results showed that, after decapitation, the NPA- and TIBA-induced lateral buds germinated due to rapidly decreasing auxin levels, caused by upregulation of the dioxygenase for auxin oxidation gene (DAO). Decreased auxin levels promoted the expression of isopentenyl transferase (IPT) and cytochrome P450 monooxygenase, family 735, subfamily A (CYP735A) genes and inhibited two carotenoid cleavage dioxygenases (CCD7 and CCD8). Zeatin levels significantly increased after the treatments. The increased cytokinin levels promoted the expression of WUSCHEL (WUS) and inhibited expression of BRANCHED1 (BRC1) in the cytokinin signal transduction pathway and initiated lateral bud outgrowth. Our data suggest that our theories concerning the regulation of shoot branching and apical dominance is really similar to those observed in annual plants. Auxin inhibits bud outgrowth and tends to inhibit cytokinin levels. The pseudobulb in the plant behaves in a similar manner to that of a shoot above the ground.

Arbuscular mycorrhizal fungi alleviate root damage stress induced by simulated coal mining subsidence ground fissures

Coal mining results in surface subsidence and induces the development of ground fissures that damage surrounding plant roots. Very few studies have explored the stress of root damage caused by ground fissures and whether arbuscular mycorrhizal fungi (AMF) can relieve root damage stress induced by ground fissures. In the present study we simulated ground fissure induced root damage, examined the resultant changes in endogenous hormones, root system morphology, leaf area, leaf chlorophyll content, nutrient content and biomass of maize, and examined the ameliorative effects of AMF on maize with root damage. Ground fissures led to significantly higher levels of endogenous abscisic acid (ABA) but significantly reduced levels of indole-3-acetic acid (IAA), gibberellins (GA) and cytokinin (CTK). In addition, ground fissures led to significantly reduced root biomass, total root length, root tip number, total root volume, plant nutrient content, leaf chlorophyll content and leaf area. The shoot biomass of root damaged maize decreased significantly by 46%. By contrast, AMF increased IAA and CTK levels in maize roots, reduced ABA levels, improved the hormone balance of damaged plants, increased total root length, root tip number, total root volume, leaf area and leaf chlorophyll content, increased nutrient content and increased shoot biomass by 34%. Overall, by simulating coal mining subsidence ground fissures, the study investigated the effects of root damage stress on plant biomass, found that AMF can alleviate the mechanical damages to the root system, and provided a theoretical basis for microbial remediation in areas subject to subsidence due to coal mining.

Effect of arsenate on endogenous levels of cytokinins with different existing forms in two Pteris species

Our previous results showed that content of trans-zeatin (tZ) increases in leaves of heavy metal hyperaccumulators but decreases in non-hyperaccumulators growing in multiple heavy metal polluted soils. However, the relationship between arsenic (As) accumulation and endogenous forms of cytokinins (CTKs) in As hyperaccumulators remains unknown. Here a hydroponic experiment was conducted to compare the CTK forms in the As hyperaccumulator Pteris cretica var. nervosa and non-hyperaccumulator Pteris ensiformis under arsenate stress (0, 2, 5, and 10 mg L−1). A simple and cost-effective procedure for the determination of CTK forms in plants was established, and a stepwise regression analysis was used to study the relationship among total As contents and different forms of endogenous CTKs in fronds of two plants. The results showed that the optimized chromatographic parameters were Zobax SB-C18 column (5 μm × 4.6 mm × 250 mm), UV detection detector at 269 nm, a flow rate of 0.6 mL min−1, constant temperature of 45 °C and gradient elution with methanol-acetonitrile-1% acetic acid. Contents of chlorophylls in the fronds of P. ensiformis were significantly decreased with addition of As compared to P. cretica var. nervosa. Furthermore, the total As content in fronds of P. cretica var. nervosa was positively correlated to the contents of N6-(2-isopentenyl) adenine-7-β-D-glucoside (iP7G) and N6-(2-isopentenyl) adenosine (iPR). However, the total As content in fronds of P. ensiformis was negatively correlated to its trans-zeatin riboside (ZR) content. Therefore, iP7G and iPR could positively improve As accumulation by P. cretica var. nervosa.

