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Tetrazolium Testing Handbook

Contribution No. 29

To the Handbook on Seed Testing

Prepared by

The Tetrazolium Subcommittee of the

Association of Official Seed Analysts

Jack Peters, Editor Bill Lanham, Illustrator

Published by the Association 1970

First Revision 2000

2001, 2002, 2004, 2005 updates: see www.aosaseed.com

Copyright © 2000 by Association of Official Seed Analysts

All rights reserved; no part of this publication may be reproduced, stored in retrieval system, or transmitted in any form or by any means: electronic, mechanical, photocopying, recording, or

otherwise, without the prior written permission of the AOSA Executive Board.

Dedication

This handbook is dedicated to the memory of Philena Heckert, a botanist, seed analyst, and colleague with the Federal Seed Laboratory.

Her diligence and hard work were a source of inspiration for the completion of this seed testing resource.

Updated 2001: v, 7, 18, Cactaceae, Cucurbitaceae, Cupressaceae, Lamiaceae, Malvaceae, Poaceae II, Tiliaceae, Index A and Index B: addition of Cynodon, Cucumis, and Nassella. Entire lab reference section. Bibliography corrections on Abdul...page and Troneberry... page.

Updated 8/2002: iv, v, 3,4, Amaranthaceae, Apiaceae, Poaceae I, Poaceae II,

Poaceae IV (new), Lab references, Index A and Index B: addition of Petroselinum. Updated 2004: Asteraceae, Fabaceae I, Hydrophyllaceae, Malvaceae, Lab

references. Updated 2005: iv, 9, Fabaceae II, Tropaeolaceae, Index A, Index B. New pages for

2005: Sparganiaceae, Solanaceae (Petunia and Nicotiana).

i

TABLE OF CONTENTS Table of Contents i, ii, iii Plant families iv Acknowledgements v Preface vi PART I. PRINCIPLES OF TETRAZOLIUM TESTING 1. INTRODUCTION 1 2. NATURE OF THE TZ TEST 1 3. HISTORY 2 4. CLASSIFICATION AND STRUCTURE 3 4.1 Monocotyledon 3 4.1.1 Simple embryonic axis with scutellum and endosperm 3 4.1.2 Elongated, single cotyledon embryo and endosperm 3 4.2 Dicotyledon 3 4.2.1 Low level embryonic differentiation 3 4.2.2 High level embryonic differentiation 4 4.3 Gymnosperm 4 5. SEED DEFINITION AND BASIC STRUCTURE 5 5.1 Embryos 5 5.1.1 Basal 5 5.1.2 Axile 5 5.1.3 Peripheral 5 5.1.4 Entire 5 5.2 Nutritive tissue 6 5.2.1 Nutritive tissues apart from the embryo 6 5.2.1.1 Endosperm 6 5.2.1.2 Perisperm 6 5.2.1.3 Female gametophyte 6 5.2.2 Nutritive tissues within the embryo 6 5.2.2.1 Cotyledons 6 5.3 Seed coat, testa, pericarp 6 6. DORMANT SEED 6 6.1 Types of dormancy 7 6.1.1 Exogenous 7 6.1.2 Endogenous 7 6.2 Degrees of dormancy 7 6.3 Deep dormancy 7 6.4 Hard seed and scarification 7

ii

7. EQUIPMENT AND SUPPLIES 8 7.1 Staining dishes 8 7.2 Cutting, piercing, cracking devices 8 7.3 Forceps/ tweezers 8 7.4 Visual aids 8 7.5 Conditioning (seed moistening) media 8 7.6 Temperature control units 8 7.7 Miscellaneous items 8 7.8 Check sample to monitor TZ solution 8 8. SOLUTIONS 9 8.1 Tetrazolium solution 9 8.2 Buffer solution 10 8.3 Other solutions used in TZ testing 10 8.3.1 Lactic acid and glycerol 10 8.3.2 Gibberellic acid (GA3) 10 8.3.3 Hydrogen peroxide (H2O2) 10 8.3.4 Mucilage hardening reagents 11 8.3.5 Acids and bases used for the scarification of hard seed 11 9. TEMPERATURE 11 10. LIGHT 11 11. PRESSURE (Partial Vacuum) 11 12. SOURCE OF SEEDS TO TEST 12 13. NUMBER OF SEEDS TO TEST 12 14. PREPARATION OF SEED FOR TESTING 12 14.1 Preconditioning 12 14.1.1 Slow moistening 13 14.1.2 Soaking 13 14.1.3 No conditioning 13 14.2 Preparation and staining 13 14.3 Post-staining 14 15. EVALUATION OF TETRAZOLIUM TESTS 14 15.1 TZ as a viability test 14 15.1.1 Speed 15 15.1.2 Accuracy 15 15.1.3 Evaluation subtleties 16 15.1.3.1 False positives 17 15.1.3.2 False negatives 17 15.1.3.3 Multiple seed units and multiple embryos 18 15.1.3.4 Immature seed 18 15.2 TZ as a vigor test 18

