Phytochemistry 71 (2010) 937–947
Contents lists available at ScienceDirect
Phytochemistry
journal homepage: www.elsevier.com/locate/phytochem
Chemodiversity of exudate flavonoids in Dionysia (Primulaceae): A
comparative study
Karin M. Valant-Vetschera a,*, Tshering D. Bhutia a, Eckhard Wollenweber b
a
b
Chemodiversity Research Group, Department of Systematic and Evolutionary Botany, University of Vienna, Rennweg 14, A-1030 Wien, Austria
Botanisches Institut der TU Darmstadt, Schnittspahnstrasse 3, D-64287 Darmstadt, Germany
a r t i c l e
i n f o
Article history:
Received 8 July 2009
Received in revised form 29 January 2010
Accepted 3 March 2010
Available online 8 April 2010
Keywords:
Dionysia
Primulaceae
Exudate constituents: flavones, flavonols,
flavanones
Diversification
Taxonomy and phylogeny
a b s t r a c t
More than 60 accessions of various Dionysia spp. were analysed for their exudate flavonoid composition.
Many Dionysia spp. accumulate the typical Primula flavonoids with irregular substitution (unsubstituted
flavone, its 20 ,50 -substituted derivatives and corresponding 5-OH-flavones), but flavones, flavonols and
flavanones with regular 5,7-diOH-substitution are also encountered in their exudates. The formation of
both types of flavonoids is not mutually exclusive. This paper analyses the chemodiversity of Dionysia
exudates with respect to infraspecific variability, infrageneric distribution, patterns in hybrid taxa, and
comparisons of biogenetic tendencies between Dionysia and closest related species of Primula. The
uniqueness of occurrence of Primula-type flavonoids in the family Primulaceae, and their presumed different biosynthetic origin, suggest significance as further character in the Primula–Dionysia assemblage.
Principal component analysis was applied to test the significance of variation of flavonoid composition
across Dionysia. Comparative analysis of flavonoid profiles against the current taxonomic views yielded
correlations, confined to the level of smaller groups, and only in parts at level of the current infrageneric
concept. Flavonoid data are further discussed against the background of morphological and biogeographic differentiation of the genus. Increased diversification of flavonoid profiles may be interpreted
as a derived status in Dionysia, which agrees with current views on the phylogeny of Dionysia as a specialised group within Primula. Functional aspects of exudate flavonoid formation are shortly addressed.
Ó 2010 Elsevier Ltd. All rights reserved.
1. Introduction
About 50 species are currently known under the generic name
Dionysia, which according to recent molecular data, form a monophyletic group deeply nested within groups of the genus Primula
(Mast et al., 2001; Trift et al., 2004). Even earlier views consider
‘‘Dionysia represents a blind alley in the scheme of evolution – it
is more or less unsuccessful, but so far the best attempt of Primula
to enter the domaine of the xeromorphs” (Wendelbo, 1961). This
quotation refers to the growth habit of most of the Dionysias: fruticulose, loose or dense tufts and cushions, reduced leaves – all
adaptations to grow under extreme conditions such as in vertical
niches or overhanging cliff walls at higher altitudes. However, Dionysia spp. are not really xerophytic, but need a certain amount of
water supply and shade that is provided in these niches (Wendelbo, 1961). Apart from the extreme growth form, typical characters of Dionysia are the base chromosome number of 10, the
occurrence of polycolpate pollen, and the five-valved capsule, all
uncommon in Primula (Trift et al., 2004). The distribution area is Irano-Turanian, with the majority of species found in the Zagros
* Corresponding author. Tel.: +43 1 4277 54070; fax: +43 1 4277 9541.
E-mail address: karin.vetschera@univie.ac.at (K.M. Valant-Vetschera).
0031-9422/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.phytochem.2010.03.004
Mountains, partly as endemics. The Iranian Alburz mountain range
harbours fewer species, and others occur exclusively in Afghanistan. Only a few taxa extend into Turkey, Turkmenistan, Oman
and Pakistan. Recently, the phylogeny of this genus was analysed
by sequence data, which were not fully in accordance with morphological differentiations, and there is difficulty finding diagnostic characters that are related to the evolution of this genus (Trift
et al., 2004). Analysis of leaf sclereid characters, combined with
molecular and biogeographic data (Trift and Anderberg, 2006),
gave the basis for a recent revision by Lidén (2007), who maintained Dionysia as a genus for convenience.
The production of waxy or farinose coatings on aerial parts
including the floral region is a major character of Dionysia and
Primula. In Dionysia, farina may have either a woolly or powdery
consistency, but some of the species are efarinose, a character
sometimes used for infrageneric classification (Trift et al., 2004).
In contrast to Primula, Dionysia leaves exhibit an aromatic smell
(Melchior, 1943). In Primula, farina and oily exudates consist of
unsubstituted flavone and some further biogenetically rare types
of flavones (Valant-Vetschera et al., 2009 and references cited
therein). Earlier, the presence of some of these flavones was
reported in hydrolysates of some Primula and Dionysia species
(Harborne, 1968, 1971). Whether the anti-tumour activity of the
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K.M. Valant-Vetschera et al. / Phytochemistry 71 (2010) 937–947
Iranian medicinal plant Dionysia termeana Wendelbo is related to
the presence of flavonoids, has not been specified (Zahra et al.,
2007). Preliminary analyses of exudate flavonoid composition in
Dionysia indicated chemical similarities to Primula exudates
concerning major accumulation tendencies. Larger sampling, consisting in parts of the same material that was studied phylogenetically by Trift et al. (2002, 2004) and Trift and Anderberg (2006)
became available for comparative studies at the infra- and interspecific level. The results are discussed in relation to morphological
features and current taxonomic views, also with regard to the closely related genus Primula.
2. Results and discussion
2.1. Exudate flavonoid composition
Sixty-seven accessions of Dionysia species including subspecies
and some hybrid taxa have been analysed for the chemodiversity
of exudate flavonoids. The species studied belong to different subgeneric units as described by Lidén (2007). Flavonoid data are summarized in Table 1, in which the species are arranged according to
their position based on current phylogenetic data (Trift and Anderberg, 2006). Clades are marked as A, B, C, D, E, F, G and combinations there of, while section names and their abbreviations
follow Lidén (2007). Details are explained in the legend to Table 1.
This table also includes data on infraspecific variation in cases
where several accessions per taxon were available.
The formation of flavones with the irregular substitution pattern typical of Primula exudate flavonoids is also a general feature
of Dionysia exudates. These flavonoids are marked as ‘‘Primula-type
flavonoids” (in Table 1) and include one chalcone derivative (20 ,bdiOH-chalcone), further unsubstituted flavone, its 20 - and 50 -Osubstituted derivatives (5-deoxyflavones), and the structurally related 5-OH-flavones. Occasionally, typical Primula-structures such
as 30 ,40 -di-OH-flavone, earlier reported from a few Dionysia spp.
