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Plant Pathologist’s Pocketbook 3rd Edition

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  • Plant Pathologists Pocketbook3rd Edition

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  • Plant Pathologists Pocketbook3rd Edition

    Edited by

    J.M. Waller, J.M. Lenn and S.J. Waller

    CABI Publishing

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  • CABI Publishing is a division of CAB International

    CABI PublishingCAB InternationalWallingfordOxon OX10 8DEUK

    Tel: +44 (0)1491 832111Fax: +44 (0)1491 833508Email: [email protected] site: www.cabi-publishing.org

    CABI Publishing10 E 40th Street

    Suite 3203New York, NY 10016

    USA

    Tel: +1 212 481 7018Fax: +1 212 686 7993

    Email: [email protected]

    CAB International 2002. All rights reserved. No part of this publicationmay be reproduced in any form or by any means, electronically,mechanically, by photocopying, recording or otherwise, without the priorpermission of the copyright owners.

    A catalogue record for this book is available from the British Library, London, UK.

    Library of Congress Cataloging-in-Publication DataPlant pathologists pocketbook / edited by J.M. Waller, J.M. Lenn and S.J.Waller. --3rd ed.

    p. cm.Includes bibliographical references (p. ).ISBN 0-85199-458-X (hardcover) -- ISBN 0-85199-459-8 (pbk.)

    1. Plant diseases--Handbooks, manuals, etc. 2. Plant diseases--Bibliography. I. Waller,J. M. II. Lenn, Jillian M. III. Waller, S. J. (Sarah J.)

    SB601 .P57 2001632.3--dc21

    2001020356Pbk ISBN 0 85199 459 8Hbk ISBN 0 85199 458 X

    Typeset by AMA DataSet Ltd, UK.Printed and bound in the UK by Biddles Ltd, Guildford and Kings Lynn.

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  • ContentsContentsContents

    Contents

    Contributors viii

    Preface to the 3rd Edition x

    Part I: Recognition and Evaluation of Plant Diseases

    1 Landmarks in Plant Pathology 12 Some Major Plant Diseases 4

    J.M. Lenn3 Surveys and Sampling 19

    M. Holderness4 Disease and Yield Loss Assessment 25

    P.S. Teng and W.C. James5 Postharvest Diseases 39

    Revised by J.M. Waller6 General Bibliography of Plant Pathology 55

    Compiled by J.M. Waller7 Bibliography of Crop and Plant Diseases 60

    Compiled by J.M. Waller8 Collection and Dispatch of Plant Material 72

    J.M. Waller and B.J. Ritchie

    Part 2: Causes of Plant Disease

    9 Fungi as Plant Pathogens 75J.M. Waller and P.F. Cannon

    v

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  • 10 Bacteria and Plant Disease 94G. Saddler

    11 Virus Diseases 108Revised by J.M. Waller

    12 Phytoplasma Plant Pathogens 126P. Jones

    13 Plant Parasitic Nematodes 140J. Bridge and T.D. Williams

    14 Insect and Other Arthropod Pests 163G.W. Watson

    15 Weeds 175P.J. Terry and C. Parker

    16 Parasitic Higher Plants 183C. Parker

    17 Non-infectious Disorders 190J.M. Lenn

    18 Air Pollution Effects and Injury 195A.R. Wellburn

    19 Effects of Climate Change 203S. Chakraborty

    Part 3: Disease Diagnosis and Investigation

    20 Detection and Isolation of Fungal and Bacterial Pathogens 208J.M. Waller

    21 Microscopy 216J.M. Waller

    22 Immunological Techniques 221F.M. Dewey

    23 Biochemical and Molecular Techniques 229P. Bridge

    24 Inoculation 245J.M. Waller

    25 Epidemic Modelling and Disease Forecasting 252M.W. Shaw

    26 Design of Experiments 266J. Riley

    Part 4: Disease Management

    27 Plant Health and Quarantine 279J.M. Waller

    28 Regional and Country Lists of Plant Diseases 287Revised by J.M. Waller

    29 Seed Health 309Revised by J.M. Waller

    30 Cultural Control 318A.J. Termorshuizen

    vi Contents

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  • 31 Disease Resistance 328J.M. Waller and J.M. Lenn

