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Ph Syst. Evol. 151,223-228 (1986)

9 by Springer-Verlag 1986

Systematics of Nama (Hydrophyllaceae): Flavonoids and

Phyletic Position of sect. Arachnoidea and sect. Cinerascentia
By

John D. Bacon, Nianbai Fang, and Tom J. Mabry

(Received July 10, 1985)
Key Words: Angiosperms, Hydrophyllaceae, Nama.-Chemotaxonomy,
flavones, 6-methoxy flavones, C-glycosyl flavones.
Abstract: Eight flavonoids, four 6-oxygenated flavones, two methyl ethers of
luteolin, apigenin 6,8-C-diglucoside and quercetin 3-O-glucoside, were isolated
from Nama lobbii and N. rothrockii, sole members of sects. Arachnoidea and
Cinerascentia, respectively. Both taxa diverge markedly from other namas in
morphology and chromosome number and their placement in Nama has been
questioned. The occurrence of 6-oxygenated flavones in these taxa adds to their
already distinctive nature. Flavonoid evidence argues that both are more closely
allied to Eriodictyon than either is to Nama.
As second largest genus in the Hydrophyllaceae, Nama comprises
about 50 species distributed primarily in the drier areas of the southwestern United States and northern Mexico. Some species, nevertheless,
occur in more mesic habitats in the sierras of the western United States and
the highlands of central and southern Mexico. Morphologically diverse,
species range in habit from small, short-lived annuals to robust, woody
subshrubs. Following the treatment of HITCHCOCK (1933), species are
structured into five sections: sects. Arachnoidea, Cinerascentia,
Conanthus, Zonolacus, and Nama. The latter section is much the largest
and houses the bulk of the species while the former four shelter only six
species, with three of those resident in sect. Conanthus. Although each of
the smaller sections is possessed of clearly unique features, the species of
sect. Arachnoidea and sect. Cinerascentia are so distinctive that their
placement within the boundaries of Nama has been questioned (BAcoN
1974, CHANCE& BACON 1984, RAVEN & AXZLROD 1978). Nama rothrockii
GRAy, the single species of sect. Chwrascentia, exhibits capitate inflorescences and crenate leaves, features found in no other namas: N.
lobbii GRAY, sole member of sect. Arachnoidea, exhibits loculicidally and
septicidally dehiscent capsules, while capsules of other namas are

224

J.D. BACON& al.:

loculicidally dehiscent only. Indeed, N. Iobbii was treated as an Eriodictyon by GREENE (1885: p. 202) and HALE (1906: p. 265), the two taxa
exhibit the same capsular dehiscence pattern, and it is through N. lobbii
that the nearest relative of Nama traditionally has been identified as
Eriodictyon (H~TcHCOCK 1933). Therefore, placement of N. lobbii, as well
as N. rothrockii, is critical for establishing not only generic limits but also
relationships of Nama. As part of an ongoing systematic revision of Nama,
the flavonoids of these two species have been isolated and identified. We
report herein the flavonoids accumulated by the two species and consider
their placement in light of these and other data.

Experimental
Air-dried leaf, stem and flower material (300g for N. Iobbii [BAcoN 1701,
U.S.A.: Nevada: Douglas Co.]; 390g for N. rothrockii [BAcoN 1812, U.S.A.:
California: lnyo Co.], vouchers to be deposited at TEX) was ground to a fine
powder and extracted 3
hours with dichloromethane: ethyl acetate,
70:30 v/v, air drying between extractions. Material was then extracted 2 24
hours with 85% aqueous methanol. Like extracts were combined, reduced to a
small volume under water vacuum and subjected to column chromatography over
Polyclar (GAF Corp.) using various mixtures of dichloromethane, ethyl acetate
and methanol as solvents. Flavonoids were monitored with UV light (366 nm) as
they were eluted from columns. Eluted fractions were purified by further column
chromatography or by paper chromatography on Whatman 3 MM paper using
15% or 40% acetic acid or tertiary butanol : glacial acetic acid : H20, 4 : 3 : 1, v/v,
as solvents. UV spectra were recorded following procedures as set forth in MABRY
& al. (1970). IH NMR spectra were recorded for trimethylsilyl ether derivatives of
all compounds on a Varian EM 390 spectrometer.