(Harborne, 1968, 1971), and some of its derivatives are accumulated in trace amounts. While 5-OH-flavone is found in most of
the species, 5,8-diOH-flavone is of more restricted distribution,
and corresponding 6-O-substituted derivatives have not been detected so far. Treatment of these flavones as a separate group (Table 1) is based upon their irregular substitution pattern suggesting
a different biosynthetic origin (Valant-Vetschera et al., 2009; for
structures see Fig. 1). The formation of flavonoid types with ‘‘regular” substitution pattern (i.e., 5,7-OH-substitution in Ring A; 3’ and/
or 40 -substitution in Ring B) in exudates of Dionysia therefore is
remarkable. These substitution patterns characterise the occurring
flavanones, flavones and flavonols, indicated separately in Table 1.
It must be mentioned that both biosynthetic routes are expressed
in single species (e.g., Dionysia diapensifolia Boiss.), thus being not
mutually exclusive. Structural variation within flavanones and flavone derivatives of apigenin and luteolin is limited. Flavonols
based upon quercetin and kaempferol occur rather infrequent,
but their structural diversity is greater than that of corresponding
flavones. A series of unidentified flavonoids (u1–u6 in Table 1)
have been found in addition, but their structures could not be fully
determined yet due to lack of material. Nevertheless, they have
been included because of their informative value.
Distribution of single flavonoids in exudates of Dionysia was
quantified to be able to visualize the Dionysia flavonoid profile
composition at large. Detected compounds were coded either as
‘‘1” (minor amounts) or ‘‘2” (large amounts). Occurrences were
summarized and divided by the number of studied taxa, and calculated values were subsequently transformed to a bar chart as
shown in Fig. 2. Unsubstituted flavone represents the main accumulation tendency in Dionysia exudates, followed by a series of
the typical Primula-flavonoids. Regular substituted flavanones,
flavones and flavonols are minor accumulation tendencies as are
the yet unidentified flavonoids (u1–u6).
2.2. Infraspecific variation
The stability of accumulation tendencies at the infraspecific level was studied in different taxonomic groups across the genus
(Table 1). Plant source numbers appear in parentheses in the following text; for sources see Table 3. Only two accessions were
available for comparison of exudate profiles of Dionysia esfandiarii
Wendelbo (20, 21) and Dionysia mozaffarrianii Lidén (46, 47),
which did not exhibit any differences. By contrast, those of Dionysia hausknechtii Bornm. et Strauss (51, 52), of Dionysia freitagii
Wendelbo (7, 8), of Dionysia lamingtonii Stapf (55, 56) and of Dionysia paradoxa Wendelbo (38, 39) differed with respect to minor
compounds. With exception of D. lamingtonii, all material originated from cultivation in Gothenburg Botanic Garden. Accession
numbers 30–35 of Dionysia aretioides (Lehm.) Boiss. (cultivated
material), proved to be quite stable, with only one of the six samples deviating lightly. Some variation was noted among six samples of Dionysia tapetodes Bunge (9–14), but also at a rather low
level, and two samples coming from one clone being cultivated
in different institutions, showed identical profiles (10, 11). Four
out of five accessions of Dionysia archibaldii Wendelbo (15–18;
all cultivated material) proved to be quite uniform, while one collection, which was received as Dionysia bazoftica (19; natural habitat), differed by production of 5,8,20 -substituted flavones and of 3
of the unknown compounds. D. bazoftica, earlier described by Jamzad (1996) was later considered to be synonym to D. archibaldii.
Maybe the separation as a distinct taxonomic entity would be rectified, but certainly more material needs to be studied chemically
and morphologically.
Larger discrepancies were noted between the two collections of
Dionysia microphylla Wendelbo (4, 5), concerning the formation of
5-OH-flavones and 3 of the unidentified compounds in one collection. This is the more remarkable since both collections are of the
same clone (GWH-1302; Grey-Wilson, 1989), albeit cultivated in
different institutions. Similarly, one accession of Dionysia michauxii
(Duby) Boiss. (62) was found to accumulate the flavanone naringenin-7-Me in addition, while the other accession (63) yielded quercetin-7,30 ,40 -triMe together with u3, u5, and u6. Both accessions
came from cultivation in the Gothenburg Botanic Garden, but
material originated from different collections in the wild. The
two collections of Dionysia teucrioides Davis et Wendelbo (27, 28)
also yielded quite diversified profiles. They are supposed to be
clones, being under cultivation in different institutions.
The reasons for the observed variation even among clones are
still obscure. It may only be speculated that culture conditions
could play a role. It is known that Dionysia species are quite susceptible to fungal infections and also to insecticidal attack (GreyWilson, 1989). Thus, it is conceivable that some material was treated with chemicals which could affect the composition of exudates.
It would be interesting to test material from the wild and their offsprings under controlled cultivation conditions, for better assessment of variation. On the other hand, Primula species exhibited
stable exudate flavonoid profiles even when old herbarium material was analysed (Bhutia, pers. comm.).
2.3. Hybrid taxa
Hybrid taxa are rarely observed to occur in nature due to geographic isolation (Grey-Wilson, 1989). Hybrids produced under
cultivation frequently have D. aretioides as one parent species.
The samples analysed now (Table 2) represent artificial crossings
between species of sect. Dionysiopsis with those of sect. Dionysia-
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K.M. Valant-Vetschera et al. / Phytochemistry 71 (2010) 937–947
Table 1
Distribution of exudate flavonoids in sections and clades of Dionysia.
Source Clade Dionysia
number
Sectional
alignment
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
A
A
BC
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BF
BF
BF
BF
BF
Non-aligned (NA)
Non-aligned (NA)
Dionysiopsis (B-3)
Dionysiastrum (A2-b)
Dionysiastrum (A2-b)
Dionysiastrum (A2-b)
Dionysiastrum (A2-b)
Dionysiastrum (A2-b)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
20
21
22
23
BF
BF
BF
BF
24
BF
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
BF
BF
BF
BF
BF
BF
BF
BF
BF
BF
BF
BF
BF
BE
BE
BEG
BEG
42
43
44
BEG
BEG
BEG
45
46
47
48
49
50
BEG
BEG
BEG
BEG
BEG
BEG
51
52
53
54
55
56
57
58
59
60
61
BEG
BEG
BEG
BEG
BEG
BEG
BEG
BEG
BEG
BEG
BEG
62
63
BEG
BEG
balsamea
hissarica
mira
microphylla
microphylla
involucrata
freitagii
freitagii
tapetodes
tapetodes
tapetodes
tapetodes
tapetodes
tapetodes
archibaldii
archibaldii
archibaldii
archibaldii
archibaldii
(sub bazoftica)
esfandiarii
esfandiarii
viva
revoluta
subsp.
revoluta
revoluta
subsp.
