Photochemistry, 1969, Vol. 8,
PP .
1963 to 1984.
Pergamon Press.
Printed in England.
CHEMOSYSTEMATICS OF THE UMBELLIFERAEA GENERAL SURVEY
R. K.
CROWDEN
Department of Botany, University of Tasmania, Hobart, Australia
J. B.
H A R BO R N E
and V. H.
HEYWOOD
Phytochemical Unit, Department of Botany, University of Reading, U.K.
(Received 7 May 1969)
Abstract-Some 300 umbellifer species, representing 52 per cent of the genera of the family, have been surveyed
for their leaf phenolics, using both fresh and herbarium tissue. The results show that, with few exceptions,
species can be divided into two groups, those with flavone (usually luteolin) and those with flavonol (kaempferol and/or quercetin). These groupings are mainly of interest at the generic level but are also related to tribal
divisions and may be of phylogenetic significance in the family. Other classes of flavonoid are rare: leucocyanidin was detected once in Apiastrum, and the glucoxanthone mangiferin once in Heptaptera. Furanocoumarins were found in the leaves mainly of Angelica, Peucedanum and Seseli species but a survey of seeds
of 130 species showed that these compounds were widespread in the family, some correlation with tribal
divisions being apparent. Examination of the Umbelliferae for presence of polyacetylenes, simple hydroxycoumarins and the rare sugars, apiose and umbelliferose, has shown that these substances are widespread and
consequently of little systematic interest within the family. Soluble proteins and the enzymes peroxidase and
esterase present in the seed of selected species from all tribes in the Apioideae were studied by acrylamide gel
electrophoresis. Distinct differences in patterns were found to be present at the tribal and generic levels. In
some cases, the macromolecular supported the micromolecular data in confirming generic separations. The
general value of the various chemical characters in the systematics of the family is discussed.
INTRODUCTION
A MULT I VAR I AT E approach to the classification of the tribe Caucalideae, family Umbelliferae
was initially undertaken at the University of Liverpool and is now being continued at the
University of Reading in order to compare chemical with the conventional morphological
characters and to test numerical methods in higher plant systematics.l In Bentham’s
classification of the Umbelliferae, the tribe Caucalideae comprises a group of genera with
spiny fruits that Drude3 distributed between the tribes Dauceae and Scandiceae (subtribe
Caucalineae) which are widely separated in his system. It was therefore of some interest,
before attempting a detailed chemical investigation of the Caucalideae, to carry out a general
survey of the whole family in order to see if the plant group in question was in any way
chemically distinct. Such a survey might also reveal, more generally, whether there were
chemical differences at the tribal and generic level throughout the family.
From the chemotaxonomic view-point, the Umbelliferae is difficult to survey exhaustively,
since it is a large family with 240-300 genera and over 3000 species, normally arranged in three
subfamilies Hydrocotyloideae, Saniculoideae and Apioideae. It is, however, relatively rich
1 J. MCNEILL, P. F. PARKER and V. H. HEYWOOD, in Numerical Taxonomy (edited bi A. J. COLE), Academic
Press, London and New York (1969).
2 G. BENTHAM, in Genera Phnturum (edited by G. BENTHAM and J. D. H OOK ER ) 1,859 (1867).
3 0. DRUDE, in Die natiirlichen Pflunzenfamilien (edited by A. E NGLER and K. P RANTL) 3 (S), 63 (1897-8).
1963
1964
R. K. CROWDEN, J. B.
H ARBORNE
and V. H.
HEYW~~D
in secondary constituents and much chemical work has been carried out, especially on the
furanocoumarins, terpenoids and polyacetylenes in these plants. In the present survey, the
difficulty of obtaining fresh plant material of a representative sample of taxa was circumvented
largely by analysing either leaf material from identified herbarium specimens or plant seed.
This has limited work mainly to characters which are easily detected using small amounts of
plants and to chemicals which are not destroyed during the drying process. However,
analyses of dried material have been supported by examination of fresh plant tissue of a
reasonable percentage of species and in the case of the polyacetylenes the survey has been
limited to roots of living plants. The seeds or fruits were first used exclusively for examination
for flavonoids and furanocoumarins but it was then discovered that, although small in size
compared to legume seeds, they were relatively rich in protein and enzyme content. Few
earlier chemotaxonomic surveys have been devoted to studying such a range of chemical
characters and the present examination of the Umbelliferae is one of the first attempts to
employ both secondary and macromolecular constituents in such studies.
RESULTS
Leaf Phenolics
The results of surveying 300 umbellifer species for leaf phenolics are given in Table 1. In
the case of flavonoids, the results considerably extend earlier chemical studies on some dozen
species4 and the earlier leaf survey of Bate-Smith of ten species.5 The survey was carried out
mainly on herbarium material, some specimens dating from 1840 or earlier. The majority
gave a positive result for flavonoids and the few species that were negative (mainly Angelica
and Seseli spp.) generally had large amounts of other phenolics, e.g. coumarins, in their leaves.
The flavonoids present in most umbellifers are kaempferol and quercetin; glycosides of these
two compounds have previously been reported in five species.4 Other unidentified flavonols
are occasionally present (Table 1); one is probably isorhamnetin, already identified in flowers
of O enanthe stolonifera by Matsushita and Iseda. 6 Significantly, neither myricetin nor ellagic
acid was detected anywhere; furthermore, a leucoanthocyanidin was only found once, in
Apiastrum. Hacquetia epipactis, reported by Bate-Smith5 as having leucocyanidin, was
negative (two dried specimens examined) in our tests. These results are in line with the
predominantly herbaceous character of the family. 4. 5 Luteolin was the main flavone
detected in many umbellifers; apigenin, although its first isolation was as the 7-apiosylglucoside apiin in celery seed Apium graveolens,’ is in fact rather uncommon. Some unidentified flavones were detected in a few species; in some cases these may well be methylated
derivatives. Indeed, the presence of diosmetin (luteolin 4’-methyl ether) was confirmed in
Cnidium silaifolium by spectral studies. The occurrence of diosmetin in the leaf of the poisonous hemlock, Conium maculatum, reported earlier, * however, could not be confirmed and
only luteolin was found in the specimens available for study.*
From the systematic point of view, the most significant discovery is that nearly all species
have either flavonols or flavones, but not both, some Daucus and Laserpitium species being
* This is not surprising, since this record is based on the presence of crystals with characteristics attributed
to those of “ diosmetin”.
4 J. B. HARBORNE , Comparative Biochemistry of the Flavonoids, Academic Press, London (1967).
5 E. C. BATE-SMITH, J. Linn. Sot. (Botany) 58,371 (1962).
6 A. MATXJSHITA and S. ISEDA , Nippon Nogei Kagaku Kaishi 39, 3 17 (1965).
7 E. VON G ERICHTEN , Liebig’s Ann. 318, 121 (1901).
* 0. A. O ESTERLE and G. W AND ER , Helv. Chim. Acta 8, 519 (1925).
1965
Chemosystematics of the Umbelliferae-a general survey
T ABLE 1. F LAVONOID
AND
COUMARIN
SURVEY OF LEAVES AND SEED OF THE
FAMILIES AND TRIBES ACCORDING TO
Leaf flavone or
flavonol
Species
Subfamily HYDR~COTYLOIDEAE
Tribe 1. Hydrocotyleae
Actinotus helianthi Labill,
Hydrocotyle asiatica L., H. americana L.,
H. bonariensis Lam., H. centella L.,
H. interrupta Muhl, H. javanica Thunb.,
H. prolifera Kellogg, H. repanda Pers.,
H. ranunculoides L. fil., H. umbellata L.,
H. vulgaris L.
Micropleura renifolia Lag.?
Platysace ovalis (DC.)
Trachymene pilosa Sm.
Xanthosia pilosa Rudge
Tribe 2. Mulineae
Asteriscium chilense Cham. & Schiechtt
Bowlesia incana Ruiz & Pavon$
Diposis saniculifolia (Lam.) DC.?
Domeykoa amplexicaulis (Wolff) Mathias
& Constance?
Hermas villosa (L.) Thunb.?
Huanaca andina Phil.?
Laretia acaulis (Cav.) Gill & Hooker?
Mulinum nivale (Phil.) Constancei
Spananthepaniculata Jacq.i
Subfamily SANICUL~IDEAE
Tribe 1. Saniculeae
Alepidea amatymbica Eckl. 8~ Zeyh.
A. ciliaris La Roche
Astrantia bavarica F. W. Schultz
A. carniolica Jacq.
A. major L.
A. minor L.
Eryngium alpinum L.
E. amethystinum L.
E. bourgatii Gouan
E. campestre L.
E. campestre (E. virens Link)
E. dilatatum Lam.
E. planum L.
E. glaciale Boiss
E. ilicifolium Lam.
E. leavenworthii Torrey & Gray
E. maritimum L.
E. petiolatum Hooker
E. spinalba Vill.
E. tricuspidatum L.
E, yuccifolium Michx.
Hacquetia epipactis (Stop.) DC.