    32 Fungicides 336D. Hollomon

    33 The Application of Chemicals for Plant Disease Control 345G.A. Matthews

    34 Biological Control of Fungal Plant Pathogens 354S.S. Navi and R. Bandyopadhyay

    35 Plant Pathogens for Biological Control of Weeds 366H.C. Evans

    Part 5: General Techniques and Information

    36 Safety in the Laboratory 379B.J. Ritchie

    37 Culturing, Preservation and Maintenance of Fungi 384D. Smith

    38 Mycological Media and Methods 410B.J. Ritchie

    39 Photography 432E. Boa

    40 Standards and Measurements 446Compiled by J.M. Waller

    41 Publication 454J.M. Waller

    42 Electronic Databases and Information Technology in PlantPathology 461H.L. Crowson and L.A. McGillivray

    43 Glossary of Plant Pathological Terms 470Compiled by J.M. Lenn

    44 Useful Addresses 487

    General Index 491

    Diseases and Pathogens Index 505

    Contents vii

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  • ContributorsContributorsContributors

    Contributors

    R. Bandyopadhyay, International Crops Research Institute for the Semi-AridTropics (ICRISAT), Patancheru, Andhra Pradesh 502 324, India

    E. Boa, CABI Bioscience UK Centre, Bakeham Lane, Egham, Surrey TW20 9TY,UK

    J. Bridge, CABI Bioscience UK Centre, Bakeham Lane, Egham, Surrey TW209TY, UK

    P. Bridge, Mycology Section, Royal Botanic Gardens, Kew, Richmond, SurreyTW9 3AE, UK and School of Biological and Chemical Sciences, Birkbeck,University of London, Malet St, London WC1E 7HX, UK

    P.F. Cannon, CABI Bioscience UK Centre, Bakeham Lane, Egham, Surrey TW209TY, UK

    S. Chakraborty, CSIRO Plant Industry, CRC Tropical Plant Protection,University of Queensland, Queensland 4072, Australia

    H.L. Crowson, CAB International, Wallingford, Oxfordshire OX10 8DE, UKF.M. Dewey, Plant Sciences Department, University of Oxford, South Parks

    Road, Oxford OX1 3RB, UKH.C. Evans, CABI Bioscience UK Centre, Silwood Park, Ascot, Berkshire SL5

    7PY, UKM. Holderness, CABI Bioscience UK Centre, Bakeham Lane, Egham, Surrey

    TW20 9TY, UKD. Hollomon, Long Ashton Research Station, University of Bristol, Long

    Ashton, Bristol BS18 9AF, UKW.C. James, ISAAA AmeriCenter, c/o Cornell University, Ithaca, NY 14853,

    USAP. Jones, Plant Pathogen Interactions (PPI), IACR-Rothamsted, Harpenden,

    Hertfordshire AL5 2JQ, UK

    viii

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  • J.M. Lenn, International Crops Research Institute for the Semi-Arid Tropics(ICRISAT), Patancheru, Andhra Pradesh 502 324, India

    G.A. Matthews, International Pesticide Application Research Centre, ImperialCollege at Silwood Park, Ascot, Berkshire SL5 7PY, UK

    L.A. McGillivray, CAB International, Wallingford, Oxfordshire OX10 8DE, UKS.S. Navi, International Crops Research Institute for the Semi-Arid Tropics

    (ICRISAT), Patancheru, Andhra Pradesh 502 324, IndiaC. Parker, 5 Royal York Crescent, Bristol BS8 4JZ, UKJ. Riley, IACR-Rothamsted, Harpenden, Hertfordshire AL5 2JQ, UKB.J. Ritchie, CABI Bioscience UK Centre, Bakeham Lane, Egham, Surrey TW20

    9TY, UKG. Saddler, CABI Bioscience UK Centre, Bakeham Lane, Egham, Surrey TW20

    9TY, UKM.W. Shaw, Department of Agricultural Botany, School of Plant Sciences, The

    University of Reading, Reading RG6 6AS, UKD. Smith, CABI Bioscience UK Centre, Bakeham Lane, Egham, Surrey TW20

    9TY, UKP.S. Teng, Monsanto Company, Kings Court II Building, 2129 Chino Roces

    Avenue, Makati City, PhilippinesA.J. Termorshuizen, Biological Farming Systems, Wageningen University,

    Marijkeweg 22, 6709 PG Wageningen, The NetherlandsP.J. Terry, Long Ashton Research Station, University of Bristol, Long Ashton,

    Bristol BS18 9AF, UKJ.M. Waller, CABI Bioscience UK Centre, Bakeham Lane, Egham, Surrey TW20