canescens
oreodoxa
rhaptodes
teucrioides
teucrioides
bornmuelleri
aretioides
aretioides
aretioides
aretioides
aretioides
aretioides
khatamii
janthina
paradoxa
paradoxa
lurorum
lurorum (sub
aubrietioides)
iranica
caespitosa
caespitosa
subsp.bolivarii
zagrica
mozaffarrianii
mozaffarrianii
odora
gaubae
gaubae var.
megalantha
hausknechtii
hausknechtii
cristagalli
zetterlundii
lamingtonii
lamingtonii
termeana
bryoides
diapensifolia
sarvestanica
sarvestanica
subsp.
spathulata
michauxii
michauxii
Primula-type flavones
20 ,bFlavone 20 -OH- 20 20 ,50 diOHflavone OMe- diOHchalcone
flavone flavone
t
O
t
O
O
O
O
t
O
O
O
O
O
t
O
O
O
O
O
O
t
t
t
t
O
O
O
t
O
O
t
t
Dionysiopsis (B-3)
t
t
t
O
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Zoroasteranthos (B-4)
Zoroasteranthos (B-4)
Non-aligned (NA)
Non-aligned (NA)
Mucida (B-5)
Mucida (B-5)
O
O
O
t
t
O
O
Dionysiopsis
Dionysiopsis
Dionysiopsis
Dionysiopsis
(B-3)
(B-3)
(B-3)
(B-3)
Dionysia (B-6)
Dionysia (B-6)
Dionysia (B-6)
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
Dionysia (B-6)
Dionysia (B-6)
O
t
t
t
t
t
O
O
O
O
O
O
O
O
O
20 50 diOMeflavone
O
O
t
t
t
t
t
t
O
O
O
O
O
O
O
O
20 -OH-50 OAcflavone
30 ,40 diOHflavone
t
O
O
O
t
t
t
t
O
t
5-OH-20 OMeflavone
5,20 ,50 triOHflavone
5,8diOHflavone
O
O
t
O
t
t
O
O
O
O
O
t
t
t
t
t
t
t
t
t
O
O
t
O
O
t
O
t
t
t
t
t
t
t
t
t
t
O
O
O
O
O
O
O
O
O
O
O
O
O
O
t
t
t
t
t
O
O
O
O
O
t
t
O
O
O
O
O
O
O
O
O
t
t
t
t
t
t
t
O
O
O
t
t
t
O
t
t
O
O
t
t
t
t
t
t
t
t
t
t
t
t
t
t
O
t
t
t
t
t
t
O
O
t
t
O
O
O
O
O
O
O
t
t
O
O
t
O
O
t
t
O
t
t
t
t
t
t
t
O
O
t
O
t
O
O
O
t
O
t
O
O
O
t
O
O
O
O
O
O
t
t
t
t
t
t
t
t
t
O
t
t
O
O
O
O
t
t
t
O
t
O
O
t
t
t
t
t
O
t
t
t
t
O
O
t
t
t
30 -OMe-40 ,50 - 5-OH- 5,20 O2CH2flavone diOHflavone*
flavone
t
O
O
O
O
t
O
t
O
t
O
O
t
t
O
t
t
t
t
t
(continued on next page)
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K.M. Valant-Vetschera et al. / Phytochemistry 71 (2010) 937–947
Table 1 (continued)
Source
number
Clade
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
A
A
BC
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BF
BF
BF
BF
BF
20
21
22
23
BF
BF
BF
BF
24
BF
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
BF
BF
BF
BF
BF
BF
BF
BF
BF
BF
BF
BF
BF
BE
BE
BEG
BEG
42
43
44
BEG
BEG
BEG
45
46
47
48
49
50
BEG
BEG
BEG
BEG
BEG
BEG
51
52
53
54
55
56
57
58
59
60
61
BEG
BEG
BEG
BEG
BEG
BEG
BEG
BEG
BEG
BEG
BEG
62
63
BEG
BEG
Dionysia
balsamea
hissarica
mira
microphylla
microphylla
involucrata
freitagii
freitagii
tapetodes
tapetodes
tapetodes
tapetodes
tapetodes
tapetodes
archibaldii
archibaldii
archibaldii
archibaldii
archibaldii (sub
bazoftica)
esfandiarii
esfandiarii
viva
revoluta subsp.
revoluta
revoluta subsp.
canescens
oreodoxa
rhaptodes
teucrioides
teucrioides
bornmuelleri
aretioides
aretioides
aretioides
aretioides
aretioides
aretioides
khatamii
janthina
paradoxa
paradoxa
lurorum
lurorum (sub
aubrietioides)
iranica
caespitosa
caespitosa
subsp.bolivarii
zagrica
mozaffarrianii
mozaffarrianii
odora
gaubae
gaubae var.
megalantha
hausknechtii
hausknechtii
cristagalli
zetterlundii
lamingtonii
lamingtonii
termeana
bryoides
diapensifolia
sarvestanica
sarvestanica
subsp.
spathulata
michauxii
michauxii
Sectional
alignment
Non-aligned (NA)
Non-aligned (NA)
Dionysiopsis (B-3)
Dionysiastrum (A2-b)
Dionysiastrum (A2-b)
Dionysiastrum (A2-b)
Dionysiastrum (A2-b)
Dionysiastrum (A2-b)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis
Dionysiopsis
Dionysiopsis
Dionysiopsis
Primula-type flavones
Flavanones
5-OH8OMeflavone
Pinocembrin
5,8,20 triOH
flavone
5-OH8,20 diOMeflavone
Naringenin
t
t
t
O
t
t
O
O
t
O
t
t
t
Ap40 Me
Luteolin7,30 diMe
Kaempferol
Kae3Me
Kae7Me
O
t
t
t
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Zoroasteranthos (B-4)
Zoroasteranthos (B-4)
Non-aligned (NA)
Non-aligned (NA)
Mucida (B-5)
Mucida (B-5)
t
t
t
t
O
O
O
O
O
O
O
O
O
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
O
Dionysia (B-6)
Dionysia (B-6)
Dionysia (B-6)
Dionysia (B-6)
Dionysia (B-6)
Ap7Me
t
t
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
Apigenin
t
t
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Eriod7,30 diMe
O
Dionysiopsis (B-3)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
Eriodictyol7-Me
t
(B-3)
(B-3)
(B-3)
(B-3)
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Flavones/Flavonols
Nar7Me
t
O
O
O
t
O
t
t
t
t
t
t
t
t
t
t
t
t
t
O
O
O
t
t
t
t
t
t
t
t
t
t
t
t
t
t
O
t
t
O
t
t
t
t
t
t
O
t
t
t
t
O
t
t
t
t
O
t
t
t
t
941
K.M. Valant-Vetschera et al. / Phytochemistry 71 (2010) 937–947
Table 1 (continued)
Source
number
Clade
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
A
A
BC
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BCD
BF
BF
BF
BF
BF
20
21
22
23
BF
BF
BF
BF
24
BF
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
BF
BF
BF
BF
BF
BF
BF
BF
BF
BF
BF
BF
BF
BE
BE
BEG
BEG
42
43
44
BEG
BEG
BEG
45
46
47
48
49
50
BEG
BEG
BEG
BEG
BEG
BEG
51
52
53
54
55
56
57
58
59
60
61
BEG
BEG
BEG
BEG
BEG
BEG
BEG
BEG
BEG
BEG
BEG
62
63
BEG
BEG
Dionysia
balsamea
hissarica
mira
microphylla
microphylla
involucrata
freitagii
freitagii
tapetodes
tapetodes
tapetodes
tapetodes
tapetodes
tapetodes
archibaldii
archibaldii
archibaldii
archibaldii
archibaldii
(sub bazoftica)
esfandiarii
esfandiarii
viva
revoluta subsp.
