儿茶素对蝴蝶兰叶片离体培养褐变发生的影响

用0.3 g/L儿茶素处理蝴蝶兰叶片外植体,在MS+3 mg/L 6-BA(pH 5.8)培养基上培养4 d,外植体褐变率高于对照227.6%.用香草醛-盐酸溶液染色反应法发现鞣质分布在维管束和细胞间隙.叶片剪切的外植体(培养0 d)鞣质含量较高,培养4 d降低,8 d后再升高.儿茶素处理的外植体鞣质含量明显高于对照.在培养期间,儿茶素处理的外植体PPO、POD和PAL活性高于对照外植体,其中POD活性在第8天达到最大值,PPO和PAL活性在培养第12天达到高峰.     

Plant regeneration of Nerium oleander L. from alginate-encapsulated shoot explants after short-term cold storage

In this study, an efficient protocol for the regeneration of encapsulated explants of oleander (Nerium oleander L.) has been developed. Shoot tips and 1st nodal segments below the shoot tip, from in vitro-derived oleander microshoots, were encapsulated in 2.5% sodium alginate prepared in liquid MS sucrose-free nutrient medium and hardened in 50 mM of calcium chloride producing solid beads, uniform in shape. These artificial seeds, irrespective of their maintenance under light or in darkness, germinated at frequencies of 38.8–42.2%, producing 3.0–3.3 microshoots per bead. In the case of using 100 mM of calcium chloride for hardening, the beads were firm, of uniform globular shape and suitable for handling, exhibiting a germination response of 68.9%. Encapsulated shoot tip explants, following storage at 4°C for 8 weeks, exhibited a higher regeneration response (60.0%) than non-encapsulated similar explants stored under the same conditions (11.1%). Microshoots, excised from cold-stored encapsulated explants after germination, rooted easily in agar-solidified MS medium with 2 μΜ IBA and after their transplantation into a peat-perlite substrate (3:1, v/v), were acclimatised successfully and established in the greenhouse with minimal losses. The present encapsulation procedure could be applied as an alternative method of micropropagation of desirable elite clones of oleander.     

铁皮石斛幼苗壮苗培养的研究

以铁皮石斛无菌幼苗为外植体,研究不同基本培养基、生长激素、天然添加物、蔗糖浓度和活性炭浓度对幼苗壮苗的影响。结果表明:苗龄4个月的铁皮石斛幼苗最佳的壮苗培养基为:1/2MS+1.0mg/L NAA+30%椰汁+30g/L蔗糖+2g/L活性炭+8g/L琼脂,pH 5.8。     

香榧茎段离体培养再生植株的研究

对香榧茎段不定芽诱导研究的结果表明,不定芽诱导的最佳培养条件为:培养基为B5+KT0.1mg/L+IBA0.5mg/L+0.08%活性炭+2%葡萄糖,培养温度为20℃,光强不高于400lx,该条件下的不定芽诱导率可达55%。将不定芽培养在1/2B5+IBA0.1mg/L+0.08%活性炭+2%葡萄糖的生根培养基上,生根率最高达到40%。通过器官发生途径国内外首次建立了香榧离体再生体系和快繁体系,并获得了完整的再生植株,为今后开展香榧的遗传转化和品种改良奠定了基础。     

Micropropagation of Cymbidium Sleeping Nymph through protocorm-like bodies production by thin cell layer culture

Transverse thin cell layers (tTCLs) of protocorm-like bodies of two stages of PLBs (30 d and 60 d old) of Cymbidium Sleeping Nymph were used as explants to induce PLBs by using coconut water (CW) as a natural additive. 5% (v/v) CW supplemented to KC medium induced an average of 5 PLBs per responding tTCL of 30 d old PLBs with 83% of responding tTCLs. A low percentage of responding tTCLs were observed in 60 d old PLBs’ tTCLs. Anatomical and confocal microscopic studies traced the origin of PLBs to subepidermal layers of the tTCL. A significantly high percentage of shoot regeneration was obtained from PLBs formed on 1–10% (v/v) CW from tTCLs of 30 d old PLBs in comparison to PLBs induced on control after first subculture on KC medium (without CW). The induced PLBs regenerated into plantlets with velamenous roots and these plantlets were transferred to greenhouse conditions on cocopeat:perlite (9:1) with nearly 100% survival. Post-transfer performance of the plantlets was monitored. The results suggest tTCLs as potential explants (with respect to economy of precious hybrid materials) which can overcome the slow growth of hybrid PLBs and coconut water as a single natural additive for the mass multiplication of commercially important orchids.     