iii

PART II. TETRAZOLIUM TESTING PROCEDURES USING THE HANDBOOK AS A REFERENCE GUIDE 19 HANDBOOK CONTENTS 19 SYMBOLS AND ABBREVIATIONS 20 Icons 20 Illustration labels 20 TETRAZOLIUM SEED PREPARATION METHODS AND TECHNIQUES (illustrations) 21 ______________________________________________________________________________ [ Tabbed Section] Alphabetized Plant Families (see next page for the plant families included in this handbook) ______________________________________________________________________________ APPENDIX GLOSSARY INDEX - A [Family/Genus] INDEX - B [Genus/Family] LABORATORY REFERENCES BIBLIOGRAPHY

iv

Plant Families Included in Handbook A

ACANTHACEAE ACERACEAE AGAVACEAE AIZOACEAE AMARANTHACEAE ANACARDIACEAE APIACEAE APOCYNACEAE ASCLEPIADACEAE ASTERACEAE B BALSAMINACEAE BEGONIACEAE BERBERIDACEAE BETULACEAE BIGNONIACEAE BORAGINACEAE BRASSICACEAE I BRASSICACEAE II C CACTACEAE CAMPANULACEAE CAPPARACEAE CAPRIFOLIACEAE CARYOPHYLLACEAE CASUARINACEAE CELASTRACEAE CHENOPODIACEAE CISTACEAE COMMELINACEAE CONVOLVULACEAE CORNACEAE CRASSULACEAE CUCURBITACEAE CUPRESSACEAE CYPERACEAE

D DIPSACACEAE E EBENACEAE ELAEAGNACEAE EPHEDRACEAE ERICACEAE EUPHORBIACEAE F FABACEAE I FABACEAE II FABACEAE III FAGACEAE

G

GENTIANACEAE GERANIACEAE GESNERIACEAE GINKGOACEAE GROSSULARIACEAE H HAMAMELIDACEAE HIPPOCASTANACEAE HYDROPHYLLACEAE I IRIDACEAE J JUGLANDACEAE JUNCACEAE K L LAMIACEAE LILIACEAE I LILIACEAE II LIMNANTHACEAE LINACEAE M MAGNOLIACEAE MALVACEAE I MALVACEAE II MYRTACEAE N NYCTAGINACEAE NYSSACEAE O OLEACEAE ONAGRACEAE P PAPAVERACEAE PASSIFLORACEAE PEDALIACEAE PINACEAE PLANTAGINACEAE PLATANACEAE PLUMBAGINACEAE POACEAE I POACEAE II POACEAE III POLEMONIACEAE POLYGONACEAE

PORTULACACEAE PRIMULACEAE PROTEACEAE

Q R RANUNCULACEAE RESEDACEAE RHAMNACEAE ROSACEAE I ROSACEAE II ROSACEAE III RUTACEAE S SALICACEAE SAPINDACEAE SCROPHULARIACEAE SIMAROUBACEAE SIMMONDSIACEAE SOLANACEAE Nicotiana, Petunia SPARGANIACEAE T TAXACEAE TAXODIACEAE TILIACEAE TROPAEOLACEAE U ULMACEAE V VALERIANACEAE VERBENACEAE VIOLACEAE VIATACEAE W – X – Y Z ZYGOPHYLLACEAE

2005

5

Acknowledgements

The preparation of this handbook began in earnest in the early 1990’s

by the AOSA Tetrazolium Subcommittee members who compiled the information.

Major contributors include:

Stan Akagi Barbara Atkins Charles Baskin

Bill Ebener Amy Ferry Mike Gill

Susana Goggi Tim Gutormson

Jane Hall Philena Heckert

Kristi Huber Mering Hurd Gary Johnson Bill Lanham

Annette Miller Marilyn Milhous

Jack Peters Nancy Shaw

Marian Stephenson Victor Vankus Nancy Vivrette

The following pages of tetrazolium testing principles and procedures (by plant family) represent the combined work and collaborative effort of many seed analysts of the Association of Official Seed Analysts and the Society of Commercial Seed Technologists. I extend a sincere debt of gratitude to all those individuals, seed laboratories, and seed companies who contributed to this effort. Thank you. Jack Peters, Editor Purchase books directly from:

Association of Official Seed Analysts PMB #411 1763 E. University Blvd. Suite A, Las Cruces, NM 88001 USA Phone/FAX: 505-522-1437 Email: [email protected] website: www.aosaseed.com Direct questions or comments about the contents of this handbook to the AOSA Tetrazolium Subcommittee (see website) or the appropriate laboratory references listed for each family. Newly revised pages and procedures for submitting changes and additions are also available at www.aosaseed.com.

vi

PREFACE

This handbook has been the result of several years of collaboration by many seed analysts in the Society of Commercial Seed Technologists and the Association of Official Seed Analysts. Many are listed in the acknowledgements. While it has taken many years to develop this edition, the handbook is by no means complete. Additional procedures for plant families not listed in this handbook may be added in future editions. The intent of the handbook is to offer guidelines and methodologies in tetrazolium testing. The ultimate goal is to provide uniformity in tetrazolium seed testing, with regard to procedures and TZ staining evaluation. Illustrations in the handbook are used to describe seed/embryo morphology, preparation/cutting techniques, and TZ staining patterns. A bibliography at the end of the handbook lists references associated with tetrazolium seed testing. The handbook’s notebook format is intended to encourage further refinement and experimentation with the TZ methods and techniques. Ample space for notes allows analysts to personalize the instructions. Part II is unpaginated to allow easy updates and additions of plant family procedures. It is the committee’s hope that analysts will continue to share experiences and improve the handbook by submitting suggested changes to the committee. Periodic additions and changes are the best way to ensure that the handbook will continue to meet the needs of analysts. Furrows be glad, though earth is bare — one more seed is planted there. Give up your strength the seed to nourish — that in course the flower may flourish. Besançon Carol

