Stem girdling influences concentrations of endogenous cytokinins and abscisic acid in relation to leaf senescence in cotton

Many studies have shown that root–shoot imbalance influences vegetative growth and development of cotton (Gossypium hirsutum L.), but few have examined changes in leaf senescence and endogenous hormones due to stem girdling. The objective of this study was to determine the correlation between some endogenous phytohormones, particularly cytokinins and abscisic acid (ABA), and leaf senescence following stem girdling. Field-grown cotton plants were girdled on the main stem 5 days after squaring (DAS), while the non-girdled plants served as control. Plant biomass, seed cotton yield, main-stem leaf photosynthetic (Pn) rate, chlorophyll (Chl) and malondialdehyde (MDA) concentrations, as well as levels of cytokinins and ABA in main-stem leaves and xylem sap were determined after girdling or at harvest. Main-stem girdling decreased the dry root weight and root/shoot ratio from 5 to 70 days after girdling (DAG) and reduced seed cotton yield at harvest. Main-stem leaf Pn and Chl concentration in girdled plants were significantly lower than in control plants. Much higher levels of MDA were observed in main-stem leaves from 5 to 70 DAG, suggesting that stem girdling accelerated leaf senescence. Girdled plants contained less trans-zeatin and its riboside (t-Z + t-ZR), dihydrozeatin and its riboside (DHZ + DHZR), and isopentenyladenine and its riboside (iP + iPA), but more ABA than control plants in both main-stem leaves and xylem sap. These results suggested that main-stem girdling accelerated leaf senescence due to reduced levels of cytokinin and/or increased ABA. Cytokinin and ABA are involved in leaf senescence following main-stem girdling.     

Effects of early-fruit removal on endogenous cytokinins and abscisic acid in relation to leaf senescence in cotton

Numerous studies have shown that early-fruit removal enhances vegetative growth and development of cotton (Gossypium hirsutum L.). However, few studies have examined changes in leaf senescence and endogenous hormones due to fruit removal. The objective of this study was to determine the correlation between some endogenous phytohormones, particularly the cytokinins and abscisic acid (ABA), and leaf senescence following fruit removal. Cotton was grown in pots and in the field during 2005 and 2006. Two early-fruiting branches were excised from plants at squaring to form the fruit removal treatment while the non-excised plants served as control. Plant biomass, seed cotton yield, cytokinins and ABA levels in main-stem leaves and xylem sap as well as main-stem leaf photosynthetic rate (Pn) and chlorophyll (Chl) concentration were determined after removal or at harvest. Fruit removals increased the leaf area, root and shoot dry weight and plant biomass at 35 days after removal (DAR), whether in potted or field-grown cotton; under field conditions, it also improved plant biomass and seed cotton yield at harvest. The Pn and Chl concentration in excised plants were significantly higher than in control plants from 5 to 35 DAR, suggesting that fruit removal considerably delayed leaf senescence. Fruit-excised plants contained more trans-zeatin and its riboside (t-Z + t-ZR), dihydrozeatin and its riboside (DHZ + DHZR), and isopentenyladenine and its riboside (iP + iPA) but less ABA in both main-stem leaves and xylem sap than control plants from 5 to 35 DAR. These results suggest that removal of early fruiting branches delays main-stem leaf senescence, which can be attributed to increased cytokinin and/or reduced ABA. Cytokinin and ABA are involved in leaf senescence following early fruit removal.     

Comparison of Whole System of Carbon and Nitrogen Accumulation Between Two Maize Hybrids Differing in Leaf Senescence

A field experiment was conducted to investigate the carbon (C) and nitrogen (N) balance in relation to grain formation and leaf senescence in two different senescent types of maize (Zea mays L.), one stay-green (cv. P3845) and one earlier senescent (cv. Hokkou 55). In comparison with Hokkou 55, P3845 had a higher N concentration (N_c) in the leaves and a higher specific N absorption rate by roots (SAR_N), which indicated that a large amount of N was supplied to the leaves from the roots during maturation. This resulted in a higher photosynthetic rate, which supports saccharide distribution to roots. Thus, stay-green plants maintained a more balanced C and N metabolism between shoots and roots. Moreover, the coefficients of the relationship between the relative growth rate (RGR) and N_c, and between the photon-saturated photo-synthetic rate (P _sat) and N_c were lower in P3845. The P _sat per unit N_c in leaves was lower in the stay-green cultivars, which indicated that high yield was attained by longer green area duration and not by a high P _sat per unit N_c in the leaf. Consequently, a high P_sat caused a high leaf senescence rate because C and N compounds will translocate actively from the leaves.     

