Identification of Seeds to Genus and Species Level
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12.06.2009 ISTA Purity Seminar 15. June 2009 Zürich Identification of seeds to genus and species level by Norbert Leist and Andrea Jonitz Prof. Dr. Norbert Leist Dr. Andrea Jonitz Brahmsstr.25 LTZ Augustenberg 76669 Bad Schönborn Neßlerstr.23 Germany 76227 Karlsruhe [email protected] Germany [email protected] From flower to seed Pulsatilla vernalis 1 12.06.2009 From flower to seed Pulsatilla Can we find botanical characters which allow a systematic search leading to the families first, than to the genera and at least to the species? Mostly you can find the following structures, more or less well visible: Testa developed from the integument. The hole between the integument remains as micropyle, in every seed this is the point where the radicle emerges. The funiculus growth a shorter or larger part together with the integument, this part is the raphe. The breaking point of the funiculus at ripeness is the hilum. The region where the funiculus enters the integument for nutritition of the embryo is the chalaza. Hilum --Integument Cardiospermum halicacabum 2 12.06.2009 Definition of seed • A seed is a matured ovule that contains an embryo and often nutritive tissue (endosperm, perisperm) Judd et al. Plant Systematics • Achene: Asteraceae (Carex, Ranunculus, Rosa, Rumex) • Caryopsis: Poaceae • Drupe: Celtis, Cocos, Prunus • Nut: Castanea, Corylus, Fagus, Quercus • Samara: Ailanthus, Betula, Fraxinus, Liriodendron, Ulmus Carya ovata • ShiSchizocarp: Acer, AiApium, Daucus, EdiErodium, EhbiEuphorbia, LiLamium, MlMalva • Utricle: Amaranthus, Chenopodium • Seed units are commonly found dispersal units ISTA Rules 2009 Abrus precatorius Caryopsis Schizocarp Alopecurus Euphorbia gerardii verrucosa Aethusa Echinochloa cynapium crus-galli Abelmoschus Bromus esculentus mollis 3 12.06.2009 Achaene Bidens Leontodon pilosa hispidus Taraxacum Rumex officinale crispus Seed Unit Tragopogon or Taraxacum ? 4 12.06.2009 Seed characters Color : black, brown, yellowish, white, different colored, shiny, dull Shape, size : round, edged, flat, triangular, fourthsized, barrelshaped Testa surface: smooth, ribbed, striped, edged, winged, furrowed, wrinkled, dimplelike, reticular, humplike, spiny, dotted, flaking, finehirsute, woolly hirsute, shaggy Appendages : arillus, elaiosom, caruncula, crista, strophiolum, awn, arista, glumes, beak, wing, pappus Hilum : tiny, spotshaped, with other color Raphe : Strophiolum Micropyle : Caruncula Types of seed units as used in the seed key of Brouwer Stählin 1. Seed unit edged or with border, spiny point (35 families) 2. Seed unit with caruncula, often with clear raphe (2 families) 3. Seed unit winged (9 families) 4. Seed unit with hairs, partly as crown, tuft or scale (6 families) 5. Seed unit bottleshaped (2 families) 6. Seed unit stickshaped, at least 4 times as long as wide (4 families) 7. Seed unit with shiny testa or special structures of surface (6 families) 8. Seed unit enrolled, therefore kidney – or hornshaped (3 families) 9. Seed unit thin and flat, partly bended and/or with small wing ring (4 families) 10. Seed unit ball-to oval rounded, hilum visible (2 families) 11. Seed unit flat and smooth, often like amphora (1 family) 5 12.06.2009 1. Seed unit edged or with border, point spine (35 families) Amranthaceae Apiaceae Aristolochiaceae Asclepiadaceae Asteraceae Boraginaceae Brassicaceae Campanulaceae Caryophyllaceae Chenopodiaceae Convolvulaceae Coriariaceae Cucurbitaceae Cyperaceae Dipsacaceae Euphorbiaceae Fabaceae Fumariaceae Geraniaceae Iridaceae Lamiaceae Liliaceae Malvaceae Onagraceae Papaveraceae Plantaginaceae Plumbaginaceae Poaceae Polygonaceae Primulaceae Rosaceae Rubiaceae Rutaceae Santalaceae Scrophulariaceae Solanaceae Tropaeolaceae Valerianaceae Zygophyllaceae 2. Seed unit with caruncula, often with clear raphe Euphorbiaceae Polygalaceae Violaceae Chelidonium 3. Seed unit winged Asclepiadaceae Asteraceae Brassicaceae Caryophyllaceae Chenopodiaceae Cyperaceae Liliaceae Gentianaceae Iridaceae 4. Seed unit enrolled, therefore kidney – or hornshaped Asteraceae Caryophyllaceae Chenopodiaceae Fabaceae 6 12.06.2009 How stable are these seed characteristics? Depending on the type of reproduction: Apogamie =Apomixis, parthenogenesis without pollination Alchemilla,,,,,,, Amelanchier, Hieracium, Poa, Rubus, Sorbus, Taraxacum (2000 microspecies) Autogamie – selfpollination, cleistogamy Viola Heterogamie - self incompatible, pollination by other plant of the same species Primula, Astragalus Polyploidie - multiplication of the chromosome number Biscutella Hybridisation – pollination between species Aquilegia, Avena, Betula, Carex, Epilobium, Paeonia, Poa, Rumex, Salix (worldwide 70.000 naturally occurring interspecific plant hybrids are