The effects of foliar fungicides on beneficial arthropods in wheat fields

Large scale replicated field trials were carried out over a 2 year period to quantify the insecticidal properties of 3 foliar fungicides commonly used in wheat fields. Pyrazophos significantly reduced the numbers of many groups of the natural enemies of cereal aphids and the insects known to be vital in the diet of gamebird chicks for up to 4 weeks after spraying. No such reductions were found after the use of either triadimefon or propiconazole. Aphid numbers on plots sprayed with pyrazophos were twice the numbers found on unsprayed plots either 6 weeks after spraying in 1985 or on unsprayed plots and plots sprayed with the other fungicides 20 days after treatment in 1984. In 1985, this necessitated the additional use of an aphicide only on those plots previously sprayed with pyrazophos. The removal of chick food insects following the use of pyrazophos was considered to be sufficient to reduce survival rates of wild gamebird chicks.      

In vitro Mutation of Chrysanthemum (Dendranthema grandiflora Tzvelev) with Ethylmethanesulphonate (EMS) in Immature Floral Pedicels

Many new cultivars of chrysanthemum were derived from induced mutation. However, chemical mutagenic agents are not widely used due to their low penetration into plant tissues. The objective of the present study was to induce mutation in chrysanthemum using ethylmethanesulphonate (EMS) in immature floral pedicels, followed by the induction and production of adventitious buds in vitro. Preliminary studies on the sensitivity of pedicels to EMS revealed that the LD50 was close to 0.82% (v/v). Immature pedicels of chrysanthemum cv. Ingrid (dark pink color) were treated with 0.77% (0.075 M) EMS solution for 1 h and 45 min, which was followed by rinsing in water for 15 min and surface disinfection. Afterwards, they were cultivated in Murashige and Skoog medium (salts and vitamins) amended with 1 g l⁻¹ of hydrolyzed casein, 1 mg l⁻¹ 6-benzylaminopurine (BAP) and 2 mg l⁻¹ indole-3-acetic acid (IAA). A total of 910 plants were obtained from the pedicels treated with EMS and were evaluated at the flowering stage. Forty-eight mutants (5.2%) were obtained, deviating in petal color (pink-salmon, light-pink, bronze, white, yellow and salmon color). Most of them (89.6% of the total) were phenotypically uniform. The results showed the efficiency of EMS to induce in vitro mutation of chrysanthemum.     

Species composition of thrips (Thysanoptera: Thripidae) and spider mites (Acari: Tetranychidae) on cultivated chrysanthemum (Asteraceae) in Okinawa,southwestern Japan

We investigated the species composition of phytophagous thrips and spider mites on cultivated chrysanthemum in Okinawa, southwestern Japan. Eight thrips species belonging to the genera Frankliniella, Microcephalothrips, Scirtothrips and Thrips were found on chrysanthemum leaves. Among them, Thrips nigropilosus was the predominant species irrespective of season, island or cultivation environment (meshed greenhouse or open field), with its infestation frequency being 89% of the fields in which thrips occurred. This high frequency of occurrence suggests that T. nigropilosus is a major pest of chrysanthemum in Okinawa, even though this species has rarely been regarded as an important pest of chrysanthemum or other crops in any other areas. Thrips palmi was the second most dominant (infestation frequency 36%) and other species were scarce (<14%). Tetranychus urticae (green form) was the only tetranychid species on chrysanthemum in our survey. This lack of diversity among spider mites on chrysanthemum is peculiar considering that eight Tetranychus species have been found on vegetables in the same area. Since T. urticae (green form) has been shown to be resistant to a number of pesticides, severe pesticide applications might have simplified the spider mite fauna on chrysanthemum in Okinawa.     

