Postharvest Decay: Control Strategies

Postharvest Decay

Control Strategies

Editor

Silvia Bautista-Baños

Instituto Politécnico Nacional. Centro de Desarrollo de, Productos Bióticos, Yautepec Morelos, Mexico

Table of Contents

Cover image

Title page

Copyright

Preface

List of Contributors

Chapter 1. Rhizopus stolonifer (Soft Rot)

Overall Importance of the Fungus

Taxonomy and Morphology

Biology of the R. Stolonifer Infection Process

Environmental and Agronomic Factors Favoring R. Stolonifer Infection

Postharvest Factors Favoring R. Stolonifer Infection

Control

Concluding Remarks

Chapter 2. Penicillium digitatum, Penicillium italicum (Green Mold, Blue Mold)

Overall Importance of Fungi

Taxonomy, Morphology and Genomics

Biology of Penicillium Digitatum and P. Italicum Infection Process

Postharvest Factors Influencing Penicillium Digitatum and P. Italicum Infection

Control

Concluding Remarks

Chapter 3. Crown Rot Disease of Bananas

Overall Importance of the Disease

Taxonomy and Morphology

Biology of Crown rot Infection Process

Preharvest Factors Favoring Crown Rot Disease

Postharvest Factors Favoring Crown Rot Disease

Control Methods and Alternatives to Chemical Control

Concluding Remarks

Chapter 4. Botrytis cinerea (Gray Mold)

Overall Importance of the Fungus

Taxonomy

Morphology

Biology of Botrytis Cinerea Infection Process

Agronomic and Environmental Factors Influencing Botrytis Cinerea Infection

Postharvest Factors Favoring Botrytis Cinerea Infection

Control

Concluding Remarks

Chapter 5. Alternaria alternata (Black Rot, Black Spot)

Overall Importance of the Fungus

Taxonomy and Morphology

Biology of A. Alternata Infection Process

Agronomic and Environmental Factors Favoring A. Alternata Infection

Postharvest Factors Favoring A. Alternata Infection

Control

Concluding Remarks

Chapter 6. Penicillium expansum (Blue Mold)

Overall Importance of the Fungus

Taxonomy and Morphology

Biology of P. expansum Infection Process

Agronomic and Environmental Factors Favoring P. Expansum Infection

Postharvest Factors Favoring P. Expansum Infection

Control

Concluding Remarks

Chapter 7. Monilinia fructicola, Monilinia laxa (Monilinia Rot, Brown Rot)

Overall Importance of Fungi

Taxonomy and Morphology

Biology of Monilinia Fructicola and M. Laxa Infection Process

Environmental and Agronomic Factors Favoring Monilinia Fructicola and M. Laxa Infection

Postharvest Factors Favoring Monilinia Fructicola and M. Laxa Infection

Control

Concluding Remarks

Chapter 8. Aspergillus spp. (Black Mold)

Overall Importance of Fungi

Taxonomy and Morphology

Agronomic and Environmental Factors Favoring Aspergillus Infection

Postharvest Factors Favoring Aspergillus Infection

Control

Concluding Remarks

Chapter 9. Thielaviopsis paradoxa, Thielaviopsis basicola (Black Rot, Black Root Rot)

Introduction

Overall Importance of Fungi

Taxonomy and Morphology

Biology of Fungal Infection Process

Agronomic and Environmental Factors Favoring T. paradoxa and T. basicola Infection

Postharvest Factors Favoring T. paradoxa and T. basicola Infection

Control

Concluding Remarks

Chapter 10. Lasiodiplodia theobromae in Citrus Fruit (Diplodia Stem-End Rot)

Overall Importance of the Fungus

Taxonomy and Morphology

Biology of L. Theobromae Infection Process

Agronomic and Environmental Factors Favoring L. Theobromae Infection

Control

Concluding Remarks

Chapter 11. Colletotrichum gloeosporioides (Anthracnose)