Rapid in vitro germination of zygotic embryos of fluted pumpkin (Telfairia occidentalis Hook. f.)

Summary

Fluted pumpkin, Telfairia occidentalis, is becoming an important regional vegetable for its food and medicinal uses. The recalcitrant nature of its seed makes conservation difficult and in vitro techniques may be a viable option for conservation. A pilot study was conducted on the effects of different concentrations of a commercial bleach [3.85% (w/v) sodium hypochlorite] for surface sterilisation of T. occidentalis seed. The optimum concentration [25% (v/v)] was then used as a basis to investigate the responses of mature embryonic axes of T. occidentalis to different concentrations of kinetin (Kin; 0, 1.0, or 2.0 mg l^–1) and 1-naphthaleneacetic acid (NAA; 0, 0.5, or 1.0 mg l^–1) combined in a factorial design. The results of the first experiments indicated that commercial bleach at 25% (v/v) resulted in the lowest contamination of explants (10%), with no evident injury to the embryonic axes. The results revealed that root emergence started 3 d after initiation (DAI) only on Murashige and Skoog medium (MS) with no added plant growth regulator (PGR), and that, by 12 DAI, all media supported the rooting of explants. The highest rooting percentage (69%) was observed at 15 DAI on MS medium with 0.5 mg l^–1 NAA, without Kin. However, shoot emergence started at 9 DAI on PGR-free MS medium, on MS with 0.5 mg l^–1 NAA, or on MS plus 1.0 mg l^–1 Kin. The highest shooting percentage (91%) of explants was observed with 0.5 mg l^–1 NAA at 21 DAI. Considering all other growth parameters, MS medium supplemented with 0.5 mg l^–1 NAA was found to be best for the germination of embryonic axes of T. occidentalis.     

Factors affecting tissue culture of Damask rose (Rosa damascena Mill.)

The present study was conducted to evaluate the regeneration ability of Damask rose. Single-node explants were surface sterilised with 10% chlorox for 15 min and cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of N⁶-benzyladenine (BA) or kinetin (Kin) separately or in combination with different concentrations of indole-3-butyric acid (IBA). Combination of 2.5–3 mg/l BA and 0.1 mg/l IBA was the most suitable treatment for proliferation. In vitro derived shoots were subcultured four times on the fresh medium within a 4-week period. Other treatments such as explant orientation (horizontal, vertical and oblique) and explant wounding were also examined but did not affect shoot multiplication rate significantly. Several experiments were carried out to stimulate in vitro rooting of Damask rose. Application of different media such as MS, 1/2 MS, 1/3 MS and 1/4 MS with different concentrations of indole-3-acetic acid (IAA), IBA and naphthaleneacetic acid (NAA) did not produce satisfactory results. Quick-dip method using sterilised 0–2000 mg/l IAA, IBA and NAA solutions was also studied. Other treatments such as using various concentrations of abscisic acid (ABA) in combination with various concentrations of IAA, IBA and NAA, and using various concentrations of sucrose and agar did not produce any roots. The best treatment for rooting of shoots was 2.5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) for 2 weeks in MS medium and then transferring the explants to MS medium without any growth regulator. Plantlets were acclimatised in a soil mixture consisting of peat moss and sand 1:1 (v/v) and successfully transferred to the greenhouse after 3 weeks.     

High frequency plantlet regeneration from callus and artificial seed production of rock plant Pogonatherum paniceum (Lam.) Hack. (Poaceae)

