S'0-M•^ Genetics and breeding of Agaricus Proceedings of the First International Seminar on Mushroom Science, Mushroom Experimental Station, Horst, the Netherlands, 14-17 May 1991 LJ.LD. van Griensven (Editor) a Pudoc Wageningen 199 KVwÜNIVERSITKO CIP-data Koninklijke Bibliotheek, Den Haag Genetics Genetics and breeding of Agaricus :proceeding s of the First International Seminar on Mushroom Science, Mushroom experimental Station, Horst, the Netherlands, 14-17 May 1991 / L.J.L.Dva n Griensven (ed.) - Wageningen : Pudoc. - III. ISBN 90-220-1045-7 bound NUGI 835 Subject heading: mushroom breeding. ISBN 90-220-1045-7 NUGI 835 ©Centr e for Agricultural Publishing and Documentation (Pudoc), Wageningen, Netherlands, 1991. All rights reserved. Nothing from this publication may be reproduced, stored in acomputerize d system or published inan yfor m or inan ymanner , includingelectronic , mechanical,reprographi c or photographic, without prior written permission from the publisher, Pudoc, P.O. 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CONTENTS Preface vii Keynote lecture A personal view on mushroom breeding from 1957 - 1991 Gerda Fritsche Analysis of the genome of Agaricus bisporus A short introduction to the ecology, taxonomy and nomenclature of the genusAgaricus. C. Bas 21 Freeze-drying of fungal hyphae and stability of the product. CS. Tan, C.W. Van Ingen & J.A. Stalpers 25 A genetic linkage map for Agaricus bisporus. R.W. Kerrigan, P.A. Horgen & J.B. Anderson 31 Mitochondrial plasmids and homologous sequences inAgaricus. Mary M. Robinson & Paul A. Horgen 37 Mitochondrial genotypes and their inheritance in the cultivated mushroom Agaricus bisporus. Anton S.M. Sonnenberg, P.C.C. Van Loon & L.J.L.D. Van Griensven 42 Efficient protoplast formation and regeneration and electrophoretic karyotype analysis of Agaricusbisporus. J.C. Royer, W.E. Hintz & P.A. Horgen 52 Chromosome separation and assignment of DNA probes inAgaricus bisporus. Anton S.M. Sonnenberg, K. Den Hollander, A.P.J. Van De Munckhof & L.J.L.D. Van Griensven 57 The use of protoplast production, protoplast regeneration and restriction fragment length polymorphisms in developing a systematic and highly reproducible breeding strategy for Agaricus bisporus. P.A. Horgen, T. Jin & J.B. Anderson 62 Use of the polymerase chain reaction (PCR) inA. bisporusbreedin g programs. R.S. Khush, L. Morgan, E. Becker & M. Wach 73 Progress in the molecular analysis ofAgaricus enzymes. D.A. Wood & CF. Thurston 81 Genetics and Disease Control Fungi in the cultivation ofAgaricus bisporus-a n updated list of species. Albert Eicker & Martmari Van Greuning 89 Nature of disease resistance to compost-borne and airborne pathogens ofAgaricus bisporus. P.J . Wuest 97 Molecular dissection and control of virus disease inAgaricus bisporus. M.C. Harmsen & J.G.H. Wessels 103 Evidence for transmission of La France disease inAgaricus bisporus by ds RNA. Anton M. Sonnenberg & L.J.L.D. Van Griensven 109 Hydrophobin genes in mushroom development. J.G.H. Wessels 114 A DNA sequence inducing mushroom development in Schizophylum. Carlene A. Raper & J. Stephen Horten 120 Strategies for breeding and preparation of spawn Transformation strategies for Agaricus.M.P . Challen, B.G. Rao & T.J. Elliott 129 Towards a transformation system for Agaricus bisporus. John C. Royer & P.A. Horgen 135 The isolation of two tandemly linked glyceraldehyde-3-phosphate dehydrogenase genes from Agaricusbisporus. M.C. Harmsen, J. Scheer, T.A. Schuurs & J.G.H. Wessels 140 Maintenance, rejuvenation and improvement of Horst® Ul. Gerda Fritsche 145 The development of a set of characteristics for D.U.S. tests of cultivated mushroom varieties. A. Van Der Neut 153 List of authors 161 PREFACE The First International Seminar on Mushroom Science was held in The Netherlands from May 14 to May 17, 1991 at the occasion of the retirement of Dr. Gerda Fritsche. A unique feature of the seminar was that it brought together all scientists working in the field of molecular genetics of Agaricus. The international gathering and free and mutual exchange of ideas has led to an intensified international cooperation. This will bring mushroom genetics where it belongs: at the forefront of science. This book has become a comprehensive condensation of Agaricus genetics. It contains the scientific knowledge of the genome and has practical use in breeding and spawn preparation. The invaluable help of Ms. Ineke Dohmen in the preparation of the manuscript is gratefully