Overall Importance of the Fungus

Taxonomy and Morphology

Biology of C. Gloeosporioides Infection Process

Agronomic and Environmental Factors Favoring C. Gloeosporioides Infection

Postharvest Factors Favoring C. Gloeosporioides Infection

Control

Concluding Remarks

Index

Copyright

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Preface

Fruits and vegetables are living organisms that after harvest obtain their energy through the respiration process. This postharvest metabolism causes the commodity to ripen leading eventually to senescence. The positive characteristics that make fruit suitable for consumption also make them susceptible to disease. During this ripening period, fruit is prone to develop rots caused by microorganisms that hasten commodity ripening, damage their internal and/or external appearance, cause off-odors, produce mycotoxins and contaminate adjacent commodities. Although the economic losses due to fungal infection in fruits and vegetables during the postharvest chain are variable and not well documented, they usually reach anywhere from 30 to 50% and, on some occasions, rots can lead to total loss of the produce. Both fungi and bacteria cause rots; however, in general, fungal infections are reported to have a greater ability to infect a broader range of hosts throughout the whole postharvest chain. Several fungal species affect commercially important fruits and vegetables, such as Alternaria alternata, which is present in tomatoes, peppers, mangoes and other crops. The genus Penicillium, including species such as P. italicum, P. digitatum and P. expansum, has a wide range of hosts such as citrus fruits and apples. Aspergillus is also considered a serious postharvest fungus of temperate and tropical fruits and vegetables. Probably the most important aspect of the above-mentioned fungi is that they are also considered allergenic. Monilia fructicola, M. laxa and Botrytis cinerea are important pathogens of various economically important temperate fruits such as apples, grapes, kiwifruit and peaches, among others. Other important fungi are Colletotrichum gloeosporioides, Botryiodiplodia theobromae and various species of the genus Thielaviopsis, which can be highly infectious for fruits grown under tropical and subtropical conditions, such as mangoes, papayas, citrus and pineapples. Crown rot disease, caused by a complex fungal association, is also a serious disease that affects bananas and certain vegetables while Rhizopus stolonifer is a saprophytic fungus that can totally invade fruits and vegetables in less than three days.

Infection by these fungi can occur during crop blossom, at harvest or during handling operations. Control of these fungi has always included the use of fungicides. However, in spite of developing new chemicals with apparently less side effects, producers are under heavy pressure to reduce or eliminate their application. To control these fungi, other commercially viable alternatives or possible future options that work integrated with synthetics or alone have been put forward. They include physical means including heat, irradiation, and low temperatures, hypobaric, modified and controlled atmospheres, biological control with antagonistic microorganisms and GRAS (generally regarded as safe) compounds such as sanitizers, salts, volatiles, and so forth. To date, postharvest pathology research is also focusing on enhancing the individual potential of the host to respond to the attack of the pathogen – resistance inducers. Products such as jasmonates, chitosan, salycilic acid and harpin among others are under study. Depending on the particular pathogen, control may vary; however, all the contributors to this book agree that integration of one or more different strategies to achieve higher levels of disease control should be proposed.

The information in this book covers a wide range of topics related to each selected fungus, such as economic importance, morphology and taxonomy, biology of the infection process, the influence of preharvest agronomic practices and the environment, and the effect of handling operations for the infection and disease development to occur and the strategic controls for each host–pathogen, including traditional and non-traditional alternatives.

This book is aimed at a wide audience including researchers involved in the study of postharvest handling of agricultural commodities and undergraduate and graduate students whose area of research deals with postharvest topics. Growers, managers and operators working at packinghouses and storage, retail and wholesale facilities can also benefit from this book.

The collaborators of this book are experienced researchers in the field of postharvest pathology of fruits and vegetables. Contributors are high-level scientists with many years of research experience related to each selected fungus. The authors belong to scientific associations, participate and collaborate as authors of articles published in international journals and serve as advisors on scientific committees in the postharvest pathology area. I am deeply grateful to all of them for being willing to spare time in their busy agendas as lecturers and researchers to attend to my invitation. I am sure that the information stated in this book will serve present and future generations involved in the postharvest pathology field.

I am also grateful to the editors Patricia Osborn, Carrie Bolger and the Elsevier Production Department for their guidance from beginning to the end of this project.