Pogonatherum paniceum (Lam.) Hack. is a rock plant with good potential for vegetative recovery on naked lands. A high frequency in vitro regeneration system was developed for P. paniceum. Calli were induced from explants of mature seeds, seedlings, young leaves, and stem segments on Murashige and Skoog (MS) medium supplemented with 1.0 mg L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D), 2.0 mg L⁻¹ α-naphthalene acetic acid (NAA) and 0.2 mg L⁻¹ 6-benzylaminopurine (BAP). High induction rates (59.57%) and regeneration rates (100%) were obtained from mature seed explants; calli were sub-cultured for over 2 years and still retained a high regenerative capacity. One seed explant resulted in 69,997 plants in 1 year. Shoot buds derived from calli were used for encapsulation in liquid MS medium containing 3% sucrose and two different alginate matrices (3% sodium alginate (w/v) + MS medium containing 3% sucrose and 3% sodium alginate + 1% activated carbon (w/v) + MS medium containing 3% sucrose) with a 20-min exposure to 2% CaCl₂ and 0.3% bavistin (w/v). The capsule with 3.0% sodium alginate (w/v) and 1% activated carbon (w/v) showed a higher conversion rate (61.58%) and stronger plantlets under non-aseptic conditions. These systems are useful for the rapid clonal propagation and dissemination of artificial seed material of P. paniceum for eco-recovery.     

In vitro colchicine-induced polyploid plantlet production and regeneration from leaf explants of the diploid pear (Pyrus communis L.) cultivar, ‘Fertility’

Summary

Polyploid plantlets, including triploid, tetraploid, and mixoploid, were induced from the European pear (Pyrus communis L.) cultivar ‘Fertility’ by in vitro colchicine treatment of leaf explants. The leaf explants were incubated in 0.4% (w/v) colchicine for 24, 48, or 72 h, then transferred to adventitious shoot-induction medium. Regenerated shoots were pre-selected according to their morphological characteristics when compared to control shoots from untreated shoot proliferation cultures. Shoots with putative polyploid morphological characteristics were maintained and proliferated. The ploidy levels of all putative polyploid individuals were analysed by flow cytometry and identified by chromosome counts of shoot tip tissue squashes. Polyploid shoots were rooted, and the resulting plantlets were transferred to the field. Polyploid plantlets had a higher specific leaf mass and larger stomata than those of diploid plantlets.     

Induction of novel variants through physical and chemical mutagenesis in Barbeton daisy (Gerbera jamesonii Hook.)

Summary

Gerbera (Gerbera jamesonii) is an attractive ornamental flower of high economic importance. The present investigation was aimed at generating novel flower colour variants in the gerbera cultivar ‘Harley’ through physical and chemical mutagenesis. In vitro-raised shoot cultures of gerbera, established from petiole explants, were exposed to different doses of γ-rays (1.5, 2.0, 2.5, 5.0, 10.0, 15.0, 20.0, or 30.0 Gy) using a Cobalt-60 source emitting 2.51 kGy h^–1. To induce mutations through chemical mutagenesis, different concentrations of ethylmethane sulphonate [EMS; 0.1, 0.2, 0.5, 0.8, or 1.0% (v/v)] were administered for 10 min or for 20 min. The LD50 values calculated for shoot survival and the induction of mutations were approx. 6.5 Gy for γ-rays and 0.65% (v/v) EMS for 10 min, or ≤ 0.1% (v/v) EMS for 20 min. Investigations revealed a negative correlation between mutagen dose and plant survival, both in vitro and after acclimatisation. Morphological variants showing changes in leaf shape, leaf size, scape length, flower diameter, and flower colour were obtained. Significantly, early flowering was induced in all mutated plants compared to non-mutated plants.The high frequencies of colour variants obtained using Bγ-rays, or the application of EMS to in vitro-raised shoot cultures could be an effective way to improve gerbera cultivars.     

Evaluation of the efficiency of somatic embryogenesis in guava (Psidium guajava L.)