Changes of Photosynthetic Characteristics in Relation to Leaf Senescence in Two Maize Hybrids with Different Senescent Appearance

A field experiment was conducted to investigate the changes in chlorophyll (Chl) and nitrogen (N) contents, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) and phosphoenolpyruvate carboxylase (PEPC) contents and PEPC activity, and the photon-saturated net photosynthetic rate (P _Nsat), and their relationships with leaf senescence in two maize hybrids with different senescent appearance. One stay-green (cv. P3845) and one earlier senescent (cv. Hokkou 55) hybrid were used in this study, and we found that Chl and N contents and the P _Nsat in individual leaves of P3845 were greater than those in corresponding leaves of Hokkou 55 at the successive growth stages. In addition, larger contents of RuBPCO and PEPC, and a greater activity of PEPC were observed in P3845. Due to the lower rates of decrease of Chl, RuBPCO, and PEPC amounts per unit of N, and the lower net C translocation rate per unit of N in the stay-green hybrid, leaf senescence was delayed in comparison to the earlier senescent hybrid.     

Effects of cotton rootstock on endogenous cytokinins and abscisic acid in xylem sap and leaves in relation to leaf senescence

Leaf senescence varies greatly among cotton cultivars, possiblydue to their root characteristics, particularly the root-sourcedcytokinins and abscisic acid (ABA). Early-senescence (K1) andlate-senescence (K2) lines, were reciprocally or self-graftedto examine the effects of rootstock on leaf senescence and endogenoushormones in both leaves and xylem sap. The results indicatethat the graft of K1 scion onto K2 rootstock (K1/K2) alleviatedleaf senescence with enhanced photosynthetic (Pn) rate, increasedlevels of chlorophyll (Chl) and total soluble protein (TSP),concurrently with reduced malondialdehyde (MDA) contents inthe fourth leaf on the main-stem. The graft of K2 scion ontoK1 rootstock enhanced leaf senescence with reduced Pn, Chl,and TSP, and increased MDA, compared with their respective self-graftedcontrol plants (K1/K1 and K2/K2). Reciprocally grafted plantsdiffered significantly from their self-grafted control plantsin levels of zeatin and its riboside (Z+ZR), isopentenyl andits adenine (iP+iPA), and ABA, but not in those of dihydrozeatinand its riboside (DHZ+DHZR) in leaves in late season, whichwas consistent with variations in leaf senescence between reciprocallyand self-grafted plants. The results suggest that leaf senescenceis closely associated with reduced accumulation of Z+ZR, andiP+iPA rather than DHZ+DHZR, or enhanced ABA in leaves of cotton.Genotypic variation in leaf senescence may result from the differencein root characteristics, particularly in Z+ZR, iP+iPA, and ABAwhich are regulated by the root system directly or indirectly. Key words: Abscisic acid, cotton, cytokinins, grafting, leaf senescence Received 23 October 2007; Revised 17 January 2008 Accepted 23 January 2008     

源库关系改变对棉叶内源激素的影响

以中棉18号为材料,在大田条件下研究了摘蕾引起源库关系改变对主要源叶（对应的主茎叶和对位果枝叶）中内源激素的影响。结果表明;去蕾处理推迟了相应主茎叶和果枝叶中吲哚乙酸（IAA）峰出现,降低了玉米素及其核苷（Z ZR）含量,推迟了果枝叶中异戊烯基腺嘌呤及其核苷（iP iPA)的积累,提前了二氢玉米素及其核苷（DHZ DHZR）的积累,降低了脱落酸（ABA）含量。去除一个蕾对相应叶片内源激素的影响是短期的,随整株源库关系的协调,与对照趋同。激素的这种变化与叶片功能变化是一致的,表明棉花源库关系的协调不仅是物质上的关系,植物激素可能是其中重要的信息调控系统。     

Plant Growth Regulators and Induction of Leaf Senescence in Nitrogen-Deprived Wheat Plants

