Biological control of crown rot of bananas with Pichia anomala strain K and Candida oleophila strain O

The antagonistic activity of two yeast strains (Pichia anomala (E.C. Hansen) Kurtzman, strain K and Candida oleophila Montrocher, strain O) against the parasitic complex responsible for banana crown rot was evaluated. The strains were applied at three different concentrations (10⁶, 10⁷, 10⁸ cfu/ml) and their efficacy tested in vivo on three separate fungi (Colletotrichum musae (Berk. & Curt.) Arx, Fusarium moniliforme Sheldon, and Cephalosporium sp.) and on a parasitic complex formed by association of these three fungi. At the concentrations used C. musae appeared to be the most pathogenic. The complex showed intermediate aggressiveness between C. musae and both other fungi.Statistically significant antagonistic effects were observed on C. musae, F. moniliforme, and the fungal complex. The highest protection level (54.4%) was observed with strain O added at 10⁸ cfu/ml on crowns previously inoculated with the fungal complex. The level was lower when the fungi were inoculated separately.Furthermore, the antagonistic effect was strongly reinforced when strain O at 10⁸ cfu/ml was applied 24 h before fungal complex inoculation (59.9%), as compared to its application 15 min (24.3%) or 3 h (27.3%) after fungal complex inoculation. Bananas showed increased susceptibility to the fungal complex from March to June, and this influenced the level of protection by yeast, which decreased over the same period. A strict negative correlation (R² = 0.83) was highlighted between susceptibility of banana to crown rot and protection provided by yeast.     

Endophytic fungi from Musa acuminata and their reintroduction into axenic plants

Sixteen fungal taxa were isolated from 2400 leaf fragments of mature Musa acuminata plants collected from three different localities in São Paulo State, Brazil. The most frequently found endophytic fungi were Xylaria sp., Colletotrichum musae and Cordana musae. The standard distribution of endophytes was similar in the three localities. Spontaneous resistant mutants to the fungicides thiabendazole (thi-1) and benomyl (ben-5), were obtained from Colletotrichum musae wild-type isolates. Equal amounts of conidia mixtures of combinations of wild-types and mutants were reintroduced in axenic banana plantlets derived from tissue culture; thi-1 showed a selective advantage when compared to both wild-types used. For ben-5, no significant difference was found. Results showed that wild-types and spontaneous mutants of endophytic fungi as Colletotrichum musae, can be successfully reintroduced in plantlets derived from tissue culture.     

Control of tomato grey mould on unheated crops using non- systemic fungicide sprays

High volume sprays of dichlofluanid (0.1 % a.i.) reduced total fruit numbers but gave better control of Botrytis cinerea infections of tomato stems, leaves and fruit than sprays at 0.05 or 0.025% a.i. Surface residues from the sprays at 0–1, 0.05 or 0.025% a.i. were respectively 5.4, 1.6, 0.7 μg/g fresh weight of ripe fruit at harvest. Tank-mixed zineb (0.12% a.i.) and captan (0.2% a.i.) were less effective and increasing the spray intervals from 1 wk to 2 and 3 wk reduced the fungitoxicity of captan and zineb more than that of dichlofluanid. Harvest residues on ripe fruit were 7.1, 2.8 or 2.4 μg/g when dichlofluanid (0.1 % a.i.) sprays were applied at 1, 2 and 3 wk intervals respectively. Good control of B. cinerea was achieved if the whole plant was sprayed initially with dichlofluanid as soon as the second truss was flowering and subsequent sprays were restricted to the upper section of stem including the four or five youngest trusses of buds, flowers and fruit. When used as a post-infection spray there was a period of c. 8 wk before dichlofluanid markedly reduced the incidence of grey mould. Tank-mixed zineb (0.12% a.i.) controlled Botrytis fruit spotting but not leaf and stem infections. Botrytis stem lesions extended more rapidly on zineb-sprayed plants than on unsprayed plants or on plants treated with captan or dichlofluanid.     