Sanicula europaea L.
S. laciniata Hooker & Arnott
S. uradirius Watson
S. bipinnatiJda Douglas
D RUDE IN E NGLER
UMBELLIFERAE
PRANTL
GROUPED IN SUB-
AND
Other leaf
phenolics
Seed*
furanocoumarins
Qu, Km, Ir
Qu (as 3-glucoside), KmzyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJIHGFED
Qu, Km
Qu (as J-rutinoside), Km
au, Km
Qu, Km
+
$g Km
Qu
Qu, Km
Glycoflavone(?)
Qu, Km, Ir
Qu, Km
Aes
-
Qu, Km
Km
Km
Km
$1 EE, Fi
Km
Fi
Km, Fi
Et
Km
Km
Km
Km
Km
Aes
Aes
-
zz
g:Km
Qu
Qu
&cm
Qu (as 3-glucoside)
Ros
-
R. K. zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJIHGFEDCBA
CROWDEN, J. B. H ARBORNE and V. H. HEYWOOD
1966
T A BL E
-
-
l-continued
_
Leaf flavone or
flavonol
Species
Tribe 2. Lagoecicae
Arctopus echinatus L.fLagoecia cuminoides L.
Qu
Qu
Other leaf
phenolics
Seed*
furanocoumarins
-
Subfamily APIOIDEAE
Tribe 1. Echinophoreae
Echinophora tenuifolia L.
Qu
Pycnocycla ledermannii Wolff?zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJIHGFEDCBA
Qu
Tribe 2. Scandiceae (Scandicineae)
Antkriscus sylcestris (L.) Hoffm.
Lu
Chaeroph.yllum aromaticum L., C. aureum
1.~1 (as 7-glucoside)
L., C. coloratum L., C. heldreickii Orph.
EY Boiss., C. kirsutum L.. C. temulentum L..
Huetiu cyn&ioides subsp. &acrocarpa
Lu
(Boiss. & Spruncr) P. W. Ball (Freyera
parnassica Boiss. & Heldr.)
~Molopospertnumpeloponnesicrcr~n~ (L.)
Qu (as 3-glucoside), Km
Koch.:
_
-
_
_.
_
Tinguarra cerrariae@ia (DC). Parl:f
r. montana Benthaml
LU
LU
_
-
Tribe 2. Srandiceae (Caucalineae)
Astrodaucus littoralis (Bieb.) Drude,
A. orientalis (L.) Drude, A. persicus
Lu (as 7-glucoside)
Is0
Lu
_
Lu (as 5- and 7glucoside)
Lu
-
(C. A..Meyer) Rouy & Camus
(Boiss.) Drude.
Caucalis plafycarpos L., C. microcarpa
Hooker & Am.
Ckaetosciadium trichospermum (L.)
Boiss.f$
Lisaea hetrrocarpa (DC.) Boiss, L. syriaca
-
Lu (as 7-glucoside)
Lu
Lu (as 7-glucoside)
Lu (as 7-glucoside)
Myrrhis odorata (L.) Stop.
Osmorhiza claytonii (Michx.) C. B. Clarke
0. chilensis Hooker & Arn.
Scandix australis L., S. balansae Reuter,
S. pecten-ceneris subsp. brachycarpa
(Cuss.) Thell., subsp. marrorhynca
_
I
-
Boiss.$
Orlaya grandiflora (L.) Hoffm., 0. kochii
Lu (as 7-glucoside)
Heywood
Psamnzogeton canescens (DC.) Vatke
P. setifolium (Boiss.) Boiss.
Torilis arrensis (Hudson) Link, subsp.
arcerzsis, subsp. purpurea (Ten.) Hayek,
(T. heterophylla GUS.), subsp.
neglecta (Schultes) Thell. T. japonica
(Houtt.) DC., T. nodosa (L.) Gaertner,
T. stocksiana (Boiss.) Drude, T. ucranica
Qu
LU
Lu (as 7-glucoside)
Coumarins
_
-
Sprengel, T. fenella (Delile) Reichenb. fil.
Turgenia latifolia (L.) Hoffm.
Turgeniopsis foeniculacea (Fenzl) Boiss.?
Tribe 3. Coriandreae
Bifora radians Bieb.
Lu
Qu
Qu, Km
+
Chemosystematics of the Umbelliferae-a general survey
1967
T ABLE l-continued
Leaf flavone or
flavonol
Other leaf
phenolics
Seed*
furanocoumarins
Qu, Km (as 3-rutinoside)
Qu (Km in flower)
-
+
-
Species
B. testiculata (L.) Roth.
Coriandrum sativum L.
Tribe 4. Smyrnieae
Apiastrum angustifolium Nutt.7zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJIHGFEDCBA
Umb, leucocyanidin
Qu
Arracacia arguta (Rose) Matthias &
Umb
Qu
Constance
Astoma seselifolium DC.?
Lu
Cachrys Iibanotis L.
Coumarins
Qu
C. ferulacea (L.) Calestani
Aes
Qu, Km
C. trifida Miller
Umb, coumarins
Qu
Conium maculatum L.
Lu
Donnellsmithia hintonii Mathias & ConstancetQu, Km
Erigenia bulbosa (Michx.) Nutt.
Gr&a golaka (H&q.) R&henb.
Heptaptera triquetra (Vent.) Tutin
Mangiferin,
Coumarins
Hladnikiapastinacifolia Reichenb.
Qu (as 3-rutinoside)
Magydarispanacifolia (Vahl) Lange
Umb, coumarins
Flavonol
Musenium divaricatum Nutt.
Umb
Qu, Km
Oreomyrrhis eriopoda (DC.) Hooker?
Lu
Orogenia linearifolia S. Watson
Physospermum verticillatum (Waldst. &
$, Km
Kit.) Vis.
P. cornubiense (L.) DC.
Pleurospermum austriacum (L.) Hoffm.
Coumarins
&&l
Aes
Prangos uloptera DC.
Qu (as 3-glucoside)
Smyrnium olusatrum L. S. perfoliatum L.,
Qu, Km
S. rotundifolium Miller
Umb
Tauschia texana A. Gray?
Km
Trachydium depressun Boiss.?
Km
Vicatia millefolia (Kltzsch.) C. B. Clarke?
LU
Tribe 5. Apieae
Aegopodium podagraria L.
Aethusa cynapium L.
Ammi majus L.
A. visnaga (L.) Lam.
Annesorhiza hirsuta Eckl. & Zeyh.t
Apium graveolens L.
A. inundatum (L.) Reichenb. fil.
A. nodifiorum (L.) Lag.
Athamanta turbith (L.) Brot. subsp.
haynaldii (Borbas) Tutin
A. turbith subsp. hungarica (Borbas) Tutin
A. macedonica (L.) Sprengel
Berula erecta (Hudson) Coville
Bupleurum afine Sadler
B. angulosum L.
B. dianthifolium Guss.
B. falcatum L.
B. gibraltaricum Lam.
Carum carvi L.
C. rigidulum (Viv.) Koch ex DC.
C. heldreichii Boiss.
C. verticillatum (L.) Koch
124
Qu
Qu, Km
,“u” Km
Qu’
Lu
Qu
Qu (as 3-glucoside)
Lu (Lu in flowers)
Lu (Lu in flowers)
Qu, Km (Km in flowers)
Qu
Qu (as 3-rutinoside)
Qu (as 3-rutinoside), Km
Qu (as 3-rutinoside)
Qu (as 3-rutinoside)
Km (as 3-rutinoside)
Qu, Km (also in flowers)
Qu, Km (also in flowers)
Qu (as 3-rutinoside)
Qu (as 3-rutinoside), Km
Umb
Umb
Isoflavone(?)
Coumarin
-
R. K. zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJIHGFEDCBA
CROWDEN, J. B. HARBORNE and V. H. HEYW~~D
1968
T ABLE l- continued
Leaf flavone or
flavonol
Species
Other leaf
phenolics
Seed*
furanocoumarins
Cenolophium denudatum (Hornem.) Tutin
Qu, Km
Chaemaesciadum acaule C. A. Meyer?
Qu
_
Qu (as 3-rutinoside), Km
Cicuta buibiferu L.
C. n7aculata L.
Qu
C. virosa L.
Qu, Km
Coumarins
Cnidium silaifolium (Jacq.) Simonkai
Qu, Km
Conopodium bunioides (Boiss.) Calestani,
Diosmetin, Lu, Flavone
C. majus (Gouan) Loret, C. thalictrifilium (Boiss.) Calestani
Coumarin
Crithmum maritimum L.
Qu, Km
Cryptotaenia canadensis (L.) DC.?
Glycoflavone
Lu (as 7-giucoside)
C. elegans Webb ex Bolle
Cuminum cyminum L.
LU
Cyn7opterus terebinthinus Torrey & Gray?zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJIHGFEDCBA
Umb
Qu
Cy77osciadium digitatum D C .