Production and characterisation of the intergeneric hybrids between Dendranthema morifolium and Artemisia vulgaris exhibiting enhanced resistance to chrysanthemum aphid (Macrosiphoniella sanbourni)

Aphids represent the most destructive of chrysanthemum pests to cultivation. Reliable variety sources of resistance and control methods are limited, so development of highly resistant breeding lines is desirable. An intergeneric hybrid between Dendranthema morifolium (chrysanthemum) variety ‘Zhongshanjingui’ and Artemisia vulgaris (mugwort) ‘Variegata’ was attempted. Most of the hybrid embryos aborted at an early developmental stage. Embryo rescue allowed the generation of hybrid plants, whose hybridity was confirmed by a combination of morphological, cytological and GISH analysis. The hybrids were vigorous, flowered normally, and their flower and leaf shape resembled those of the chrysanthemum more than those of the mugwort parent. The hybrids showed much higher resistance to chrysanthemum aphid (Macrosiphoniella sanbourni) than maternal chrysanthemum by inoculation test. The leaves of the hybrid developed a higher density of trichomes and secretory glands compared to the maternal chrysanthemum. GC–MS analysis revealed that ~51% of the essential oil in the hybrid leaves were monoterpenoids and sesquiterpenoids, while the proportion in the chrysanthemum was ~37%, and in the mugwort was ~90%. It is inferred that higher aphid resistance in the hybrid mainly owed to the leaf micromorphology and bioactive essential oil content.     

Fungal utilization of organophosphate pesticides and their degradation byAspergillus flavus andA. sydowii in soil

Fungal species were isolated which utilize organophosphate pesticides,viz. phosphorothioic (pirimiphos-methyl and pyrazophos), phosphorodithioic (dimethoate and malathion), phosphonic (lancer) and phosphoric (profenfos) acid derivatives. Pesticide degradation was studiedin vitro andin vivo (soil).Aspergillus flavus, A. fumigatus, A. niger, A. sydowii, A. terreus, Emericella nidulans, Fusarium oxysporum andPenicillium chrysogenum were isolated from pesticide-treated wheat straw. The number ofA. sydowii colonies was significantly promoted by 1 mmol/L pirimiphos-methyl, pyrazophos, lancer, dimethoate and malathion when used as phosphorus sources and by pirimiphos-methyl and pyrazophos when used as carbon sources. The number ofA. flavus colonies increased with 0.5 mmol/L lancer and malathion used as the only carbon sources.A. sydowii, A. niger, A. flavus, E. nidulans andF. oxysporum grew on, and utilized, 5 pesticides as phosphorus source and showed more than 50% mass growth.A. sydowii, A. flavus andF. oxysporum phosphatase hydrolyzed the pesticides suggesting that these species are important pesticide degraders.A. sydowii produced higher amounts of the phosphatase thanA. flavus andF. oxysporum. The enzyme was highly active against pyrazophos, lancer and malathion used as the only sources of organic phosphate.A. flavus andA. sydowii phosphatases efficiently hydrolyzed pesticides at 300 ppm in soil, the degradation at 1000 ppm was lower. Mineralization of 1000 ppm pesticides in soil amended with wheat straw was higher than in nonamended soil. All added pesticides except profenfos were degraded within 3 weeks. Lyophilized adapted biomass ofA. flavus andA. sydowii could thus be used for field biodegradation of these pesticides.     

Control of Western flower thrips (Frankliniella occidentalis Pergande) pupae in compost

Western flower thrips (Frankliniella occidentalis) develops from an active larval stage through to a non-feeding, almost immobile, pre-pupal and pupal stage. This generally occurs in the compost or soil below the plant on which the larvae fed. Control of thrips at this stage in their development offers a chance of utilising pathogens or pesticides not suitable for use in an integrated control programme aimed at adult and larval stages. Trials were done with F. occidentalis pupae and pre-pupae in a soil/peat based compost using 11 pesticides, three fungal pathogens and four species of insect parasitic nematodes. The pesticides malathion and chlorpyrifos-methyl gave the most promising result with 97.5% and 96.5% control, respectively. The fungal pathogen Metarhizium anisopliae proved better when applied as a pre-pupation rather than as a post-pupation treatment (74.5% : 26.6% control). The insect parasitic nematode Steinernema carpocapsae gave 76.6% control when applied at 25 times 10⁴ nematodes litre^-1 of compost. The results are discussed in relation to control of thrips in glasshouses.     