The sequence of events and the signals that regulate the remobilization of nitrogen (N) reserves during senescence induced by N starvation were studied in leaf 3, the last fully expanded leaf, in 17-day-old wheat (Triticum aestivum L.) plants. The first event observed was a rapid decrease in the isopentenyl adenosine (iPA) concentration during the first 24 h of N starvation. No differences in t-zeatin riboside and dihydrozeatin riboside concentrations were observed until the end of the assay. During the following 6 days, a decrease in soluble amino acids, chlorophyll, and protein, as well as an increase in soluble sugar concentration and endoproteolytic activity, could be observed. At day 3 of the experiment, the abscisic acid (ABA) concentration in the leaves of N-deprived plants started to increase. After 6 days of N deprivation there was a rise in oxidative stress, as indicated by the increase in malondialdehyde concentration, as well as a decrease in the activities of antioxidant enzymes catalase and ascorbate peroxidase. To analyze interactions with leaf development, the first, second, third, and fourth leaves were studied. iPA concentration decreased in all the leaf stages, including leaf 4, which was not fully expanded. A linear correlation between iPA and protein concentration was determined. These results suggest that the sharp fall in iPA could be the earliest event that induces protein degradation during the development of senescence induced by N deficiency, and that only later is ABA accumulated and oxidative stress developed.     

Ability of bacterium Bacillus subtilis to produce cytokinins and to influence the growth and endogenous hormone content of lettuce plants

Hormone production by micro-organisms selected as antagonists of pathogenic fungi and the effect of their introduction into soil on hormone content and growth of lettuce plants were studied. Hormones in bacterial cultural media and in plant extracts were immunopurified and assayed using specific antibodies to indolyl-3-acetic acid (IAA), abscisic acid (ABA), and different cytokinins (zeatin riboside (ZR), dihydrozeatinriboside (DHZR) and isopentenyladenosine (iPA)). ZR was shown to be the main cytokinin present in bacterial cultural media as a complex with a high molecular weight component. Inoculation of lettuce plants with bacteria increased the cytokinin content of both shoots and roots. Accumulation of zeatin and its riboside was greatest in roots shortly 2days after inoculation, when their content was 10 times higher than in control. Changes in the content of other hormones (ABA and IAA) were observed at the end of experiments only. Accumulation of cytokinins in inoculated lettuce plants was associated with an increase in plant shoot and root weight of approximately 30% over 8days.     

The Importance of Roots in Regulating the Senescence of Soybean Primary Leaves

The role of roots in regulating primary leaf senescence of 14-day-old soybean seedlings was investigated. Compared with intact seedlings, the senescence of primary leaves is accelerated by removal of the root system but delayed if apical bud and the first trifoliate leaf are removed. No difference in senescence was found between intact seedlings and seedlings without roots, apical bud, and first trifoliate leaf. Lateral roots seem to play a predominant role in regulating primary leaf senescence. However, neither root nodules nor primary root play any function in senescence. Results indicate that benzyladenine (BA) at optimal concentration (2 mg/1) completely replaces the roots to prevent the senescence of primary leaves, whereas gibberellic acid (GA) and abscisic acid (ABA) accelerate. The effect of indole-3-acetic acid (IAA) to replace roots in preventing senescence depends on the season the young seedlings are grown. Additional, though indirect, information of acropetal transport of ABA is provided. In conclusion, it seems that cytokinins in lateral roots play a predominant role in leaf senescence and the normal supply of root cytokinins is important in leaf metabolism.     

Effects of elevated carbon dioxide and/or ozone on endogenous plant hormones in the leaves of <Emphasis Type="Italic">Ginkgo biloba</Emphasis>

Four-year-old Gingko (Ginkgo biloba L.) trees were exposed to ambient and elevated concentrations of CO₂ and O₃, and their combination for 1 year, using open-top chambers in Shenyang, China in 2006. Growth parameters and endogenous plant hormones were measured simultaneously over the experiment period. Elevated CO₂ increased leaf area and leaf dry weight but had no effect on shoot length, increased indole-3-acetic acid (IAA), gibberellins A₃ (GA₃), zeatin riboside (ZR), dihydrozeatin (DHZR) and isopentenyl-adenosine (iPA) content but decreased abscisic acid (ABA) content. Elevated O₃ significantly decreased leaf area, leaf dry weight, shoot length, and decreased IAA, GA₃, ZR content but increased ABA content and had a little effect on iPA, DHZR content. Elevated CO₂ + O₃ decreased IAA, iPA and DHZR content while increased ABA and GA₃ content in the early stage of the exposure and then decreased in the late stage. The evidence from this study indicates that elevated CO₂ ameliorated the effects of elevated ozone on tree growth, and elevated CO₂ may have a largely positive impact on forest tree growth while elevated O₃ will likely have a negative impact.     