_
Qu
Dethawia tenuifalia (Ramond ex DC.) GodronLu
Diplolophium iambesianum Hiernt ’
_
Endressia pyrenaica (Gay ex DC.) Gay
_
Eulophus bolanderi Coulter & Rose?
Eurytaenia texana Torrey & Gray?
Fatcaria vulgaris Bernh.
Foe77iculum vulgare Miller
Aes
Heteromorpha arborescens Cham. &
Schlecht
Hohenackeria polyodon Cosson & Dur.
Kundmannia s&la (L.) DC.
Lereschia thomasii (Ten.) Boiss.
Lichtenstehtia burchellii Hooker fil.
Ligusticum lucidum Miller
L. mutellina (L.) Crantz.
L. scoticum L.
Lilaeopsis carolinensis Coulter & Rose?
Meum athamanticum Jacq.
Oenantke aquatica (L.) Poiret
0. banatica Heuffel
0. crocata L.
0. fistulosa L.
0. foucaudii Tesseron
0. globulosa L.
0. lachenalii C.C. Gmelin
0. peucedanifolia Pollich
0. aimnincttoides L.
0. sarmentosa Presl ex DC.
0. silaifolia Bieb.
Oliveria decumbensi
Petroselinum crispum (Miller) A. W. Hill
P. segetunz (L.) Koch
Pimpinella anisum L., P. major (L.)
Hudson, P. siifolia Leresche, P. tragium
Qu (as 3-rutinoside)
Qu
Qu (as
3-glucoside)
QLl
Qu (as 3-rutinoside)
Qu, Lu (as 7-glucoside)
Lu
Qu
Qu (as
3-rutinoside),
Qu (in flower)
Qu. Km
Qu; Km
Qu
Lu
$, Km
Qu, W)
Qu, Km
Qu
Qu
_
_
_
Ott. KmzyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPO
Qu
@_I’
Lu
Qu (as 3-rutinoside)
WI.
P. saxifraga L.
P. procumbens (Boiss.) H. Wolff
P. stadensis D. Dietr.
Pleurospermum austriarun (L.) Hoffm.
Polemannia gros.rulariifolia Eckl. & Zeyh.?_
Coumarin
Coumarins
-
Km
Qu (as 3-rhamnoside)
Lu (as 7-glucoside)
Qu
Qu, Km
-
Chemosystematics of the Umbelliferae-a general survey
1969
T ABLE l-continued
Leaf flavone or
flavonol
Species
Portenschlagiella ramosissima
(Portenschl.) Tutint
Ptychotis saxifiaga (L.) Loret & Barr.
Rhyticarpus difirmis Benthamt
Ridol’a segetum Moris
Schuhzia crinita Sprengelt
Selinum calidense
S. carvifolia (L.) L.
S. capitellatum Bentham ex S. Watson
S. pyrenaeum (L.) Gouan
Seseligracile Waldst. & Kit.
S. hippomarathrum Jacq.
S. Iibanotis (L.) Koch
Silaum silaus (L.) Schinz. & Thell.
Sison amomum L.
Sium latifolium L., S. sisarum L.,
S. thunbergii DC.
Thaspium trifoliatum (L.) Grayt
Trinia glauca (L.) Dum.
T.fiigida (Boiss. & Heldr.) Drude
Trochtscanthes nodifora (Vill.) Koch
Tribe 6. Peucedaneae
Angelica sylvestris L.
Astydamia canariensis DC.?
Capnophyllum peregrinum (L.) Langet
Choritaenia capensis Benthamt
Conioselinum tataricum Hoffm.
Dorema aureum Stocks?
Eriosynaphe longifolia (Fischer ex
Sprengel) DC.*
Fe&a sadlerana Ledeb.
Fe&ago campestris (Besser) Grec.
F. granatensis Boiss.
F. sartorii Boiss.
F. sylvatica (Bess.) Reichenb.
Heracleum austriacum L. subsp. austriacum,
H. austriacum subsp. siifolium (Stop.)
Nyman
H. sphondylium L., H. minimum Lam.,
H. lanatum Michx., H. rigens Wall.
Johrenia distans (Griseb.) Halacsy
Levisticum oficinale Koch
Lomatium californicum Nutt.t
Malabaila aurea (Sibth. & Sm.) Boiss.
Opopanax chironium (L.) Koch,
0. hispidus (Friv.) Griseb.
Ormosciadium aucheri Boiss.
Ostruthium spp.
Pastinaca sativa L. subsp. sativa
Other leaf
phenolics
Seed*
furanocoumarins
+
Qu, Km
Qu, Lu (as 3-glucoside)
$:E
Lu
Qu
Lu (as 7-glucoside)
Qu
Qu
Qu (as 3-glucoside), Km
Qu (as 3-glucoside), Km
Qu
$?(!?-glucoside) Km
Qu (as 3-rutinoside;
-zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQ
-
Umb, Aes
14 other spp. all
had Umb, Aes
and other
coumarins
+
Qu (as 3-rutinoside and
3-glucoside), Km
Qu, Km
Qu (as 3-rutinoside)
Lu (as ‘I-glucoside)
Qu
Qu
Km
Km
Qu
Km
-*
+
-
9 other spp. lacked
flavonoids but
had coumarins,
inc. Umb.
Umb
-
+
+
+
Flavonol
Qu
Qu, Km
Qu
Umb
$?(%-rutinoside)
Qu (as 3-rutinoside)
Aes
Km
Umb
Qu
-
Qu (as 3-rutinoside)
$ (as 3-glucoside), Km
Glycoflavones(?)
Qu (Qu in flowers)
Qu
Qu (as 3glucoside),
Qu & Km in both
flowers and seeds
+
Coumarins
Coumarins
+
R. K. zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJIHGFEDCBA
CROWDEN, J. B. HARBORNE and V. H. HEYW~~D
1970
T ABLE l-continued
-
-
Species
Peucedanum alsaticum L., P. arenarium
Waldst. & Kit., P. ausrriacum (Jacq.)
Koch., P. carvifolia Vill., P. certiaria (L.)
Lapeyr., P. gallicum Latourr., P.
lancifolium Lange, P. latifolium (Bieb.)
DC., P. leiocarpum Nutt., P. macrocarpum Nutt., P. oficinale L., P. oreose&urn (L.) Moench, P. palustre (L.)
Moench, P. simplex, P. officinale subsp.
stenocarpum (Boiss. & Reut.) Font Quer,
P. certicillare (L.) Koch ex DC.
P. rillosum Nutt.
Polytaenia nuttallii DC.?
Tordylium maximum L.
T. oficinale L.
Zosima orientalis Hoffm:i_
Tribe 7. Laserpitieae
Elaeoselinum foetidum (L.) Boiss:F
E. asclepium (L.) Bertol. subsp. meoides
(Desf.) Fiori and subsp. asclepium
E. gummiferum (Desf.) Tutin
E. tenuifolium (Lag.) Lange
Laser trilobum (L.) Borkh.t$
Laserpitium krapfii Crantz
L. gallicum L.
L. latifolium L.
L. nestleri Soyer-Willemet
L. nitidum Zanted.
L. peucedanoides L.
L. prutenicum L.
L. siler L.
Thapsia garganica L.
T. Gllosa L.
Tribe 8. Dauceae
Artedia squamata L.
Daucus aureus Desf.
D. broteri Ten.
D. carota L.
D. crinitw Desf.
D. durieua Lange
D. glochidiatus (Labill.) Fischer &
C. A. Meyer
D. involucratus Sibth. & Sm.
D. muricatus (L.) L.
D. setifolius Desf.
Pseudorlaya pumila (L.) Grande
Seed*
Other leaf
furanophenolics
coumarins
_-_-_--__~_ _
Qu (as 3-rutinoside),
Most spp. have
f
coumarins, some
Km
with Aes or Umb
Leaf flavone or
flavonol
Ap (as 7-glucoside)
Qu
_
Coumarins
Umb
_
Coumarins
+
-
Lu
Lu
Qu
Km
Lu (as 7-glucoside)
Lu (as 7-glucoside), Ap
Lu, AP
_
-
-
Aes
c
Qu
Lu, Km
Lu, AP
Qu (as 3-glucoside), Km
+
c
-
Qu, Lu
Qu, Lu, AP
Lu (as 7-glucoside)
Lu (as 7-glucoside)
Qu (as 3-glucoside)
Lu
Lu
Lu (as 7-glucoside)
G, Km, Lu
Qu
Qu (as 3-glucoside)
Lu
Qu
Umb
+_
-
-
Lu
* Key: Km, Kaempferol; Qu, Quercetin; Ir, Isorhamnetin(?); Lu, Luteolin; My, Myricetin; Ap, Apigenin;
flavonol=unidentified flavonol; flavone=unidentitied flavone; Aes, aesculetin; Umb, umbelliferone;
coumarin(s) = unidentified furanocoumarin(s); Ros, rosmarinic acid; Iso, isochlorogenic acid ; + , present;
- not detected (blank=not examined).
j’ Source of material: Herbarium, Royal Botanic Gardens, Kew.