Chemical alteration in wheat (Triticum aestivum) shoot induced by mefluidide and defoliation

Mefluidide [N-2(2,4-dimethyl-5-((trifluoromethyl)sulfonyl)amino)phenyl)acetamide] was applied to winter wheat (Triticum aestivum) to determine the alterations in shoot chemical composition and the value of the changes to a wheat-based forage system in field and controlled-environment chamber experiments. Mefluidide, applied at rates between 0.1 and 0.25 kg/ha during full tiller stage (Feekes stage 4), slowed down the rate of cellulose deposition in wheat shoot and reduced neutral-detergent fiber fractions. Mefluidide decreased the rate of nitrogen decline with advancing maturity of treated plants when compared to untreated ones. Mefluidide also reduced cellulose deposition and maintained high nitrogen contents in regrowth shoot tissues in addition to the effect produced by mechanical defoliation. Mefluidide, applied during jointing stage (stage 5), also retarded wheat maturation as the chemical maintained shoot chemical characteristics equivalent to those of earlier stages of development. Mefluidide did not alter significantly shoot chemical composition when applied at boot stage (stage 10), compared to untreated wheat. Based on the experimental results, mefluidide may be a management tool to alter timing of plant development. The delay in maturation and maintenance of vegetative quality longer into the spring attained with mefluidide may suggest a beneficial role of the chemical in a dual-purpose wheat production system that favors forage utilization.Mention of a trademark, proprietary product or vendor does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products that may also be suitable.     

Fate of nitrogen fertilizer applied on two main arables crops,winter wheat (Triticum aestivum) and sugar beet (Beta vulgaris) in the loam region of Belgium

Since 1986, the fate of fertilizer N (NH₄NO₃ or NaNO₃) applied in field conditions on two main arable crops, winter wheat (Triticum aestivum) and sugar beet (Beta vulgaris), has been studied using ¹⁵N. Up to a rate of 200 kg ha^-1 of N, mean recovery of fertilizer by winter wheat was 70%, provided it had been split applied. Single application (with or without dicyandiamid) was less effective. For sugar beet, in 1990, 1991 and 1992, 40% of fertilizer N was found in the crop at harvest when NH₄NO₃ had been broadcast at 100 to 160 kg N ha^-1 at sowing time. For the same N rate, recovery was 50% when row applied near the seeds and 60% for 80 kg N ha^-1. For the two experimental crops, residual fertilizer N in soil was exclusively organic. It ranged from 15 to 30% of applied N and was located in the 30 cm upper layer. Losses were generally lower with winter wheat (12%) than with sugar beet (20–40%) and could be ascribed to volatilization and denitrification. Soil derived N taken up by the plant was site and year dependent.     

ANALYSIS OF GAMETIC PROTOPLAST RELEASE IN THE CLOSTERIUM PERACEROSUM-STRIGOSUM-LITTORALE COMPLEX (CHLOROPHYTA)1

A biologically active glycoprotein (protoplast-release-inducing protein; PR-IP), which induces the release of gametic protoplasts from mating type minus (mt^-) cells of the Closterium peracerosum-strigosum-littorale complex, was prepared from a medium in which mt^- and mt⁺ cells had been previously incubated together. The process of PR-IP-inducing protoplast release was analyzed. Induction of protoplast release was dependent upon the duration of both PR-IP treatment and preincubation in nitrogen-deficient mating medium before PR-IP treatment. Low cell density in the preculture stage had a significant stimulative effect upon the induction of protoplast release. Light was necessary for protoplast release, especially just before PR-IP treatment. Chloramphenicol and 3-(4-chlorophenyl)-1,1-dimethylurea (CMU) exerted inhibitory effects on protoplast release, especially when they were applied to the preculture stage but not when they were applied to the protoplast-releasing stage after the PR-IP treatment. We suggest that preculture at a low cell density under continuous light conditions that may cause metabolic changes in the chloroplast is a very important stage for gametic protoplast release in this Closterium.     