revoluta
revoluta subsp.
canescens
oreodoxa
rhaptodes
teucrioides
teucrioides
bornmuelleri
aretioides
aretioides
aretioides
aretioides
aretioides
aretioides
khatamii
janthina
paradoxa
paradoxa
lurorum
lurorum
(sub aubrietioides)
iranica
caespitosa
caespitosa subsp.
bolivarii
zagrica
mozaffarrianii
mozaffarrianii
odora
gaubae
gaubae var.
megalantha
hausknechtii
hausknechtii
cristagalli
zetterlundii
lamingtonii
lamingtonii
termeana
bryoides
diapensifolia
sarvestanica
sarvestanica
subsp. spathulata
michauxii
michauxii
Sectional
alignment
Flavones/Flavonols
Kae-3,7diMe
Kae-3,40 diMe
Unidentified
Kae-7,40 diMe
Kae3,7,40 triMe
Non-aligned (NA)
Non-aligned (NA)
Dionysiopsis (B-3)
Dionysiastrum (A2-b)
Dionysiastrum (A2-b)
Dionysiastrum (A2-b)
Dionysiastrum (A2-b)
Dionysiastrum (A2-b)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiastrum (A2-a)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis
Dionysiopsis
Dionysiopsis
Dionysiopsis
Quercetin
Qu7Me
Qu7,30 diMe
Qu7,30 ,40 triMe
t
u1
u2
t
O
t
Dionysia (B-6)
Dionysia (B-6)
Dionysia (B-6)
t
Dionysia (B-6)
Dionysia (B-6)
O
O
O
O
t
t
t
O
t
O
O
O
O
O
O
O
t
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
O
u6
t
t
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Dionysiopsis (B-3)
Zoroasteranthos (B-4)
Zoroasteranthos (B-4)
Non-aligned (NA)
Non-aligned (NA)
Mucida (B-5)
Mucida (B-5)
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
u5
O
Dionysiopsis (B-3)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
(B-6)
u4
t
(B-3)
(B-3)
(B-3)
(B-3)
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
Dionysia
u3
t
t
t
O
O
O
t
t
O
u7
O
t
t
O
t
O
O
O
O
O
O
O
O
O
O
O
t
O
O
O
t
O
O
O
O
t
t
O
O
O
O
O
O
O
O
O
O
O
t
t
t
O, major compounds.
t, minor compounds.
Sources: see Table 3.
Clades according to Trift and Anderberg (2006).
Sectional alignment following Lidén (2007): NA, non-aligned; Eastern species: A2-a, sect. Dionysiastrum subsect. Tapetodes; A2-b, sect. Dionysiastrum subsect. Involucratae;
Western species: B-3, sect. Dionysiopsis; B-4, sect. Zoroasteranthos; B-5, sect. Mucida; B-6, sect. Dionysia.
*
In earlier papers (Budzianowski and Wollenweber, 2007; Valant-Vetschera et al., 2009) 30 -hydroxy-40 ,50 -methylendioxyflavone was erroneously reported instead of 30 methoxy-40 ,50 methylendioxyflavone from Primula elatior.
942
K.M. Valant-Vetschera et al. / Phytochemistry 71 (2010) 937–947
It appears that additive flavonoid patterns are not the rule within
Dionysia, which correlates also to earlier observations in the genus
Achillea (Valant-Vetschera and Wollenweber, 1988).
2.4. Flavonoid diversification in taxonomic groups of Dionysia
Fig. 1. Main structural types of Dionysia flavonoids.
strum (Lidén, 2007). Interpretation of exudate flavonoid profiles is
limited since the actual parent species were not available for analysis. Different flavonoid profiles were observed for two accessions
of D. aretioides Dionysia bornmuelleri (Pax) Clay, but both shared
the formation of flavanones with the parent species D. bornmuelleri. Also, the hybrid D. aretioides D. microphylla exhibited the flavanone naringenin-7-Me found in all accessions of D. aretioides, but
exhibited a reduced profile when compared to the parent species.
No fundamental differences were obvious between D. archibaldii D. microphylla and D. microphylla, and some of the typical
compounds from D. archibaldii were not detected in this hybrid.
Principal component analysis (PCo-analysis) was applied in
search of distribution patterns and variation of exudate flavonoid
distribution within Dionysia. Artificial hybrid taxa were excluded,
and so was Dionysia khatamii Mozaffarian because of coding ‘‘zero”
in all data. Quantities of accumulation were not taken into account
in the binary presence/absence data matrix, as was performed recently (Wollenweber et al., 2008). Variation was sufficiently indicated by values of the first 5 components extracted. Fig. 3 results
from plotting component 1 against component 2. Cases are represented by species, which were coded with respect to their sectional
alignment (Lidén, 2007), corresponding to Table 1. Three main
groups are defined: group 1 consists of some species from sect.
Dionysia (B-6, part 1), with Dionysia janthina Bornm. et Wink from
sect. Zoroasteranthos (B-4) falling into this group. The second group
comprises the remaining species from sect. Dionysia (B-6, part 2),
mixed with several species from sect. Dionysiopsis (B-3) and from
sect. Dionysiastrum subsect. Involucratae (A2-b), respectively.
Non-aligned taxa (NA) such as Dionysia hissarica fall within taxa
of sect. Dionysiopsis (B-3), while D. paradoxa bridges between species of sect. Dionysiopsis (B-3) and sect. Dionysiastrum subsect.
Involucratae (A2-b). While group 1 consists of Western species
only, group 2 is a geographic mix of Eastern and Western taxa. This
is also true for group 3, in which species of sect. Dionysiopsis subsect. Tapetodes (A2-a) and the remaining species of sect. Dionysiastrum are grouped. The other non-aligned taxa are now associated
with other groups: both accessions of D. paradoxa are positioned
between sect. Dionysiopsis and sect. Dionysiastrum subsect. Involucratae (A2-b) in group 2, while Dionysia balsamea shows up within
Dionysiopsis (B-3) in group 3. Species of sect. Dionysia, which are all
efarinose, group together best, which is not true for species from
Fig. 2. Frequences of single flavonoids in exudates of Dionysia spp. x-axis: all detected compounds (abbreviations and sequence as in Table 1). y-axis: absolute calculated
value (sum of occurrences/number of taxa analysed). Occurrences of single compounds were coded as (2) for large amounts, and as (1) for small amounts. Undetectable
amounts were coded as (0).
