Biological control of postharvest fungal rot of yam (<Emphasis Type="Italic">Dioscorea</Emphasis> spp.) with<Emphasis Type="Italic">Bacillus subtilis</Emphasis>

The potential of isolates of Bacillus subtilis from yam farm soil to control rot of yam in storage barns was investigated. Yam tubers inoculated in vivo with B. subtilis showed no rot while those inoculated with Aspergillus niger, Botryodiploidia theobromae or Penicillium oxalicum showed considerable rot. The set of yams in which B. subtilis and the fungi were simultaneously inoculated produced rot whereas those in which B. subtilis was inoculated a day before the fungi was inoculated were totally reduced or free of rot. Many fewer fungi were isolated from the surface of tubers treated with B. subtilis than from the untreated (control) and there was high recovery of B. subtilis (99–100%) throughout the period of storage. Rot build up was faster in uninoculated control tubers or those inoculated with a spoilage fungus, while those treated with the antagonist were totally reduced or free of rot. The culture filtrate of B. subtilis prevented spore germination in some spoilage fungi. The importance of this study in relation to farmers in developing countries is discussed.     

Evaluation of isolates of Trichoderma,Pseudomonas and Bacillus species as treatment for the control of post-harvest fungal rot disease of yam (Dioscorea spp.)

The efficacy of four biological control agents (BCAs): Trichoderma asperellum strain NGT158, T. longibrachiatum strain NGT167, Bacillus subtilis and Pseudomonas fluorescens for the management of post-harvest tuber rot among four yam species, Dioscorea rotundata, D. cayenensis, D. alata, and D. dumetorum was evaluated. Rotted yam tubers were collected across three agroecological zones in Nigeria to isolate six infecting fungal pathogens: Aspergillus niger, Lasiodiplodia theobromae, Rhizoctonia solani, Penicillium oxalicum, Fusarium oxysporum and Sclerotium rolfsii. The BCAs were isolated by serial dilution and rot inhibition of treated tubers was evaluated using destructive sampling method in vivo after six months of storage. Bacillus subtilis was generally most effective, especially when applied 24 h before the inoculation of test pathogens across the four yam species, with percent inhibition that ranged between 47.8 and 81.2%. However, the four BCAs showed good potential in the control of the fungal pathogens causing post-harvest yam rot.     

Mycoflora of tuber surface of white yam (Dioscorea rotundata poir) and postharvest control of pathogens with Bacillus subtilis

Bacillus subtilis (Enrenberg) Cohn was investigated for its antagonistic properties against surface mycoflora of yam (Dioscorea rotundata Poir) tubers in storage. Yam tubers inoculated with a spore suspension of B. subtilis in potato dextrose broth using a knapsack sprayer showed a drastic reduction in the range and number of mycoflora, including pathogens of the tuber surface in contrast to the control tubers, during the five-month storage period in a traditional yam barn. However, B. subtilis maintained a high frequency of occurrence during the same period. Botryodiploidia theobromae Pat, Fusarium moniliforme Wollen and Reink., Penicillium sclerotigenum Yamamoto, and Rhizoctonia sp. were displaced completely on the treated tubers. The antagonism of B. subtilis was so effective that the normal tuber surface mycoflora was greatly reduced throughout the storage period of five months by a simple initial application of the antagonist.This revised version was published online in October 2005 with corrections to the Cover Date.     

Severity of spoilage storage rots of white yam (Dioscorea rotundata Poir.)

Severity of storage rots in different sections of white yam tubers (Dioscorea rotundata Poir.) was investigated. Yam samples with rots were collected from a yam barn and from selected markets in Accra, Ghana, to identify the most predominant pathogens associated with the rots. Nine fungal spoilage microorganisms, including Aspergillus flavus, Aspergillus niger, Botryodiplodia theobromae, Fusarium culmorum, Fusarium oxysporium, Fusarium sp., Penicillium brevi‐compactum, Penicillium sp. and Rhizopus stolonifer and a bacterium species Erwinia carotovora were identified. The mean incidence of occurrence of the organisms on rotten tissues ranged from 1.2% to 28.5%. Of the 10 microorganisms isolated, B. theobromae, F. oxysporium and R. stolonifer were the most frequently encountered spoilage microorganisms in the markets. E. carotovora, Fusarium solani and Penicillium sp. were relatively sparse (incidence not exceeding 3%) compared to the other yam spoilage microorganisms. The surface area and weight of necrotic tissue induced by the spoilage fungi in the various zones of the tubers over a 28‐day storage period were assessed. All the spoilage microorganisms produced rots in the yams, although to different degrees. The severity of the rots increased in weight and area over the period when the tubers were in store but were normally not significantly different in the zones of tubers. There was, however, a linear progression of rots in the various zones of the yam tubers. Although there was generally no significant (P ≥ 0.05) difference in the severity of rots induced by the different microorganisms in the tubers, R. stolonifer commonly induced more rot in the zones of the tubers compared to B. theobromae and F. oxysporium.     