Results
A total of eight flavonoids, six aglycones and two glycosides, were
isolated From the two species o f Nama (Table 1), Included a m o n g the
isolated compounds were the aglycones 6-methoxyapigenin (I), 6-methoxyapigenin 7-methyl ether (II), 6-methoxyapigenin 4'-methyl ether (III),
luteolin Y-methyl ether (IV), luteolin 7,Y-dimethyl ether (V), 6-methoxyluteolin 4'-methyl ether (VI) and the glycosides quercetin 3-O-glucoside
(VII) and apigenin 6,8-C-diglucoside (VIII). In addition to isolated
flavonoids, crude chromatograms of each species exhibited trace amounts
of yet to be isolated glycosides; N. lobbii exhibited two faint spots with Rf
values in keeping with those of flavonol 3-O-monoglycosides. Nama
rothrockii exhibited a single faint spot with Rf values approximating those
of (VIII) and suggesting a C-glycosyl flavone or a 3,7-O-glycoside.
While the flavonoid profiles of each species are clearly different, the
aglycone component of both are similar and dominated by 6-oxygenated
flavones. It appears that N. rothrockii lacks the ability to methylate
position 7 of 6-methoxyapigenin while N. lobbii is incapable o f m e t h y l a t -

Flavonoids and Systematics of Nama (Hydrophyllaceae)

225

Table 1. Structure of elucidated flavonoids of Nama lobbii and N. rothrockii

R~

OH

Flavonoid

I
II
llI
IV
V
VI
VII
VIII

Substitution pattern +

6-Methoxyapigenin
6"Methoxyapigenin 7-methyl ether
6-Methoxyapigenin 4'-methyl ether
~uteolin 3'-methyl ether
Luteolin 7,3'-dimethyl ether
6-Methoxyluteolin 4'-methyl ether
Apigenin 6,8-C-diglucoside
Quercetin 3-O-glucoside

RI

R2

R3

R4

R~

R6

H
H
H
H
H
H
H
O-glc

OMe
OMe
OMe
H
H
OMe
C-glc
H

OH
OMe
OH
OH
OMe
OH
OH
OH

H
H
H
H
H
H
C-glc
H

H
H
H
OMe
OMe
OH
H
OH

OH
OH
OMe
OH
OH
OMc
OH
OH

+glc = glucose; OMe = OCH 3.

Table 2. Distribution of flavonoids in Nama lobbii and N. rothrockii
Taxon

N. lobbff
N. rothrockii

Flavonoid number
I

I1

III

IV

VI

VII

+
+

+
+

+
+

VIII
+

ing either position 3' or 7, or both, on the luteolin skeleton (Table 2). The
glycosidic components of each species, in contrast, are quite different.
Even assuming that the un-isolated glycosides of each are identical and the
same as those identified, there remains a remarkable quantitative
difference in the major glycoside accumulated; the C-glycosyl flavone of
N. lobbii is the major flavonoid, quantitatively, in the taxon, while
quercetin 3-O-glucoside is a minor component in N. rothrockii.
Discussion

In segregating N. lobbii and N. rothrockii into monotypic sections,

HITCHCOCK (1933) was giving due recognition to tile singularly notable
16

PI. Syst. Evol., Vol. 151, No. 3 ~ 4

226

J.D. BACON& al.:

characters which each displays. Indeed, GREENE (1885) and HALL (1906)
ascribed even more significance to the features exhibited by N. Iobbii and
treated that taxon as an Eriodictyon. HITCnCOCK'Sdisposition o f N . lobbii
was influenced, at least in part, by his disposition of another problematical
taxon, Turricula parryii (GRAY)MACBRIDE. This taxon had been shuffled
back and forth between Nama (GRAY 1876: 621, PETER 1897: 69, BRANr~
1913: 160) and Eriodictyon (GREENE 1889: 22, HALL 1902: 106) or
recognized as the monotypic Turricula (MACBR1DE 1917: 42). HITCHCOCK
(1933), noting similarities in seed features, corolla features, habit and
capsular dehiscence of the latter taxon and Eriodictyon species, chose to
recognize it as Eriodictyon parryii. Considering N. lobbii, he concluded
that, with the exception of capsular dehiscence, its characters were those of
Nama. although divergent enough to merit recognition as a monotypic
section. He then went on to point out that it was through E. parryii (= T.
parryii) and N. lobbii that the true relationships of Eriodictyon and Nama
were manifest. However, additional data forthcoming since 1933 portray
the relationship between Nama and Eriodictyon as less direct than
concluded by HJTCHCOCKand suggest that N. lobbii and N. rothrockii are
more distant from the bulk of Nama than he reckoned.
CAVE & CONSTANCE (1942, 1944, 1947, 1957, 1959) and CONSTANCE
(1963) have extensively surveyed the Hydrophyllaceae for chromosome
numbers and additional surveys have been conducted for Nama (BACON
1974, 1984). All species o f EriodicO,on have been counted and all possess
n = 14. In contrast, Turricula exhibits n = 13, supporting its treatment as
a monotypic genus; accepting this placement, any direct link between
Nama and Eriodictyon, as envisioned by HITCHCOCK, is severed. With
about 90% of the species of Nama counted, the emergent chromosomal
picture is that o f a strikingly diploid taxon with n = 7, with the exception
of sects. Arachnoidea and C#wrascentia; N. Iobbii exhibits n = 14, while N.
rothrockii has n = 17. Although three taxa of Nama are consistently
tetraploid, excluding the aforementioned sections, all have readily
identifiable relatives within Nama proper; neither N. rothrockii nor N.
Iobbii can be so positioned. Also, CHANCE & BACON (1984) have shown
that seed features of N. lobbii and N. rothrockii are distinctive within
Nama; both species produce much larger seeds with a thicker seed coat and
both exhibit distinctive sculpturing patterns when compared to other
namas. Moreover, the habit o f these two species is not that of Nama, for
both are rhizomatous (BacoN, pers. obs., JEPSON 1943, MuNz & KECK
1959); other namas are tap-rooted. It is significant in this respect that
Eriodictyon is a rhizomatous taxon (MuNz & KECK 1959).
The presence o f &oxygenated flavones in N. Iobbii and N. rothrockii
adds further to the unique nature of these taxa within Nama. Additional