$ Source of material: fresh plants grown from spontaneous seed. All other material from the University of
Liverpool Herbarium. Most species in the Scandiceae (Caucalineae) and the Dauceae were examined both
as fresh and herbarium material.
Chemosystematies of the Umbelliferae-a general survey
1971
rare exceptions. Furthermore, flavones were found almost entirely in taxa generally
considered to be more specialized or advanced (e.g. zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONML
Daucus, Torilis), while flavonols predominated in the less advanced genera (e.g. Hydrocotyle). Thus, the replacement of flavonol
T A BLE 2. UHBELLIFER
SPECIES KNOW N
Tribe, genus and species
1
SAN~ULEAE
Eryngium planum L.
1,2
SCANDICEAE (Scandicineae)
Chaerophyllum tenellum
Osmorhiza aristata
Myrrhoides nodosa (L.) Cannon
1
3
2
S CANDICEAE (Cauealineae)
Astrodaucus orientalis (L.) Drude
Chaetosciadum trichospermum (L.)
2
2
Boiss.
Orlaya daucorlaya M urb.
Torilis arvensis (Hudson) Link,
T. nodosa (L.) Gaertner
T. japonica
Turgenia latifolia (L.) Hoffm.
2
2
3
2
S~~YRNIEAE
Conium maculatum L.
2
APIEAE
Aegopodium podagraria
Aethusa cynapium
Ammi visnaga
Apium graveolens
Bupleurum rotundifolium
B. longeradiatum
Carum carvi
Cicuta virosa
Crithmum maritimum
Cryptotaenia canadensis
Falcaria vulgaris
CONTAIN
Reference*
HYDROCOTYLEAE
Trachymene australis
TO
1
1
1
1
1,3
3
1
1
1
1
1
POLYACETYLENES
IN
THE ROOTS
Tribe, genus and species
Ligusticum mucronatum Hort.
Libanotis transcaucasica Schisk.
Oenanthe crocata, 0. javanica
0. pimpinelloides, 0. peucedanifolia
Petroselinum satiuum
Pimpinella koreana
P. saxifraga L.
Seseh ugoense
S. gummiferum Pall. as ex Sm.
S. dichotomum Pallas
S. buchtormense Koch
S. libanotis (L.) Koch
Sium suave W alter, S. sisarum L.
Thaspium trifoliatum (L.) Gray
Reference*
2
I23
113
1
3
2
3
2
2.3
2
2
192
2
PEu~EnAm54s
“Angelica deculsiva”,
“A. edulis”
A. miqueliana, A. polymorpha
A. sylvestris L.
Conioselinum chinense
Opopanax chironium (L.) Koch, 0.
hispidus (Friv.) Griseb.
Peucedanum verticillare
Pastinaca sativa
Tordylium maximum L.
3
3
2
3
192
1
4
2
LAS~RPI~~EAE
Thapsia villosa L.
2
D A UCEA E
Daucus aureus Desf. ,D. carotat L.
D. crinitus l&f., D. pusillus
2
2
M ichx.
D. guttatus Sibth. & Sm.
Pseudorlaya pumila (L.) Grande
2
2
* 1. BOHLM ANN et al., Chem. Ber. 94,958 (1961).
2. This paper.
3. I. YOSHIOKA et al., Yakugaku Zasshi 86,1216 (1966).
4. S. SAFE and V. THA LLER , .7. Chem. Sot. c, 1220 (1966).
t Both wild and cultivated forms were examined.
by flavone appears to have an evolutionary significance within the family, as it probably has
among the angiosperms generally.4, 5 In general, flavonoid type is very consistent at the
generic level (Table 1). Thus flavonols occur in all species that have been examined of
Carum (4), Ery ngium (15), Hydrocotyle (1 I), Peucedunum (16), Pimpinella (7) and Sanicula
(4). Similarly, flavones are present throughout Chaerophy llum (6), Conopodium (3) and
1972
R. K. CROWDEN, J. B. HARBORNE and V. H. HEYW~~DzyxwvutsrqponmlkjihgfedcbaZYXWVUTS
Tori& (8). In the few genera where this is not so, differences can be related to some morphological characteristics. For example, in Oenanthe, ten of the eleven species studied have leaf
flavonols; the exception with flavone, O.$stulosa, is a species with a very highly specialized
floral morphology. Again, differences in the genera Daucus and Laserpitium show some
correlation with taxonomic heterogeneity. At the sub-family level (Table 2) presence of
flavones separates the Apioideae from the other two sub-families. Within the Apioideae,
tribes vary according to whether flavones are rare (Peucedaneae, in one of forty-six species),
common (Dauceae, in seven of eleven species) or predominant (e.g. Caucalineae, twenty-one
of twenty-three species).
The flavonoid pattern in the flowers of umbellifers has not been examined so far extensively
because of lack of material but a sample analysis (Table 1) indicates that the pattern is similar
to that of the leaves. Anthocyanin is too rare in either flower or leaf to be of any systematic
interest; the pigments that have been studied in leaf and Bowers of the Caucalideae are uniformly based on the commonest anthocyanidin cyanidin and it is clear that there has been
little selection for an anthocyanin-based flower colour in much of the family.
A study of the flavonoid glycosidic pattern in the Umbelliferae indicates that in the leaf
it is generally a simple one (Table 1). Luteolin is commonly present as the 7-glucoside and
quercetin as the 3-glucoside and 3-rutinoside. More complex patterns are present in the
Caucalideae; luteolin occurs in leaves of some genera, e.g. Tori& as a range of other glycosides, including the 5-glucoside and 7-rutinoside, and a complex mixture of Aavone glycosides,
which are under examination, occurs, for example, in the fruits of wild specimens of Daucus
carota. C-Glycosylflavones are rare or absent and they have not been found with certainty
in any plant; the leaf flavones in Opopanax chironium (Peucedaneae) appear to be C-glycosides of apigenin and luteolin but do not correspond in R,with known glycoflavones. Glycoflavones are found in a number of unrelated plant groups in close association with glycoxanthones and it is interesting that mangiferin (2-C-glucosyl-1,3,6,7-tetrahydroxyxanthone)
has been found in a single umbellifer species Heptaptera triquetra (Colladonia triquetra)
(Smyrnieae). This substance is more often associated with flavonols than flavones, as in the
present case.
Other leaf phenolics detected during the present survey were the two hydroxycoumarins,
umbelliferone and aesculetin, already well known as constituents of the fruits and roots of
this family. They were of widespread but sporadic occurrence throughout the tribes. Other
coumarins, presumably furanocoumarins, were detected particularly in the leaves of Angelica,
Peucedanum and Seseli species, but were not further identified.
Polyacetylene Distribution
Polyacetylenes were first identified in the family as toxic principles of the water dropwort,
Oenanthe crocata, and in Cicuta virosa and Aethusa cynapium.9a lo Bohlmann and his
co-workers’l later surveyed roots of forty-one species from thirty-five genera and found
falcarinone or related compounds in fourteen species and unidentified acetylenes in three
further species. However, the remainder, including the domestic carrot, Daucus carota, were
reported as apparently lacking in polyacetylenes. Subsequently, the acetylene carotatoxin
was isolated l2 from D. carota and shown to be identical to falcarinol, CH2=CH-CHOH9 E. F. L. J. ANET, B. LYTHGOE , M. H. SILK and S. TRIPPETT , J. Chem. Sot. 309 (1953).
10 F. BOHLMANN , C. ARNDT , H. B~RNOWSKI and P. HERBST , Chem. Ber. 93,981 (1960).
11 F. B~HLMANN, C. ARNDT, H. B~RNOWSKI and K. M. KLEINE, Chem. Ber. 94,958 (1961).
12 D. G. CROSBY and N. AHARONSON, Tetrahedron 23,465 (1967).
.
Chemosystematics of the Umbelliferae-a general survey
1973
[C-C]2-CH2-CH=CH-(CH2)6CH3. 13, l4 Very recently, Yoshioka et al.l5 obtained
positive tests for polyacetylenes in thirteen umbellifer species from ten genera.
The above surveys were mainly limited to plants of the Apieae and Peucedaneae and the
present work was developed to see if the acetylenic character is uniformly distributed throughout the family and in particular to see if there was any structural variation in acetylenic type
in the tribe Caucalineae. Ether extracts of fresh roots of thirty-five species, from twenty-five
genera, most of which had not been previously surveyed, were analysed for polyacetylenes by
TLC and U.V. spectroscopy. Every plant examined (see Table 2) showed the presence of two
or more polyacetylenes, one of which corresponded in Rfand U.V. spectral maxima with the
major domestic carrot polyacetylene, and is presumably falcarinol. It is clear from these
results that polyacetylenes are widespread, if not universal, in the family. The earlier negative
findings of Bohlmann et al. were probably due to insensitive methods of detection or to
quantitative variations; several of the species reported by them as negative have now been
shown to be positive. The present survey reports the first record of polyacetylenes in the
Laserpiteae (in Thapsia). They are clearly widespread in the Caucalineae and the Dauceae,
occurring in both wild and domestic forms ofD. carota and in other Daucus such as D. aureus
and D. crinitus. Some variations in polyacetylene pattern were noted among Daucus species
and these are being further investigated. It must be concluded from this study that, as a
general character in the family, polyacetylenes, because of their ubiquitous occurrence, have
little systematic interest at the tribal level.