Natural variation in cytokinin maintenance improves salt tolerance in apple rootstocks

Plants experiencing salt-induced stress often reduce cytokinin levels during the early phases of stress-response. Interestingly, we found that the cytokinin content in the apple rootstock "robusta" was maintained at a high level under salt stress. Through screening genes involved in cytokinin biosynthesis and catabolism, we found that the high expression levels of IPT5b in robusta roots were involved in maintaining the high cytokinin content. We identified a 42bp deletion in the promoter region of IPT5b, which elevated IPT5b expression levels, and this deletion was linked to salt tolerance in robusta×M.9 segregating population. The 42bp deletion resulted in the deletion of a Proline Response Element (ProRE), and our results suggest that ProRE negatively regulates IPT5b expression in response to proline. Under salt stress, the robusta cultivar maintains high cytokinin levels as IPT5b expression cannot be inhibited by proline due to the deletion of ProRE, leading to improve salt tolerance.

Inhibition of adventitious root development in apple rootstocks by cytokinin is based on its suppression of adventitious root primordia formation.

Cytokinin (CK) inhibits adventitious root (AR) formation in stem cuttings. Little is known, however, about the mechanism underlying the inhibitory effect. In this study, 2mg l-1 of exogenous 6-benzyl adenine (6-BA) was administered to 3 and 7-day-old apple rootstocks 'M.26' cuttings (3 and 7 days 6-BA) by transferring them from a rooting medium containing indole-3-butanoic acid to the medium containing 6-BA. Anatomical and morphological observations revealed that the exogenous application of 6-BA inhibited primordia formation in the 3 days 6-BA but not the 7 days 6-BA group. The concentration of auxin (IAA), the ratios of IAA/CK and IAA/abscisic acid were lower in 3 days 6-BA than in 7 days 6-BA. Expression analysis of genes known to be associated with AR formation was also analyzed. In the 3 days 6-BA group, high level of CK inhibited the synthesis and transport of auxin, as a result, low endogenous auxin level suppressed the auxin signaling pathway genes, as were other AR development and cell cycle related genes; all of which had an inhibitory impact on AR primordium formation. On the contrary, low CK level in the 7 days 6-BA, reduced the inhibitory impact on auxin levels, leading to an upregulated expression of genes known to promote AR primordia formation. Collectively, our data indicated that 3-7 days is the time period in which AR primordia formation occurs in cuttings of 'M.26' and that the inhibition of AR development by CK is due to the suppression of AR primordia development over 3-7 days period after culturing in rooting medium.

Effect of constitutive expression of Arabidopsis CLAVATA3 on cell growth and possible role of cytokinins in leaf size control in transgenic tobacco plants

We generated transgenic tobacco plants (Nicotiana tabacum L.) with overexpression of the Arabidopsis thaliana CLAVATA3 (CLV3) gene which is known to be a negative regulator of cell division. Surprisingly, most of the 35S::CLV3 transgenic plants showed no phenotypic differences with the wild type plants. However, there were considerable changes in the morphological parameters between 35S::CLV3 overexpressors and wild type plants. As expected, the number of meristematic cells in the shoot apical meristem was reduced in 35S::CLV3 plants as compared to the wild type plants. Moreover, overexpression of CLV3 exerted morphological changes not only to shoot apical meristem but also to leaves and flowers. Thus, transgenic plants were characterized by reduced number of epidermal and mesophyll cells as well as stomatal pores in mature leaves. However, there was a compensatory increase in leaf cell size of 35S::CLV3 plants that contributed to maintenance of organ size within the normal range. We observed that expression of cell expansion-promoted genes, expansin NtEXPA4 and endo-xyloglucan transferase NtEXGT, were elevated in mature leaves. In contrast, there was a decrease in the transcript level of the cell division-related AINTEGUMENTA-like (NtANTL) gene in 35S::CLV3 transgenic plants. In addition, we detected an increase in cytokinin level without any changes in the contents of IAA and ABA in 35S::CLV3 overexpressors. Interestingly, cytokinin treatment was shown to stimulate the expression of NtEXPA4 and NtEXGT genes in 35S::CLV3 transgenic plants. We propose that observed compensatory cell expansion in leaves of 35S::CLV3 transgenic plants may be due, at least in part, to a possible link between cytokinin signalling and cell expansion-related genes.