9TETRAZOLIUM TESTING HANDBOOK

2005 8. SOLUTIONS The essential reagent is the tetrazolium staining solution. Other reagents may be used in facilitating seed conditioning, staining, or evaluation. These reagents include: gibberellic acid, pH buffers, hydrogen peroxide, mucilage hardeners, lactic acid, and glycerol. Material Safety Data Sheets (MSDS) produced by the chemical manufacturer are available for most reagents to alert consumers to toxicity, flammability, other hazards, and the proper disposal of used chemical solutions. 8.1 Tetrazolium solution

The recommended tetrazolium derivative is 2, 3, 5-triphenyl tetrazolium chloride (TTC). This salt is sold in powder form and is available from several chemical suppliers. Three sources are listed below:

MP Biomedicals, Inc. (formerly ICB) 1263 S. Chillicothe Road, Aurora, OH 44202 1-800-854-0530 Fisher Scientific Company 1458 N. Lamon Avenue, Chicago, IL 60651 1-800-766-7000 United States Biochemicals Corporation 26201 Miles Road, Cleveland, OH 44128 1-800-321-9322

The tetrazolium staining solution consists of the TTC and water. A buffer can be added to the TTC and water solution when there is need to correct a pH problem. Several concentrations of tetrazolium may be used with comparable results. For convenience, solutions of 1.0% and 0.1% TZ can be prepared and stored in cold (5° C) conditions. The TZ concentration used for the test is generally dependent on how the seed is prepared. A 1.0% solution is used for seeds that are not bisected through the embryo, while a 0.1% solution is used for seeds in which the embryo is bisected. Other low concentrations such as 0.2% and 0.5% are sometimes used. To prepare a 1.0% solution of TZ, dissolve 1.0 gram of tetrazolium powder in 100 ml of distilled or tap water. A 0.1% solution is prepared by dissolving 0.1 gram of tetrazolium powder in 100 ml of water, or by mixing one part of 1.0% solution with nine parts of water. Store all prepared TZ solutions in the dark or in amber- or brown-colored bottles to prevent deterioration from light. The TZ solution can be kept for several months if kept in a tightly capped container, in the dark, and refrigerated at 5° C. The 2, 3, 5-triphenyl tetrazolium chloride MSDS describes and lists the known toxicological properties of the chemical. Although TZ is not considered to be a highly toxic or dangerous biological hazard (especially when diluted to liquid concentrations of 1.0% or less), there are proper protocols for the handling, usage, and disposal of the TZ solution (APPENDIX ).

Laboratories must practice safety procedures to ensure the health and well-being of their employees and the safety of their lab facility. Latex and rubber gloves, safety eyewear, exhaust/fume hoods and chemical storage cabinets should all be part of the laboratory set-up. Labs should follow the MSDS guidelines and properly dispose of the chemical through incineration or chemical storage and transport methods. Disposal methods are dependent on state and local regulations. Safety and concern for employees and the environment are essential considerations.

10ASSOCIATION OF OFFICIAL SEED ANALYSTS

2005 8.2 Buffer solution

The pH for the aqueous TZ solution should be between 6.5 and 7.5 for optimum staining results. A pH meter or litmus paper is used to check the pH of the solution (low pH is acidic and high pH is basic). TZ may be made with water or a buffer solution. Hard or soft water may also affect the TZ solution and chemistry. If this is suspected, use distilled water or a buffer solution. If making the solution with water, check the pH of the water before adding the TTC powder. If the pH is between 6.5 and 7.5, mix in the TTC powder at the desired concentration. Check the pH again. If the pH is out of the appropriate range, discard the solution and prepare a new batch of TZ using a buffer solution instead of water. Many different chemical compounds can be used to make buffer solutions. In TZ testing, sodium phosphate and potassium phosphate are commonly used, prepared according to the following recipe:

Solution 1 - Dissolve 9.078 grams KH2PO4 in 1000 ml water.

Solution 2 - Dissolve 9.472 grams Na2HPO4 in 1000 ml water.

Mix two parts of solution 1 to three parts of solution 2. Then add TTC powder at the desired concentration. No further pH measurement or adjustment is necessary. The shelf life of buffer stock solution is approximately eight months when stored in a clean amber-colored bottle at 5° C.

8.3 Other solutions used in TZ testing 8.3.1 Lactic acid and glycerol

Lactic acid (85%) or glycerol can be used to remove pigmentation from the seed coats of certain small-seeded grasses, legumes, and other small-seeded species that are pierced or immersed whole. Thesesolutions make the light-pigmented seed coats translucent, so that the stained embryo can be evaluated with an adequate light source and dissecting microscope. The average length of time for this ‘clearing’ treatment is one half hour. Clearing time is dependent on the tissue composition and pigmentation and may take several hours for some species. These solutions are easily obtained from most chemical companies and pharmacies. Lactic acid or glycerol can be used to preserve a stained sample for a limited time. (Lactophenol was once used because the phenol component helped preserve prepared seed; however, the toxicity of phenol compound makes this a hazardous option.)