Chitosan (biochikol 020 PC) in the control of some ornamental foliage diseases

Chitosan, (Biochikol 020 PC) a potential elicitor of plant defence and also an active inhibitor of fungal growth was used in experiments. The compound was used at concentrations 0.01 to 0.2% as a plant spray for rose protection against Sphaerotheca pannosa var. rosae, Peronospora sparsa and Diplocarpon rosae, Oidium chrysanthemi and Puccinia horiana on chrysanthemum, against Melampsora epitea on willow. Myrothecium roridum on dieffenbachia and against Lophodermium spp on Scots pine. Effectiveness of this product, applied curatively 2-times at 7-day-intervals after appearance of rose powdery mildew symptoms on most of plant parts, was about 32%. When compound was applied just after the first disease symptoms appearance, after 2-week-protection its effectiveness was about 43-60%. After 4 weeks, effectiveness of chitosan (Biochikol 020 PC) increased and ranged from 67 to 77%. In the control of P. sparsa on rose shrubs, growing in plastic tunnel, chitosan at conc. 0.025% was applied 4 times at weekly intervals. Effectiveness of chitosan against this pathogen was over 72%, similar as standard fungicide. Increase of chitosan (Biochikol 020 PC) concentration resulted in decrease of its effectiveness. In case of D. rosae control, chitosan used 9 times at weekly intervals was applied after first disease symptoms. After 3-week-protection effectiveness of the compound varied from 18 to 60% according to used concentration. After 9 weeks of protection effectiveness of tested product ranged from 16 to 23% and was connected with used concentration. Increase of chitosan concentration resulted in better protection of rose shrubs. Effectiveness of chitosan (Biochikol 020 PC) used 4-times at conc. 0.01 to 0.05% as chrysanthemum spray in the control of Oidium chrysanthemi ranged from 69 to 79%, whereas against Puccinia horiana from 54-97%. Two-time-spraying of willow with rust symptoms (Melampsora epitea) with chitosan at conc. 01-0.04% caused decrease of disease severity from 26 to 47%. Additionally about 10 to 25% of uredinia were dried up. Chitosan (Biochikol 020 PC) applied for leaf spraying inhibited the development of Myrothecium leaf spot on dieffenbachia more than 85%. Curative application of the compound controlled Myrotecium leaf spot at about 40%. Chitosan efficacy was also tested against one-year-old Pinus sylvestris seedlings growing in field conditions. Plants were sprayed fortnightly 14-times (long programme) or 5-times (short programme) against Lophodermium spp. Observations made during next spring showed that effectiveness of chitosan (Biochikol 020 PC) was more than 50% (long programme) and below 50% (short programme).     

Tillers Do Not Influence Phytotoxicity of Imazamethabenz in Wild Oat (Avena fatua)