Furanocoumarins and Simple Sugars
Furanocoumarins such as bergaptene (I) and xanthotoxin (II) and their isoprenoid
derivatives have been isolated from the roots or fruits of some thirty-five umbellifer species
and provide a chemical character linking the Umbelliferae with the Leguminosae and
Rutaceae.16,i7 Furanocoumarins often occur as complex mixtures and as many as fifteen
such compounds have been isolated from a single umbellifer species. Previous work has been
restricted entirely to species of the Smyrnieae (e.g. Prangos), the Apieae (Ammi, Pimpinella)
and the Peucedaneae (e.g. Peucedanum) and the present survey was carried out on seed
material to see if these substances occur throughout the family.
(I) Bergaptene
(II) Xanthotoxin
Presence of furanocoumarins in the seeds was determined by appearance of mauve, blue
or yellow fluorescent spots on thin-layer chromatograms run in appropriate solvent systems.l*
In many cases the simple hydroxycoumarins umbelliferone or aesculetin were detected,
13 R. K. BENTLEY and V. THALLER, Chem. Commun. 439 (1967).
14 R. K. BENTLEY , D. BHATTACHARJEE, E. R. H. J ONES and V. THALLER, J. Chem. Sot. (c) 685 (1969).
1s I. Y OSHIOKA , T. TIMURA, H. I MAGAWA and K. TAKARA, Yukuguku Zeisshi 86,1216 (1966).
16 F. M. DEAN, Naturally Occurring Oxygen Ring Compounds, Butterworth’s, London (1963).
Chem. Abst. 1960-1968.
18 L. H&HAMME R , H. WAGNER and D. KRAEMER-HEYDWELLER, Deutsche Apoth.-Ztg. 106,267 (1966).
17
TABLEI DISTRIBUTIONOF CHEMICAL CHARACTERSINTFIE UMBELLIFERAE
Subfamily and tribe
Hydrocotyloideae
1. Hydrocotyleae
2. Mulineae
Saniculoideae
1. Saniculeae
2. Lagoecieae
Apioideae
1. Echinophoreae
2. Scandiceae
(a) Scandicineae
(b) Caucalineae
3. Coriandreae
4. Smyrnieae
5. Apieae
6. Peucedaneae
7. Laserpitieae
8. Dauceae
No. species with
,----Generic
flavonol
flavone
ascertainment
Presence of
Patterns of*
Furocoumarin r-------h___---.c--~~
Umbelliferose Polyacetylenes
proteins
enzymes
distribution
l/2
l/l
+
-
16
0
7
0
1
26
2
0
O
1
> 7j9
2
0
215
1
2
3
21
79
16
19
0
5
20
> 17121
215
22129
52185
6/11
2/10
l/3
2i3
14129
+
+
+
+
+
+
+
+
+
+
IV
II
III
III
46
7
5
1
11
7
23141
418
314
15119
316
117
+
+
f
+
+
+
IV
V
1
13/34
J
l/3
-
-
-
-I+
-
Esterase III
* The numbers refer to particularly distinctive, intense protein or enzyme bands of different R, value (see Figs l-3).
1
Esterase III
Esterase I
Esterase II
Esterase II
Esterase I
(no peroxidase)
Esterase IV
Strong peroxidase
Esterase I
(medium
peroxidase)
Chemosystematics of the Umbelliferae-a general survey
1975
together with the more complex coumarins. Analyses were carried out on comparable
amounts of seed material and species that were positive had an average of five components
with as many as eight appearing in a few taxa.
The results of surveying some 130 species for seed furanocoumarins are recorded in Table 1
and show that these substances are characteristic of the family, being present in every tribe.
The distribution at the tribal level is summarized in Table 3 and it is clear that the sampling
is too low to draw any definite systematic conclusion. However, the results confirm that the
Peucedaneae and the Smyrnieae are very rich in such constituents. By contrast, both the
Caucalineae (2/10 spp.) and the Dauceae (l/7 spp.) are relatively poor in furanocoumarins
and there is some indication here supporting the union of these taxa in one tribe, as in the
system of Bentham and Hooker.
An unusual oligosaccharide a-D-galactosido-2G-sucrose, called umbelliferose, was
reported in roots of thirteen genera of the Umbelliferae by Wickstrom and BoerheimSvendsen in 1956.ig* *O It was present in five of the eight tribes of the subfamily Apioideae but
was not recorded in the Caucalineae, Coriandreae, Laserpiteae or Dauceae. A survey was
therefore initiated to see if this trisaccharide, apparently peculiar to the Umbelliferae, was
uniformly distributed throughout the family. It was indeed, readily detected in fresh root
extracts of Torilis nodosa, Turgenia latifolia (both Caucalineae), Coriandrum sativum (Coriandreae), Thapsia villosa (Laserpiteae) and Daucus carota (in all of three subspecies)
(Dauceae). It was detected in all of eighteen umbellifer taxa examined and seems to be
universal in the family and thus valueless as a tribal character.
Examination of the free sugars in roots, leaves and fruits of representative taxa of the
family also failed to show any other characters of systematic interest. The rare pentose
sugar apiose, first reported7 in combined form as a flavone glycoside in Apium graveolens
seed, was detected free in a number of species but its distribution was quite sporadic. Apiose
was also present occasionally in the polysaccharide fraction of umbellifer species but again
its distribution was of no taxonomic significance.
Seed Proteins
In recent years, a number of investigators have shown a correlation between protein
composition and systematics in higher plants. Using electrophoretic techniques, Johnson
and Ha1121 investigated relationships in the Triticineae (Gramineae) and Boulter et a1.22-24
examined separately the systematic relationships of albumin and globulin fractions in seeds
of certain legumes, and of two dehydrogenase enzymes within the same family while Vaughan
and Denford surveyed the albumin and globulin fractions of the seeds of a number of
Brassica and Sinapis species, correlating the results with the established taxonomy. Other
workers *& *’ have used immunochemical techniques to demonstrate taxonomic affinities
between species (see review by Fairbrothers**). Rarely, however, has a sufficient number of
19 A. WICKSTROM and A. BOERHEIM-SVENDSON, Acta Chem. Scund. lo,1199 (1956).
20 A. BOERHEM-SVENDSEN, Med. Norsk. Farm. S&k. 20,l (1958).
21 B. L. JOHNSON and 0. HALL, Am. J. Botany 52,506 (1965).
22 D. J. Fox, D. A. THURMAN and D. BOULTER, Phytochem. 3,417 (1964).
23 D. B~ULTER, D. A. THURMAN and E. DERBYSHIRE , New Phytol. 66,27 (1967).
24 D. A. THURMAN , D. BOIJLTER E. DERBYSHIRE and B. L. TURNER. New Phytol. 66,37 (1967).
25 J. G. VAUGHAN and K. E. DENFORD, J. Exp. Botany 19,724 (1968).
26 P. G. H. GELL. J. G. HAWKES and S. T. C. WRIGHT . Proc. R. Sot. B 151.364 (1960).
27 k. k. L&ER, ‘R. E. ALSTON and B. L. TURNER , Am. J. Botany 52,165 (1965).
28 D. E. FAIRBROTHERS, in Modern Methods in Plant Taxonomy (edited by V. H. HEYWOOD),~. 141, Academic
Press, London and New York (1968).
1976
R. K. CROWDEN, J. B. HARBORNE and V. H. HEYWWD
species been examined to show the true potential of seed proteins as general taxonomic
characters. In this present investigation, extracts from 174 samples of seed, covering ninety-zyxwvutsrqp
+-ro %
0.6
0
czzx
in”
.= I
: 0.2 _
0 - -~ - ----02.
+.1.0- _
cz
E” 06 n3
s;a”
h 0.2 Ez
0-E
Es
gg
-
--o-2
b
0
FIGS .
c
d
e
f
g
h
i
j
k
I
m
n
o
p
q
r
l-3. IN T E R P R E T A T IVE DIAGRAMS ILLUSTRATING THE ELECTROPHORETIC DISTRIBUTIONS O F
ESTERASE AND PEROXIDASE COMPONENTS IN SEED EXTRACTS OF SPECIES OF U MBELLIFERAE
PROTEIN ,
SUBFAM .
F IG. I. Tribe I. Echinophoreae
Tribe II. Scandiceae
1. Scandicineae
2. Caucalineae
APIOIDEAE .
a. Echinophora sibthorpiana Guss.
b. Anthriscus nemorosa (Bieb.) Sprengel
C. A. sylvestris (L.) Hoffm.
d. Chaerophyllum aromaticurn L.
C. hirsutum L.
;: Molopospermum peleponnesiacum (L.) Koch
g. Scandix iberica Bieb.
h. S. pecten-veneris L.