Some effects of benomyl and carbendazim on Aphis fabae and Acyrthosiphon pisum on field bean (Vicia faba)

Field bean plants were treated with benomyl in a glasshouse, then nymphs of Aphis fabae and Acyrthosiphon pisum were caged on the second pinnate leaf. Soil drenches at concentrations of 150 and 75 μg benomyl/ml or above increased mortality of A. fabae and A. pisum respectively; 250 μg a.i./ml increased mortality and decreased progeny production of alate A. fabae. The effect on mortality persisted for at least 16 days after treatment. Foliar sprays increased mortality at concentrations of 75 μg a.i./ml and above (A. fabae). Field populations of A. pisum were reduced when bean plants were drenched or sprayed at a concentration of 250 μg a.i./ml and A. fabae populations were reduced by drenches but not by foliar sprays. A commercial formulation of carbendazim (Bavistin) increased aphid mortality whereas the formulation medium did not. Under the experimental conditions, benomyl affected the distribution of both species on young bean plants but did not induce a repellent effect; aphids preferred untreated leaves. Mortality and preference tests, and a field experiment, indicated that A. pisum was affected more than A. fabae.     

Black leaf streak of bananas in Fiji

Black leaf streak of bananas, caused by Mycosphaerella sp., prevented fruit of export quality forming and bunches maturing. Some infected leaves lived less than 50 days and were seldom retained until harvest. Maneb or benomyl applied in oil/water emulsions gave good control and benomyl was so effective that plants had ten leaves at harvest and some leaves survived for 245 days. Plants sprayed with maneb or benomyl flowered I month early. No benomyl residues were detected in the fruit exported to New Zealand. The control of black leaf streak by sprays containing oil has caused other leaf diseases to become more prevalent and the ensuing complex disease situation is discussed.     

Characterizing the production of a wild-type and benomyl-resistant Fusarium lateritium for biocontrol of Eutypa lata on grapevine

Benomyl-resistant (BR) and wild-type (WT) strains of Fusarium lateritium were examined for their tolerance to benomyl on potato dextrose agar (PDA) containing benomyl and control of the Eutypa lata in grapevine bioassays. The WT strain grew on PDA containing 1 μg/ml benomyl at 13, 26 and 29°C. The BR strain grew on PDA containing 10 μg/ml benomyl at 4°C, on PDA containing 100 μg/ml benomyl at 29°C, and on PDA containing 1000 μg/ml benomyl at 13 and 26°C. The BR strain was also able to colonize grapevine segments and control E. lata in the presence of 1000 μg/ml benomyl. Both strains were amenable to production via liquid fermentation and both achieved 100% control of E. lata in grapevine bioassays. Neither the duration of fermentation nor incubation temperature during grapevine bioassays influenced the efficacy of either strain against E. lata. The results suggest that application of BR F. lateritium alone or in combination with benomyl may provide good control of E. lata. Journal of Industrial Microbiology & Biotechnology (2001) 26, 151–155. Received 22 December 1999/ Accepted in revised form 20 October 2000     

Postharvest disease control in mangoes using high humidity hot air and fungicide treatments

The disease control efficacy of quarantine heat treatments developed for fruit fly disinfestation in mangoes cv. Kensington Pride was evaluated in this study. Heat was applied using high humidity (>95% r.h.) hot air (HHHA) at temperatures ranging from 47–49°C. Anthracnose, caused by Colletotrichum gloeosporioides, was well controlled in mangoes heated to a core temperature of 46°C, 47°C or 48°C for 24, 10 or 8 min respectively, prior to ripening at 23°C for 16 days. Stem end rot, caused by Dothiorella dominicana and Lasiodiplodia theobromae, was not satisfactorily controlled by these treatments. In a subsequent experiment, fruit were immersed in a hot benomyl (0.5 g a.i. litre“¹ at 52°C for 5 min) or unheated prochloraz (0.25 ml a.i. litre¹ at 28°C for 30 s) dip before or after the application of HHHA (core temperature of 47°C for 10 min). During storage at 23°C for 15 days, the incidence of stem end rot was reduced by HHHA alone, although immersion in hot benomyl either before or after HHHA treatment greatly improved stem end rot control. HHHA treatment (core temperature of 46.5°C for 10 min) alone reduced the incidence of anthracnose in mangoes stored at 13°C for 14 days prior to ripening at 22°C, although a combination treatment consisting of HHHA and either hot benomyl or unheated prochloraz gave complete control of anthracnose under these storage conditions. HHHA treatment alone gave no control of stem end rot in mangoes stored at 13°C prior to ripening at 22°C. A supplementary hot benomyl treatment was required for acceptable control of this disease in cool-stored mangoes. The development of yellow skin colour in fruit was accelerated by HHHA treatment.     