Field Studies on Cross-protection against Japanese yam mosaic virus in Chinese yam (Dioscorea opposita) with an Attenuated Strain of the Virus

An attenuated strain of Japanese yam mosaic virus (JYMV), designated T‐3, was evaluated for its cross‐protection efficacy against virulent (native) strains of JYMV in Chinese yam (Dioscorea opposita) grown in farmers’ fields in Japan. Native strains of JYMV were detected by a polymerase chain reaction‐based assay in all the Chinese yam plants grown from virus‐free tubers in the first growing season in the fields. In contrast, the virus was detected in only one of fifty plants grown from tubers preinoculated with T‐3 during the experiments for 6 years, suggesting that T‐3 consistently cross‐protected against native JYMV in Chinese yam in the field. Chinese yam plants preinoculated with T‐3 produced significantly greater yield of tubers per plant compared with non‐inoculated plants.     

Evaluation of Clove Essential Oil as a Mycobiocide Against Rhizopus stolonifer and Fusarium solani,Tuber Rot Causing Fungi in Yam (Dioscorea rotundata Poir.)

To exploit natural products for plant disease control, the essential oil of Syzygium aromaticum (L.) Merr. & Perr. (clove) was investigated for its antifungal activity against Rhizopus stolonifer and Fusarium solani, the postharvested yam tuber rot pathogens. The essential oil was obtained by hydrodistillation using a Clevenger‐type apparatus. The chemical composition of the oil was determined by gas chromatography and gas chromatography coupled with mass spectrometry. Antifungal activities of the oil were tested in vitro against mycelia growth and spores germination. In situ tests were conducted on healthy yam tubers, and necrosis symptoms were assessed. Results showed that eugenol (79.4%), eugenylacetate (9.2%) and isocaryophyllene (7.0%) were the major components. The oil exerted antifungal activities with total inhibition (TI) of the mycelia growth of R. stolonifer and F. solani was recorded at 200 and 300 ppm, respectively, while TI of spores germination was recorded at 31.25 and 250 ppm, respectively. For the standard fungicide (Ridomil^®), TI value of mycelia growth was 1600 ppm for the both pathogens, while TI of spores germination were 200 ppm and 1600 ppm, respectively, for Rhizopus and Fusarium. In situ tests showed complete inhibition of yam tuber rot when the essential oil was applied at 2000 ppm for preventive tests. This oil also reduced significantly (P ≤ 0.05) necrosis development on yam tuber for curative test at the same concentration. Total inhibition of the necrosis by Ridomil (3000 ppm) was observed only for Rhizopus on preventive test. There were positive correlations between the oil concentration and the reduction of necrosis cause by R. stolonifer and F. solani. These findings showed that clove essential oil may serve as environmental friendly bio‐fungicide for the management of postharvest yam tuber rot.     

Studies on the Biological Control of Phytophthora cryptogea Pethybr. et Laff. II. Effectiveness of Trichoderma and Gliocladium spp. in the Control of Phytophthora Foot Rot of Gerbera

Bip T containing about 10⁹ spores of Trichoderma viride, applied to peat 10 days before inoculation of substrate with Phytophthora cryptogea, effectively controlled Phytophthora foot rot of gerbera. The biocide in dosage of 300 and 600 g/m³ inhibited the development of the pathogen in substrate. The other potential antagonists T. hamatum and T. viride applied into peat at a dosage of 600 g/m³, decreased Phytophthora foot rot development.     