Immunoaffinity Chromatographic and ELISA determination of Cytokinins in Germinating Pollens in Lilium davidii Dach.

Liliurn davidii Duch. pollen was germinated in PEG-400. A rapid immunoaffinity chromatograpy system with high efficiency and specificity was adopted to isolate and purify trans-zeatin riboside (t-ZR) and isopentenyladenosine (ipA). Cytokinin determination was carried out by a highly sensitive enzyme linked immunosolvent assay (ELISA) for t-ZR and iPA respectively, t-ZR and iPA were present at the level of 10-8 g/g fr. wt in Lilium davidii pollen. After hydration, t-ZR content of pollen decreased slightly, while iPA increased remarkably, and the total amount of t-ZR and iPA underwent almost no change. Pollen tubes grew fast in the first 3 h after germination, and the amount of t-ZR and iPA also increased greatly both in pollen tubes and in germination medium. The increasing trend of t-ZR and iPA and growth rate of pollen tubes was basically synchronous.     

Cytokinin oxidase activity and cytokinin content in roots of sunflower under water stress

Contents of trans-zeatin riboside (ZR), dihydrozeatin riboside (DZR) and N6-(delta2-isopentenyl) adenosine (iPA) was quantified by an indirect ELISA using polyclonal antibodies, in the roots, xylem sap and leaves of pot grown sunflower plants subjected to water stress (RWC of leaves approximately 65 per cent). The delivery rates of all three cytokinins decreased significantly under stress. Cytokinin levels also decreased in roots and in leaves of stressed plants. Three-fold increase in cytokinin oxidase activity was observed in stressed roots after polymin P-ammonium sulphate fractionation. Further purification using Con A agarose resulted in elution of protein with cytokinin oxidase activity and was found to be 30 kDa protein on SDS-PAGE.     

Fluctuation in levels of endogenous plant hormones in ovules of normal and mutant cotton during flowering and their relation to fiber development

The contents of indole-3-acetic acid (IAA), gibberellins (GAs), abscisic acid (ABA), and cytokinins were determined in ovules of normal cotton (Tm-1) and a kind of fiber differentiation mutant (Xin) before and after flowering by enzyme-linked immunosorbent assays. It was found that 24 h before flowering, a peak of IAA content was observed in ovules of Tm-1, whereas in ovules of Xin, a low level of IAA was determined. From –1 day (1 day before flowering) to +3 days (3 days after flowering), GA₁₊₃ levels in ovules of Xin were 40–70% lower than those of Tm-1; GA₄₊₇ levels were very low, and there was no visible difference in GA₄₊₇ content between normal and mutant cotton. The ABA content in ovule of Tm-1 decreased by 70% 3 days after flowering, whereas that of Xin only decreased by 20%. The levels of cytokinins in ovules of Tm-1 decreased after flowering, and those of Xin kept up a steady increase.Abbreviations IAA indole-3-acetic acid - GA gibberellin - ABA abscisic acid - ELISA enzyme-linked immunosorbent assay - FW fresh weight - PBS phosphate-buffered saline - iPA isopentenyladenosine - ZR zeatin riboside - DHZR dihydrozeatin riboside - CTK cytokinin     

The effect of aluminum on cytokinins in the mycelia of Amanita muscaria

High performance liquid chromatography analysis of immunoaffinity-purified extracts of mycelia of Amanita muscaria, and the Amaranthus bioassay of the eluted fractions, revealed the following seven cytokinins: zeatin, zeatin riboside, zeatin N-9-glucoside, dihydrozeatin, dihydrozeatin riboside, isopentenyl adenine, and isopentenyl adenosine. The decreased growth of aluminum-treated mycelia correlated with a 35% decrease in the total amount of the cytokinins. Among individual cytokinins, zeatin was the most affected, exhibiting a reduction of about 90%. The results are compared with previous investigations of aluminum effects on cytokinins in the mycelia of Lactarius piperatus, whose growth is stimulated by aluminum.Abbreviations ZR zeatin riboside - iPA isopentenyl adenosine - Z zeatin - DHZ dihydrozeatin - iP isopentenyl adenine - DHZR dihydrozeatin riboside - Z-9G zeatin N-9-glucoside - iP-9G isopentenyl N-9-glucoside - HPLC high performance liquid chromatography - DHZRMP dihydrozeatin riboside monophosphate - ZRMP zeatin riboside monophosphate     