The mixture of tertiary and quaternary alkaloids isolated from Argemone ochroleuca inhibits spore germination of some fungi

The mixture of tertiary and quaternary alkaloids isolated from Argemone ochroleuca was separately assessed against spore germination of some plant pathogenic fungi, e.g. Alternaria alternata, Alternaria brassicae, Alternaria cajani, Bipolaris sp., Curvularia lunata, Curvularia sp., Colletotrichum musae, Fusarium udum, Helminthosporium sp., Helminthosporium pennisetti and Helminthosporium speciferum. Spore germination of Fusarium udum and Helminthosporium sp. was completely inhibited at very low concentration (200 ppm). A similar effect was observed on A. alternata, C. musae and H. pennisetti at 600, 800 and 1000 ppm. With quaternary alkaloids, Curvularia sp. and Colletotrichum musae were most sensitive as complete inhibition of spore germination was observed at 400, 600, 800 and 1000 ppm and a similar effect was observed with A. brassicae and A. cajani at 600, 800 and 1000 ppm. The remaining fungi were also highly sensitive to the mixture at different concentrations.     

An integrated approach with Trichoderma harzianum DGA01 and hot water treatment on control of crown rot disease and retention of overall quality in banana

A biological control of crown rot disease of banana fruit was analysed using an integrated approach combining hot water treatment and Trichoderma harzianum strain DGA01. Treated fruits were stored at 22–25 °C and 90–95% relative humidity for 2 weeks. The bioefficacy of fungal antagonist in vitro towards crown rot-causing pathogens, namely Lasiodiplodia theobromae, Thielaviopsis paradoxa, Colletotrichum musae and Fusarium verticillioides, was enhanced by 11.41% following hot water treatment (50 °C, 20 min). DGA01 germinated on the fruit 48 h after inoculation and parasitised the pathogen. Postharvest application showed that hot water treatment and conidial suspension of DGA01 (10⁶ ml^?1) applied singly performed significantly better than the untreated control in reducing the incidence of crown rot, but were not as effective as the fungicide. The combination of hot water treatment and DGA01 gave 93% control of fruit decay which was comparable with fungicide treatment of 95%. The quality of fruit was markedly improved in hot water treatment + DGA01 as compared to those dipped in fungicide solution. The inconsistencies of single treatments, by DGA01 or hot water dips, in controlling crown rot such as variation in severity of disease among treatments and within a treatment, were lessened by dipping the fruit in DGA01 conidial suspension following hot water treatment.     

Benomyl Tolerance of Ten Fungi Antagonistic to Plant-parasitic Nematodes

Ten strains of fungi were tested for tolerance to the fungicide benomyl. Verticillium chlamydosporium strain 2 did not grow in the presence of benomyl; Drechraeria coniospora strains 1 and 2 and Chaetomium sp. tolerated only 0.1 μg benomyl/ml medium; Acremonium bacillisporum, an unidentified fungus, and Phoma chrysanthemicola uniformly grew at 1 μg/ml, but some hyphae grew at higher benomyl concentrations; Fusarium sp. tolerated 475 μg/ml, but some hyphae grew on medium amended with 1,000 μg/ml; Verticillium lecanii and V. chlamydosporium strain 1 routinely tolerated 1,000 μg/ml. Fungi generally grew more slowly at higher than at lower benomyl concentrations. Strains with elevated tolerance to benomyl were selected from Acremonium bacillisporum, Drechmeria coniospora, Fusarium sp., and an unidentified fungus. These strains retained the increased tolerance after repeated transfers on unamended medium.     

An assessment of some new treatments for the control of rotting of stored apples

Benomyl (0.025% a.i.) and thiabendazole (0.05 % a.i.) applied in July, August and September to apple trees cvs. Sunset and Cox's Orange Pippin gave slightly better control of Gloeosporium spp. rots in stored fruit than captan (0.1% a.i.) applied at similar intervals. On Cox's Orange Pippin, treatment with captan at 0.1 % a.i. in July and at 0.2% a.i. in August was as effective as three sprays at 0.1 % a.i. in July, August and September in controlling both Gloeosporium spp. and Nectria galligena rots. Benomyl applied in the spring and early summer to control apple scab on orchard trees reduced the incidence of Gloeosporium spp. and Monilia fructigena in stored fruit. Pre-harvest sprays of benomyl and captan gave good control of rotting caused by N. galligena but thiabendazole was ineffective. Post-harvest dips of thiabendazole controlled rotting by Gloeosporium spp. butnot byN. galligena.     