Silvia Bautista-Baños

List of Contributors

Asgar Ali,     Centre of Excellence for Postharvest Biotechnology (CEPB), School of Biosciences, The University of Nottingham Malaysia Campus, Semenyih, Selangor D. E., Malaysia

Laura L. Barrera-Necha,     Centro de Desarrollo de Productos Bióticos, Instituto Politécnico Nacional Carretera Yautepec-Jojutla km. 8.5, Col. San Isidro, Yautepec Morelos, Mexico

Silvia Bautista-Baños,     Centro de Desarrollo de Productos Bióticos, Instituto Politécnico Nacional Carretera Yautepec-Jojutla km. 8.5, Col. San Isidro, Yautepec Morelos, Mexico

Elsa Bosquez-Molina,     Universidad Autónoma Metropolitana-Iztapalapa, Col. Vicentina, Mexico

Luc de Lapeyre de Bellaire,     CIRAD, Research Unit Banana, Plantain and Pineapple Cropping Systems, Montpellier, France

Deena Errampalli,     Agriculture and Agri-Food Canada, SCPFRC, Vineland Station, ON, Canada

Erica Feliziani,     Marche Polytechnic University, Department of Agricultural, Food, and Environmental Sciences, Ancona, Italy

I.G.N. Hewajulige,     Senior Deputy Director/Principal Research Scientist, Food Technology Section, Industrial Technology Institute, Colombo, Sri Lanka

Ludivine Lassois,     Forest, Nature and Lanscape Department, Liège University, Gembloux Agro-Bio Tech, Gembloux, Belgium

Marta Mari,     CRIOF, DipSA, University of Bologna, Cadriano, Bologna

Camilla Martini,     CRIOF, DipSA, University of Bologna, Cadriano, Bologna

Lluís Palou,     Laboratori de Patologia, Centre de Tecnologia Postcollita (CTP), Institut Valencià d’Investigacions Agràries (IVIA), Montcada, València, Spain

Maribel Plascencia-Jatomea,     Departamento de Investigación y Posgrado en Alimentos, Universidad de Sonora, Hermosillo, Sonora, México

Gianfranco Romanazzi,     Marche Polytechnic University, Department of Agricultural, Food, and Environmental Sciences, Ancona, Italy

Yasmeen Siddiqui,     Laboratory of Food Crops, Institute of Tropical Agriculture, Universiti Putra Malaysia, Serdang Selangor D. E., Malaysia

Martín Ernesto Tiznado-Hernández,     Coordinación de Tecnología de Alimentos de Origen Vegetal, Centro de Investigación en Alimentación y Desarrollo, Hermosillo, Sonora, México

Rosalba Troncoso-Rojas,     Coordinación de Tecnología de Alimentos de Origen Vegetal, Centro de Investigación en Alimentación y Desarrollo, Hermosillo, Sonora, México

John Martin Velez-Haro,     Departamento de Investigación y Posgrado en Alimentos, Universidad de Sonora, Hermosillo, Sonora, México

R.L.C. Wijesundera,     Senior Professor of Plant Pathology, Department of Plant Sciences, University of Colombo, Colombo, Sri Lanka

María Susana Yépiz-Gómez,     Departamento de Investigación y Posgrado en Alimentos, Universidad de Sonora, Hermosillo, Sonora, México

Jiuxu Zhang,     Florida Department of Citrus, Lake Alfred, Florida, USA

Chapter 1

Rhizopus stolonifer (Soft Rot)

Silvia Bautista-Baños¹, Elsa Bosquez-Molina²,  and Laura L. Barrera-Necha¹     ¹Centro de Desarrollo de Productos Bióticos, Instituto Politécnico Nacional Carretera Yautepec-Jojutla km. 8.5, Col. San Isidro, Yautepec Morelos, Mexico     ²Universidad Autónoma Metropolitana-Iztapalapa, Col. Vicentina, Mexico