O
O
O
t
t
t
t
O
O
O
O
O
O
O
O
O
O
t
t
t
t
O
t
O
O
O
O
O
t
t
t
t
t
t
t
t
t
O
O
O
O
O, major compounds.
t, minor compounds.
Sources: see Table 3.
Grey, hybrid taxa.
O
O
O
O
O
O
O
O
O
O
29
66
65
30
31
32
33
34
35
64
4
5
67
15
16
17
18
19
microphylla
t
microphylla
archibaldii microphylla
archibaldii
archibaldii
archibaldii
archibaldii
archibaldii (sub
t
bazoftica)
bornmuelleri
bornmuelleri aretioides
aretioides bornmuelleri
aretioides
aretioides
aretioides
aretioides
aretioides
aretioides
aretioides microphylla
O
O
O
O
O
O
O
O
t
t
O
O
O
O
O
O
O
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
t
t
t
O
O
O
O
O
O
t
O
O
O
O
5,8,20 triOH
flavone
5,85-OH-8diOHOMeflavone flavone
5-OH-20 OMeflavone
5-OH- 5,20 flavone diOHflavone
30 -OMe40 ,50 O2CH2flavone
20 -OH50 -OAcflavone
Flavone 20 -OH- 20 20 ,50 20 ,bdiOHflavone OMediOHchalcone
flavone flavone
Source Dionysia
number
Table 2
Exudate flavonoids of artificial hybrids and parent species.
O
t
5-OH-8,20 diOMeflavone
Naringenin- Eriodictyol- u1 u2 u4 u5 u6
7-Me
7-Me
K.M. Valant-Vetschera et al. / Phytochemistry 71 (2010) 937–947
943
other taxonomic units exhibiting varying types of farina and/or
glandular hairs. Flavonoid diversification hardly shows correlation
with biogeographic diversification which is basal to the current
concept of Lidén (2007). However, the species groups are well supported by their flavonoid accumulation tendencies. Thus, species of
group 1 accumulate more of the regular flavones and flavonols and
hardly any of the typical Primula flavones. Group 2 accumulates
both types on a more or less equal level, while species of group 3
tend to accumulate mainly the typical Primula flavonoids. Interestingly, the flavanones occur scattered throughout all three groups.
The present data suggest that unsubstituted flavone and specific
groups of biogenetically defined derivatives represent accumulation tendencies which could be used for interpretation.
The observed diversifications of accumulation tendencies were
expected to be possibly associated with characters related to flavonoid production, i.e., production of woolly versus powdery farina,
or lack of farina (but presence of glandular hairs). Therefore, farina
characters were mapped on the scatterplot (Fig. 3). Whereas species of group 1 (Fig. 3) are all efarinose (sect. Dionysia, B-6), combined with D. janthina of sect. Zoroasteranthos (B-4; reduced
presence of glandular hairs; Lidén, 2007), the other two groups
are mixed with respect to farina production. Group 2 consists of
the remaining species of sect. Dionysia (B-6; similarly efarinose),
but they are mixed with taxa of woolly and, in one case, of mealy
(powdery) farina from sect. Dionysiopsis, as well as with two nonaligned taxa D. hissarica Lipsky, D. paradoxa (NA) both showing
woolly farina. Group 3 contains species which are described as
having both states (woolly farina or efarinose), along with species
of the mealy farina type. According to Trift et al. (2004), woolly farina represents an ancestral state, and it is as yet unclear whether
‘‘efarinose” represents an advanced character state or not. It appears that the degree of correlation between glandular morphology, production of farina, and flavonoid diversification is equally
obscure as was recently found for Primula (Valant-Vetschera
et al., 2009).
In the following, flavonoid diversification tendencies (Table 1)
are discussed against infrageneric (Lidén, 2007) and cladistic concepts (Trift et al., 2004; Trift and Anderberg, 2006) more in detail
(Table 1). If not cited otherwise, botanical information mentioned
here originates from these publications. Clade A comprises D.
balsamea Wendelbo & Rech. f. and D. hissarica, with strong support
as a sister clade to the rest of the genus and considered subsequently as non-aligned. D. hissarica (group 2, Fig. 3) differs from
D. balsamea (group 3, Fig. 3) by the formation of flavonols and of
some of the unidentified flavonoids. Thus they are chemically well
differentiated and the same applies in parts to their morphological
characters (Trift et al., 2004). Dionysia mira, another basal taxon
being considered to be the closest relative to the genus Primula
(Wendelbo, 1961), is found as a sister taxon to subclade BCD on
the molecular tree (low support). Flavonoid chemistry places this
taxon together with D. paradoxa and D. hissarica in group 2
(Fig. 3). The earlier assumed affinities between D. balsamea and
D. paradoxa (Grey-Wilson, 1989) are neither obvious from chemical nor from molecular analysis. Generally, flavanones and flavonols are quite rare in subclade BCD, and only some of the
unknown compounds were found to be occasionally accumulated.
The two chemically different accessions of D. microphylla are found
in group 2 (No. 5, Table 1) and group 3 (No. 4, Table 1), respectively, because of their different flavonoid complement.
In clade BF, Primula-type flavones dominate in all of the species,
but flavanones and some flavonols were found in a few species, together with some of the unidentified flavonoids. In this clade, Dionysia oreodoxa Bornm. and Dionysia rhaptodes Bunge form one pair
of species, and another group consists of D. khatamii, D. janthina,
and Dionysia curviflora of a separate section named Zoroasteranthos
(B-4). This section is morphologically well defined, and as far as
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K.M. Valant-Vetschera et al. / Phytochemistry 71 (2010) 937–947
Fig. 3. Principal component analysis of Dionysia flavonoids and comparison with farina characters. Sectional alignment following Lidén (2007): NA, non-aligned; Eastern
species: A2-a, sect. Dionysiastrum subsect. Tapetodes; A2-b, sect. Dionysiastrum subsect. Involucratae; Western species: B-3, sect. Dionysiopsis; B-4, sect. Zoroasteranthos; B-5,
sect. Mucida; B-6, sect. Dionysia. Farina characters: e: efarinose, lack of farina, but presence of glandular hairs; w, woolly farina; m, mealy (powdery) farina; we, either woolly
farina or efarinose; rgh, glandular hairs reduced to specific organ parts in low number. Species of group 1: zetterlundii, sarvestanica, gaubae var. megalantha, termeana,
lamingtonii (all accessions), odora, gaubae, iranica, cristagalli, zagrica, janthina. Species of group 2: bryoides, diapensifolia, sarvestanica subsp. spathulata, michauxii (all
accessions), caespitosa and subsp. bolivarii, mozaffarrianii (all accessions), haussknechtii (all accessions), revoluta subsp. revoluta, hissarica, oreodoxa, rhaptodes, esfandiarii (all
accessions), mira, paradoxa (all accessions), microphylla (No. 5 in Table 1), involucrata, freitagii (all accessions). Species of group 3: viva, revoluta subsp. canescens, lurorum (incl.