Fungicide programmes for the control of spur blight, powdery mildew and grey mould on raspberry

Five sprays of dichlofluanid (0–1% a. i.) reduced spur blight (Didymella applanata) on canes and powdery mildew (Sphaerotheca macularis) on fruit more than five sprays of captan (0–1% a. i.). Omission of the first three applications resulted in little increase in disease but when the fourth (full bloom) application also was omitted the incidence of disease was comparable with that on untreated plants. The final (fruitlet stage) application did not further reduce disease incidence. Five sprays of dichlofluanid reduced grey mould (Botrytis cinerea) on stored fruit more than five sprays of captan. There was an increase in grey mould with progressive reductions in the number of sprays. Incidence on canes was slight and was unaffected by treatments. A programme of five dichlofluanid sprays applied at 14-day intervals until c. 2 wk before harvest and supplemented by a single early full bloom spray of either dinocap (0–025% a. i.), binapacryl (0–05% a. i.), triforine (0–025% a. i.), benomyl (0–025% a. i.) or replaced by benomyl (0–025% a. i.) all halved both the number of fruits infected by powdery mildew and the number of spur blight lesions on canes, compared with unsprayed controls. Six applications of any of the following fungicides at 7-day intervals from 23 May until 29 June 1977 reduced powdery mildew incidence on fruit, the last fungicide being particularly effective: dinocap (0–0125%), dichlofluanid (0–075%), binapacryl (0–025%), triforine (0–0125%), bupirimate (0–0075%), ditalimfos (0–01875%) and fenarimol (0–0036%). Only triforine and bupirimate reduced the incidence of mildew on shoots. Dichlofluanid, binapacryl, triforine and fenarimol all reduced the amount of spur blight on canes. The largest fruit occurred on plants sprayed with bupirimate, ditalimfos and fenarimol, and late ripening was associated with triforine treatments.     

Studies on the fungicidal control of stem lesions caused by Botrytis cinerea on glasshouse tomatoes

In glasshouse pot experiments, uptake of benomyl, thiophanate-methyl and carbendazim from equivalent soil applications (in the range 0–003– 0–035 %^a-i- ^atarateof ²&4 ml/plant) gave equal levels of control of Botrytis lesions developing from inoculations of freshly exposed leaf scars on tomato stems. Spray applications of benomyl to exposed leaf scars controlled infection at concentrations down to 0025 % a.i. The effect of lower concentrations of the stem spray could be markedly enhanced either by the addition of a mineral oil (2 % Actipron) or by a prior soil application of benomyl at a low rate which on its own had little effect on lesion development. Protectant spray applications of glycophene and vinclozolin gave levels of control quite comparable to that of benomyl at equivalent concentrations. Evidence was obtained that the lesions formed at the artificially-inoculated leaf scars at the top of the stems of young pot-grown tomato plants were larger than those lower down. In spite of this, the level of disease control with soil applications of fungicides containing or generating carbendazim (MBC) was greater at the top than at the bottom, probably because of the normal migration of the fungicides and their accumulation at the extremities of the plant. In an observation trial in a commercial crop of tomatoes, benomyl applied either as five soil drenches at approximately monthly intervals, or as two drenches followed by five sprays at three-weekly intervals, or as five sprays alone gave marked reductions in plant loss and number of Botrytis stem lesions in both cvs Eurocross BB and Cudlow Cross. Those stem lesions which did develop, however, were generally as large as those on untreated control plants. Five sprays of dichlofiuanid gave similar levels of disease control. All the treatments gave apparently higher yields (statistically untested) in Eurocross BB, but less consistent responses were recorded in Cudlow Cross.     