1. Torilis nodosa (L.) Gaertner
J. T. japonica (Houtth.) DC.
k. T. tenella (Delile) Reichenb. fil.
1. Caucalis platycarpos L.
m. Pseudorlayapumila (L.) Grande
n. Orlaya kochii Heyw.
0. 0. grandifora (L.) Hoffm.
P. Turgenia latifolia (L.) Hoffm.
9. Astrodaucus littoralis (Bieb.) Drude
r. A. orientalis (L.) Drude
nine species and thirty-nine genera, with representation from all eight tribes of the subfamily
Apioideae, were examined by acrylamide gel electrophoresis. Examination was made of the
general protein pattern, using amido black and light green as staining reagents, and also of
enzymatic activity with respect to esterase, peroxidase, catalase, and in some instances
Chemosystematics of the Umbelliferae-a general survey
1977
amylase. The block technique was used because this method affords a more direct means of
comparison between a number of samples under strictly comparable conditions in a single gel
preparation. Also, by slicing the gel horizontally, it is possible to compare each sample for
a variety of properties, by differential staining of the slices.
I +
-
lo-
szyxwvutsrqponmlkjihgfedcbaZY
06.
:a
LI
b
a
0.2
O_-
~
-
-
- -0 2 .
+ _ I-O -
+r
k” -
-
IO
b
-
c
-
d
_
_
e
-
_
f
g
FIG. 2. Tribe III. Coriandreae
Tribe IV. Smyrnieae
Tribe V. Apieae
Tribe VIII. Dauceae
h
i
-
j
k
I
m
n
o
p
q
r
a. Coriandrum melphitense Ten. & Guss.
b. C. sativum L.
Bifora radians Bieb.
“d: Conium maculatum L.
e. Prangos lophoptera Boiss.
f. P. ferulacea Lindley (L.) Calestani
Smyrnium perfoliatum L.
i? Cachrys trifida Miller
J. Ammi visnaga (L.) Lam.
k. Apium graveolens L.
1. A. inundatum (L.) Reichenb. fil.
m. Bupleurum prealtum L.
n. Falcaria vulgaris Bernh.
o. Daucus carota L. subsp. sativus (Hoffm.) Arcangeli
P. D. carota L. spont.
4. D. aureus Desf.
r. D. carota L. subsp. gadecaei (Rouy & Camus)
Heywood
The investigation has revealed a number of differences between the species examined,
some of which are undoubtedly of taxonomic significance. Staining with amido black showed
that most species of the Caucalineae, except Turgenia Iatifolia (three accessions), O rlay a
grandlflora (two act.) and Pseudorlaya (two sp., five act.), contained cationic proteins which
gave heavily staining bands. These cathodic-migrating bands were not seen with amidoblack staining in any of the samples from other tribes. However, the variation is quantitative
only, since in many cases cationic peroxidases were detected in other species, the enzymatic
R. K. CROWDEN, J. B. HARBORNE and V. H. HEYW~~D
1978
assay being a more sensitive means of protein detection than the amido-black reagent. With
regard to the other tribes, the Dauceae and Apieae gave essentially similar protein patterns as
did the Coriandreae with the Smyrnieae, and the Scandiceae with the Peucedaneae. The
Laserpiteae were distinct in that their electrophoretograms showed a number of strong
-
iIz
ZZ
(I
b
z!
1
-
c
d
-
-
is
--
Et
-
e
f
FIG. 3. Tribe VI. Peucedaneae
-
B
”
-
=
-
P
-
g
h
a.
b.
C.
d.
7
g.
h.
1.
Tribe VII. Laserpitieae j.
k.
1.
m.
n.
0.
P.
9.
r.
i
klmnopqr
jzyxwvutsrq
Ferula tenuisecta Korov.
Ferula tschimganica Lipsky
Opopanax hispidus (Friv.) Griseb.
0. chironium (L.) Koch
Heracleum stevenii Manden.
H. ponticum (Lipsky) Schischk. ex Grossh.
Pastinaca sativa L.
Peucedanum lubimenkoanum Kotov
P. oreoselinum (L.) Moench
Laserpitium latifolium I...
L. siler L.
L. halleri Crantz
L. archangelica Wulfen
Laser trilobum (L.) Borkh.
L. trilobum (L.) Borkh.
Elaeoselinum thapsoides DC.
E. gummiferum (Desf.) Tutin
Thapsia cillosa L.
Rp bands were also seen in several
bands of high electrophoretic mobility (R,). 22 Some high zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJIHGFE
species of the Caucalineae (e.g. Tori& Astrodaucus). Examples of the typical protein patterns
for the various tribes are shown in Figs. l-3. Staining for protein with the light-green reagent
was much weaker than amido-black, but was useful in that it allowed better resolution of some
close-running bands.
Chemosystematics of the Umbellifera+a general survey
1979
All species were shown to contain a variable number of esterase enzymes. However, in
only three species, Conium maculatum, Cachrys ferulacea (Smyrnieae) and Pastinaca sativa
(Peucedaneae) was a cationic esterase observed. In respect of the general esterase patterns,
the Apieae and Dauceae were very similar to one another, and to Tori& and Pseudorlaya of
the Caucalineae. The Coriandreae again bore close resemblance to the Smyrnieae, as did the
Scandiceae with the Laserpitieae and Echinophoreae. In each of these groups there was a
single major esterase component and a variable number (2-5) of minor bands, the main
difference being the Rp of the major component. The Peucedaneae presented a rather mixed
esterase pattern, sometimes with more than one major component and of variable Rp, whilst
for the most part the Caucalineae, particularly Astrodaucus, Caucalis and O rlay a, had
distinctive esterase patterns with several very strong bands.
A strong peroxidase reaction was shown by many species of the tribe Laserpitieae (though
not Laser trilobum, Laserpitium archangelica and L. gallicum) and by Chaerophyllum species
(Scandiceae). Other species of the Scandiceae and the Dauceae gave a very modest peroxidase
reaction, but in other tribes the reaction was weak (Caucalineae) or absent altogether. In all
cases both cationic and anionic peroxidases were observed.
Oddly, most of the species with weak or no evident peroxidase activity gave a strong
catalase reaction. Catalase activity was negligible in the Laserpitieae. However, in the whole
survey only one catalase band per species was observed, and this always occupied the same
region on the electrophoretogram (Rp approx. O-1). Amylase activity was examined for many
species but it appeared to be a very poor taxonomic character, and it did not allow any
differentiation between species.
DISCUSSION
1. Taxonomic History of the Umbell~erae
As an introduction to the discussion of the chemical data presented in this paper we have
felt it useful to give a brief survey of the taxonomic background of the Umbelliferae. The
Umbelliferae have been recognized as a natural group since the earliest studies of plants
dating back to Theophrastus (see Rodriguezzg). So striking are the features in which the
members agree, such as the typical unbellate inflorescence, pentamerous perianth and
androecium, and characteristic fruits, that the group occupies a special role in the development of the systematist’s concept of natural groups and affinity (cf. Sachs30). In practice this
is mainly true as regards the Apioid group of Umbelliferae to which most North Temperate
members belong.31 This very uniformity in structural features makes for considerable
difficulties in subdividing the family and there has been disagreement between authors from
the time of Linnaeus until the present day as to the number and circumscription of the tribes
to be recognized. In this respect it is similar to other “ natural” families such as the Compositae and Cruciferae where recognition of tribes and genera is often highly unsatisfactory
or frankly artificial.
At the subfamily level, morphological divisions are fairly clear-cut and most people
nowadays follow Drude (in Engler and Prant13) in recognizing three subfamilies based
largely on anatomical features : Hydrocotyloideae, Apioideae and Saniculoideae. While the
R. L. RODRIGUEZ, Univ. Calif. Publ. Bot. 29, 145 (1946).
30 J. VON SACHS, History of Botany (transl. H. E. F. GARNSEY and I. B. BALFOUR), Clarendon Press, Oxford
29
(1890).
31 M. E. MATHIAS and L. CONSTANCE , Univ. CaliJ Publ. Bot. 33, No. 2 (1962).
1980
R.K. CROWDEN, J.
B. H ARBORNE
andV .H. HEYW~~D
first of these is sometimes recognized as a separate family, the Hydrocotylaceae, Calestani 32
did not agree that the main feature separating the Hydrocotyloideae from the Saniculoideae,
the woody endocarp, is sufficient for even subfamilial recognition and united the two groups.
On the other hand, he considered Lagoecia (of the subfamily Saniculoideae) to be sufficiently
different to merit treatment as a subfamily of its own, the Lagoecineae.
At tribal and subtribal level there is much greater diversity of treatment, depending on
which characters (usually of the fruit) are given greater importance (cf. Ref. 1). Thus Boissier,
following Bentham (in Bentham and Hooker*), recognized eleven tribes, Drude (in Engler
and Prant13) twelve which were often considerably different in circumscription from those of
Boissier, Calestani32 ten, again often very different from those of the other authors, and
Cerceau-Larriva133 twenty-seven based largely on fruit seedling and pollen characters, later
modified34 to thirty-one following a detailed study of the pollen-types Tori/is-CaucalisDaucus group of genera.