Abstract

Rhizopus stolonifer is considered the most important species in the genus Rhizopus. Disease symptoms that characterize R. stolonifer infection are watery areas quickly covered by coarse, gray hairy mycelia forming a mass of black sporangia at their tips. Infection usually occurs during harvest and handling. Due to the wide array of hosts that R. stolonifer can infect and its fast penetration and colonization, it has become an important target to control. Overall, R. stolonifer is controlled using synthetic fungicides; among the various registered fungicides are thiabendazole, benomyl, imazalil, propiconazole and thiocarbamates. The fungicides fluodioxonyl, tebuconazole, fenhexamide and azoxystrobin have also achieved notable control of this fungus. Sanitizers like chlorine, sodium hypochlorite, calcium hypochlorite, ozone and peracetic acid have shown to have good control on Rhizopus. Of the various non-conventional methods under experimentation, antagonists have attracted considerable attention. Bacteria including Pantoe agglomerans, Enterobacter cloacae, Pseudomonas syringae, and Bacillus spp., as well as yeasts of the genera Pichia, Kloeckera, Candida, Cryptoccoccus, Metschnikowia and Acremonium, have exerted a significant control on this fungus. Aqueous plant extracts belonging to the botanical families Amaryllidaceae, Annonaceae, Arecaceae, Bromeliaceae, Caricaceae, Lamiaceae, Lauraceae, Leguminosae, Meliacea, Myrtaceae, Pinaceae, Sapotaceae, Solanaceae and Urticaceae, along with the essential oils of the genera Thymus, Cinnamom, Origanum and Citrus, have also produced excellent results under experimentation. UVC irradiation levels of up to 3.6  kJm² showed levels of control up to 50% on R. stolonifer. Chitosan, a widely known chitin derivative, alone or in combination with other natural products, has been able to reduce significantly Rhizopus soft rot on various horticultural commodities at various concentrations. Volatiles, hypobaric atmospheres, salts, minerals, plant phenolics, hot water, wax and salicylic acid have resulted in good control methods with promising applications.

Keywords

black mold; rhizopus rot; postharvest control; non-conventional methods

Contents

Overall Importance of the Fungus 1

Taxonomy and Morphology 5

Biology of the R. stolonifer Infection Process 6

Environmental and Agronomic Factors Favoring R. stolonifer Infection 8

Postharvest Factors Favoring R. stolonifer Infection 9

Control 9

Conventional Alternatives 9

Fungicides 9

Sanitizers 14

Alternatives Under Experimentation 18

Antagonists 18

Plant Derivatives 24

Irradiation 29

Chitosan 31

Hypobaric Atmosphere 31

Generally Regarded as Safe (GRAS) Compounds 34

Concluding Remarks 35

References 37

Overall Importance of the Fungus

Rhizopus stolonifer is one of the most common and fastest-growing species in the Zygomycota phylum. Disease caused by this fungus is known as soft rot, black mold and Rhizopus rot. It often grows within a few days in moist and humid conditions because its spores are quite common in the air. Rhizopus stolonifer is an important postharvest pathogen that contributes to considerable spoilage of a great variety of fruits and vegetables (Table 1.1). Genera susceptible to this fungus include Allium, Ananas, Brassica, Cucumis, Cucurbita, Fragaria, Lycopersicon, Phaseolus, Pisum, Solanum, and several more. Other Rhizopus species such as R. oryzae and R. artocarpi are also reported to cause serious rots on fruits such as papaya, banana, mulberry roots and jackfruit, among others (McMillan, 1986; Yoshisda et al., 2003; Kwon et al., 2012). Due to its fast growth, it is considered one of the most devastating fungi during the storage period of various horticultural commodities.

Table 1.1

Summary of some Plant Species and Organs Affected by Rhizopus stolonifer and Overall Expressed Symptomatology∗

-; Not reported.

∗ Symptoms for all the above-mentioned fruits and vegetables also include host covered by coarse, gray and hairy white mycelia that usually form a mass of visible black sporangia at their tips.