Summary

An embryogenic protocol for plant regeneration of guava (Psidium guajava L.) was established using 10-week post-anthesis, zygotic embryo explants. Somatic embryogenesis was induced on Murashige and Skoog medium (MS) containing 3% (w/v) sucrose, 0.8% (w/v) agar and various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) by continuous treatment of the zygotic embryo explants. Somatic embryos appeared as globular structures at the end of the third week from culture initiation, and heart-shaped, cotyledonary-stage, and torpedo-stage embryos appeared within the next few weeks. The development of somatic embryos was asynchronous and showed five-to-seven discernible stages. Depending upon the response of the somatic embryos during their maturation, germination, acclimatisation, and encapsulation, they were grouped into one of three categories. The preferred type of somatic embryos (≥ 1.5 mm) were called the “elongated torpedo” (ET) category. The slightly less-preferred type of stomatic embryos (from 1.0 – 1.5 mm) were termed the “short torpedo” (ST) category. The least preferred types of somatic embryos, at the cotyledonary, heart-shaped, and/or globular stages of development (< 1.0 mm), were grouped into a third category designated “CHG”. The suitability and efficacy of various growth regulators and other treatments were assessed based on six different embryogenic parameters: (i) the frequency of embryogenesis; (ii) the intensity of embryogenesis, defined as the average number of somatic embryos produced per culture (“ANEPC”); (iii) the frequency of ET somatic embryos; (iv) the frequency of ST somatic embryos; (v) the frequency of CHG somatic embryos; and (vi) the overall efficiency of embryogenesis, defined as the potential of a treament to produce somatic embryos at the ET or ST stages, or at both stages of development, that could be converted into plantlets. In the present report, we found that 0.01 mg l^–1 2,4-D gave the maximum frequency and intensity of embryogenesis. But the highest frequencies of ET and ST somatic embryos were produced on MS medium containing 3% (w/v) sucrose and 0.001 mg l^–1 2,4-D, while CHG embryos were produced at the highest frequency on the same medium, but containing 0.5 mg l^–1 2,4-D. It was difficult to calculate the most effective concentration of 2,4-D for somatic embryogenesis based on parameters (i) – (v) above. Hence, quantitative estimations of the efficiency of embryogenesis (sixth parameter) were imperative in order to analyse the potential of the different treatments. The highest efficiency of somatic embryogenesis was achieved by continuous treatment of 10-week post-anthesis, zygotic embryo explants with 0.01 mg l^–1 2,4-D on full-strength MS agar medium containing 3% (w/v) sucrose. These somatic embryos matured normally on the same medium, and germinated well both on half-strength solid and in half-strength liquid MS medium containing 3% (w/v) sucrose. They grew in full-strength liquid MS medium with 3% (w/v) sucrose and showed maximum survival upon transfer to soil and hardening. Evaluations of the efficiency of somatic embryogenesis in guava, based on the six parameters defined above, have also helped us to understand and evaluate processes for high efficiency micropropagation in other species.     

High efficiency shoot regeneration from calluses of strawberry (Fragaria X ananassa Duch.) stipules of in vitro shoot cultures

A highly efficient method of shoot regeneration was developed from calluses of four culti- vars of strawberries (Fragaria x ananassa Duch.), ‘Addie’, ‘Dana’, ‘Gea’ and ‘Santana’, grown in vitro. Optimum shoot regeneration (84-96% of the explants), with four to eight shoots was obtained from calluses developed from stipules near the internal zone between petiole and stipule, on Murashige and Skoog (1962) or Gamborg et al. (1968) media, supplemented with 3% (w/v) glucose, 10 [iM BAP and 2.5 IBA and 0.8% agar. The calluses continued to produce shoots for at least six subsequent subcultures. This has been reported, up to now, only in juvenile explants. Microscopic observation showed no preformed buds or meristematic groups of cells in the connecting zone of petiole and stipule prior to culture. However, there were several layers of cells in this area containing higher amounts of starch, which were not observed in the cells of the bottom or in the external side of the stipule. Regeneration from stipules occurred five to six days earlier in whole leaves (stipule + petiole + lamina) than in leaves without laminae, but the final percentage was the same in the cases of all explants. The percentage of regenerating calli from the other explant sources (leaf lamina, petiole and root) was low and dependent on cultivar. Cv. Gea, which showed the highest regeneration capacity, regenerated 32% from leaf laminae, 16% from petiole and root calluses, followed by cv. Addie with 12% from leaf laminae only; the others failed to regenerate from calluses derived from these tissues. The regenerated shoots were successfully rooted and hardened for further observations on eventual somaclonal variation.     

激素对白菜GMS系不同育性外植体再生植株的影响

 以白菜核雄性不育两用系的可育株和不育株带有子房及花柄的花托为外植体, 比较了不同育性外植体再生体系的异同; 并结合内源激素含量测定, 探讨了内源激素、外源植物生长调节剂间的相互作用对不同育性外植体再生的影响。结果表明: 不育和可育再生体系所需的最佳NAA浓度分别为0.2 mg·L^-1和0.1 mg·L ^-1; 在最适的NAA浓度下补给不同浓度的6-BA, 不育外植体不定芽诱导率差异不大, 而可育外植体不定芽诱导率则随62BA浓度的增加表现不规律的变化。不同育性的外植体内源激素含量测定结果表明, 可育外植体内源GA₃、IAA和ZT含量分别比不育外植体高出37.8%、28.0%和224.9%; ABA含量则是不育外植体比可育外植体高出20.4%。花柄及花托再生得到的不定芽在不继代或继代次数少的情况下生根培养时高频现蕾, 生根率极低; 随着继代培养时间的延长, 生殖生长趋势逐渐减弱, 生根率明显提高。     

