Nematode collections at the NPPO (NL) Gerrit Karssen
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1 Nematode collections at the NPPO (NL) Gerrit Karssen
2 Introduction Why do we have biological collections? Collections at the NPPO (NL) What collections do we have? What is in it? How do we maintain them? Spin-off NCE Delivery on demand Courses. 2
3 Introduction Why do we have biological collections at the NPPO (NL)? *Serve as a reference standard for taxa or symptoms: identification & comparison. *Needed for research studies: taxa description & revisions host-plant relation molecular test development. *Training & education. 3
4 Collections What kind of collections do we have? Symptoms: pictures (slides) & plant (parts) in jars Permanent slides: regular & types Green house cultures in vitro cultures Micro-plots. 4
5 Collections: symptoms What is in it? How do we maintain them? Photo s of plant disease symptoms caused by nematodes: colar slides stored in a closed cabinet at room temp have been digitalized so far. Plant or parts with symptoms in jars: 185 sealed jars 6% formaline stored in a closed cabinet in (aerated) lab maintained according to protocol. 5
6 Collection: permanent slides What is in it? How do we maintain them? Regular slides: Cobb slides stored in fireproof cabinets at room temperature Excel database maintained by technician. 6 Type slides: nearly 3800 slides (Cobb & glass) stored in fireproof cabinets at room temperature Excel database on-line available at slides maintained by reseacher & technician website maintained by reseacher & Wiesweb.
7 Collection: green house cultures (1) What is in it? How do we maintain it? Root-knot nematodes: populations + 22 species (see next slide) temp. (20-15C) & tropical compartments (21C) main host: tomato (cv Dometica) sandy soil mixture (melo soil) all year round renewed & ID check 1x year some on other hosts or in micro-plot maintained according to protocols. 7
8 Melo species in culture: Collection: green house cultures (2) Temperate species: M. chitwoodi M. fallax M. minor M. hapla M. artiellia (on cabbage) M. naasi (on wheat) M. kralli (on Carex acuta) M. ardenensis (micro-plot on Ligustrum vulgare) M. mali (micro-plot on Ulmus hollandica Wredei ). Tropical species: M. javanica M. incognita M. arenaria M. hispanica M. enterolobii M. ethiopica M. inornata M. luci M. exigua M. ichinohei (on Iris leavigata) M. graminicola (on Echinochloa crus-galli) M. africana (on Coffea arabica) M. camelliae (on Camellia japonica). 8
9 Collection: green house cultures (3) Cyst nematodes: + 30 populations (incl. pathotypes) 12 species (see next slide) temperate compartment tropical : H. glycines, H. ciceri & C. cacti host: several soil: sandy/clay mixture (cyst soil) not all year round (stored at 4C) renewed & ID check 1x year maintained according to protocols. 9
10 Collection: green house cultures (4) Cyst species in culture: -Globodera pallida (Pa2 & 3) potato cv Bintje - G. rostochiensis (Ro 1, 2/3, 4 & 5),, - G. tabacum tobacco -Heterodera glycines sojabean -H. schachtii cabbage -H. betae cabbage -H. trifolii clover -H. daverti clover -H. cruciferae cabbage -H. ciceri lentil -H. carotae carrot -Cactodera cacti cactus -Testing: H. koreana, H. bifenestra, H. elachista, Globodera ellingtonae & G. mexicana. Micro-plot: H. humuli, H. riparia, H. urticae, H. ustinovi & Punctodera punctata. 10
11 Collection: green house cultures (5) Other nematodes 16 populations 12 species o.a.: -Nacobbus abberans -Radopholus similis -Pratylenchus goodeyi -P. zeae -P. bolivianus -Rotylenchulus reniformis -Tylenchulus semipenetrans tomato banana banana grasses tomato jasmine citrus all maintained as for melo s some are highly experimental! 11
12 Collection: in vitro cultures (1) What is in it? How do we maintain them? In petri dish on fungi 28 populations 14 species (next slide) Botrytis cinerea on malt extract agar incubator at room temperature renewed & ID check + 12 weeks maintained according to protocols. 12
13 Species in vitro culture: Collection: in vitro cultures (2) Bursaphelenchus xylophilus B. mucronatus (EU-type & East-Asian type) B. fungivorus Cryptaphelenchus sp. Laimaphelenchus pannocaudatus Aphelenchoides besseyi A. subtenuis A. saprophilus A. fragariae A. blastophthorus A. composticola A. sinodendroni Ditylenchus arachis D. myceliophagus. 13
14 Collection: in vitro cultures (3) In petri dish on carrot 7 populations 3 species on Daucus carota disc incubator at room temperature renewed & ID check + 8 weeks maintained according to protocols. in culture: Aphelenchoides ritzemabosi Ditylenchus dipsaci D. destructor. 14
15 Collection: micro plots What is in it? 40 micro plots restricted area different soil types clover/grass main hosts all PPN checked 6 years ago PPN: 14 genera & 46 species includes some q s PPN mainly used for courses maintained by greenhouse technicians o.a. M. ardenensis on Ligustrum vulgare M. chitwoodi on grasses/clover M. mali on Ulmus hollandica Wredei M. minor on grasses. 15
16 Spin-off Nematode Collection Europe (NCE) on-line database for type collections from D, UK, B & NL nomenclatorial database for Nematoda (excl. animal parasites) maintained by Wiesweb and reseacher. Delivery on demand Courses about 75 populations delivered/per year mostly used for DNA tests (reference or ID test development) free for NPPO s. slides & living nematodes used for teaching UGent & WUR-NPPO ID course once a year several courses on request each year (1-2 persons). 16
17 The team Meloidogyne: Evelyn van Heese & Patricia Bor In vitro & others: Anne Sophie van Bruggen, Evelyn van Heese & Patricia Bor Cysts & micro-plots: Anne-Marie Heij & Patricia Bor Slides: Evelyn van Heese & Rens van de Biggelaar WaNeCo: Evert Wiesenekker (Wiesweb) & G. Karssen. 17
18 18 THANKS!
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