acknowledged. L.J.L.D. Van Griensven vu KEYNOTE LECTURE A PERSONAL VIEW ON MUSHROOM BREEDING FROM 1957-1991 Gerda Fritsche Professor van Griensven has invited me to tell you about my views on 34 years of personal involvement in the field of mushroom breeding. I was pleased to accept, since it has been a wonderful time during which many things have changed. Encouraged by Van Griensven, I should also like to tell you something about the period preceeding my mushroom career. From 1929 -1955 I was born in 1929 in Merseburg, a medium-sized town in central Germany near Leipzig. When the Nazi's came to power, I was three years old. At ten years old, I saw the outbreak of the Second World War. After the War, my home town was allocated to the Russian zone of occupation, later East-Germany. In the meantime I was 16 years old and went to a highschool for girls. Relatives of my father, he himself originating from the countryside, found refuge in my parental home. They were evicted from hearth and home by the new authorities, because they owned more than 250 acres of land. I went to school until 1948. It was never a problem for me, what I was going to be one day. I wanted to be engaged in nature in the widest sense. My mother, watching me nursing our pot plants with great care, suggested botany as a field of study. A vocational bureau informed us, that I could become a teacher after studying in botany. Yet, I wanted to go into research. A study in horticulture was recommended. This study was preceeded by a horticultural training. I could practise this in a nursery, owned by relatives of a schoolfriend and charmingly situated in the beautiful south Harz countryside. After this training was finished, I applied in 1950 for a course of study in horticulture at the Humbold University in East-Berlin. I was refused without reason. Probably this was because my father was a physician and at that time children of university graduates were not allowed to study. Another reason was perhaps that I was not a member of the communist youth organization 'Free German Youth'. At the time, the refusal was not catastrophic for me, since the borders to the free West were still open. You only had to take the underground in East-Berlin and to travel to West-Berlin. But in West-Berlin, courses in horticulture started only in 1951. From 1950 till 1952, I was a horticultural assistant in a seed breeding nursery in the 'Flowertown' Erfurt. Plant breeding already fascinated me at that time. This period in practical plant breeding was very beneficial. In November 1952, I started my horticultural study at the Technical University in West-Berlin, together with some 20 fellow-students, most of them also originating from the East. We received a scholarship, one warm meal daily distributed by the Americans, and the opportunity to earn money from the 'TUSMA'. TUSMA stands in German for the sentence: 'Call And Students Do Everything'. Common problems united all the students together. Wonderful friendships were made. After a few years break, we have met every two years for a professional excursion and to talk about old memories. From 1956 - 1958 In 1956 I finished both my study in Horticulture at the Technical University of West-Berlin and training as a technical assistant at the Institute for Genetics and Breeding at the same University. The training was supposed to be a jumping-board to an appointment. My fellow-student Gertraud Lemke had the same training. As Professor Von Sengbusch had promised me that I could start in January 1957 to breed mushrooms, Professor Kappert, Director of the Berlin Institute, organized another short training course for me in a microbiological laboratory. Later, I have realized how important this additional training was and I am still grateful to Kappert for his support. In Berlin I was also helped by Professor Riethus, our teacher in vegetable growing. Guided by him, my fellow-student Hans-Joachim Tschierpe graduated on the role of C02 in mushroom growing (Tschierpe, 1959). Riethus organized a visit for me to a mushroom farm. Of course, the owner of the farm refused to show his spawn laboratory. This should remain a secret. Professor Dr Reinhold Von Sengbush, director of the Max Planck Institute for Plant Breeding in Hamburg When I started in January 1957 with my work, Von Sengbusch was already a famous plant breeder, from whom I could learn a lot. In 1928 he had developed sweet lupin (Von Sengbusch, 1931), which made it possible to use lupin as fodder.