High-frequency embryogenesis,regeneration of broccoli (Brassica oleracea var. italica) and analysis of genetic stability by RAPD

High-frequency somatic embryogenesis and shoot regeneration of broccoli (Brassica oleracea var. italica) were achieved. Cotyledon and hypocotyl explants from four varieties of broccoli were cultured on MS and modified MS media (mMS, supplemented with PG-96 organic components) with different combinations of growth regulator. The effects of genotypes, different explants, growth regulator combinations, organic components and AgNO₃ on induction of calli and shoots were evaluated. The optimal media for inducting calli/shoots and roots were mMS medium containing 3% (w/v) sucrose and 0.8% (w/v) agar supplemented with NAA at 0.5 mg l⁻¹, 6-BA at 3.0 mg l⁻¹, AgNO₃ at 4.0 mg l⁻¹ and MS medium containing 3% sucrose and 0.8% (w/v) agar supplemented with NAA at 0.2 mg l⁻¹, respectively. The callus induction percentages were over 90% in all four varieties; shoot induction percentage was 92.5% and the average number of shoot per explant was 4.1 from cotyledon explant in variety Bishan. In this study, we established high-efficient embryogenesis and shoot regeneration system of broccoli and analyzed genetic stability of regenerants at DNA level using RAPD molecular marker. Out of 62 arbitrary primers screened using PCR amplification, 79 polymorphic bands were amplified from 20 primers. The results demonstrated the genetic stability of regenerants from the same variety.     

Mineral nutrition of jojoba explants in vitro under sodium chloride salinity

Jojoba (Simmondsia chinensis (Link) Schneider) explants were cultured in vitro on a basal medium supplemented with sodium chloride up to 169 mM during the proliferation stage. At the second and third month of salinity stress, the mineral nutrition (macro- and micro-elements) of the explants was assessed. Explants accumulated significant amounts of sodium and chloride (jojoba is an ‘includer’) while potassium, manganese, phosphorus and nitrate concentration was reduced. The concentration of the other elements did not exhibit significant changes. Each level of salinity stress affected the nutrient status of the explants distinctively. Jojoba explants tolerate salinity up to a level of sodium chloride concentration (113 mM), without showing any stress symptoms. Above this level, the salinity stress impact was observed as succulence and chlorosis of leaves and shoots.     

In vitro cloning of two cumin landrace lines via shoot-tip culture

Summary

Shoot-tip explants were excised from axenic and non-axenic plant cultures of two cumin (Cuminum cyminum L.) landrace lines from Assiut (ASS) and Qina (QIN), Egypt. Explants were cultured on MS (Murashige and Skoog, 1962) medium supplemented with different concentrations of benzyladenine (BA) or kinetin. The two landraces performed similarly throughout the study. Shoot-tip explants from axenic cultures were superior to those prepared from non-axenically grown plants regarding the percentage of explants that produced micro-shoots and the number of micro-shoots that proliferated. The maximum number of excisable micro-shoots was produced on medium with 1 µM BA. Up to 20 micro-shoots per explant were excised from cultures on this medium. The largest number of micro-shoots obtained on medium containing kinetin was five. Most (80–90%) micro-shoots formed roots on medium with 1 µM indole-3-butyric acid (IBA) and 2% (w/v) polyethylene glycol (PEG-6000). The survival rate ex vitro was as high as 70%. A high concentration of BA (4 µM) induced calli, retarded elongation of the micro-shoots and reduced both the number of roots formed on subsequent rooting medium, and plant survival ex vitro. This study supports the feasibility of in vitro cloning of cumin using shoot tips for germplasm collection, conservation and exchange.     

Droplet-vitrification of apical shoot tips of Rubus fruticosus L. and Prunus cerasifera Ehrh.