8.3.2 Gibberellic acid (GA3) Gibberellic acid may be used to precondition dormant seed. (To make a 400 ppm solution, mix 400 mg GA3 in 1.0 liter of distilled H2O.) 8.3.3 Hydrogen peroxide (H2O2)

3.0% concentration or less (0.5%) of H2O2 is used as an oxygenating and bleaching agent during preconditioning (3.0% concentration can be purchased at pharmacies, then diluted with distilled water to obtain the desired concentration).

TETRAZOLIUM TESTING HANDBOOK 2005

FAMILY: FABACEAE II Genera: Acacia, Calliandra, Dalea, Desmanthus, Hedysarum, Indigofera, Kummerowia, Lupinus, Medicago, Melilotus, Trifolium

1. PRECONDITIONING:

METHOD TIME (hrs) TEMP (°C)imbibe between moist blotters overnight 20-25 Notes: Hard seed may be present (see section 5.3). Seed may need scarification (see sections 6.4 and 8.3.5).

2. PREPARATION AND STAINING:

METHOD TZ Conc(%) TIME (hrs) TEMP (°C) generally no cutting is required, however a slight cut (clip or nick) in cotyledon may be necessary. 1.0 2-72 30-35 Notes: Staining time is dependent upon degree of hard-seededness.

Cot

Rad

SC

or or

ASSOCIATION OF OFFICIAL SEED ANALYSTS 2005 FAMILY: FABACEAE II Post Staining Notes: Clear with glycerol or lactic acid for approximately one hour at 35 °C (Trifolium hybridum will not clear due to dark pigmentation). It may be necessary to remove seed from seed coat to evaluate all tissue.

3. EVALUATION

VIABLE (NORMAL STAINING) - entire embryo evenly stained, turgid, and unfractured - slight damage to radicle acceptable - radicle stained slightly darker - small, shallow, unstained or intensely stained areas on periphery of hypocotyl and cotyledons - half or more of cotyledons attached to the embryo axis and evenly stained NON-VIABLE (ABNORMAL OR NO STAINING) - embryo completely unstained - embryo flaccid - unstained, deteriorated, or fractured radicle above tip of central conducting tissue - unstained or watery, darkly-stained areas extending to inner surfaces of cotyledons - less than half of cotyledons remaining functional and attached to embryo axis - damage to embryo axis OTHER TISSUE/NOTES Seed must be examined critically for mechanical damage, especially at juncture of cotyledons and hypocotyl. Immature seed may stain unevenly, or remain green. If the embryo tissue is flaccid, the seed is non-viable regardless of the color.

Fig 4 Seed stain evaluation REFERENCES: 1, 2, 3, 5, 6, 7


FAMILY: SOLANACEAE Genus: Petunia, Nicotiana

1. PRECONDITIONING:

METHOD TIME (hrs) TEMP (°C)imbibe on moist blotters, filter paper,or paper towels

overnight

20-25

Morphology Nicotiana

Fig 1 External Fig 2 Embryo 2. PREPARATION AND STAINING:

METHOD TZ Conc(%) TIME (hrs) TEMP (°C) cut deeply leaving part of the seed coat uncut to hold the two halves together 0.1 overnight to 24 35

Petunia Nicotiana Fig 3 Preparation method

SC

Cot

Rad/Hyp End

ASSOCIATION OF OFFICIAL SEED ANALYSTS 2005 FAMILY: SOLANACEAE GENUS: Petunia, Nicotiana Post Staining Notes: None

3. EVALUATION:

VIABLE (NORMAL STAINING) - entire embryo evenly stained - entire endosperm evenly stained NON-VIABLE (ABNORMAL OR NO STAINING) - any part of embryo or endosperm unstained OTHER TISSUE/NOTES Normal endosperm may appear granular. Petunia + -

Nicotiana + +

- - - -

Fig 4 Seed stain evaluation


FAMILY: SPARGANIACEAE Genus: Sparganium (bur-reed)

1. PRECONDITIONING:

METHOD TIME (hrs) TEMP (°C)soak overnight 20-25

Morphology longitudinal cross section lateral cross section

Fig 1 External Fig 2 Embryo Notes: Seed unit may contain one or two seeds.


METHOD TZ Conc(%) TIME (hrs) TEMP (°C) 1. Cut longitudinal section to expose endosperm. 1.0 overnight 20-30

2. Laterally cut and remove distal end. 0.1 overnight 20-30

Fig 3 Preparation methods

Pcrp Plu Rad

End

2 1 1 2

distal

basal

ASSOCIATION OF OFFICIAL SEED ANALYSTS 2005

FAMILY: SPARGANIACEAE Post Staining Notes: Remove embryo from seed unit. Longitudinally cut seeds: cut another longitudinal section to expose the embryo. Laterally cut seeds: Embryos will protrude slightly from the cut end. Grasp embryo with forceps and gently pull the embryo out from the top of laterally cut seed units. longitudinal cut

3. EVALUATION:

VIABLE (NORMAL STAINING) - entire embryo evenly stained - endosperm will not stain NON-VIABLE (ABNORMAL OR NO STAINING) - any part of the plumule or radicle, unstained - soft or flaccid embryo + + -