    9TY, UKS.J. Waller, 45 St Michaels, Longstanton, Cambridge, UKG.W. Watson, Entomology Department, The Natural History Museum,

    Cromwell Road, London SW7 5BD, UKA.R. Wellburn, formerly of Division of Biological Sciences, Lancaster

    University, Lancaster LA1 4YQ, UKT.D. Williams, 4 Warren Park, West Hill, Ottery St Mary, Devon EX11 1TN, UK

    Contributors ix

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  • Preface to the 3rd EditionPreface to the 3rd EditionPreface

    Preface to the 3rd Edition

    The second edition of the Plant Pathologists Pocketbook published in 1983 isnow well out of date and this edition has been completely reorganized to createa format more suitable to the current needs of plant pathologists. Thetraditional and well-known title has been retained although the book is nowrather larger than pocketbook size. The contents have been arranged into fivesections covering the main activity groups in plant pathology. Several newtopics have been added including chapters covering epidemiology and diseaseforecasting, disease resistance, biochemical and molecular techniques, andelectronic databases and information technology. Other topics covered in thesecond edition have all been revised or completely rewritten and the associatedbibliographies updated. In order to keep the pocket book to size it has beennecessary to restrict accounts of most subjects to succinct overviews, but hope-fully the attached bibliographies will provide sufficient information to assistpathologists with their work. Inevitably decisions have had to be made on howmuch to include on particular aspects; the pocketbook is not intended as ageneral text on plant pathology but as a reference for information and relevantliterature. As such the book will be used more in the laboratory than carried inthe pocket and the larger size and rearranged format will hopefully enablebetter access to the contents.

    Many chapters have been written or revised by authors both from CABIBioscience and from outside; others have been revised or written by the editors.We acknowledge particular assistance with the chapter on viruses fromProfessor Mike Thresh of NRI, University of Greenwich.

    Jim WallerJill Lenn

    Sarah Waller

    x

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  • Landmarks in Plant PathologyLandmarks in Plant Pathology1

    1Landmarks in Plant Pathology

    Early Recognition of Plant Diseases

    c. 300 BC Theophrastus of Lesbos references to plant diseases in Historiaplantarum and De causis plantarum.

    c. AD 50 Caius Plinius Secundus references to common diseases such asmildew and rusts of cereals in Historia naturalis.

    c. AD 700 Cassianus Bassus Geoponica a compilation of Byzantineagriculture with many references to plant diseases.

    c. AD 1200 Ibn-al Awam Kitab al-Felahah Arabic treatise from Sevillewith a chapter on problems of regional fruit crops.

    Beginnings of Plant Pathology

    1665 Robert Hooke Micrographia contains first illustration of amicroscopic plant pathogen (rose rust).

    1755 Mathieu Tillet demonstrates seed-borne nature of wheat bunt(Tilletia caries).

    1794 J.J. Plenk publishes Physiologia et Pathologia Plantarum con-taining a classification of plant diseases based on symptoms.

    1802 William Forsyth introduces lime sulphur for control of mildewon fruit trees first example of generally used fungicide.

    1807 I.-B. Prevost publishes first experimental proof of fungal patho-genicity on plants.

    18451849 Potato blight (Phytophthora infestans) epidemics in Ireland.

    CAB International 2002. Plant Pathologists Pocketbook(eds J.M. Waller, J.M. Lenn and S.J. Waller) 1

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  • 1853 Anton de Bary publishes Unterschungen ber die brandpilzeand established the role of fungi as plant pathogens.

    1858 Julius Kuhn publishes Die Krankheiten der Kulturgewachse the first plant pathology text.

    1865/66 De Bary demonstrates heteroecism in Puccinia graminis.18681882 Coffee rust (Hemileia vastatrix) epidemics in Sri Lanka and

    classic studies on its aetiology by Marshal Ward.1874 Paul Soraur publishes first edition of Handbuch fr Pflanzen-

    krankheiten of which there have been many subsequent editions.18781885 Vine downy mildew (Plasmopara viticola) epidemics in France

    precipitate introduction of Bordeaux mixture by Millardet.1885 J.C. Arthur provides first proof of bacterial pathogenicity to

    plants with fire blight (Erwinia amylovora).18861898 Tobacco mosaic recognized as a virus disease transmitted by

    contagious living fluid by Mayer, Ivanovski and Beijerinck.1889 Hot water treatment of seed introduced for control of loose smut

    of wheat (Ustilago tritici).