2. Relationships with the Araliaceae
Despite its naturalness, systematists have long recognized a close relationship between
the Umbelliferae and the Araliaceae (and to a lesser extent the Cornaceae) : the three families
are frequently placed in a single higher group (e.g. the cohort Umbellales, Bentham in
Bentham and Hooker2) and indeed the Umbelliferae and Araliaceae are united as one family
by some workers such as Baillon,35 Calestani32 and recently by Thorne.36 Calestani discusses
and tabulates the differences between the two families and concludes that although several
characters separate them none of them is without exceptions and none has “real systematic
importance”. If lack of exceptions is a criterion, then both must be united as one family
which he calls the Apiaceae. Anatomical studies have stressed the similarities between the
families and an important role is played by such genera as M y odocarpus of the Araliaceae,
which although sometimes placed in the Umbelliferae, is similar in its fruit structure to the
schizocarpous condition of the Umbelliferae. A special study of this genus was made by
Baumann,37 who showed that while its fruits showed certain features such as primary ribs and
resin canals which are characteristic of, although more strongly developed in, the Apioid
group of the Umbelliferae, in vegetative features, inflorescence and flower structure the
genus agreed more closely with the Araliaceae. Baumann concluded that the Umbelliferae
were derived from a pre-Araliaceous stock which had become more highly evolved than the
Araliaceae.
Rodriguez,2g following a study of the woody genus M y rrhidodendron and the histology
of secondary xylem throughout the Umbelliferae which he compared with a number of
Araliaceae and other Umbellales, agreed with Baumann’s concept of a number of divergent
lines arising from a theoretical pre-Araliaceae. As regards wood anatomy and other morphological considerations, he considered the Araliaceae as forming an uninterrupted series with
the Umbelliferae but standing at a more primitive level, although exhibiting at the same time
a greater versatility of characters. Accordingly he felt it to be immaterial whether they were
labelled as separate families or as a single one. On the other hand he found the relationship
of these two families to the others placed in the same order, i.e. the Cornaceae, Nyssaceae
32V. CALESTANI, Webbiu 1,89 (1905).
33 M. T. CERCEAU-LARRIVAL, M&n. MUS. Nat. Hist. ser. B, 14,l (1962).
34 M . T. CERCEAU-LAP-RIVAL, Pollen et Spores 7 (l), 35 (1965).
35 H. E. B AILLON, Histoire des Plantes 7, 84-256 (1879-1880).
36 R. F. THORNE , Aliso 6,57 (1968).
37 M. G. B AUMANN , Ber. Schw. Bot. Ges. 56, 13 (1946).
Chemosystematics of the Umbelliferae-a general survey
1981
and Garryaceae less obvious, so agreeing with most other workers (cf. Cronquist 38) although
Tamamschian (reported in Cronquist 38) considers there to be a close relationship between
the Cornales and Umbellales. It is clear, therefore, that a full assessment of the phytochemical
characteristics of the Umbelliferae would have to take into account the Araliaceae as well.
The survey reported here has, however, been restricted so far to the former family. It is
evident from what is already known of the chemical constituents of the Araliaceae39 that
they are similar to the Umbelliferae. In terms of phenolic constituents the data agree in broad
terms with the evidence from wood anatomy (see Rodriguez,2g Fig. 69) as regards the relative
advancement of the two families but this is largely to be expected due to the degree of
necessary correlation between these two features.
This then is the taxonomic background against which the chemical data have to be considered. To summarize, the family shows considerable uniformity in several conspicuous
morphological features which lead to its instant recognition, and superimposed on this
uniformity there is a reticulate distribution of characters which makes tribal and generic
delimitation difficult and which leads to the conclusion that no one subfamily is overall more
advanced in evolutionary terms than any other, although in respect of individual features
character trends can be detected. This is well illustrated by Rodriguez29 who pointed out
that in order to list the species in a continuous sequence according to mean vessel-length,
the systematic division into subfamilies and tribes had to be ignored. In other words the
family illustrates in a very clear fashion the concept of evolution of characters (semophyleses)
as opposed to overall evolutionary advancement which is usually an average condition since
individual characters evolve at different rates.40
3. Chemotaxonomic Relationships
Much of the chemical data obtained in the present survey, summarized in Table 3, support
the above view of the family; thus, surveys for acetylenes, free sugars, simple and furanocoumarins show rather clearly the chemical homogeneity of these plants. How far these
characters distinguish umbellifers from related families is not at present completely known,
except that polyacetylenes have also been reported in species of the Araliaceae.
Once again, the only secondary constituents showing significant variation within the
family are the flavonoids. Sufficient numbers of species and genera have been surveyed to
show that presence of flavone versus flavonol is correlated with systematics. Tribes can be
divided (Table 3) into two broad groups: the majority of nine in which flavones are rare or
lacking and four in which flavones are common or predominant (Apieae, Dauceae, Laserpitieae and Scandiceae). The significance of this division, which may have some systematic
and evolutionary significance, is difficult to assess. Certainly, the subfamily Hydrocotyloideae
which is completely lacking in flavones in less typically umbelliferous than, say, the Apioideae,
and closer to the Araliaceae in possessing a woodykndocarp, no separate carpophoer and
frequent absence of vittae, but cannot be regarded as in any way ancestral to present-day
Apioideae or Saniculoideae but rather as one of several separate independently developed
lines from some pro-Araliaceous-Umbelliferous stock. Again, the Saniculoideae, which in
our sample completely lacks flavones, is not to be regarded as primitive in view of the many
specializations of features it shows but has probably evolved separately from the other groups
38 A. CRONQUIST, The Evolution and Classification of Flowering Plants, Nelson, London and Edinburgh (1968).
39 R. HEGNAWER, Chemotaxonomie der Pfanzen, Vol. 2. (1964).
40 P. H. DAVIS and V. H. HEYWOOD, Principles of Angiosperm Taxonomy, Oliver and Boyd, Edinburgh and
London (1963).
1982
R. K. CROWDEN, J. B. HA RBORNE and V. H. zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPON
J~EYWGOD
of the family for a long period of time. The A pioideae in respect of presence or absence of
flavones is very mixed : although most tribes possess them, in some they are w ell represented
w hile in others they are rare or absent altogether in our sample.
A t the tribal level the most distinctive differences observed during this survey are in the
protein and enzyme patterns. This is largely because these patterns contain many items of
information w hich can be assessed visually, although it is at present difficult to analyse and
interpret this information in terms of individual characters, even w ith the use of computers
(cf. Ref. 25). For example, it is difficult to know the degree of homology w hich can be accorded
to bands appearing in the same position but in different samples. This difficulty has been
largely overcome by the use of enzyme stains, the results of w hich confirm the cruder protein
patterns obtained w ith amido-black. It is generally possible by the combined use of protein
and enzyme patterns to distinguish all tribes w ithin the subfamily A pioideae, w ith the exception of the Smyrnieae and Coriandreae. This ties in w ith preliminary results reported by
Pickering and Fairbrothers 42 w ho indicate that by using serological and disc electrophoretic
methods the three subfamilies can be separated on the basis of their protein chemistry. The
sample of species examined in the protein survey has, however, been limited by the seed
material available and differences observed may possibly reflect differences at the generic
rather than at the tribal level.
Certainly, the macromolecular data obtained are most immediately applicable to
problems at the generic level, where in fact most difficulties are experienced by taxonomists.
In this connexion, it is satisfying that the protein and the flavonoid data support each other in
drawing generic limits. Two examples may be mentioned. Thus the monotypic g enu s
Turgenia latifolia, w hich has often in the past been classified w ith Caucalis, is readily separated
from the latter genus on the basis of the specific occurrence of luteolin 4’-glucoside and
chrysoeriol 7-glucoside in the seed and of the absence of peroxidase enzymes and of backmigrating bands in its seed protein pattern. The recognition of Turgenia as a separate genus
is also supported by details of the microstructure of the fruit, as revealed by scanning electron
microscopy (Heywood 43 and unpublished) and in a pilot multivariate numerical assessment.
Again, Daucus and Astrodaucus, two genera w hich as their names imply are closely allied
morphologically, can be separated on a leaf phenolic character, the complete replacement of
chlorogenic acid as the principle caffeic ester by isochlorogenic acid in the latter genus, and
also on their protein patterns, only Astrodaucus show ing the presence of a cation migrating
band.
Finally, turning to the taxonomic problem of w hether the spiny-fruited umbellifers should
be separated into tw o groups as Drude w ould have it, or combined in a single tribe according
to Bentham, the present chemical survey does provide a partial answ er. On balance, the data
obtained, particularly regarding the flavone and furanocoumarin characters, are in favour of
treating the taxa as a single unit. Furthermore, the Caucalineae and Dauceae have very
similar protein patterns and are united in a common esterase pattern. By contrast, protein
and enzyme patterns clearly distinguish the Caucalineae from the Scandicineae and if one
wished to maintain the tribe Scandiceae as in the classification of Drude, one would have to
accept considerable chemical heterogeneity within its bounds.