Trichoderma viride—Mediated Modulation of Oxidative Stress Network in Potato Challenged with Alternaria solani

Potato is a staple food crop cultivated globally. Heavy losses to potato production are reported annually due to soil borne phytopathogens. Trichoderma viride is a potential biocontrol agent that improves host defense. In the present study, potato tubers bio-primed with T. viride were studied for its effect on growth promotion and modulation of antioxidant system as well as defense-related enzymes in potato plants when challenged with Alternaria solani. Potato tubers treated with T. viride and after 45 days of sowing, plants were challenged with pathogen. Significant improvement in various growth parameters was recorded in bio-primed plants. While, in pathogen-challenged plants, an enhanced intracellular concentration of H₂O₂ and O₂^? was observed. Interestingly, T. viride when applied with pathogen, significantly improved the redox homeostasis by modulating the antioxidant enzyme activities. The significant induction of defense enzymes and free phenolic content suggested that T. viride-treated plants provide enhanced protection from oxidative stress induced during A. solani challenge via. elevated accumulation of antioxidant enzymes, polyphenolic compounds, and defense-related enzymes.

     

Concentrated Distribution of Wild Yam Patches: Historical Ecology and the Subsistence of African Rainforest Hunter-Gatherers

Availability of annually reproducing wild yam species (Dioscorea praehensilis and Dioscorea semperflorens) is considered to be a key factor for the viability of forest hunter-gatherer subsistence in the African rainforests. I here describe the habitat and distribution of wild yam patches, and discuss possible human impacts on the formation of wild yam patches, which currently support long-term forest expeditions by Baka hunter-gatherers in southeastern Cameroon. An analysis of canopy photographs shows that annual yams favor a habitat of canopy gaps which receive sufficient sunlight. However, the fact that annual yams patches are limited to specific locations even though gaps in the canopy occur throughout the forest, combined with the mostly vegetative reproductive system of the yams, suggests annual yams do not propagate extensively in the wild. Moreover, areas with abundant annual yam patches that are presently situated far from any villages contain several abandoned village sites. Since the Baka often transplant parts of yam tubers to encourage their spread, it seems that human activities may have contributed to the formation of locally concentrated annual yam patches.     

Biocontrol of onion white rot by application of Trichoderma species formulated on wheat bran powder

In vitro, Trichoderma album, Trichoderma harzianum, Trichoderma koningii, Trichoderma viride and Trichoderma virens showed antagonistic effect against the most pathogenic isolate (Sc2) of Sclerotium cepivorum, the cause of onion white rot disease. Five Trichoderma preparations of each Trichoderma sp. were prepared on wheat bran powder to be used for controlling white rot disease of onion. Greenhouse and field experiments followed the same trend where T. harzianum and T. koningii were the most effective in reducing the incidence and severity of white rot disease compared with the control. Trichoderma species preparations caused promotion to vegetative parameters of onion plants in pots and increase bulb productivity in filed. In this regard, T. harzianum and T. koningii were the most effective. A positive correlation was found between the biocontrol activity of Trichoderma species preparations and enhancement of peroxidase, polyphenoloxidase and chitinase enzymes in onion plants to resist infection with S. cepivorum.     

Preponderance and distribution of plant-parasitic nematodes in Nassarawa State,North Central Nigeria: Consequences for healthy tuber production and export

One of the most cosmopolitan pests responsible for considerable negative economic impacts on yam is the plant-parasitic nematodes. They are also capable of facilitating fungal, bacterial and viral infections of yams. In order to produce healthy yam seeds to meet local demand and tubers for export, there is a need to identify nematode-free areas in Nassarawa, one of the hubs of yam production in Nigeria. To this end, 30 major yam growing communities of the state were surveyed to identify nematode-free areas. Structured questionnaires were administered to three farmers in each of the 10 local government areas selected. Results showed that 97% of the study areas investigated was seriously infested with the three key genera of plant-parasitic nematodes namely Scutellonema, Pratylenchus and Meloidogyne. Incidence and severity of tuber deterioration in transit threatened acceptability of the tubers when it eventually reached England. Healthy yams would attract more income than damaged ones.     