So far, there are limited data on the exact losses attributed to R. stolonifer. In this respect, Ceponis and Butterfield (1979) reported that 80% of the total losses in the New York City tomato market was due to R. stolonifer and Alternaria alternata. In Nigeria, Fajola (1979) conducted a survey in five states of postharvest fruit rot diseases of tomato, finding that 85% of the total loss was due to soft rot diseases with R. stolonifer among the major pathogens in this country. Shehu and Muhammad (2011) and Bayewu et al. (2007) identified R. stolonifer among the most pathogenic in the mycoflora associated with storage disease of onion bulbs and papaya fruit, respectively. Results of a 3-year survey carried out in the Central province of Sri Lanka showed total postharvest losses from 5 to 50% due to microorganism infection. Among the identified fungi, R. stolonifer was considered as the one to cause fast and extensive rotting of a wide range of vegetable and fruit hosts (Adikaram, 1993). In markets in Asiut, Egypt, Abdel-Mallek et al. (1995) and Ziedan and Farrag (2008) reported that R. stolonifer from diseased tomatoes and peaches was isolated at a frequency of 35.9% and 56.5%, respectively. This last value is only exceeded by Aspergillus niger and A. alternata. This fungus is also reported to infect severely roots such as white yam (Dioscorea rotundata) in Ghana. In this study, out of the 10 microorganisms isolated from this tuber, R. stolonifer incidence was in a range of 20 to 55% depending on the market sampled (Aboagye-Nuamah et al., 2005). In other studies, Bautista-Baños et al. (2008) reported that in tomatoes collected from 15 different local markets in the state of Morelos, Mexico, R. stolonifer was the most predominant fungus, isolated from almost 50% of the samples. Out of the 15 fungal genera identified on imported fruits in Shangai, China markets (Teng-fei et al., 2009), Rhizopus was identified in durian and mango; however, it was not the most predominant. In guava fruits harvested in Aksum, Ethiopia, Pestalotia psidii had a frequency of 32.1% followed by R. stolonifer with 16% and Aspergillus niger 10.4% (Mathew, 2010). Out of 18 fungal species isolated from banana, grape, litchi and mango collected at a market in Dhaka, Bangladesh, R. stolonifer was the second most common fungus with a frequency of 37% (Bashar et al., 2012). A monthly market survey in Gorakhpur, India, found the presence of R. stolonifer in papaya fruit throughout the year (Singh et al., 2012).

Taxonomy and Morphology

Rhizopus stolonifer belongs to the class Zygomycetes, order Mucorales, family Mucoraceae, genus Rhizopus and species R. stolonifer. Based on the morphological classification carried out by Shipper (1984), the common features of the R. stolonifer group are: complex and well-developed rhizoids, sporangiophores, sporangium and sporangia. The shape of the main columnella (mycelium) is conical-cylindrical; it is gray or brownish, up to 140  μm in height and branches until a tangled mycelium is formed (Fig. 1.1A). This mycelium sends up from various points aerial erect hyphae forming whitish growth in cultures turning eventually into black spots, named sporangiophores, about 1–3  mm long and up to 20–25  μm in diameter (Fig. 1.1B). Each sporangiophore bears a single spherical sporangium of up to about 250  μm in diameter that contains numerous sporangiospores of up to 13  μm in length and diameter in a range of 100–275  μm. Rhizopus spores are of different shapes: angular, subglobose and ellipsoidal, with well- marked ridges on the surface (Fig. 1.1C) depending on maturity (Swingle, 1903). However, in this respect, Hernández-Lauzardo et al. (2005) confirmed the globose type as the principal shape of R. stolonifer, which varies in size, diameter and area according to isolate. Further taxa characterization of various species of the genus Rhizopus (Bengston Frye and Reinhardt, 1993), based on DNA differences, gave R. stolonifer a mol % G + C value of 37.8.

Figure 1.1   Rhizopus stolonifer morphology: (A) in Petri plate, (B) optical photomicrography showing (B1) branched rhizoid, (B2) columnella, (B3) sporangiophores and sporangiospores, and (C) scanning electron micrography showing shape diversity of sporangiospores.