estimated) How stable are these seed characteristics? These characteristics are genetically controlled, but some factors may lead to abnormal or untypical seeds: nutritition of the plant insect damage of the seed unit fungi infection environmental stress conditions 7 12.06.2009 Equipment for seed identification • Pincette • Scalpel • Small boxes • Tubes, adhesive papel, pencil • Magnifying lens Tools for seed identification • Binocular 10‐60 fold • Microscope – Lig ht microscope – Electron microscope – Special types like Keyence 8 12.06.2009 The KEYENCE‐Microscope (VHX‐500) Key of Science Irreplaceable Training Experience Patience 9 12.06.2009 If Seed identification with morphological characters is not successful ? Check for alternative methods: • Anatomy of the seed (type of endosperm) • Seedling (Rumex) • Chemical reactions (phenol, sulphuric acid..) – Lolium, Poa, Phleum, Cuscuta, Orobanche, Vicia – Ultraviolet radiation • Seedproteins, electrophoresis Anatomy of seed units to be observed by a transverse cut Characteristics :With or without endosperm, perisperm,( endosperm oily (ol), with starch (st), fleshy (fl)); embryos place on the side, inner part; embryo straight or bended Perisperm Endosperm large Endosperm small None or nearly no endosperm Amaranthaceae Apiaceae Asclepiadaceae Asteraceae Caryophyllaceae Aristolochiaceae (fl) Boraginaceae Brassicaceae Chenopodiaceae Euphorbiaceae Campanulaceae (fl) Cucurbitaceae Plantaginaceae Poaceae (st) Dipsacaceae Fabaceae Papaveraceae (oil) Fabaceae Geraniaceae Plumbaginaceae (st) Lamiaceae Lamiaceae Polygonaceae (st) Rosaceae Onagraceae Primulaceae (fl) Rutaceae Tropaeolaceae Rubiaceae Santalaceae Valerianaceae Scrophulariaceae (fl) Polygalaceae Solanaceae (fl) Convolvulaceae Iridaceae Liliaceae 10 12.06.2009 If Seed identification with morphological characters is not successful ? Check for alternative methods: • Anatomy of the seed (type of endosperm) • Seedling (Rumex) • Chemical reactions (phenol, sulphuric acid..) – Lolium, Poa, Phleum, Cuscuta, Orobanche, Vicia – Ultraviolet radiation • Seedproteins, electrophoresis Fabaceae (Leguminosae) 643 Genera, 18.000 Species Caesalpinioideae 153 Genera, 2. 175 Species Mimosoideae 64 Genera, 2.950 Species Papilionoideae 426 Genera, 12.150 Species Lathyrus 160 Vicia 140 11 12.06.2009 Seeds showing one typical characteristic for one certain family only well expressed hilum, micropyle, raphne, chalaza Fabaceae: Medicago, Sophora, Melilotus, Robinia, Vicia ..... Medicago trunculata Sophora japonica Melilotus alba Seed characteristics Micropyle Hilum Phaseolus vulgaris; Micropyle, Hilum, Testa structure 12 12.06.2009 Seedshape: a) globular d) rhomboid b) ovoid e) hatchlike c) edged f) cylindrical g)hd) pearshaped . Testa surface Smoothy shining, smoothy dull, rough, velvet, like hoarfrost, warty structures, pories structures, labyrinth structures. These characters can be observed with magnification 13 12.06.2009 Testa colour and pattern Variation from white to light brown, dark to black, greenish and from uniform colour to different pattern This pattern can be named as: marked - small area spot, speckle, reptile pattern, marble pattern. The micropyle can be well expressed to nearly not visible. The hilum is more informative with its size, shape, colour, structure and the distance to the micropyle. The hilum can be: round-oval, less than 2 x as long as wide (eg. Lathyrus gorgoni Parl.) elongated-oval, at least 2 x, but not more than 5 x as long as wide (eg. Vicia dumetorum L.); linear, more than 5 x as long as wide (eg. Lathyrus articulatus L.) 14 12.06.2009 Furthermore, the hilum itself can show the same width in its whole length, it can narrow at one or both ends and became also lanceolate at one or both ends. Once more the are a of the hilum can differ in colour. The middle part (M) can be white-light, sometimes yellowish, with the border region (R) darkened, in some species a very light line separates the hilum lengthwise (L). M R L Genus Lathyrus 19. Lathyrus maritimus (L.) Bigelow 20. Lathyrus montanus Bernh. 1. Lathyrus amphicarpos L. 21. Lathyrus neurolobus Boiss. Et Heldr. in Boiss. 2. Lathyrus angulatus L. 22. Lathyrus nissolia L. 3. Lathyrus annuus L. 23. Lathyrus niger (L.) Bernh. 4. Lathyrus aphaca L. 24. Lathyrus ochrus (()L.) DC. 5. Lathyrus articulatus L. 25. Lathyrus odoratus L. 6. Lathyrus aureus (Steven) Brandza 26. Lathyrus Pannonicus (Jacq.) Garcke 7. Lathyrus bauhinii Genty ssp. varius 8. Lathyrus cicera L. 27. Lathyrus pisiformis L. 9. Lathyrus clymenum L. 28. Lathyrus pratensis L. 10. Lathyrus davidii Hance 29. Lathyrus roseus Steven 11. Lathyrus digitatus (Bieb.) Fiori 30. Lathyrus rotundifolius Willd. 12. Lathyrus gorgoni, Parl. 31. Lathyrus sativus L. 13. Lathyrus heterophyllos L. 32. Lathyrus sphaericus Retz. 14. Lathyrus hierosolymitanus