sub aubrietioides), teucrioides (all accessions), microphylla (No. 4 in Table 1), tapetodes (all accessions), bornmuelleri, archibaldii (all accessions and ‘‘bazoftica”), balsamea,
aretioides (all accessions).
exudates are concerned, also exceptional. Only one compound of
unknown structure (u7 in Table 1), not detected in any of the other
Dionysia species studied, was found in the exudate of D. khatamii,
while D. curviflora did not yield any exudate at all. Flavonoid profiles had a low degree of similarity in all species from this clade,
but infraspecific variation is not known. The closely related D.
aretioides, the Turkish D. teucrioides, and D. bornmuelleri exhibit
similar flavonoid profiles, and are found in group 3 (Fig. 3).
The following part of Clade B splits up into Dionysia lurorum
Wendelbo (BE), the sister taxon to the larger group of species of
subclade BEG. This clade conforms as a whole to sect. Dionysia
(B-6 in Fig. 3). D. lurorum, being now placed in the new section
Mucida (B-5; Lidén, 2007), is found in group 3 (Fig. 3) according
to its flavonoid accumulation tendencies. Within subclade BEG, a
tendency to accumulate flavanones, flavones and flavonols in addition or instead of the Primula flavonoids is noted (Table 1). Also,
unidentified flavonoids are encountered in some of the taxa of this
group. These diversifications lead to a split into two distinct groups
in PCo-analysis as already discussed (see also Fig. 3). Exudates of
Dionysia odora Fenzl, Dionysia gaubae Bornm., Dionysia zetterlundii
Lidén, D. lamingtonii, and Dionysia sarvestanica Jamzad et Grey-Wilson, were found to be devoid of the typical Primula flavones. However, D. odora occupies quite a large distribution area in Iran, and
infraspecific variation may occur. The rare flavanone pinocembrin
was detected in exudates of D. odora, D. gaubae var. megalantha,
and D. zetterlundii, respectively. Particularly D. odora is seen as
close relative to D. gaubae. The existing chemical differences between D. gaubae and its var. megalantha contrast to their small
morphological differences which resulted in taxonomic recognition on the level of variety only. Unfortunately, this collection
has not yet been included in molecular phylogenetic analysis.
The phytochemical differences between D. sarvestanica and its
subsp. spathulata are even greater: D. sarvestanica differs by the
lack of Primula flavones in its exudate and also shows a different
composition of flavones and flavonols. While samples of D. gaubae
originated from the wild, both D. sarvestanica collections were
grown under identical conditions in Gothenburg, thus excluding
different growth conditions as cause for variation. D. termeana is
the only species accumulating kaempferol derivatives (3,7-diMe;
7,40 -diMe; 3,7,4’-triMe) in its exudate. In this clade, D. termeana
is quite close to D. michauxii, which is not reflected by the exudate
flavonoid composition.
2.5. Exudate flavonoid diversification between Primula and Dionysia
Accumulation tendencies of Dionysia are further compared with
those of the designated Primula outgroup (species of subgen.
Sphondylia; Trift et al., 2004; see Fig. 4). 5-Deoxyflavones (DF in
Fig. 4) are the major accumulation tendency both in Dionysia and
in the Primula outgroup, followed by the corresponding 5-hydroxyflavones (5F in Fig. 4). Dionysia differs by the lack of 5,6-disubstituted 5-hydroxyflavones, and by the accumulation of 20 ,b-diOHchalcone, flavanones, regular substituted flavones and flavonols,
and the unknown flavonoids from the Primula outgroup. Also,
quantitative expression of single groups of typical Primula flavonoids is different. The degree of exudate diversification in biogenetic terms is larger within Dionysia as in the Primula outgroup
and in other Primula spp. studied so far (Valant-Vetschera et al.,
2009). However, Primula spp. exhibit more complex derivatives
of only one biogenetic group of flavones (Primula-type flavonoids),
and some of these complex derivatives have so far not been detected in the studied species of Dionysia.
Based upon current phylogenetic data, D. tapetodes, D. microphylla, Dionysia involucrata Zapr. and D. janthina (Trift et al.,
2002), or D. tapetodes and D. aretioides (Mast et al., 2001) are seen
as closely allied to Primula. Within Primula, species of subgen.
Sphondylia were earlier considered to be the most primitive
(Wendelbo, 1961). Their flavonoid profile appears less complex
K.M. Valant-Vetschera et al. / Phytochemistry 71 (2010) 937–947
945
Fig. 4. Comparison of accumulation tendencies between Primula (outgroup) and Dionysia. x-axis, compounds and derivatives; y-axis, absolute value (calculated: sum of
occurrences/number of taxa analysed). Black bars, Dionysia; hatched bars, Primula outgroup; for calculated values see supplementary data.
when compared to other members of the genus (Valant-Vetschera
et al., 2009). The most ‘‘Primula-like” profiles were now found in
exudates of D. mira, D. microphylla, D. freitagii (p.p.), D. tapetodes,
D. archibaldii, D. esfandiarii, and D. paradoxa, of different taxonomic
groups within Dionysia. If chemical diversity is applied as a character, these species should be regarded as the most primitive ones of
the genus. Earlier, Grey-Wilson (1989) had suggested that species
of Primula sect. Bullatae (subgen. Auganthus) should be the closest
relatives to Dionysia. So far, only Primula forrestii from this group is
known for its exudate flavonoid profile (Valant-Vetschera et al.,
2009), which would not contradict this assumption.
Correlating exudate flavonoid chemistry with phylogenetic
positions derived from DNA-data does not yield satisfactory results for all groups of Dionysia. The significance of this observation
is not yet clear. It is hard to believe that distinct biosynthetic
routes are expressed at random. This assumption is backed by
the fact that Cortusa matthioli, which is deeply nested within
Primula, corresponds chemically quite well. Thus, this taxon is
more Primula-like (Valant-Vetschera et al., 2009) than many of
the Dionysia species studied. We expect to better understand
the evolution of exudate flavonoid pathways once the biosynthesis of the Primula-type flavonoids is fully known. In our view, the
parallel expression of various accumulation tendencies in Dionysia
indicates a larger degree of diversification (complexity) and probably a derived state. This is paralleled by the current view of the
phylogenetic position of Dionysia in the Primulaceae (Trift et al.,
2004), and suggests that the genus Dionysia represents an evolutionary newer lineage. It certainly will be necessary to study also
the genus Primula more in detail for a better understanding of
relationships in this assemblage. Whether the extreme habitat
occupied by Dionysia species is causal to the biosynthetic complexity of exudate flavonoid profiles is currently open to speculation. However, anti-oxidative and radical-scavenging activities of
these flavonoids may explain their accumulation in plants of arid
and/or alpine regions, where increased UV-radiation prevails (Valant-Vetschera and Brem, 2006; Valant-Vetschera et al., 2009).