Biology of the R. Stolonifer Infection Process

For infection to occur, R. stolonifer commonly requires, among others, injuries caused by hail, cracking and mechanical wounds (Vicente et al., 2005). In this respect, Nguyen-The et al. (1989) demonstrated that microcrackings on the surface of nectarine fruits are the main entrance for R. stolonifer spores; however, they were unable to germinate unless a nutrient solution was available. On sweet potatoes, rhizopus rot development was higher in the bruise wound-type, with infection susceptibility also being associated with cultivar and storage period (Holmes and Stange, 2002). According to Clark and Hoy (1994), susceptibility to R. stolonifer infection was clearly associated with sweet potato genotypes. For most fruits and vegetables, symptoms of infection caused by this fungus may be dramatically seen in matter of 3–6 days (Fig. 1.2A–C). Overall, rhizopus rot appears water soaked and exudes a clear leachate. Mena-Nevarez et al. (2012) reported that on infected mango fruit, leachate production was four times higher than on non-inoculated fruit after 35 days of storage, while in orange fruits it was ten times higher. During the initial day of infection, the surface may be covered with thin, fluffy, cotton-like fungal structures. Once R. stolonifer has gained entry into wounded tissue, its mycelia spread around the site of infection. Eventually, dark sporulation forms a mass of black sporangia at their tips, covering the entire surface of the commodity. Lisker et al. (1996) reported that germinating Rhizopus spores in inoculated grapes were observed after 12–15 h; one h later, germ tubes started to develop and, after 24 h, fruit was covered by hyphae. For tomatoes, no germination took place by 5 h (Fig. 1.3A) postinoculation but, after this time, mycelia grew inter- and intracellularly (Fig. 1.3B); after 24 h, mycelia spread all over the fruit cell walls with a concomitant disappearance of the spore (Fig. 1.3C).

Infection by R. stolonifer is closely associated with enzymatic activities, which play an important role in its colonization of fruit. Polygalacturonase (PG) is considered the primary hydrolase involved in the softening process of tomatoes, netted muskmelons and dates during Rhizopus infection (Bruton et al., 1998; Stevens et al., 2004; Al-Hindi et al., 2011) (Table 1.2). In addition, a variety of other macerating enzymes have been assigned roles in fruit cell wall degradation in infected dates such as xylanase, cellulose and amylase (Al-Hindi et al., 2011). As reported by Villanueva (2004) and Vicente (2004), other factors associated with the enzymatic activity are the fungus structure and spore load. In those studies, the activity of PG on infected tomato markedly increased after 48 h in tomatoes inoculated with mycelia (8.8  μL galacturonic acid/min/mg of protein) compared with those inoculated with spore suspension (6.7  μL of galacturonic acid/min/mg of protein). Likewise, tomatoes inoculated with 10⁷ spores mL−¹ concentration showed the highest pectin methylesterase activity after 24 h (1.8  μL/acid/min/mg of protein) compared to tomatoes inoculated at 10⁵ spores mL−¹ (1.4  μLl/acid/min/mg of protein).

Figure 1.2   Rhizopus rot development on (A) strawberry (1, 2 and 3 days), (B) tomato (1, 2 and 4 days), and (C) papaya (1, 3 and 6 days) fruit during ambient storage (20–25°C).

Figure 1.3   Scanning electron micrographs showing the infection process of wounded tomato inoculated by Rhizopus stolonifer: (A) non-germinated spores after 0–4 h post-inoculation, (B) germinating spores after 8–12 h, and (C) tomato surface completely covered by mycelia after 24 h.

Table 1.2

Rhizopus stolonifer Cell Wall Degrading Enzyme Production on Different Horticultural Commodities

Environmental and Agronomic Factors Favoring R. Stolonifer Infection

Overall, Rhizopus species including R. stolonifer are naturally found in soil, debris and air. Fungal dispersal mechanisms of R. stolonifer are wind, air currents and some invertebrates such as mites and insects, among others. In this respect, in a study about detection of R. stolonifer in various vineyards, Rhizopus was the main fungus identified in soils collected between rows or beneath plants, and a high incidence of airborne spores coincided with the high incidence of fruit infection in the vineyard and the proximity of nearby stone-fruit orchards (Lisker et al., 1996). A notable incidence of R. stolonifer has also been reported in flowers. Harris and Dennis (1980) reported 30% isolation of this fungus in strawberry flowers. Few studies exist about the preharvest factors and their influence on the incidence of Rhizopus rot. For the reports published about agronomic practices, irrigation may increase or reduce the susceptibility of fruits to be infected by this fungus (Teviotdale et al., 1995; 2001). Studies carried out on the effect of this practice on hull rot disease of almond, reported a marked reduction when irrigation of the tree crops was limited before harvest. During two consecutive years, hull rot disease was almost undetected when preharvest irrigation terminated 52 days before harvest. In this study, an increase in R. stolonifer incidence was clearly observed as the number of days between last irrigation and harvest was reduced. Additional studies also demonstrated that deficit irrigations, manner and magnitude of water reduction before harvest, also had a direct effect on hull rot disease.