Correlations between Phytohormones and Drought Tolerance in Selected Brassica Crops: Chinese Cabbage, White Cabbage and Kale.

Drought is one of the major abiotic stresses affecting the productivity of Brassica crops. To understand the role of phytohormones in drought tolerance, we subjected Chinese cabbage (B. rapa ssp. pekinensis), white cabbage (B. oleracea var. capitata), and kale (B. oleracea var. acephala) to drought and examined the stress response on the physiological, biochemical and hormonal levels. The phytohormones abscisic acid (ABA), auxin indole-3-acetic acid (IAA), brassinosteroids (BRs), cytokinins (CKs), jasmonates (JAs), and salicylic acid (SA) were analyzed by ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS). Based on the physiological and biochemical markers the Chinese cabbage exhibited the lowest tolerance, followed by the white cabbage, while the kale appeared to be the most tolerant to drought. The drought tolerance of the kale correlated with increased levels of SA, ABA, IAA, CKs iP(R) and cZ(R), and typhasterol (TY), a precursor of active BRs. In contrast, the drought sensitivity of the Chinese cabbage correlated with a significant increase in ABA, JAs and the active BRs castasterol (CS) and brassinolide (BL). The moderately tolerant white cabbage, positioned between the kale and Chinese cabbage, showed more similarity in terms of the phytohormone patterns with the kale. We concluded that the drought tolerance in Brassicaceae is mostly determined by the increased endogenous levels of IAA, CKs, ABA and SA and the decreased levels of active BRs.

Hormonal interactions during early physiological partenocarpic fruitlet abscission in persimmon (Diospyros Kaki Thunb.) ‘Triumph’ and ‘Shinshu’ cultivars

Fruitlet abscission is typical to many fruit trees, including persimmon, (Diopyros kaki Thunb.), and often results in unstable fruit production and yield. We documented massive parthenocarpic fruitlet abscission in the persimmon cultivars 'Triumph' and 'Shinshu', which initiated soon after anthesis and peaked at 45 days post anthesis. In contrast, abscission of mature fruitlets 45–65 days after anthesis was minor. The early parthenocarpic fruitlet abscission in 'Shinshu' and 'Triumph' took place in the abscission zone located at the calyx-pedicel junction. Differentiation of the abscission layer occurred during flower development and preceded fruitlet abscission. Endogenous analysis of gibberellins, cytokinins and auxin levels in the abscission zone of 'Triumph' and 'Shinshu' during the developmental stages preceding abscission revealed a reduction in cytokinin endogenous levels in both cultivars. In addition, a decline in GA3 and IAA levels was found in 'Triumph' and 'Shinsu' respectively during flower development. Gibberellin, auxin and cytokinin application to inhibit early fruitlet abscission were analyzed and gibberellin treatment exhibited the highest and most consistent inhibition effect in 'Triumph' and 'Shinshu'. An increase in endogenous levels of gibberellins, cytokinins and auxin in the flower abscission zone was found following GA application in both cultivars. Our results suggest that gibberellin application inhibits early fruitlet abscission by elevating cytokinins, and auxin levels and that a reduction in gibberellins, cytokinins and auxin levels may induce abscission during parthenocarpic fruitlet development in 'Shinshu' and 'Triumph' cultivars.