Effects of Preharvest Applications of CaCl2, 2,4-D and Benomyl and Postharvest Hot Water, Yeast and Fungicide Treatments on Development of Decay on Satsuma Mandarins

In this study, an integrated approach was evaluated for the control of postharvest decays of mandarin including some pre‐ and post‐harvest treatments under storage conditions. The efficacy of the treatments both as alone and in combination was evaluated during 3 years. Preharvest application of benomyl resulted in significantly less decay of mandarin fruit after storage in 3‐year tests. Calcium chloride (CaCl₂), 2,4‐dichlorophenoxyacetic acid (2,4‐D) and gibberellic acid (GA₃) as stand‐alone treatments or combinations were not effective in controlling Penicillium and total decay infections on inoculated mandarin. Postharvest application of imazalil (200 μg/ml) in solution heated to 54°C for controlling postharvest green and total decay of mandarin was significantly effective for 3 months under storage conditions. The biocontrol activity of yeast (Metschnikowia pulcherrima) was improved when yeast treatment was combined with imazalil (200 μg/ml) at postharvest. The data suggest that preharvest application of benomyl and postharvest treatments of imazalil, hot water and yeast may reduce postharvest green mould and total decay of mandarin under storage conditions.     

Hand position on the bunch and source–sink ratio influence the banana fruit susceptibility to crown rot disease

The postharvest development of crown rot of bananas depends notably on the fruit susceptibility to this disease at harvest. It has been shown that fruit susceptibility to crown rot is variable and it was suggested that this depends on environmental preharvest factors. However, little is known about the preharvest factors influencing this susceptibility. The aim of this work was to evaluate the extent to which fruit filling characteristics during growth and the fruit development stage influence the banana susceptibility to crown rot. This involved evaluating the influence of (a) the fruit position at different levels of the banana bunch (hands) and (b) changing the source–sink ratio (So–Si ratio), on the fruit susceptibility to crown rot. The fruit susceptibility was determined by measuring the internal necrotic surface (INS) after artificial inoculation of Colletotrichum musae. A linear correlation (r = −0.95) was found between the hand position on the bunch and the INS. The So–Si ratio was found to influence the pomological characteristics of the fruits and their susceptibility to crown rot. Fruits of bunches from which six hands were removed (two hands remaining on the bunch) proved to be significantly less susceptible to crown rot (INS = 138.3 mm ²) than those from bunches with eight hands (INS = 237.9 mm ²). The banana susceptibility to crown rot is thus likely to be influenced by the fruit development stage and filling characteristics. The present results highlight the importance of standardising hand sampling on a bunch when testing fruit susceptibility to crown rot. They also show that hand removal in the field has advantages in the context of integrated pest management, making it possible to reduce fruit susceptibility to crown rot while increasing fruit size.     

Fungitoxic and phytotoxic effects of fungicides controlling powdery mildew on apple

Benomyl, pyrazophos (HOE 2873), triarimol, triforine and the non-systemic fungicide diethyl phthalimidophosphonothionate (Dowco 199) were sprayed at c. 200 gal/acre (2250 1/ha) at 14-day intervals from pink bud (late April/early May) on seven apple cultivars including Cox's Orange Pippin, Golden Delicious and Jonathan. Compared with binapacryl or dino-cap (and with elemental sulphur on the sulphur-tolerant cv. Golden Delicious), these compounds were more phytotoxic without substantially or consistently improving the control of Podosphaera leucotricha; fruit russet was increased and the crop and fruit size of some cultivars were reduced in some trials. In 1969 the inclusion of bis(dimethylthiocarbamoylthio)-methylarsine (44 ppm) as Urbacid^(R) in the sprays reduced russeting by benomyl and Dowco 199 so that these fungicides became as safe as captan on several cultivars; Dowco 199 was also safened by captan (500 ppm). Urbacid^(R) (88 ppm a.i.) safened benomyl and triarimol on fewer cultivars in 1970 when russet was less severe. The addition of Urbacid^(R) did not prevent yield losses caused by sprays in either year. The crop is most sensitive to spray damage from pink bud until late June. This is a crucial period for the control of mildew, and unless new fungicides are less phytotoxic, palliatives may become necessary adjuvants.     

The control of Gloeosporium storage rot of apple by reduced spray programmes

Single sprays of benomyl (0.025 % a.i.) applied to Cox's Orange Pippin apple in either June, July or August, controlled rotting in store by Gloeosporium spp. Post-harvest dips of either benomyl (0.05 % a.i.) or thiabendazole (0.025 % a.i.) effectively controlled any infection surviving orchard treatment.     