Fig 4 Seed stain evaluation Notes: The embryos on the left were removed from laterally cut seed units. The cut is at the top of the photo. The embryos in the center and right photos are intact embryos removed from longitudinally cut seed units. In seed units with more than one embryo, the seed unit is viable if at least one embryo is normally stained.

lateral cut with protruding embryos prior to removal


FAMILY: TROPAEOLACEAE Genus: Tropaeolum

1. PRECONDITIONING:

METHOD TIME (hrs) TEMP (°C)imbibe on moist blotters, filter paper,or paper towels

overnight

20-25

Morphology

Fig 1 External Fig 2 Embryo (mericarp)


METHOD TZ Conc(%) TIME (hrs) TEMP (°C) cut longitudinally leaving seed intact at distal end 0.1 overnight to 24 20-25

Fig 3 Preparation method

Pcrp SC

Cot

Rad/Hyp

ASSOCIATION OF OFFICIAL SEED ANALYSTS

FAMILY: TROPAEOLACEAE GENUS: Tropaeolum Post Staining Notes: None

3. EVALUATION:

VIABLE (NORMAL STAINING) - entire embryo evenly stained NON-VIABLE (ABNORMAL OR NO STAINING) - any essential part of embryo unstained - cotyledon unstained on cut surface OTHER TISSUE/NOTES Cotyledons stain rapidly on cut surface, but penetration into rest of tissue is quite slow. Stain may not penetrate cotyledon tissue at all. + + + -

dark stain second cut (left) shows light stain purplish discolored weak stain penetration diseased tissue Fig 4 Seed stain evaluation

2005

2005

INDEX - A

(cross-reference)

Family/Genus

For common names, species, or classification references, please refer to the AOSA Uniform Classification of Weed and Crop Seeds, Contribution No. 25 to the Handbook on Seed Testing

2005

FAMILY GENUS Acanthaceae Crossandra Hypoestes Justicia Aceraceae Acer Agavaceae Cordyline Yucca Aizoaceae Dorotheanthus Tetragonia Amaranthaceae Amaranthus Celosia Gomphrena Anacardiaceae Rhus Apiaceae Anethum Anthriscus Apium Carum Coriandrum Cuminum Daucus Pastinaca Petroselinum Apocynaceae Catharanthus Asclepiadaceae Asclepias Asteraceae Ambrosia Artemisia Aster Baccharis Baileya Balsamorhiza Carthamus

Chrysopsis Chrysothamnus Encelia Galinsoga Grindeli Haplopappus Helenium Helianthus

FAMILY GENUS Asteraceae Lactuca Rudbeckia Verbesina Balsaminaceae Impatiens Begoniaceae Begonia Berberidaceae Berberis Mahonia Betulaceae Alnus Betula Bignoniaceae Catalpa Chilopsis Paulownia Boraginaceae Cynoglossum Heliotropium Myosotis Brassicaceae Barbarea Brassica Crambe Draba Eruca Lepidium Lesquerella Physaria Raphanus Rorippa Sinapis Cactaceae Carnegiea Ferocactus Lophocereus Opuntia Pachycereus Campanulaceae Campanula Platycodon Capparaceae Cleome


2005

FAMILY GENUS Caprifoliaceae Lonicera Sambucus Symphoricarpos Viburnum Caryophyllaceae Cerastium Dianthus Gypsophila Lychnis Saponaria Silene Vaccaria Casuarinaceae Casuarina Celastraceae Celastrus Chenopodiaceae Atriplex Beta Kochia Krascheninnikovia Spinacia Cistaceae Cistus Helianthemum Commelinaceae Tradescantia Convolvulaceae Ipomoea Cornaceae Cornus Crassulaceae Kalanchoe Sedum Sempervivum Cucurbitaceae Citrullus Cucumis Cucurbita

Lagenaria Cupressaceae Calocedrus Chamaecyparis Cupressus Juniperus Cupressaceae Platycladus Thuja

FAMILY GENUS Cyperaceae Carex Cyperus Scirpus Dipsacaceae Dipsacus

Scabiosa Ebenaceae Diospyros Elaeagnaceae Elaeagnus Shepherdia Ephedraceae Ephedra Ericaceae Arbutus Arctostaphylos Rhododendron Vaccinium Euphorbiaceae Euphorbia Ricinus Fabaceae Acacia Albizia Amorpha Arachis Calliandra Cercis Cicer Dalea Desmanthus Gleditsia Glycine Gymnocladus Hedysarum Indigofera Kummerowia Lathyrus Lens Leucaena Lupinus Medicago Melilotus Phaseolus Pisum Robinia Trifolium

TETRAZOLIUM TESTING HANDBOOK

2005

FAMILY GENUS Fabaceae Vicia Vigna Wisteria Fagaceae Castanea Castanopsis Fagus Quercus Gentianaceae Centaurium Eustoma Gentiana Lisianthus Geraniaceae Erodium Geranium Pelargonium Gesneriaceae Episcia Saintpaulia Ginkgoaceae Ginkgo Grossulariaceae Ribes Hamamelidaceae Hamamelis Liquidambar Hippocastanaceae Aesculus Hydrophyllaceae Nemophila Phacelia Iridaceae Iris Juglandaceae Carya Jug Juncaceae Juncus Lamiaceae Agastache Calamintha Clinopodium Coleus Hyssopsus Melissa Mentha