Postharvest Factors Favoring R. Stolonifer Infection

Postharvest handling operations are the main reason R. stolonifer succeeds in entering and infecting most horticultural commodities. It is continuously reported that overall R. stolonifer infection gains better access through wounds, injuries and abrasions. A study carried out by Holmes and Stange (2002) reported that disease incidence on sweet potatoes may be associated with the type of injury. The bruise-wound type injury followed by the puncture-wound type caused the highest infection rate over the 2-year assessment period. With respect to the optimum conditions for infection to occur during postharvest handling, temperature is a very important factor. Overall temperature above 15°C accelerates infection occurring in a short period of time, but in lower temperatures, infection is only delayed. Pierson (1966) reported that in peaches the maximum growth rate of R. stolonifer was achieved at 25°C, while lower temperatures of 0, 4.4 and 18°C only delayed the appearance of symptoms by a few days. Spores already attached to fruits are a source of future inoculum for future infections; therefore, a source of contamination at packing plants may be the water systems used for all commodities requiring washing as a usual postharvest operation.

Control

Conventional Alternatives

Fungicides

The use of chemicals is a common method to reduce the incidence of Rhizopus stolonifer on various horticultural commodities (Table 1.3). Its chemical control has depended on a series of fungicides that belong to the benzimidazole (thiabendazole and benomyl) and azole (imazalil, propiconazole and thiocarbamates) chemical groups. Ortho-phenylphenol (OPP) and its sodium salt, sodium ortho-phenylphenate (SOPP), are also fungicides used as postharvest treatments to control R. stolonifer on fruits and vegetables; however, their action does not control new injures that may be present after their application. The fungicide iprodione of the chemical group dicarboximide is said to reduce Rhizopus rot in various horticultural commodities; however, in some countries, this fungicide is banned or never obtained registration. Dichloran, an aromatic hydrocarbon, is a broad-spectrum fungicide used to reduce Rhizopus rot; at this time, the EPA (Environmental Protection Agency) has revoked tolerances for its use. Other registered fungicides such as captan of the chemical group thiophthalamides are rarely used since visible residues are left on the surface of the treated commodity. New fungicides such as fluodioxonyl (phenylpirrol), tebuconazole (triazole), fenhexamide (hydroxyanilide) and azoxystrobin (strobulins) are mentioned to achieve notable control of this fungus in various horticultural commodities such as pomme and stone fruits postharvest.

Table 1.3

Summary of Fungicides Applied during the pre- and Postharvest Handling of Various Horticultural Commodities and their Level of Control on Rhizopus stolonifer

The use of synthetic fungicides should be part of an integrated control. Treatments with dichloran, benomyl and paraffin-based wax mixture were approximately 95% more effective than the fungicides or wax alone in peaches and nectarines (Wells, 1972). Similar results were achieved in pear fruits with the mixture of the fungicide iprodione and wax oil (Adaskaveg et al., 1992). In other studies, a significant improvement in the control of Rhizopus rot on sweet potatoes of about 90% was achieved with the mixture of boscalid, pyraclostrobin and Pseudomonas syringae (BioSave 10LP) (Edmunds and Holmes, 2009).