Cytokinin biochemistry in relation to leaf senescence. VII. Endogenous cytokinin levels and exogenous applications of cytokinins in relation to sequential leaf senescence of tobacco

The cytokinin complex in tobacco leaves of various maturities was characterized by radioimmunoassay and mass spectrometry. Zeatin was the major base, whereas zeatin riboside was identified as the main riboside. in leaves of all maturities studied. Relative to upper younger leaves, the basal yellow leaves had reduced levels of both cytokinin bases and ribosides. Exogenous applications of dihydrozeatin and zeatin to detached tobacco leaves in amounts sufficient to delay senescence, elevated cytokinin base and riboside levels 2–5 fold. Presenescent and senescent leaves of intact plants showed quantitatively similar changes in cytokinin content. which therefore appear to be of significance in control of senescence. When supplied exogenously, the principal cytokinin bases found to occur in tobacco leaves (zeatin and dihydrozeatin) were markedly more effective than auxins and gibberellic acid in retarding senescence. Localised application of cytokinins to leaf blades of detopped plants was much less effective than application to intact plants. The cytokinin induced senescence retardation in tobacco leaves was independent of effects on directed metabolite transport. Evidence that endogenous levels of active cytokinins in intact tobacco leaves are involved in control of sequential leaf senescence is discussed.     

Root physiological factors involved in cool-season grass response to high soil temperature

High soil temperature is a critical factor limiting growth of cool-season grasses. This study was designed to examine changes in water, nutritional, and hormonal status in response to high soil temperature for creeping bentgrass (Agrostis stoloniferavar. palustris) and to compare the sensitivity of those parameters to high soil temperatures. Plants of ‘Penncross’ were exposed to 35 °C soil temperature (heat stress) or 20 °C (control) in water baths while air temperature was maintained at 20 °C in growth chambers. Turfgrass quality, shoot growth rate, and root biomass decreased below the control levels at 15, 15, and 10 days of heat stress, respectively, while root mortality increased above the control level at 5 days of heat stress. Relative water content (RWC) of leaves decreased below the control level at 15 days of heat treatment. Root N content increased while P and K content did not change over time at 35 °C. Shoot N, P, and K content decreased below the control level at 15, 15, and 10 days of heat stress, respectively. Root abscisic acid (ABA) content decreased below the control level at 10 days while shoot ABA content increased above the control level at 5 days. The content of cytokinins (zeatin (Z) and zeatin riboside (ZR), dihydrogen zeatin riboside (DHZR), and isopentenyl adenosine (iPA)) decreased below their respective control levels as early as 5 days of heat stress for roots and 10 days for shoots. The decline in cytokinin content was also more dramatic than changes in other parameters. Our results suggested that cytokinin was most sensitive to high soil temperature among parameters examined, suggesting that changes in cytokinins could serve as an early stress indicator for plant responses to high soil temperature; however, decreased water, nutrient (N, P, and K), and cytokinin content, and increased ABA could all contribute to the decline in shoot and root growth for creeping bentgrass exposed to high soil temperatures.     

Root Hypoxia-lnduced Changes in the Pattern of Translatable mRNAs In Poplar Leaves

Shoot responses to root stress may be modulated by hormonestransported from root tissues. In this study, the pattern ofin vitro translatable mRNAs from leaves on plants with oxygen-deprived(hypoxic) roots was used as a molecular phenotype to test forthe action and specificity of putative root-to-shoot signalmolecules. Root hypoxia caused a decrease in the abundance ofa group of leaf mRNAs which translated to give low molecularweight (LMW) proteins (12 to 28 kDa). Root excision induceda similar reduction in a subset of the same LMW leaf mRNAs.This suggests that the effects of root hypoxia on leaves arephysiologically related to the effects of root excision. Feedingabscisic acid to leaves did not mimic the effects of root aerationor hypoxia. Applications of zeatin riboside (ZR) to leaves ofplants with hypoxic roots duplicated the effects of root aerationon the LMW translation products. The specificity of the leafresponse to ZR was tested by feeding KC1 and sucrose to leavesof root excised plants. Applications of either KC1 or sucrosealso increased the abundance of the LMW translation products.Although the results reported in this paper are consistent withthe concept that shoot responses to root stress may be modulatedby substances moving in the xylem, a thorough understandingof how hormones control cellular responses needs to be resolvedbefore the mechanism of root-shoot integration can be understood. Key words: ABA, cytokinins, hypoxia, poplar     