Apical shoot tips excised from in vitro plantlets of blackberry (Rubus fruticosus L. ‘?a?anska Bestrna’) and cherry plum (Prunus cerasifera Ehrh.) were tested for recovery after cryopreservation using the droplet-vitrification technique. Following treatment for 30 min with a loading solution comprising 1.9 M glycerol and 0.5 M sucrose, explants were dehydrated with a highly concentrated cryoprotectant solution, so called vitrification solution. Shoot tips were dehydrated for 10, 20 and 30 min at room temperature with a solution derived from the original PVS2 solution (containing 37.5% (w/v) glycerol, 15% (w/v) dimethylsulfoxide, 15% (w/v) ethylene glycol and 22.5% (w/v) sucrose) and for 60, 90 and 120 min using the PVS3 solution (containing 50% (w/v) glycerol and 50% (w/v) sucrose). Explants were cooled by direct immersion in LN in 10 μl droplets of vitrification solution placed on aluminium foil strips. Rewarming was done by direct plunging of foil strips in a preheated (37 °C) unloading solution (0.8 M sucrose) for 30 s, after which an equal volume of unloading solution (at room temperature) was added for further incubation for 30 min. As for regrowth of blackberry, PVS3 proved more effective than the modified PVS2, but the difference was significant (P < 0.05) only for the shortest treatment duration. The duration of PVS3 treatment had no significant effect on regrowth of cryopreserved shoot tips (45.8–70%). By contrast, a 30-min treatment with modified PVS2 solution resulted in a significant increase in regeneration percentage (30%), as compared with a 10-min treatment with the same solution (5%). Cherry plum shoot tips were very sensitive to both vitrification solutions and growth recovery of cryopreserved samples was generally lower (5–20%) than that of blackberry explants. No significant influence of PVS treatment (both type of solution and treatment duration) on regrowth of cryopreserved shoot tips was observed with cherry plum shoot tips. Experiments performed in France and in Serbia produced similar results, thereby showing the robustness and reproducibility of the protocols developed.     

In vitro propagation of spathiphyllum

An in vitro method for propagation of Spathiphyllum cultivar ‘Clevelandii’ is described as an alternative vegetative-propagation method.Leaves, inflorescences, peduncles, buds and stem pieces were tried as explants. Initiation and development of shoots did not occur when leaves and peduncles were used. In a few cases, it was possible to induce shoots from pieces of inflorescence explants. Buds and especially stem pieces were very suitable as explants, with shoots developing on the basal medium both with or without a cytokinin.Further multiplication of the shoots was optimal on the basal medium supplemented with 2 mg/l PBA. On this medium, the basal part of the old shoots became swollen and callus-like, and new shoots emerged from the callused area.Shoots developed roots very easily on the basal medium without hormone additives.     

优化香蕉离体培养药物消毒技术的探讨

在香蕉离体培养过程中,进行了对外植体不经表面药物消毒与传统消毒的对比试验,结果表明:两种处理的外植体萌芽诱导率分别为77.30%和76.88%,污染率分别为22.70%和23.13%,差异不显著。由此认为,以香蕉吸芽作外植体在离体培养中所产生的污染与是否进行表面药物消毒关系不大。从而可以优化香蕉离体培养的操作程序。     

An increase in citrus fruit (Kiyomi tangor) abscission induced by ABA is accompanied by an IAA increase in the abscission zone and ethylene production

Summary

Change in IAA concentrations in the peduncle, branch and intervening abscission zone were measured to clarify the involvement of IAA in citrus fruit drop in response to ABA application. Results indicating the importance of an IAA increase in the abscission zone were obtained. One day after application of ABA, the concentration of IAA in the abscission zone showed a temporary increase and then a decrease. The concentration of IAA in the abscission zone was dependent on the concentration of ABA applied. Changes in the production of ethylene, which is involved in the process of abscission and which is induced by IAA, in explants from treated leafy inflorescences, were examined. The fruit-abscission ratios were also investigated in relation to the time required for preparation. Explants sampled 0±1.d after application showed little abscission or ethylene production during the first 24 h incubation. During the next 24 h, almost all the ABA-applied explants abscinded, as did the control, but the former produced 3±4 times more ethylene. ABA-applied explants, sampled 2.d after application, abscinded and produced ethylene markedly during the first 24.h, whereas the control explants did not abscind. Control and ABA-applied explants, sampled 3 d after application, showed no differences in their abscission ratios and ethylene production. These findings indicate that a temporary IAA rise in the abscission zone, observed one day after application, is involved in ethylene production in ABA-induced citrus fruit drop.