FAMILY GENUS Lamiaceae Monarda

Nepeta Ocimum Origanum Rosmarinus Salvia Satureja Thymus Liliaceae Allium Amaryllis Asparagus Calochortus Camassia Erythronium Kniphofia Lilium Limnanthaceae Limnanthes Linaceae Linum Magnoliaceae Liriodendron Magnolia Malvaceae Alcea Gossypium Hibiscus Malva Sphaeralcea Martyniaceae (see Pedaliaceae) Myrtaceae Eucalyptus Eugenia Nyctaginaceae Abronia Bougainvillea Mirabilis Oxybaphus Nyssaceae Nyssa Oleaceae Chionanthus Forestiera Fraxinus Syringa


2005

FAMILY GENUS Onagraceae Camissonia Clarkia Epilobium Fuchsia Gaura Godetia Oenothera Papaveraceae Argemone Eschscholzia Hunnemannia Papaver Passifloraceae Passiflora Pedaliaceae Proboscidea Sesamum Pinaceae Abies Cedrus Larix Picea Pinus Pseudotsuga Tsuga Plantaginaceae Bougeria Littorella Plantago Platanaceae Platanus Plumbaginaceae Armeria Limonium Plumbago Psylliostachys Poaceae Achnatherum (syn. Oryzopsis) Agropyron Agrostis Avena Bouteloua Bromus Buchloe Calamagrostis Cenchrus

FAMILY GENUS Poaceae Cynodon Dactylis Elymus Elytrigia Eragrostis Eremochloa Eriochloa Festuca Hilaria Hordeum Leptochloa Leymus Lolium

Nassella Panicum

Pascopyrum Paspalum Piptatherum (syn. Oryzopsis) Poa Puccinellia Schismus Schizachyrium Secale Sorghastrum Sorghum Sporobolus Stipa Triticum Zea Zoysia Polemoniaceae Cobaea Gilia Linanthus Phlox Polygonaceae Antigonon Eriogonum Fagopyrum Polygonum Rheum Rumex Portulacaceae Portulaca


2005

FAMILY GENUS Primulaceae Anagallis Cyclamen Primula Proteaceae Grevillea Ranunculaceae Aconitum Adonis Anemone Aquilegia Caltha Clematis Consolida Delphinium Eranthis Helleborus Hepatica Nigella Pulsatilla Ranunculus Thalictrum Resedaceae Reseda Rhamnaceae Ceanothus Rhamnus Rosaceae Amelanchier Cercocarpus Cowania Crataegus Fallugia Rosaceae Holodiscus Malus Potentilla Prunus Purshia Pyrus Rosa Rubus Sanguisorba Sorbus Rutaceae Dictamnus Ruta

FAMILY GENUS Salicaceae Populus Salix Sapindaceae Cardiospermum Scrophulariaceae Antirrhinum Castilleja Collinsia Digitalis Linaria Mimulus Nemesia Orthocarpus Penstemon Torenia Veronica Simaroubaceae Ailanthus Simmondsiaceae Simmondsia Solanaceae Browallia Brugmansia Capsicum Datura Lycopersicon Nicotiana Nierembergia Petunia Physalis

Salpiglossis Solanum Sparganiaceae Sparganium Taxaceae Taxus Taxodiaceae Cryptomeria Cunninghamia Metasequoia Sequoia Sequoiadendron Taxodium Tiliaceae Tilia Tropaeolaceae Tropaeolum


2005

FAMILY GENUS Ulmaceae Celtis Ulmus Zelkova Valerianaceae Valeriana Valerianella Verbenaceae Lantana Verbena Violaceae Viola Vitaceae Parthenocissus Vitis Zygophyllaceae Larrea


2005

INDEX - B

(cross-reference)

Genus/Family

For common names, species, or classification references, please refer to the AOSA Uniform Classification of Weed and Crop Seeds, Contribution No. 25 to the Handbook on Seed Testing

2005

GENUS FAMILY Abies Pinaceae Abronia Nyctaginaceae Acacia Fabaceae Acer Aceraceae Achnatherum Poaceae (syn. Oryzopsis) Aconitum Ranunculaceae Adonis Ranunculaceae Aesculus Hippocastanaceae Agastache Laminaceae Agropyron Poaceae Agrostis Poaceae Ailanthus Simaroubaceae Albizia Fabaceae Alcea Malvaceae Allium Liliaceae Alnus Betulaceae Amaranthus Amaranthaceae Amaryllis Liliaceae Ambrosia Asteraceae Amelanchier Rosaceae Amorpha Fabaceae Anagallis Primulaceae Anemone Ranunculaceae Anethum Apiaceae Anthriscus Apiaceae Antigonon Polygonaceae Antirrhinum Scrophulariaceae Apium Apiaceae Aquilegia Ranunculaceae Arachis Fabaceae Arbutus Ericaceae Arctostaphylos Ericaceae Argemone Papaveraceae Armeria Plumbaginaceae Artemisia Asteraceae Asclepias Asclepiadaceae Asparagus Liliaceae Aster Asteraceae Atriplex Chenopodiaceae Avena Poaceae Baccharis Asteraceae Baileya Asteraceae Balsamorhiza Asteraceae Barbarea Brassicaceae