Cytokinin biochemistry in relation to leaf senescence. VIII. Translocation, metabolism and biosynthesis of cytokinins in relation to sequential leaf senescence of tobacco

Cytokinin bases (zeatin and dihydrozeatin) and ribosides (zeatin riboside and dihydrozeatin riboside) were identified as major cytokinins in tobacco xylem sap by radioimmunoassay. When ³H-labelled zeatin riboside or dihydrozeatin riboside were supplied to tobacco plants via the xylem, leaves of differing maturity did not differ appreciably in level of radioactivity or in metabolism of the cytokinin. The major metabolites of zeatin riboside in leaves were adenine, adenosine and adenine nucleotides, whereas that of dihydrozeatin riboside was dihydrozeatin 7-glucoside. Incorporation of [¹⁴C]adenine into zeatin was evident in upper green leaves. indicating that young leaves have the capacity to synthesize cytokinins in situ. In contrast, fully expanded green leaves and senescing tobacco leaves exhibited little or no incorporation of [¹⁴C]adenine into cytokinins. This difference in cytokinin biosynthetic capacity may contribute to the differing cytokinin levels in leaves of different matirity, and may participate in control of sequential leaf senescence in tobacco.     

Trichoderma harzianum strain T-22 induces changes in phytohormone levels in cherry rootstocks (Prunus cerasus?×?P. canescens)

The aim of this research was to explain the direct plant growth-promoting activity of Trichoderma harzianum strain T-22 (T22), hypothesizing the involvement of different classes of plant growth regulators. Seven days after the transfer to root-inducing medium, in vitro-cultured shoots of GiSeLa6^? (Prunus cerasus  × P. canescens) were inoculated with T22. Root and shoot growth were significantly affected by T22 (+76 and +61%, respectively). Ten days after inoculation, the levels of indole-3-acetic acid (IAA), trans-zeatin riboside (t-ZR), dihydrozeatin riboside (DHZR), gibberellic acid (GA3) and abscisic acid (ABA) were analyzed by high performance liquid chromatography coupled with mass spectrometry. The results showed that after T22-inoculation, IAA and GA3 significantly increased in both leaves (+49 and +71%, respectively) and roots (+40 and +143%, respectively) whereas t-ZR decreased (−51% in leaves and −37% in roots). Changes in DHZR were observed in T22-inoculated roots (−32%) but not in leaves, whereas the levels of ABA did not differ between the two treatments. The extraction method allowed the simultaneous extraction of phytohormones. There is evidence that the change in phytohormone levels is one of the direct mechanism by which T22 promotes rooting and shoot growth, with notable advantages for rootstock production during nursery processes.     

Cytokinins from terminal buds of Euphoria longana during different growth stages

The presence of endogenous cytokinins were detected in the terminal buds of longan ( Euphoria longana Lam.) after purification by ion exchange and Sephadex LH-20 chromatography, and bioassay, enzymic degradation, high-performance liquid chromatography and gas chromatography-mass spectrometry. Permethylated derivatives of two highly active cytokinin glucoside compounds from dormant buds were: 6-(4-O-β-D-glucosyl-3-methyl-but-2-enylamino) purine (zeatin-O-glucoside) and 9-β-D-ribofuranosyl-6-(4-hydroxy-3-methyl-but-2-enylamino) purine (zeatin riboside-O-glucoside). Simultaneously, four active cytokinins from buds at the stages of leaf flush and flower bud initiation were identified as 6-(4-hydroxy-3-methyl-but- trans -2-enylamino) purine (zeatin), zeatin-9-β-D-ribofuranosylpurine (zeatin riboside), 6-(3-methyl-2-butenyl) aminopurine (isopentenyladenosine, 2iPA) and N-(3-methyl-2-butenyl) adenine (isopentenyladenine, 2iP). The total cytokinin levels were low at leaf flush, with the zeatin and zeatin riboside in the buds about 70% of the total. In the transition of the terminal bud from leaf flush to dormancy, the principal cytokinins were zeatin-O-glucoside and zeatin riboside-O-glucoside. However, significant decreases in the levels of zeatin-O-glucoside and zeatin riboside-O-glucoside and increases in those of zeatin, zeatin riboside, 2iPA and 2iP were observed at flower bud initiation. It is suggested that in longan, the cytokinins that are translocated to the shoots are accumulated in the buds at the dormant stage, and that later there is a considerable increase in free cytokinins during flower bud initiation, leading to the promotion of flower bud development.