Although there is little chemical diversity of pigmentation in the flowers, schizogenous
V. H. HEYWOOD, Modern Methods in Plant Taxonomy, 1, A cademic Press, London and New York
(1968).
42 J. L. PICKERING and D. E. FAIRBROTHERS, Am. J. Botany 55,737 (1968).
43 V. H. HEYWOOD, Proc. Linn. Sot. Lund. 179,287 (1968).
41
Chemosystematics of the Umbelliferae-a general survey
1983
oil-canals are found in the root, stem and usually in the pericarp of the fruits (where they are
termed vittae). The various oils and resins secreted are largely responsible for the characteristic odour and taste of many of the species which are used for medicinal and culinary
purposes, e.g. Betts.44 These oils and resins show considerable diversity and clearly have a
high selective value. They have not yet been fully surveyed during this study and results of
work in progress 45 will be reported later.
E X P E RI M E N T A L
Plant Material
Living material, largely belonging to the Caucalideae, was grown from seed of known origin at the
University of Liverpool Botanic Gardens, Ness, Cheshire. Voucher specimens are housed at the Herbarium,
Department of Botany, University of Reading, and details of seed sources are available on request. Herbarium
material was sampled from Umbelliferae specimens made available by the courtesy of the curators of the
University of Liverpool Herbarium and the Keeper of the Herbarium, Royal Botanic Garden, Kew. In the
case of specimens in the Liverpool Herbarium, sampling labels have been affixed to the sheets of material
examined.
Flavonoids
These were extracted and identified by standard procedures. Both direct and acid-hydrolysed leaf extracts
were chromatographed in four solvents against authentic markers. In addition, co-chromatography and
spectral confirmation was carried out in representative cases. Details of the identification of individual
flavonoids in the Umbelliferae, and of mangiferin in Heptaptera, will be given in a later paper.
Furanocoumarins
The presence of furanocoumarins was recorded in leaf extracts when spots with intense mauve, light green
of light yellow fluorescence in U.V. light and of usually high Rf values in most of the solvent systems used for
flavonoids were noted on chromatograms. For the seed survey, seeds were ground into a powder with sand,
extracted twice with netrol. ether and the residue hvdrolvsed for 30 min with 2 N HCl and the coumarins
extracted into ethyl -acetate. The concentrated petrol. -ether and ethyl acetate extracts were separately
chromatographed in ether-benzene-10 y0 HOAc (1: 1: 1) on silica gel plates. Species containing furanocoumarins showed the presence of several (from three to twelve) blue, mauve, green or yellow fluorescent
spots with R, between 0.1 and 0.9 in one or both extracts. Spectral measurements on representative species
confirmed the coumarin nature of the substances. Species already known to be rich in furanocoumarin
content responded in the same way. The simple hydroxycoumarins, umbelliferone and aesculetin, were run
as markers on all plates and their occurrence (when present) confirmed by co-chromatography.
Polyacetylenes
Fresh roots were thoroughly
- washed, dried and then macerated in ether and left for a week at 4” in the
dark. The ether layer was poured off, dried and concentrated at room temperature. The extracts were screened
for oolvacetvlenes bv TLC on silica gel in benzene-CHCIT (9: 1). pentane-ether (9: 1) and CHC&-MeOH
(9: i). Polyacetylenes appeared as br&n spots or bands of appr& R,s 0.12, 0.28‘and 0.49 after the plates
had been sprayed with 0.4 % isatin in cont. H2S04 and heated for 10 min at 110” . Other constituents with blue
or red colours occasionally appeared but further spectral examination showed them not to be acetylenes. The
polyacetylene bands, on elution into ether, each gave five or six intense peaks in the short u.v., in the region
230-310 nm characteristic of known polyacetylenes. A domestic carrot extract, which contained falcarinol,
was run as a marker on all chromatograms.
Sugars
Fresh roots were macerated in MeOH, and the filtered extract concentrated and examined for the presence
of umbelliferose by chromatography on paper in the usual sugar solvents (over-developed for oligosaccharides)
and sprays. Umbelliferose appeared in the species examined (seep. 1975) as a brown spot with aniline hydrogen
phthalate of RG0%4 in butanol-acetic acid-water (4:l: .5),@62 in butanol-benzene-pyridine-water (4: 1: 1: 3)
and 0.70 in butanol-ethanol-water (4 : 1: 2 : 2) (lit. value RSucrore 0.70 in isopentanol-pyridine-water (7:7:6)).
An unknown oligosaccharide was detected in the parsnip, Pastinaca satiua, with R, 044 in butanol-acetic
acid and water. The free sugars present in the alcohol-soluble and alcohol-insoluble (after acid hydrolysis)
T. J. BETIS, J. Pharm. Pharmac. 20,469 (1968).
45 J.B. HARBORNE,V.H.HEYWOOD andC. A. W ILLIAMS , Phytochem. 8, 1729 (1969).
44
125
1984
R. K. CROWDEN, J. B.
H ARBORNE and V. H. HEYW~~D
fractions of seed, leaf and fruit of three to seven representative species from the various umbelliferae tribes were
examined by paper chromatography. All the usual monosaccharides were detected and, in addition, apiose
was found occasionally to be present, mainly in the polysaccharide fraction; its distribution was unrelated to
taxonomy. Authentic apiose marker was obtained by hydrolysing apiin, from celery seed.
Samples for Electrophoresis
Whole seeds (about 25 mg) were ground to a fine powder with an equal weight of sand, and the grinding
continued with 6-8 drops of extracting buffer to produce a thick slurry. This slurry was allowed to stand for
0.5 to 1 hr before centrifuging. The supernatant (40 ~1) was used for electrophoresis. Protein content of the
solutions prepared in this way varied usually between 2-3 per cent (Lowry estimation).46 Concentrations were
not adjusted if they fell within this range. The extracting buffer contained hydroxymethylamine methane
(35 /AM), citric acid (2.5 ELM), ascorbic acid (6 PM), cysteine-HCl(6 PM) and sucrose (0.5 M).
Preparation of
Electrophoresis
Horizontal electrophoresis was conducted in a Shandon apparatus (Kohn type) using a 7.5 % acrylamide
gel, essentially after the method of Lund.47 The gel was polymerized in a flat perspex mould (150 x 100 x 6
mm) covered with a plate-glass lid from which projected a row of thirteen celluloid slot formers (5.0 x 1.5 x 5.5
mm) approx. 25 mm from the cathode side of the gel. Each slot accepted 40 ~1 of sample. A constant current of
20 mA was maintained throughout the electrophoresis, during which time the potential rose from 90 to about
170 V. The sample slots were filled with extracting buffer, and pre-electrophoresis conducted for approx.
1.5 hr, when the buffer was replaced by the seed extract samples. Electrophoresis of the samples was continued
for a further 2-2.5 hr, or until the borate-ion boundary had migrated 50 mm towards the anode from the sample
slots.
After electrophoresis the sample slots were cleared of liquid and the gel cut horizontally, using a taut-wire
slicer (Shandon), into four slices (150 x 100 x 1,5 mm). Each slice was then stained separately to reveal the
various protein and enzyme fractions.
Sfaining Reactions
Protein. One slice was submerged for 1 hr in a solution of 0.7 % amido black in 7 % acetic acid. The residual
dye was subsequently removed by repeated washing with 7 % acetic acid. In some cases light green (0.4 % in
7-z acetic acid) was used to stain-protein. Esterusewas demonstrated by immersing a slicein iO0 ml of phosohate buffer (0.1 M. oH 6.3) containing or-naohthvlacetate. 100 mg, and diazo blue B (Michrome 250-E.
burr), 50 mg; for 5-10 min.’ The back&oundawasclarifiededby washing with water. Per&idase was demonstrated by immersing a slice in 100 ml of acetate buffer (0.1 M, pH 4.4) containing o-dianisidine, 100 mg, and
H-O2 (30 vols.), 0.5 ml. Other reagents, e.g. benzidine, guaiacol, catechol, may also be used, but O-dianisidine gives the most intense and stable reaction product. Catalase activity was observed as areas of oxygen
evolution during evaluation of peroxidase activity. Amy lase. To observe amylase, @6% soluble starch is
added to the gel during preparation. A slice is incubated for 1 hr in phosphate buffer (0.1 M, pH 6.4) and
then transferred to a solution of I1 (0.005 %) in 1.5 KI.
Acknowledgements-Two of the authors (J. B. H. and V. H. H.) are grateful to the Science Research Council
for financial support (research grant B/SR/l923). R. K. C. carried out this work in Liverpool while on
sabbatical leave from the University of Tasmania.
46 0. H. LOWRY , N. J. ROSEBOROUGH , A. L. F ARR and
47 B. M. L UND , J. Gen. Microbial. 40,413 (1965).
R. J. RANDALL , J. Biol. Chern.
193,265 (1951).