Biological treatments for the control of decay in tree wounds.: Field tests

In an attempt to control fungi causing decay a range of micro-organisms, selected after laboratory tests, was inoculated into wounds made in the trunks of beech trees. The most successful micro-organism was an isolate of Trichoderma viride, which, over a period of 4 years, maintained significantly lower colonisation by decay fungi at both sites - c. 15% of the level found in uninoculated wounds. T. viride could be reisolated from the majority of wounds into which it had been inoculated up to 4 years later. No other antagonist persisted very effectively. Inoculation of T. viride in glycerol improved its establishment but malt extract and sucrose had no such effect. Both the addition of glycerol to wounds and the wrapping of them in black PVC sheet increased natural colonisation by T. viride. The use of PVC but not glycerol reduced numbers of decay fungi but not through the agency of T. viride present by natural infection, as this proved generally ineffective in the control of decay fungi. Another isolate of T. viride persisted considerably better when covered with a wound sealant.     

Natural plant volatiles as an alternative approach to control stem‐end rot in avocado cultivars

Stem‐end rot is a postharvest disease associated with multiple important fungal pathogens including Lasiodiplodia theobromae. The incidence of stem‐end rot in avocado during postharvest storage affects the shelf life, quality and marketability of the fruit. This study is aimed at the investigation of the antifungal activities of selected natural plant volatiles (vapour phase): citral, octanal, hexanal and thymol against L. theobromae (causal pathogen of stem‐end rot) in vitro and in vivo in “Hass” and “Fuerte” avocados. Hexanal showed a lower inhibitory effect on the radial mycelial growth of L. theobromae in vitro. However, citral at a minimum concentration of 4 μl/L revealed fungicidal activity and completely inhibited the spore germination of L. theobromae. Artificially inoculated “Hass” and “Fuerte” avocados with L. theobromae were exposed to citral (768 μl) and commercial fungicide prochloraz and stored for 6 days at 20°C and 14 days at 10°C separately and thereafter held at 20°C for 3 days to simulate the retail shelf conditions. Although citral in a volatile phase effectively reduced the development of stem‐end rot in both cultivars, its effect was significant in “Fuerte” with 75% reduction in the incidence of stem‐end rot. The biochemical analysis demonstrated an increase in total phenol contents, phenylalanine ammonia‐lyase, chitinase and β‐1, 3 glucanase activity in fruit exposed to citral when compared to the reference treatment prochloraz and the untreated control for both cultivars. Furthermore, fruits exposed to citral retained the ready‐to‐eat firmness and therefore could be considered a potential alternative treatment to control stem‐end rot at the postharvest stage.     

Biocontrol potential of Trichoderma species against mango malformation pathogens

Malformation disease of Mango (Mangifera indica L.) caused by Fusarium moniliforme var. subglutinans is one of the most destructive diseases, which is a major production constraint in the mango-growing regions of India. In this study, The bioagents Trichoderma viride (Tr1), Trichoderma virens (Tr2) and Trichoderma harzianum (Tr3) were evaluated in culture with the pathogens to monitor the antagonistic effect and their volatile compound and culture filtrates (non-volatile compound). It was found that all the three isolates of bioagents significantly checked the growth of F. moniliforme var. subglutinans. In dual culture, the best result was obtained with T. harzianum followed by T. virens and T. viride. A similar result was also observed in the case of culture filtrates ofTrichoderma spp. The results clearly showed that inhibition of the growth of the fusaria isolates by T. harzianum was significantly superior to T. viride andT.virens. In case of antifungal activity of volatile compounds released by Trichoderma isolates, it was also observed that T. virens was more superior to T.harzianum and T. viride.     

Potential for Biocontrol of Lasiodiplodia theobromae (Pat.) Griff. & Maubl. in Banana Fruits by Trichoderma Species

Fifteen Trichoderma isolates were tested for their antagonistic ability against Lasiodiplodia theobromae. Trichoderma harzianum exhibited the greatest inhibition in dual culture. Microscopic investigation demonstrated direct parasitism and coiling of T. harzianum and T. viride around hyphae of L. theobromae, causing swollen, deformed, shortened, or rounded cells of the pathogen. Granulation of cytoplasm and disintegration of the hyphal walls of L. theobromae also were noted in dual culture. Trichoderma viride reduced rotting by 29.07 to 65.06% in artificially inoculated banana fruits. Treatment of banana fruits with T. viride 4 h prior to inoculation with L. theobromae provided better protection than simultaneous application or treatment 4 h after inoculation.     