    Development of Basis for Modern Plant Pathology

    1891/92 First plant pathology journals (Zietschrift fr Pflanzenkrank-heiten and Revista di Patologia Vegetale) published.

    1894 Jacob Eriksson demonstrates host-specific special forms of cerealrusts.

    1904 Rowland Biffen demonstrates Mendelian inheritance ofresistance to rust in cereals.

    1907 First university department of plant pathology founded atCornell University, New York.

    1908 American Phytopathology Society founded.1913 Recognition of physiologic races of Puccinia graminis by

    Stakman.First use of organo-mercurial fungicides for seed application inGermany.

    1914 First International Phytopathological Convention of Rome heldbut resolutions never ratified.

    1920 Imperial Bureau of Mycology founded in UK (a forebear ofCABI Bioscience) production of Review of Plant Pathology (asReview of Applied Mycology) commences 2 years later.E.F. Smith publishes Introduction to Bacterial Diseases ofPlants.

    1929 International Convention for the Protection of Plants inRome produces first limited internationally agreed protocols forrestricting the spread of plant diseases.

    1934 Dithiocarbamate fungicides developed and patented by Du Pont.1937 F.C. Bawden and N.W. Pirie establish nucleoprotein nature of

    viruses.

    2 Landmarks in Plant Pathology

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  • 1946 Flor establishes gene-for-gene hypothesis for genetic inter-action of resistance and virulence for flax rust Melampsora lini.

    1951 European and Mediterranean Plant Protection Organizationfounded as first of several regional plant protection organizations.

    1952 International Plant Protection Convention submitted to the Foodand Agriculture Organization (FAO) and agreed in principle.

    1963 Annual Review of Phytopathology begins.J.E. Vanderplank publishes Plant Diseases: Epidemics andControl.First example of biological control of a disease Heterobasidionannosum by Peniophora gigantea.

    1965 Discovery of first systemic fungicide, carboxin.1967 First recognition of phytoplasmas (mycoplasma-like organisms)

    as plant pathogens.1968 First International Congress of Plant Pathology held in London.1969 First computer simulation program for plant disease epidemics.1970 Southern corn leaf blight epidemic on hybrid (Tms) maize in

    USA.1971 Deiner establishes potato spindle tuber as first recognized viroid

    disease.1977/78 Plant Disease: An Advanced Treatise (3 Vols) by Horsfall and

    Cowling published.1986 Plants genetically transformed to express virus coat proteins

    shown to be resistant to same virus.1992 First examples of plant resistance genes isolated.

    General References

    Ainsworth, G.C. (1981) An Introduction to the History of Plant Pathology. CambridgeUniversity Press, Cambridge, UK, 315 pp.

    Carefoot, G.L. and Sprot, E.R. (1967) Famine in the Wind. Rand McNally, Chicago, USA.Large, E.C. (1940) Advance of the Fungi. Jonathan Cape, London, UK, 488 pp.Parris, G.K. (1968) A Chronology of Plant Pathology. Johnson & Sons, Mississippi, USA.

    Landmarks in Plant Pathology 3

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  • Some Major Plant DiseasesJ.M. Lenn2

    2Some Major Plant DiseasesJ.M. LennICRISAT, Patancheru, Andhra Pradesh 502 324, India

    Diseases are listed under hosts, which are arranged alphabetically by commonname.

    In column two current scientific names are given in accordance with theCABI Crop Protection Compendium (Global Module 2nd edition, 2000); thesemay differ from those given in literature references given in column four whereearlier synonyms may have been used.

    In column three a broad indication of geographical distribution is given.In most cases detailed information on distribution can be found in the

    relevant map in the series CMI/IMI/CABI Distribution Maps of Plant Diseases,indicated by Map 38 etc. and also in the CABI Crop Protection Compendium(Global Module 2nd edition, 2000).

    In column four Desc. 147 etc. refers to the series of CMI/IMI/CABIDescriptions of Pathogenic Fungi and Bacteria; Virus Desc. 68 etc. refers to theseries of AAB Descriptions of Plant Viruses. References in the form 58, 1234 areto abstracts in the Review of Plant Pathology. Further references can be found inthe CABI Crop Protection Compendium (Global Module 2nd edition, 2000).