Sanitizers

Surface and water disinfectants have also been considered for controlling Rhizopus stolonifer. Of the various sanitizers, chlorine (HOCl) is the most widely applied treatment in packing line wash water. It is used as a general disinfectant and it could be added in water solutions (dip, spray, or drench) and as a volatile (gas). Other forms of chlorine also used are sodium hypochlorite (NaHClO) (bleach) and calcium hypochlorite Ca(ClO)2. According to the literature review, the efficacy of these treatments is proportional to the type and form of application of the sanitizer, concentration, exposure time and cultivar (Table 1.4). For example, the efficacy of chlorine was approximately 50% greater on tomatoes than on nectarines, peaches and strawberries (Vigneault 2000; Mari et al., 2004; Avis et al., 2006; Adaskaveg, 2009). For the control of this fungus, calcium and sodium hypochlorite have also been tested on table grapes cv. ‘Shami’ and tomatoes ‘V2 type’, respectively (Abdell-Mallek et al. 1995; Lisker et al., 1996), resulting in an average efficiency of 100% on table grapes and 47% on tomatoes. Scientific research has proved that ozone (O3) is another method for controlling decay caused by R. stolonifer. Although this disinfectant method is generally used for controlling food-borne pathogens, experimentally, it has given a good level of control on strawberries cv. ‘Seascape’ (Nicoué et al., 2004) and, when combined with ethanol at 70%, complete control of R. stolonifer on table grapes cv. ‘Alphonse Lavalle’, ‘Thompson Seedless’ and ‘Zeiny’ was achieved (Sarig et al., 1996). Another sanitizer reportedly under experimentation is peracetic acid (C2H4O3) (PAA). Overall, this sanitizer when used as a cleanser and disinfectant, controls various pathogenic microorganisms. Experimentation on this matter has shown that postharvest application of PAA at a concentration of 250  mg L−¹ gave a total inhibition of R. stolonifer after 2 min of exposure on apricots, nectarines and peaches (Mari et al., 2004). Additionally, it was reported that preharvest application of commercial PAA (Oxidate®) during flowering and fruit set generally reduced postharvest rots of strawberries (Narciso et al., 2007).

Table 1.4

Summary of Sanitizers Applied during the Pre- and Postharvest Handling of Various Horticultural Commodities and their Level of Control on Rhizopus stolonifer

Alternatives Under Experimentation

Antagonists

Overall, the biological control of R. stolonifer involves the use of antagonistic microorganisms such as bacteria, yeast and one fungus (Table 1.5). Bacteria such as Pantoe agglomerans, Enterobacter cloacae, Pseudomonas syringae and Bacillus spp. have shown an important reduction of R. stolonifer on apricot, nectarine, peach, pear, and sweet potato (Wilson et al., 1987; Nunes et al., 2001a, 2002; Northover and Zhou, 2002; Bonaterra et al., 2003; Edmund and Holmes 2009; Wang 2013a, b), while various yeasts of the genera Pichia, Kloeckera, Candida, Cryptoccoccus, Metschnikowia and Acremonium also exert significant control of this fungus on various species of stone fruit, strawberry, sweet cherry, table grape and tomato (McLaughlin et al., 1992; Qing and Shiping, 2000; Shena et al., 2000; Qin et al., 2004; Zhang et al., 2007b; Zhao et al., 2008) and only one fungus classified as Trichoderma harziarum has been reported to control Rhizopus rot on apple, peach, and tomato during storage (Batta, 2007; El-Katatny and Emam, 2012). As reported, the efficacy of the antagonist depends on various factors including antagonistic concentration, type of strain and culture conditions. Overall, literature reports that higher concentrations of the antagonist provided a lower incidence of Rhizopus disease and lesion diameter in most of the tested cultivars. The efficacy of the antagonist was also associated with specific strains as reported with the yeast Kloeckera apiculata and the bacterium Enterobacter cloacae (Wilson et al., 1987; McLaughlin et al., 1992), Pseudomonas syringae (Edmunds and Holmes, 2009) and various unidentified yeast isolates (Shena et al., 2000). According to McLaughlin et al. (1992), the efficacy of K. apiculata for controlling R. stolonifer of the cv. ‘Thompson Seedless’ in grapes was 40% greater when cultures were not filtrated. Another study by Zhao et al. (2008) about the performance of the antagonist indicated that its efficacy is associated with fruit maturity. In this study, the maturity stage of tomatoes was directly associated with the level of efficacy against R. stolonifer, with the mature green tomatoes being where the antagonist P. guilliermondii produced 100% inhibition compared with the 80% achieved in red and pink tomatoes. In other studies, it was stated that the combination of the antagonist with short hot water dips, calcium chloride or essential oils may enhance the efficacy of the treatments by 90–100%. This was reported in tomatoes (Abd-Alla et al., 2009), peaches and strawberries (Zhang et al., 2007a,c). So far, few commercial biofungicides that control R. stolonifer based on antagonists are available in the market; however, Biosave® 10LP