Flavonoids and Systematics of Nama (Hydrophyllaceae)

227

species of Nama have been surveyed for flavonoids including representatives from all sections except Zonolacus (BACON & al., in prep.); while
producing a diverse array of compounds, none accumulates 6-oxygenated
flavonoids. However, we have recently surveyed three species of Eriodictyon and all accumulate 6-oxygenated flavones (BAcoN& al., in review). It
is apparent, then, that N. lobbii is morphologically, cytologically and
chemically more similar to Eriodictyon than it is to yet other namas.
However, Eriodico, on is a genus of shrubs with generally scorpioid
inflorescences. While N. lobbii is woody, it is not a shrub; additionally, it
exhibits axillary flowers and there are "technical" floral features which
readily separate it from Eriodico~on. Although N. lobbii might be "fitted"
within Eriodictyon as a section or subgenus, such a placement would
disrupt the morphological homogeneity of that genus. Nama rothrockii is
morphologically and cytologically anomalous in either Nama or Eriodictyon; nevertheless, flavonoid chemistry and its rhizomatous habit argue
that its affinities lie closer to N. lobbii a n d / o r Eriodictyon than to any
species of Nama.
Accumulated evidence suggests that both N. lobbii and N. rothrockii
should be viewed as distinct phyletic elements separate from Nama.
Indeed, we would argue that both should be recognized as distinct
elements within the Hydrophyllaceae and positioned nearer to Eriodictyon
than to Nama. In fact, we are uncertain as to how Nama relates to these
taxa or to Eriodictyon. The traditional close alliance of Nama with
Eriodictyon clearly is based on subjective interpretation of the significance
of morphological features of N. lobbii and subsequent retention of that
taxon within the bounds of Nama. More recent objective evicence fails to
COuntenance that view. Retention of N. rothrockii within Nama is, again,
subjectively based and not supported by recent evidence. Nevertheless,
one might argue that EriodicO,on and N. lobbii, at least, must be closely
allied with Nama, since Nama is one of only two genera in the family with
x ~ 7 (The other is Lemmonia, a monotypic, annual taxon, perhaps not
distinct from Nama; see JEPSON 1943:284 for a discussion of the status of
this taxon.) However, CARLQUlST& al. (1983) found no evidence supportive of a herbaceous ancestry in the wood anatomy of Eriodictyon.
Nama is a predominantly herbaceous genus.
Evidence tbr relationship among N. lobbii, N. rothrockii, and Eriodictyon is persuasive. While we do not deny that Nama proper is to be allied
with these taxa, present evidence argues that any relationship must be
more distant than previously held and must be basal rather than direct.
This work was supported at Arlington by NSF grant DEB-8108513 and at
Austin by The ROBERTA. WELCHFoundation (Grant F-130) and The National
Institutes of Health (Grant HD04488).
16"

228 J. D. Bacon & al.: Flavonoids and Systematics of N a m a (Hydrophyllaceae)

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Addresses of the authors: Jo~N D. BACON, Department of Biology, The
University of Texas at Arlington, Arlington, TX 76019, U . S . A . - N~^N~A~ FANC
and TOM J. MAI3RY,Department of Botany, The University of Texas at Austin,
Austin, TX 78712, U.S.A.