GENUS FAMILY Begonia Begoniaceae Berberis Berberidaceae Beta Chenopodiaceae Betula Betulaceae Bougainvillea Nyctaginaceae Bougueria Plantaginaceae Bouteloua Poaceae Brassica Brassicaceae Bromus Poaceae Browallia Solanaceae Brugmansia Solanaceae Buchloe Poaceae Calamintha Lamiaceae Calliandra Fabaceae Calocedrus Cupressaceae Calochortus Liliaceae Caltha Ranunculaceae Camassia Liliaceae Camissonia Onagraceae Campanula Campanulaceae Capsicum Solanaceae Cardiospermum Sapindaceae Carex Cyperaceae Carnegiea Cactaceae Carthamus Asteraceae Carum Apiaceae Carya Juglandaceae Castanea Fagaceae Castanopsis Fagaceae Castilleja Scrophulariaceae Casuarina Casuarinaceae Catalpa Bignoniaceae Catharanthus Apocynaceae Ceanothus Rhamnaceae Cedrus Pinaceae Celastrus Celastraceae Celosia Amaranthaceae Celtis Ulmaceae Cenchrus Poaceae Centaurium Gentianaceae Cerastium Caryophyllaceae Cercis Fabaceae Cercocarpus Rosaceae Chamaecyparis Cupressaceae Chilopsis Bignoniaceae


2005

GENUS FAMILY Chionanthus Oleaceae Chrysopsis Asteraceae Chrysothamnus Asteraceae Cicer Fabaceae Cistus Cistaceae Citrullus Cucurbitaceae Clarkia Onagraceae Clematis Ranunculaceae Cleome Capparaceae Clinopodium Lamiaceae Cobaea Polemoniaceae Coleus Laminaceae Collinsia Scrophulariaceae Consolida Ranunculaceae Cordyline Agavaceae Coriandrum Apiaceae Cornus Cornaceae Cowania Rosaceae Crambe Brassicaceae Crataegus Rosaceae Crossandra Acanthaceae Cryptomeria Taxodiaceae Cucumis Cucurbitaceae Cucurbita Cucurbitaceae Cuminum Apiaceae Cunninghamia Taxodiaceae Cupressus Cupressaceae Cyclamen Primulaceae Cynodon Poaceae Cynoglossum Boraginaceae Cyperus Cyperaceae Dactylis Poaceae Dalea Fabaceae Datura Solanaceae Daucus Apiaceae Delphinium Ranunculaceae Desmanthus Fabaceae Dianthus Caryophyllaceae Dictamnus Rutaceae Digitalis Scrophulariaceae Diospyros Ebenaceae Dorotheanthus Aizoaceae Draba Brassicaceae Elaeagnus Elaeagnaceae Elymus Poaceae

GENUS FAMILY Elytrigia Poaceae Encelia Asteraceae Ephedra Ephedraceae Epilobium Onagraceae Episcia Gesneriaceae Eragrostis Poaceae Eranthis Ranunculaceae Eremochloa Poaceae Eriochloa Poaceae Eriogonum Polygonaceae Erodium Geraniaceae Eruca Brassicaceae Erythronium Liliaceae Eschscholzia Papaveraceae Eucalyptus Myrtaceae Eugenia Myrtaceae Euphorbia Euphorbiaceae Eustoma Gentianaceae Fagopyrum Polygonaceae Fagus Fagaceae Fallugia Rosaceae Ferocactus Cactaceae Festuca Poaceae Forestiera Oleaceae Fraxinus Oleaceae Fuchsia Onagraceae Galinsoga Asteraceae Gaura Onagraceae Gentiana Gentianaceae Geranium Geraniaceae Gilia Polemoniaceae Ginkgo Ginkgoaceae Gleditsia Fabaceae Glycine Fabaceae Godetia Onagraceae Gomphrena Amaranthaceae Gossypium Malvaceae Grevillea Proteaceae Grindelia Asteraceae Gymnocladus Fabaceae Gypsophila Caryophyllaceae Hamamelis Hamamelidaceae Haplopappus Asteraceae Hedysarum Fabaceae Helenium Asteraceae


2005

GENUS FAMILY Helianthus Asteraceae Helianthemum Cistaceae Heliotropium Boraginaceae Helleborus Ranunculaceae Hepatica Ranunculaceae Hibiscus Malvaceae Hilaria Poaceae Holodiscus Rosaceae Hordeum Poaceae Hunnemannia Papaveraceae Hypoestes Acanthaceae Hyssopsus Laminaceae Impatiens Balsaminaceae Indigofera Fabaceae Ipomoea Convolvulaceae Iris Iridaceae Juglans Juglandaceae Juncus Juncaceae Juniperus Cupressaceae Justicia Acanthaceae Kalanchoe Crassulaceae Kniphofia Liliaceae Kochia Chenopodiaceae Krascheninnikovia Chenopodiaceae Kummerowia Fabaceae Lactuca Asteraceae Lagenaria Cucurbitaceae Lantana Verbenaceae Larix Pinaceae Larrea Zygophyllaceae Lathyrus Fabaceae Lens Fabaceae Lepidium Brassicaceae Leptochloa Poaceae Lesquerella Brassicaceae Leucaena Fabaceae Leymus Poaceae Lilium Liliaceae Limnanthes Limnanthaceae Limonium Plumbaginaceae Linanthus Polemoniaceae Linaria Scrophulariaceae Linum Linaceae Liquidambar Hamamelidaceae Liriodendron Magnoliaceae