The response of wild oat to imazamethabenz varies with the growth stage, but the role of tillers in this regard is unclear. Removal of tillers at the three-leaf stage before spraying with imazamethabenz did not significantly affect the total shoot fresh weight measured 3 weeks later. The leaf area and dry weight of intact plants at the three-leaf stage were 17–21% greater than for plants with coleoptilar and first leaf main shoot tillers (T0 and T1) removed. The greater leaf area may have increased herbicide interception per plant. Similar fresh weight reductions in main shoot, total tillers, and total shoots were found whether imazamethabenz was applied to the plant at the two-leaf without tillers or the three-leaf with two tillers stage. Imazamethabenz applied only to the main shoot reduced total shoot dry weight more than an equivalent amount of imazamethabenz applied only to tiller T1 or applied over the whole shoot. Imazamethabenz had the least inhibitory effect on whole plant growth when applied only to T1. When ¹⁴C-herbicide was applied to the first main shoot leaf of plants at the three-leaf stage with two tillers, the ¹⁴C translocated 38% to roots, 33% to the main shoot, and nearly 30% to all tillers. When ¹⁴C-herbicide was applied to the first leaf of T1 then the ¹⁴C translocated 50% to T1, 25% to the main shoot, 20% to roots, and 5% to all other tillers. The translocation pattern and fresh weight values suggested that the presence of early tillers during herbicide application neither increased nor decreased imazamethabenz efficacy in wild oat. Received June 4, 1997; accepted June 5, 1997     

Lygus rugulipennis on chrysanthemum: Supplemental prey effects and an evaluation of trap plants

The European tarnished plant bug, Lygus rugulipennis Poppius, is considered a major pest in chrysanthemum nurseries in The Netherlands. Adults puncture plant's apical meristem, after which the growing point splits and growth is inhibited. Flower buds and flowers can also be severely damaged. Both types of damage result in economic losses for growers. Despite the importance of this pest for chrysanthemum nurseries, there is only very limited information about L. rugulipennis development on chrysanthemum plants, Chrysanthemum × morifolium Ramat., and whether L. rugulipennis can be controlled using trap plants is not known. We investigated whether: (1) L. rugulipennis could develop from egg to adult on the vegetative and flowering stages of chrysanthemum; (2) their performance was enhanced when a supplemental prey source (Ephestia kuehniella Zeller eggs) or another common pest (the green peach aphid, Myzus persicae Sulzer) was present; and (3) there were alternative plant species more attractive than chrysanthemum for use as trap plants for local pest control or monitoring of L. rugulipennis. L. rugulipennis developed on both vegetative and flowering chrysanthemum stages without any additional food source. Nonetheless, when chrysanthemum was supplemented with E. kuehniella eggs, L. rugulipennis achieved the best performance in terms of the number of adults developed and faster developmental time. Interestingly, L. rugulipennis developed faster on chrysanthemum infested with the aphid M. persicae compared to non-infested plants, however, there was no difference in the number of adults developed. In a trap plant experiment with 16 plant species in the vegetative stage, we found that white mustard, Sinapis alba L., was significantly more attractive than chrysanthemum to both adult and nymph L. rugulipennis. Further research is needed to evaluate the potential of S. alba as a trap plant for monitoring L. rugulipennis and how the presence of prey in the crop influences L. rugulipennis.     

Survival and reproduction of Liposcelis paeta (Pearman) (Psocoptera: Liposcelididae) when irradiated by electron beams

Electron beam irradiation has been evaluated for the survival and reproduction of Liposcelis paeta Pearman (Psocoptera: Liposcelididae). All effects increased with increasing doses from 50 to 1000 Gy when the electron energy was 2 MeV. Eggs were unable to develop into adults following a dose of 100 Gy. Complete (100%) mortality was achieved about 5 weeks after adult emergence from nymphs irradiated at 300 Gy, whereas 100% mortality of adults was achieved 9 weeks after irradiation at 300 Gy. The fecundity was greatly reduced to 1.82% of that of control when L. paeta adults were irradiated at 250 Gy. A dose of 300 Gy caused complete reproductive sterility in adults. These results suggest that the most susceptible stage was the egg stage, then nymph and adult stages. A dose of 300 Gy is suggested for quarantine treatment of commodities infested by L. paeta.     