    CAB International 2002. Plant Pathologists Pocketbook4 (eds J.M. Waller, J.M. Lenn and S.J. Waller)

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  • Some Major Plant Diseases 5

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  • 6 J.M. Lenn

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    16Z:\Customer\CABI\A4084 - Waller - Plant Pathologists Pocketbook\A4163 - Waller - Plant - All.vpMonday, October 29, 2001 4:37:48 PM

    Color profile: DisabledComposite Default screen

  • Some Major Plant Diseases 7

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    17Z:\Customer\CABI\A4084 - Waller - Plant Pathologists Pocketbook\A4163 - Waller - Plant - All.vpMonday, October 29, 2001 4:37:48 PM

    Color profile: DisabledComposite Default screen

  • 8 J.M. Lenn

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    18Z:\Customer\CABI\A4084 - Waller - Plant Pathologists Pocketbook\A4163 - Waller - Plant - All.vpMonday, October 29, 2001 4:37:48 PM

    Color profile: DisabledComposite Default screen

  • Some Major Plant Diseases 9

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    19Z:\Customer\CABI\A4084 - Waller - Plant Pathologists Pocketbook\A4163 - Waller - Plant - All.vpMonday, October 29, 2001 4:37:48 PM

    Color profile: DisabledComposite Default screen

  • 10 J.M. Lenn

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    20Z:\Customer\CABI\A4084 - Waller - Plant Pathologists Pocketbook\A4163 - Waller - Plant - All.vpMonday, October 29, 2001 4:37:48 PM

    Color profile: DisabledComposite Default screen

  • Some Major Plant Diseases 11

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    21Z:\Customer\CABI\A4084 - Waller - Plant Pathologists Pocketbook\A4163 - Waller - Plant - All.vpMonday, October 29, 2001 4:37:49 PM

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  • 12 J.M. Lenn

    Com

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    22Z:\Customer\CABI\A4084 - Waller - Plant Pathologists Pocketbook\A4163 - Waller - Plant - All.vpMonday, October 29, 2001 4:37:49 PM

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  • Some Major Plant Diseases 13

    Mille

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    23Z:\Customer\CABI\A4084 - Waller - Plant Pathologists Pocketbook\A4163 - Waller - Plant - All.vpMonday, October 29, 2001 4:37:49 PM

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  • 14 J.M. Lenn

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    4

    24Z:\Customer\CABI\A4084 - Waller - Plant Pathologists Pocketbook\A4163 - Waller - Plant - All.vpMonday, October 29, 2001 4:37:49 PM

    Color profile: DisabledComposite Default screen

  • Some Major Plant Diseases 15

    Pota

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    sum

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    cter

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    rose

    ptica

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    406

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    25Z:\Customer\CABI\A4084 - Waller - Plant Pathologists Pocketbook\A4163 - Waller - Plant - All.vpMonday, October 29, 2001 4:37:49 PM

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  • 16 J.M. Lenn

    Com

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    5

    26Z:\Customer\CABI\A4084 - Waller - Plant Pathologists Pocketbook\A4163 - Waller - Plant - All.vpMonday, October 29, 2001 4:37:49 PM

    Color profile: DisabledComposite Default screen

  • Some Major Plant Diseases 17

    Phyt

    opht

    hora

    root

    and

    stem

    rot

    Bact

    eria

    lblig

    ht

    Phyt

    opht

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    5

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    271

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    iafu

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    iana

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    Map

    62

    Des

    c.43

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    00

    Suga

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    349

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    299

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    27Z:\Customer\CABI\A4084 - Waller - Plant Pathologists Pocketbook\A4163 - Waller - Plant - All.vpMonday, October 29, 2001 4:37:49 PM

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  • 18 J.M. Lenn

    Com

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  • Surveys and SamplingM. Holderness3

    3Surveys and SamplingM. HoldernessCABI Bioscience UK Centre, Bakeham Lane, Egham, SurreyTW20 9TY, UK

    General Principles

    Surveys of diseases in crops are required for many purposes: to determine gen-eral levels of crop health; or the presence of particular diseases of quarantinesignificance; prioritization of problems to enable proper allocation of cropprotection resources; and to assess the losses caused by crop disease asdiscussed in Chapter 4. However, they are all concerned with determining thetype and severity of diseases affecting crops. The structure of disease surveysand protocols for sampling of crops varies widely between different scientificneeds, e.g. quarantine officers may require subsamples of seed lots for detectionof specific pathogens, whereas extension workers may require rapid assess-ment of diseases and of the farmers priorities in regard to the disease complexin his/her fields. The diagnostic survey thus combines identification of dis-eases with a measure of their intensity. Qualitative aspects are concerned withdetermining what is there. This may be either the type of damage, symptoms,etc., or which particular species of pest or pathogen is present. Samples willoften need to be collected in order to confirm identities of pests and pathogensby laboratory examination. Quantitative aspects determine how much of aparticular pest or pathogen is present, particularly in relation to how muchdamage is being caused to the crop. Incidence and severity need to be measuredaccording to a predetermined sampling pattern. Surveys may need to be under-taken several times during the season as particular crop stages may differ intheir susceptibility or diseases may develop in characteristic ways. The datamay also need to relate to particular crop growth stages in order to determinethe effect of disease on yield. Patterns of disease spread may be critical indiagnosis or in devising control methods. Aerial surveys are sometimes used toassess crop health or patterns of disease development over large areas. These