The involvement of fungi in the breakdown of sulphited strawberries

Between 8000 and 9000 tons of strawberries are used annually for jam manufacture in the UK, c. 65 % of which are stored in sulphite liquor (6 % SO₂+lime, c. pH 3.0). Sporadic cases of disintegration of sulphited strawberries have been observed increasingly in recent years. In laboratory experiments breakdown of whole berries in sulphite liquor was achieved by including berries partly rotted with Mucor mucedo, Rhizopus sexualis or R. stolonifer, 1.5-2.5 % infected material causing complete breakdown of all of the berries. The inclusion of up to 25 % berries infected with Botrytis cinerea caused no softening of the berries. The addition of culture filtrates to whole fruit in sulphite liquor confirmed observations with the above fungi but also showed that pectolytic enzymes from Aureobasidium pullulans and Trichosporon pullulans could cause breakdown. The incidence of spoilage fungi on commercially harvested fruit indicated that M. mucedo and R. sexualis were the main cause of breakdown, but that A. pullulans increased tenfold on fruit stored at 15 °C overnight before sulphiting. Results from commercially harvested fruit (cv. Cambridge Favourite) in 1975 and 1976 showed that: (I) fruit should be sulphited as soon as possible after harvest, since the storage of harvested fruit prior to sulphiting may give a poorer quality product; (2) there is little difference in breakdown of fruit from plants treated with the pre-harvest fungicides, Elvaron, Mildothane, or Daconil, and (3) fruit grown in different areas of the UK shows varying amounts of breakdown. This may be due to differences in the infection level of Phycomycetes and contamination by other pectolytic fungi or inherent differences in the susceptibility of the fruit to enzyme attack.     

In vitro and In vivo Effects of Some Fungicides against the Chickpea Blight Pathogen, Ascochyta rabiei

The effects of various fungicides on mycelial growth and spore germination of Ascochyta rabiei were determined by incorporating them into potato dextrose agar and measuring colony diameter and observing colony growth and spore germination at 20 ± 2°C. Eight fungicides prevented spore germination of the pathogen at concentrations of 0.125–2 μg/ml, three hindered mycelial growth at 2–4 μg/ml and seven failed to inhibit mycelial growth even at 128 μg/ml. The reference fungicide for the pathogen, chlorothalonil, stopped conidial germination at low rates but did not prevent mycelial growth at 128 μg/ml. Thirteen fungicides were tested against seed infections of the pathogen, and benomyl + thiram, carbendazim and carbendazim + chlorothalonil seed treatments gave more than 85% inhibition on both vacuum‐infiltrated and naturally infected seeds. Coating the seeds with polymers did not increase the effectiveness of fungicides. Three fungicides; (azoxystrobin, chlorothalonil and mancozeb), gave the highest protection in the field but protection decreased with increased inoculum pressure. Addition of humic acid to fungicide suspensions did not affect their performance.     

Applications of bromelain from pineapple waste towards acne

Bromelain is a proteolytic mixture obtained from pineapple (Ananas comosus (L. Merr)). It has diversified clinical properties and is used in alleviation of cancer, inflammation and oxidative stress. The current study focuses on extraction of bromelain from different parts of pineapple such as core, crown, fruit, peel and stem. The extracted enzyme was precipitated using ammonium sulphate at 40% saturation followed by dialysis. The fold of purification obtained for peel, crown, core, fruit and stem were found to be 1.948, 1.536, 1,027, 1.989, and 1.232 respectively. Bromelain activity was estimated using Azocasein assay, the highest activity was seen in peel at 3.417 U/μg. Antimicrobial activity and MIC of the bromelain purified and crude fractions was studied against the test organisms. Peel crude and purified extract exhibited highest inhibitory effect towards S. aureus followed by P. acne. The antioxidant activity was evaluated using DPPH antioxidant assay. IC50 values peel, fruit, stem and crown are found to be 13.158 μg/ml, 24.13 μg/ml and 23.33 μg/ml and 113.79 μg/ml respectively. The purified bromelain from peel, stem and crown was used to create a facewash formulation towards pathogens frequently associated with skin infections. Common skin pathogens like S. aureus and P. acne were found highly sensitive to its action. The aim of this study was to evaluate the potential of bromelain isolated from waste parts of pineapple in alleviation of acne due to its diverse antimicrobial properties.