The effects of some commonly-used foliar fungicides on Collembola in winter barley: laboratory and field studies

Four commonly-used cereal foliar fungicides were screened for their laboratory toxicity against the symphypleone collembolan, Sminthurinus aureus. A proportional hazards analysis of time-survival curves following the fungicide treatments showed that carbendazim, propiconazole, pyrazophos and triadimenol significantly increased the laboratory mortality of S. aureus. The organophosphorus fungicide pyrazophos caused high levels of mortality of S. aureus in the laboratory so a field evaluation of the effects of this fungicide on a wider range of Collembola was undertaken in winter barley. Comparison of the effects of pyrazophos with those of the broad-spectrum insecticide dimethoate in the field revealed both compounds to have similar activity against some Collembola. Of the 11 species caught only the four symphypleone species exhibited these effects but the numbers of three symphypleone species were reduced to zero 4 wk after treatment with pyrazophos. The effects of pyrazophos and dimethoate were, however, not detectable in individual species after 11 wk.     

Greenhouse Evaluation of Dose– and Time–Mortality Relationships of Two Nucleopolyhedroviruses for the Control of Beet Armyworm, Spodoptera exigua, on Chrysanthemum

Dose– and time–mortality relationships of baculoviruses in pest insects are important for the determination of effective spraying regimes. A series of experiments with Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and Spodoptera exigua MNPV (SeMNPV) against synchronized populations of S. exigua larvae in greenhouse chrysanthemum was conducted. Dose– and time–mortality relationships of different virus concentrations and S. exigua target stages were determined and the area foliage consumption was measured. Crop injury was greatly reduced when S. exigua were controlled as second or third instar larvae, whereas virus applications against fourth instar larvae could not prevent considerable crop injury, even at high concentrations. SeMNPV was approximately 10 times as infectious as AcMNPV when applied on greenhouse chrysanthemum. The relative virulence of AcMNPV and SeMNPV corresponded reasonably well with previously published laboratory bioassay data. SeMNPV killed second and fourth instar S. exigua larvae approximately 12 h faster than did AcMNPV in chrysanthemum, but no difference in speed of action was found for third instar larvae. The relative speed of action of AcMNPV and SeMNPV determined in chrysanthemum and in laboratory bioassays did not correspond for third instar S. exigua larvae; laboratory bioassay data can therefore not simply be extrapolated to the crop level.     

In vitro shoot multiplication of eastern white cedar (Thuja occidentalis)

Summary A protocol for clonal propagation of eastern white cedar (Thuja occidentalis L.) was enhanced by optimizing the shoot multiplication stage using unbranched in vitro-produced shoots. This was achieved by careful selection of different medium components. An optimum range of 10 to 14 axillary shoots was obtained when shoots were cultured on half-strength Quiorin and LePoivre medium containing 10μM filter-sterilized zeatin for 3 wk. Transfer of the treated shoots to cytokinin-free medium containing 0.05% activated charcoal improved both the number and quality of the axillary shoots produced. Maximum axillary bud induction was also accomplished when shoots were pulsed in 1 mM liquid, filter-sterilized zeatin for 3 h, and then transferred to half-strength Quiorin and LePoivre, charcoal-containing medium. Inclusion of 4% sucrose improved the number of axillary shoots obtained. Half strength of the major salts produced an optimum response. Shoots obtained from different cultures (1 to 5 yr old) responded similarly to the applied cytokinin; however, newly induced shoots (4 mo. old) gave a significantly higher response.     

Effect of sorghum phenology on the control of Chilo partellus with Nosema marucae (microspora: Nosematidae)

An aqueous suspension of Nosema marucae spores was sprayed on foliage of sorghum plants that had been infested with neonate Chilo partellus larvae at 3, 5, 7 and 9 weeks after (plant) emergence (WAE). These periods corresponded to plants at the tillering, early booting, soft dough and late maturation phenological stages, respectively. The extent of borer infestation and plant damage was monitored until crop harvest, and compared with plants which had been similarly infested but not sprayed with the pathogen, and with plants in insect‐free control plots. When applied early, infestation with neonate borer larvae caused most damage to the plants and the greatest reduction in yield in the untreated plots. Early treatment with N. marucae spores at 3 and 5 WAE resulted in the greatest reduction in damage to plants and the highest improvement in yield of seeds. It is apparent, therefore, that for maximum crop protection the microsporidian needs to be applied when sorghum is in an early phenological stage.     