    CAB International 2002. Plant Pathologists Pocketbook(eds J.M. Waller, J.M. Lenn and S.J. Waller) 19

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  • may use true light or infra-red images, but usually need to be followed up onthe ground for sampling and diagnostic procedures. Diseases do not usuallyoccur in isolation and measures of the relative importance of different compo-nents of the disease complex are required to determine which would justifycontrol measures and whether such measures are likely to be economic. Pestsand abiotic factors may also be present and causing damage to the crop andthese need to be taken into account during the survey.

    Planning the Survey

    Before any survey starts, decisions are required on:

    1. The population or region to be covered;2. The characteristics to be estimated;3. The subclasses (e.g. varieties, soil types) to be covered;4. The degree of accuracy required.

    These factors determine the basic structure of the survey. Of these, point 2involves the techniques to be used and the skills required, whereas 3 and 4determine the scale of the survey.

    The survey must cover sites that are truly representative of the region andmust be of sufficient size to obtain the required accuracy. Planning the surveyin relation to the resources (people, time, money, transport) available is crucialand the collection of extraneous information should be avoided at all costs. Theplanning should decide the minimum level of information required and matchthat against the maximum level of information which could be gathered giventhe resources available.

    To ensure survey objectives can be met, the planning process should takeaccount of all four stages of the survey:

    1. Planning: decide what data are required.2. Sampling and collection: the gathering of the data.3. Analysis: conversion of the data into accessible information.4. Report: use of the information to achieve the original objective.

    If necessary, survey objectives can be modified during stages 1, 3 or 4, but suchflexibility is difficult, if not impossible, during the sampling and collectionstage. Before data collection begins (and before the training of data collectorsbegins), it is essential to have determined the survey goals including detail onthe variables for which data will be collected, how often they will be collectedand the level of accuracy which is required. One very useful way to do thisis to undertake a small pilot survey in advance of the actual survey, so thata realistic assessment can be made of what will be feasible and achievable,before extensive resources are committed.

    A survey is not an experiment; it is feasible to use a survey to determine thelikelihood of different pest and environmental factor combinations, but onecannot expect to make very accurate estimates of the effects of different factorcombinations on the yields of crops from surveys alone. The objectives of adiagnostic survey may thus perhaps best be limited to the discovery of what

    20 M. Holderness

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  • combinations of pests and hosts exist and a relatively crude assessment of thedamage caused in the cropping systems under study.

    Sampling

    Appropriate sampling procedures comprise a combination of a suitablesampling method and a representative sample size. For a sample to be repre-sentative it must relate to the spatial distribution of disease and for it to havethe desired level of accuracy, it must be of an adequate size in relation to thesurvey objectives and logistics.

    Sampling procedures should be: simple; representative; reliable; andideally applicable in subsequent standard disease surveillance procedures.

    Sample size

    The sample is usually made up of several small sampling units (e.g. leaf, plantor plot) which are often predetermined by the purpose of the study, as aresampling dates and intervals. Stratified sampling can be appropriate to thedetection of very low disease incidences, but care is needed in the bulking ofsamples and subsequent analysis to avoid losing true replication, or converselyto avoid creating spurious levels of accuracy. If the standard deviation of esti-mated pest incidence is: +/ SD and the mean is to be estimated with standarderror: +/ SE, the number (n) of independent random samples required is:

    n = (SD)2/(SE)2

    However, the standard deviation is often unknown and must be guessed, usinginformation from a pilot study or comparable pathosystems. A rough estimateof the sample sizes required can be obtained from a pilot study in whichdisease intensities are estimated for a number of sampling units and therunning means and their corresponding standard deviations plotted againstthe number of sampling units. Both such curves tend to flatten after a certainnumber of entries and this point can be taken as the minimum (and thus inmany cases optimum) sample size to obtain a