This certainly applies also to our Dionysia species, which are so
attractive to rock gardeners because of their morphological
beauty, and equally to phytochemists because of their interesting
biosynthetic capacity.
3. Experimental
3.1. Plant material
Plant material was obtained largely from collections cultivated
in green houses at the Gothenburg Botanic Garden, the collection
of M. Kammerlander (Würzburg, Germany), Botanischer Garten
München, Botanischer Garten der TU Darmstadt, and from collections in the wild from various expeditions carried out by Swedish
botanists (see Table 3, material sent by I. Trift (SE) is marked with
an asterisk). Respective herbarium specimens are kept in the institutions from which the material was received (M, GB).
3.2. Extraction and identification
Aerial parts of the collected plant species were briefly rinsed
with acetone in order to dissolve the lipophilic exudate material.
The mixture obtained after evaporation was then analysed, as described previously (Budzianowski and Wollenweber, 2007). Comparative TLC with markers was carried out on polyamide with
the solvents: (A) petrol100–140/toluene/MeCOEt/MeOH 12:6:1:1;
(B) toluene/petrol100–140/MeCOEt/MeOH 12:6:2:1; (C) toluene/petrol100–140/MeCOEt/MeOH 10:25:1:1; (D) toluene/MeCOEt/MeOH
12:5:3; (E) toluene/dioxane/MeOH 8:1:1; and on silica with solvents (F) toluene/MeCOEt 9:1; and (G) toluene/dioxane/HOAc
18:5:1. Chromatograms were viewed under UV (366 nm) before
and after spraying with ‘‘Naturstoffreagenz A” (0.2% of diphenylboric acid 2-aminoethyl ester in MeOH). Authentic markers for
the identification of the flavonoids were available in E.W.’s laboratory. Their structures have been elucidated previously. Literature
references on UV- and MS-data along with retention times from
our study are given as supplementary material. Similarly, Rf-values
(TLC polyamide DC-11; solvent system C) and colour reactions of
compounds u1–u6 may be found in supplementary material.
3.3. Principal component analysis
Principal components were extracted from a correlation matrix
of a binary original data matrix as described in section 2.4, with
946
K.M. Valant-Vetschera et al. / Phytochemistry 71 (2010) 937–947
Table 3
List of analysed material.
*
Number in Tables 1 and 2
Dionysia
Source
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
Hybrid taxa
64
65
66
67
balsamea Wendelbo & Rech. f.
hissarica Lipsky*
mira Wendelbo
microphylla Wendelbo*
microphylla Wendelbo
involucrata Zapr.*
freitagii Wendelbo*
freitagii Wendelbo*
tapetodes Bunge*
tapetodes Bunge*
tapetodes
tapetodes
tapetodes
tapetodes
archibaldii Wendelbo*
archibaldii
archibaldii
archibaldii
archibaldii (sub bazoftica)*
esfandiarii Wendelbo*
esfandiarii Wendelbo*
viva Lidén & Zetterlund*
revoluta subsp. revoluta Boiss.*
revoluta subsp. canescens (Boiss.) Wendelbo*
oreodoxa Bornm.*
rhaptodes Bunge*
teucrioides Davis et Wendelbo*
teucrioides Davis et Wendelbo
bornmuelleri (Pax) Clay
aretioides (Lehm.) Boiss.*
aretioides
aretioides
aretioides
aretioides
aretioides
khatamii Mozaffarian*
janthina Bornm. et Wink*
paradoxa Wendelbo*
paradoxa Wendelbo*
lurorum Wendelbo*
lurorum (sub aubrietioides) Jamzad et Mozaffarrian*
iranica Jamzad*
caespitosa (Duby) Boiss.*
caespitosa subsp.? bolivarii Pau*
zagrica Grey-Wilson*
mozaffarrianii Lidén*
mozaffarrianii Lidén*
odora Fenzl*
gaubae Bornm.*
gaubae var. megalantha Bornm.*
hausknechtii Bornm. et Strauss*
hausknechtii Bornm. et Strauss*
cristagalli Lidén*
zetterlundii Lidén*
lamingtonii Stapf*
lamingtonii Stapf*
termeana Wendelbo*
bryoides Boiss*
diapensifolia Boiss.*
sarvestanica Jamzad et Grey-Wilson*
sarvestanica subsp. spathulata*
michauxii (Duby) Boiss.*
michauxii (Duby) Boiss.*
GWH 580: cultivated; Kammerlander (Würzburg, Germany)
JJH 918037 Halda 92-0057, Bot. Garden Gothenburg
Cultivated; Bot. Garden Munich (Germany)
Grey-Wilson & Hewer 1302, Bot. Garden Gothenburg
GWH 1302: cultivated; Kammerlander (Würzburg, Germany)
1992: Bot. Garden Gothenburg
Watson P. 84 Grey-Wilson & Hewer 8497: Bot. Garden Gothenburg
Watson P. 93 EGW 92/21: Bot. Garden Gothenburg
T 4Z 1086-1 Iran: Khorasan, Kuh-e-Binalud NE Darrud [220-1910]
Arch. P. 81 Hewer 1164, Bot. Garden Gothenburg
H 1164: cultivated; Kammerlander (Würzburg, Germany)
Cultivated; Bot. Garden Munich (Germany)
GWH 780: cultivated; Kammerlander (Würzburg, Germany)
MK 9630, seeds derived from cultures Kammerlander
PZ Beuken [2003-865]: Bot. Garden Gothenburg
JCA3010: cultivated; Kammerlander (Würzburg, Germany)
MK 97-310/2, seeds derived from cultures Kammerlander
MK 94-02/1, seeds derived from cultures Kammerlander
87-1 JLMS 02-087 Iran: Bazoft Valley SW-Mayr [2003-193]
SLIZE 259-3
SLIZE 259-4 [98-1939]
T 4Z 035 Iran: Fars, Vallyabad between Marwdasht and Amsanjan [2002-1949]
T 4Z 030 Iran: Yazd [2002-1943]
SLIZE 192-3
D. Zschummel 101-06 Iran: Mt Takh Ali [2001-21179
T 4Z 1060 Iran: Kerman, Kuh-e-Jupar [2003-1877]
2000 0841 from Bot. Garden Munich (Germany); cultivated Bot. Garden Gothenburg
Cultivated; Bot. Garden Munich (Germany)
Rechninger 11456: Bot. Garden Gothenburg
SLIZE 035 1998-1857 Lidén [29-9-04]
Cultivated Bot. Garden, TU Darmstadt