Some properties of chrysanthemum stunt, a virus with the characteristics of an uncoated ribonucleic acid

Grafting to virus-free Mistletoe chrysanthemums was the most reliable method of detecting the chrysanthemum stunt agent, but specific light and temperature conditions were required for the diagnostic ‘measles’ symptoms to develop. Although stunt agent was highly infectious, leaf-rubbing inoculations with chrysanthemum sap gave erratic results. Colorimetric and electrophoretic tests were unreliable for indexing chrysanthemums. Stunt agent infected eight of twenty-nine species in the family Compositae, but none of 116 species in forty-seven other families. Stunt spread rapidly by foliage contact and by handling plants, but dipping the hands in 2% trisodium orthophosphate when handling plants increased the amount of spread. Stunt agent was not transmitted by four species of aphids, the glasshouse redspider mite, dodder (Cuscuta campestris) or through chrysanthemum seed. Stunt agent withstood 10 min at c. 98 °C and dilution to 10^-4, was not pelleted by ultracentrifugation, and was inactivated by RNase in weak, but not strong, buffer, suggestive of an uncoated RNA ‘viroid’. Partially purified preparations were made by homogenizing frozen chrysanthemum leaves in 0.5 m phosphate buffer with antioxidant at c. 2 °C, and clarification by n-but-anol and chloroform, followed by centrifugation. Highly infective RNA was precipitated from the supernant fluid by 2.5 vol. cold ethanol, and resuspended in a small volume of buffer. The u.v. absorption spectra of infective preparations and the u.v. absorbance profiles of density-gradients, were very similar to those of preparations from healthy chrysanthemum. Infective partially purified preparations of stunt agent withstood exposure to 2% formaldehyde or tri-sodium orthophosphate, u.v. irradiation, and sonication. Stunt preparations contained no virus particles recognizable by electron microscopy, gave no distinct peak on analytical ultracentrifugation, and did not consistently contain any specific antigen. Although similar to the ‘viroids’ potato spindle tuber and citrus exocortis, stunt agent did not infect Citrus limon, Gynura aurantiaca or tomato.     

In vitro induction of multiple shoots and plant regeneration from shoot tips of mung bean (Vigna radiata (L.) Wilczek)

Conditions for plant regeneration from excised shoot tips of Vigna radiata were studied. Complete plants were regenerated directly without an intervening callus phase from shoot tips on basal medium (MS salts+B₅vitamins). Regeneration frequency varied with genotype, explant size and growth regulator combinations in the medium. Addition of cytokinins induced a variable amount of callus at the base of the shoot tip, followed by multiple shoot formation. Benzyladenine (BA), kinetin and zeatin at 5×10^-6 M each induced multiple shoots in 100% of the explants but the highest number of regenerants per explant (9) was produced with BA. The efficacy of BA for shoot multiplication was not improved when it was supplemented with naphthaleneacetic acid (NAA) or indoleacetic acid (IAA). NAA or adenine sulphate, when applied alone, induced complete plantlets. The growth regulator requirement of explants for the induction of multiple shoots varied with explant size. The shoot tip explants maintained proliferation ability on subculture. None of the treatments was effective in inducing shoot bud differentiation from callus. Regenerated shoots were rooted on MS basal medium and MS supplemented with either IAA or indolebutyric acid. The rooted plants were transferred to the field; 60% subsequently survived and grew.Abbreviations BM basal medium [MS (Murashige & Skoog 1962) salts+B₅ (Gamborg et al. 1968) vitamins] - BA 6-benzyladenine - AdS adenine sulphate - IAA indole-3-acetic acid - NAA-1 naphthaleneacetic acid - IBA indolebutyric acid