The effect of auxin and strigolactone on ATP/ADP isopentenyltransferase expression and the regulation of apical dominance in peach

We confirmed the roles of auxin, CK, and strigolactones in apical dominance in peach and established a model of plant hormonal control of apical dominance in peach. Auxin, cytokinin, and strigolactone play important roles in apical dominance. In this study, we analyzed the effect of auxin and strigolactone on the expression of ATP/ADP isopentenyltransferase (IPT) genes (key cytokinin biosynthesis genes) and the regulation of apical dominance in peach. After decapitation, the expression levels of PpIPT1, PpIPT3, and PpIPT5a in nodal stems sharply increased. This observation is consistent with the changes in tZ-type and iP-type cytokinin levels in nodal stems and axillary buds observed after treatment; these changes are required to promote the outgrowth of axillary buds in peach. These results suggest that ATP/ADP PpIPT genes in nodal stems are key genes for cytokinin biosynthesis, as they promote the outgrowth of axillary buds. We also found that auxin and strigolactone inhibited the outgrowth of axillary buds. After decapitation, IAA treatment inhibited the expression of ATP/ADP PpIPTs in nodal stems to impede the increase in cytokinin levels. By contrast, after GR24 (GR24 strigolactone) treatment, the expression of ATP/ADP IPT genes and cytokinin levels still increased markedly, but the rate of increase in gene expression was markedly lower than that observed after decapitation in the absence of IAA (indole-3-acetic acid) treatment. In addition, GR24 inhibited basipetal auxin transport at the nodes (by limiting the expression of PpPIN1a in nodal stems), thereby inhibiting ATP/ADP PpIPT expression in nodal stems. Therefore, strigolactone inhibits the outgrowth of axillary buds in peach only when terminal buds are present.

Identification and Characterization of ATP/ADP Isopentenyltransferases (ATP/ADP PpIPTs) Genes in Peach

ATP/ADP isopentenyltransferase (IPTs) genes encode key enzymes involved in cytokinin synthesis. In this study, the functions of ATP/ADP PpIPTs in peach were investigated. According to the genome sequence, we have found and verified that there are four members of this gene family in peach, namely, PpIPT1, PpIPT3, PpIPT5, and PpIPT7. Overexpression of each of these genes in Arabidopsis resulted in increased levels of cytokinins in the transgenic plants, confirming their roles in cytokinin synthesis. Numerous altered phenotypes were observed in the transgenic plants, including vigorous growth and enhanced salt resistance. ATP/ADP PpIPTs were expressed in tissues throughout the plant, but the expression patterns differed between the genes. Only PpIPT3 was upregulated within 2 h after the application of nitrate to N-deprived peach seedlings, and the increase was resistant to pre-treatment of a specific nitrate metabolism inhibitor. Results showed that ATP/ADP PpIPT expression levels decreased significantly in pulp within 2 weeks after flowering and remained low. However, pulp cytokinin levels were quite high during this time. Only PpIPT5 in seed increased significantly within 2 weeks after flowering, which was consistent with cytokinin levels during early fruit development, suggesting that PpIPT5 in seed is the key gene for cytokinin biosynthesis during early fruit development. ATP/ADP PpIPT expression also increased significantly during later fruit development in seed.

A 22-bp deletion in OsPLS3 gene encoding a DUF266-containing protein is implicated in rice leaf senescence.

Key message The OsPLS3 locus was isolated by map-based cloning that encodes a DUF266-containing protein. OsPLS3 regulates the onset of leaf senescence in rice. Glycosyltransferases (GTs) are one of the most important enzyme groups required for the modification of plant secondary metabolites and play a crucial role in plant growth and development, however the biological functions of most GTs remain elusive. We reported here the identification and characterization of a novel Oryza sativa premature leaf senescence mutant (ospls3). Through map-based cloning strategy, we determined that 22-bp deletion in the OsPLS3 gene encoding a domain of unknown function 266 (DUF266)-containing protein, a member of GT14-like, underlies the premature leaf senescence phenotype in the ospls3 mutant. The OsPLS3 mRNA levels progressively declined with the age-dependent leaf senescence in wild-type rice, implying a negative role of OsPLS3 in regulating leaf senescence. Physiological analysis, and histochemical staining and transmission electron microscopy assays indicated that the ospls3 mutant accumulated higher levels of ethylene and reactive oxygen species than its wild type. Furthermore, the ospls3 mutant showed hypersensitivity to exogenous 1-aminocyclopropane-1-carboxylic acid, H2O2 and high level of cytokinins. Our results indicated that the DUF266-containing gene OsPLS3 plays an important role in the onset of leaf senescence, in part through cytokinin and ethylene signaling in rice.