GENUS FAMILY Lisianthus Gentianaceae Littorella Plantaginaceae Lolium Poaceae Lonicera Caprifoliaceae Lophocereus Cactaceae Lupinus Fabaceae Lychnis Caryophyllaceae Lycopersicon Solanaceae Magnolia Magnoliaceae Mahonia Berberidaceae Malus Rosaceae Malva Malvaceae Medicago Fabaceae Melilotus Fabaceae Melissa Lamiaceae Mentha Lamiaceae Metasequoia Taxodiaceae Mimulus Scrophulariaceae Mirabilis Nyctaginaceae Monarda Lamiaceae Myosotis Boraginaceae Nassella Poaceae Nemesia Scrophulariaceae Nemophila Hydrophyllaceae Nepeta Lamiaceae Nicotiana Solanaceae Nierembergia Solanaceae Nigella Ranunculaceae Nyssa Nyssaceae Ocimum Lamiaceae Oenothera Onagraceae Opuntia Cactaceae Origanum Lamiaceae Orthocarpus Scrophulariaceae Oxybaphus Nyctaginaceae Pachycereus Cactaceae Panicum Poaceae Papaver Papaveraceae Parthenocissus Vitaceae Pascopyrum Poaceae Paspalum Poaceae Passiflora Passifloraceae Pastinaca Apiaceae Paulownia Bignoniaceae Pelargonium Geraniaceae


2005

GENUS FAMILY Penstemon Scrophulariaceae Petroselinum Apiaceae Petunia Solanaceae Phacelia Hydrophyllaceae Phaseolus Fabaceae Phlox Polemoniaceae Physalis Solanaceae Physaria Brassicaceae Picea Pinaceae Pinus Pinaceae Piptatherum Poaceae (syn. Oryzopsis) Pisum Fabaceae Plantago Plantaginaceae Platanus Platanaceae Platycladus Cupressaceae Platycodon Campanulaceae Plumbago Plumbaginaceae Poa Poaceae Polygonum Polygonaceae Populus Salicaceae Portulaca Portulacaceae Potentilla Rosaceae Primula Primulaceae Proboscidea Pedaliaceae Prunus Rosaceae Pseudotsuga Pinaceae Psylliostachys Plumbaginaceae Puccinellia Poaceae Pulsatilla Ranunculaceae Purshia Rosaceae Pyrus Rosaceae Quercus Fagaceae Ranunculus Ranunculaceae Raphanus Brassicaceae Reseda Resedaceae Rhamnus Rhamnaceae Rheum Polygonaceae Rhododendron Ericaceae Rhus Anacardiaceae Ribes Grossulariaceae Ricinus Euphorbiaceae Robinia Fabaceae Rorippa Brassicaceae Rosa Rosaceae

GENUS FAMILY Rosmarinus Lamiaceae Rubus Rosaceae Rudbeckia Asteraceae Rumex Polygonaceae Ruta Rutaceae Saintpaulia Gesneriaceae Salix Salicaceae Salpiglossis Solanaceae Salvia Lamiaceae Sambucus Caprifoliaceae Sanguisorba Rosaceae Saponaria Caryophyllaceae Satureja Lamiaceae Scabiosa Dipsacaceae Schismus Poaceae Schizachyrium Poaceae Scirpus Cyperaceae Secale Poaceae Sedum Crassulaceae Sempervivum Crassulaceae Sequoia Taxodiaceae Sequoiadendron Taxodiaceae Sesamum Pedaliaceae Shepherdia Elaeagnaceae Silene Caryophyllaceae Simmondsia Simmondsiaceae Sinapis Brassicaceae Solanum Solanaceae Sorbus Rosaceae Sorghastrum Poaceae Sorghum Poaceae Sparganium Sparganiaceae Sphaeralcea Malvaceae Spinacia Chenopodiaceae Sporobolus Poaceae Stipa Poaceae Symphoricarpos Caprifoliaceae Syringa Oleaceae Taxodium Taxodiaceae Taxus Taxaceae Tetragonia Aizoaceae Thalictrum Ranunculaceae Thuja Cupressaceae Thymus Lamiaceae Tilia Tiliaceae


2005

GENUS FAMILY Torenia Scrophulariaceae Tradescantia Commelinaceae Trifolium Fabaceae Triticum Poaceae Tropaeolum Tropaeolaceae Tsuga Pinaceae Ulmus Ulmaceae Vaccaria Caryophyllaceae Vaccinium Ericaceae Valeriana Valerianaceae Valerianella Valerianaceae Verbena Verbenaceae Verbesina Asteraceae Veronica Scrophulariaceae Viburnum Caprifoliaceae Vicia Fabaceae Vigna Fabaceae Viola Violaceae Vitis Vitaceae Wisteria Fabaceae Yucca Agavaceae Zea Poaceae Zelkova Ulmaceae Zoysia Poaceae


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