Studies on the effect of gamma radiation (from Cobalt 60 source) on storage life of white yam (Dioscorea rotundata var. efon) infected with Scutellonema bradys

Studies on the effect of gamma irradiation on yam tubers infected with Scutellonema bradys showed that dosages between 5 and 15 krad did not eliminate the nematodes completely, but suppressed sprouting and signs of deterioration in the tubers. Dosages between 20 and 30 krad eliminated 70–80 % of the nematode population in yam peels. The appearance and taste of yams treated with between 7·5 and 15 krad were generally acceptable to a panel of tasters.     

Ecological management of tropical sugar beet (TSB) root rot (Sclerotium rolfsii (Sacc.) by rhizosphere Trichoderma species

Trichoderma species are collected from different location of sugarbeet growing areas of Tamil Nadu and it is effective against Sclerotium rolfsii pathogen caused by sugarbeet ecosystems. Out of thirty-one isolates of Trichoderma viride and four isolates of Trichoderma harzianum collected and tested for their antagonistic activity against S. rolfsii by dual culture technique, one isolate was found to be effective T. viride (TVB1) that recorded the maximum (73.03%) inhibition on the mycelial growth recording only 2.40 cm growth as against 8.90 cm in the control. The isolates of T. harzianum THB-1 recorded 71.19% mycelial growth reduction over control. The colonisation behaviour of T. viride (TVB1) revealed that it completely over grew on pathogen within 48 h after interaction with the pathogen, and speed of growth on pathogen was also high and it possesses a higher level of competitive saprophytic ability. The best four isolates of TVB1, TVB-2, TVB-3 and TVB31 and two isolates of T. harzianum THB-1 and THB-2 were compared with other species of Trichoderma longibrachiatum, Trichoderma reesei, Trichoderma koningii and Chaetomium globosum and tested under in vitro condition. BA of neem cake at 150 kg ha^?1 + T. viride isolate (TVB1) at 2.5 kg/ha recorded least root rot disease incidence of 17.05% which accounted for 75.37% disease reduction over control and highest recorded maximum root yield 65.73 t ha^?1 and increasing sugar content.     

Phosphite uptake and distribution in potato tubers following foliar and postharvest applications of phosphite‐based fungicides for late blight control

Foliar and postharvest applications of phosphite (Phi)‐based fungicides are used to control the oomycete Phytophthora infestans which is responsible for the occurrence of late blight in potatoes. Optimisation of the usage of Phi‐based fungicides for disease control during the growing season and in subsequent storage can lead to improved potato production and processing quality. In order to assess the efficiency of Phi translocation to tubers, following foliar and postharvest treatments of potato crops with the Phi‐based fungicides, the amount of Phi in tubers was determined by a high‐performance ion chromatography method. The quantity of Phi found in tubers increased with the total amount of Phi‐based fungicides applied during the growing season. Foliar applications of Phi resulted in an uneven distribution of Phi in the three tuber regions analysed, with high concentrations being identified in the tuber cortex (32.5–166.4 µg g^?1 fresh tissue) and medulla regions followed by the skin area. Postharvest treatment of tubers led to a different distribution of Phi, with the highest concentrations of Phi found in the skin (411.0–876.6 µg g^?1 fresh tissue) followed by the cortex and medulla regions. As foliar treatments are essential to protect the aerial parts of the plants during the growth season, the best disease management practices of tubers should include the postharvest treatment in addition to foliar applications. The use of both types of treatments ensures that concentrations of Phi in excess of 100 µg g^?1 fresh tissue are present in tuber skin and cortex areas; such concentrations are needed to suppress the growth of P. infestans on tubers during storage.     

The infection of oranges by Trichoderma viride and mixed infection by Trichoderma viride and Penicillium digitatum

Trichoderma viride spores applied in water to apparently uninjured skin of oranges do not cause lesions. Adding orange juice, rind extract, citric acid or orange essential oil did not influence infection. Oranges became infected only when the stem-end cuts or wounds deeper than 6 mm into oil vesicles were inoculated. Sound oranges in contact with decayed oranges did not become infected. Diphenyl-impregnated wrappers reduced infection. A mixed inoculum of T. viride and Penicillium digitatum caused as fast rotting as P. digitatum, which caused faster rotting than T. viride alone. Lesions infected with P. digitatum could become infected by T. viride but those caused by T. viride did not become infected by P. digitatum. T. viride was antagonistic to P. digitatum in vivo and in vitro, possibly because it produces a heat-labile diffusible substance toxic to P. digitatum.