Population a: cultivated; Kammerlander (Würzburg, Germany)
Population b: cultivated; Kammerlander (Würzburg, Germany)
Population c: cultivated; Kammerlander (Würzburg, Germany)
Population d: cultivated; Kammerlander (Würzburg, Germany)
T 4Z 13-2 [2002–2200]
T 4Z 007 Iran: Yazd, S of Mehriz, valley to Tang-e-Chenar [2002-1919]
PW 7414: Bot. Garden Gothenburg
PW 744: Bot. Garden Gothenburg
T 4Z 136-4
T 4Z 136-18 [2002-2059]
T 4Z 118 [2002-2036]
T 4Z 155 Iran: Esfahan, above Analajeh, Kuh-e-Dalan [2002-2235]
T 4Z 120 Iran: Esfahan, Karobas Pass, N of tunnel, type loc. [2002-2040]
SLIZE 176-8
T 4Z 100-8 Iran
SLIZE 232-9
99-0713 Kammerlander 0713-B, Bot. Garden Gothenburg
DZ 0038/2
T 4Z 190 Iran: Lorestan [2002-2115]
T 4Z 175 [2002-2101]
SLIZE 322-3 [1998-1973]
DZ 01-? [2003-859]
T 4Z 125 Iran: Chaharmahal Va-Bakhtiyari, Karun Valley, Shari Pass [2002-2046]
T 4Z 138 Iran: Chaharmahal Va-Bakhtiyari, Bazoft Valley [2002-2061]
SLIZE 161-2
D. Zschummel 00-46-2 [2000-1825]
T 4Z 092 Iran: Fraz [2002-2006]
SLIZE 253-2
T 4Z 040 Iran: between Kherameh and Sarvestan
T 4Z 1044 Iran: Farz, Kuh-e-Sefidar [2003-1861]
SLIZE 254-11
SLIZE 254-8 Iran
aretioides microphylla
aretioides bormuelleri
bornmuelleri aretioides*
archibaldii microphylla
MK-91-25/3, seeds derived from cultures Kammerlander
Cultivated; Kammerlander (Würzburg, Germany)
1974: Bot. Garden Gothenburg
MK92-25/1, seeds derived from cultures Kammerlander
Material received from Ida Trift.
artificial hybrids and D. khatamii (yielding zero values) being excluded. The option ‘‘Principal components of the factor analysis”,
implemented in SPSS (version 10), was applied. Fourteen components were obtained, with the first 5 explaining 60.2% of total var-
iance. Component 1 explained 30.8% of variance, followed by
component 2 (10.8%), component 3 (6.9%), component 4 (6.5%)
and component 5 (5.2%). Components 1 and 2 were used for scatterplot illustration (see Fig. 3).
K.M. Valant-Vetschera et al. / Phytochemistry 71 (2010) 937–947
Acknowledgements
Thanks are due to all those providing plant material for analysis,
particularly to Mr. Kammerlander (Würzburg, Germany), the Botanic Garden Munich (Germany), and to Ida Trift (Stockholm, Sweden), in collaboration with Gothenburg Botanic Garden. Dr. J.
Greimler (Dept. Syst. Evol. Botany, WU) kindly helped with statistical analysis and interpretation. The skilful technical assistance of
M. Dörr (Darmstadt, Germany) is greatly acknowledged.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at doi:10.1016/j.phytochem.2010.03.004.
References
Budzianowski, J., Wollenweber, E., 2007. Rare flavones from the glandular leaf
exudate of the oxlip, Primula elatior L. Nat. Prod. Commun. 2, 267–270.
Grey-Wilson, C., 1989. Dionysia – the genus in cultivation and in the wild. Alpine
Garden Society, Woking.
Harborne, J.B., 1968. Comparative biochemistry of the flavonoids-VII. Correlations
between flavonoid pigmentation and systematics in the family Primulaceae.
Phytochemistry 7, 1215–1230.
Harborne, J.B., 1971. Primulaceae. Hirsutin and gossypetin in Dionysia.
Phytochemistry 10, 472–475.
Jamzad, Z., 1996. The genus Dionysia (Primulaceae) in Iran. Iran. J. Bot. 7, 15–30.
947
Lidén, M., 2007. The genus Dionysia (Primulaceae), a synopsis and five new species.
Willdenowia 37, 37–61.
Mast, A.R., Kelso, S., Richards, A.J., Lang, D.J., Feller, D.M.S., Conti, E., 2001.
Phylogenetic relationships in Primula L. and related genera (Primulaceae)
based on noncoding chloroplast DNA. Int. J. Plant Sci. 162, 1381–1400.
Melchior, H., 1943. Entwicklungsgeschichte der Primulaceen-Gattung Dionysia.
Mitteilungen des Thüringischen Botanischen Vereins, N.F. 50, 156–174.
Trift, I., Anderberg, A., 2006. Foliar sclereids in Dionysia (Primulaceae) from a
phylogenetic perspective. Edinburgh J. Bot. 63, 21–48.
Trift, I., Lidén, M., Anderberg, A., 2004. Phylogeny and biogeography of Dionysia
(Primulaceae). Int. J. Plant Sci. 165, 845–860.
Trift, I., Källersljö, M., Anderberg, A., 2002. The monophyly of Primula (Primulaceae)
evaluated by analysis of sequences from the chloroplast gene rbcL. Syst. Bot. 27,
396–407.
Valant-Vetschera, K.M., Brem, B., 2006. Chemodiversity of exudate flavonoids as
highlighted by publications of Eckhard Wollenweber. Nat. Prod. Commun. 1,
921–926.
Valant-Vetschera, K.M., Wollenweber, E., 1988. Leaf flavonoids of the Achillea
millefolium group part II: distribution patterns of free aglycones in leaf
exudates. Biochem. Syst. Ecol. 16, 605–614.
Valant-Vetschera, K.M., Bhutia, T.D., Wollenweber, E., 2009. Exudate flavonoids of
Primula spp.: structural and biogenetic chemodiversity. Nat. Prod. Commun. 4,
365–370.
Wendelbo, P., 1961. Studies in Primulaceae I. A monograph of the genus Dionysia.
Aarbok Univ. Bergen, Mat.-Naturvitensk. Ser. (3).
Wollenweber, E., Fischer, R., Dörr, M., Irvine, K., Pereira, C., Stevens, J.F., 2008.
Chemodiversity of exudate flavonoids in Cassinia and Ozothamnus (Asteraceae,
Gnaphalieae). Z. Naturforsch. 63c, 731–739.
Zahra, A., Masoud, B., Abbas, A., Elham, A., Katayoun, J., 2007. Antitumour activity
and apoptosis induction in human cancer cell lines by Dionysia termeana.
Cancer Invest. 25, 550–554.
Eckhard Wollenweber