Chromosoma (Berl.

) 49, 299--308 (1975)

9 by Springer-Verlag 1975

The Cytology of the Genus Briza L. (Gramineae)

I. Chromosome N u m b e r s , K a r y o t y p e s and Nuclear D N A V a r i a t i o n
B. G. Murray
Department of Botany and Biochemistry, Westfield College,
University of London
Abstract. Chromosome numbers have been determined and karyotypes studied
in several species of the genus Briza L. Nuclear DNA amounts have also been
determined for some of the species and show a considerable variation between the
species. The evolution of the karyotypes, particularly that of B. minor, is discussed and an attempt made to relate nuclear DNA content to karyotypes, breeding
systems and habit in the different species.
Introduction
The species of the genus Briza are disjunctly distributed between
Eurasia and South America. The large majority of the species are found
in the temperate and sub-tropical regions of South America but six
species are native to Eurasia.
Previous reports on the cytology of the genus have been confined
mainly to counts of chromosome number (see Table 2). However Tateoka
(1955b) has analysed the karyotype of B. minor, and in B. media
K a t t e r m a n n (1930, 1933, 1938a, b) proved the existence of interchange
heterozygosity and Cauderon and Saigne (1962) of polyploidy. No
estimates of nuclear DNA content have previously been made.
There is now evidence illustrating wide variation in muelear DNA
content between numerous groups of related species (for a review see
l~ees and Jones, 1972). Various workers have attempted to correlate
these changes in nuclear DNA with other features such as plant habit,
breeding system and evo]utionary status. I n Lolium, Rees and Jones
(1967) were able to show that the inbreeding species contained about
35% more nuclear DNA t h a n the outbreeding species; however Rees
and Hazarika (1969) showed that the reverse was the c~se in Lathyrus,
where the taxonomically more advanced inbreeding species have smaller
nuclear DNA amounts t h a n the more primitive outbreeding species.
Kadir (1974) on the other hand has found that there is a trend towards
an increase in nuclear DNA with evolutionary advancement in the genus
Phalaris. Within each of the groups of species with different basic
chromosome numbers the perennials have lower nuclear DNA values
t h a n the annuals. I n Allium the most advanced groups of species are
thought to be derived by both loss and gain of nuclear DNA (Jones
and l~ees, 1967).
20

C h r o m o s o m a (Bell.), Bd. 49

300

B.G. Murray

:Bennett (1972) has shown t h a t there is a clear correlation b e t w e e n

n u c l e a r D N A c o n t e n t a n d m i n i m u m g e n e r a t i o n time i n a large n u m b e r
of species of herbaceous plants. Species with a n n u a l or ephemeral life
cycles have considerably less nuclear D N A t h a n most p e r e n n i a l species.
The species of Briza show differences i n breeding system, h a b i t a n d
chromosome n u m b e r a n d the aim of this s t u d y was to investigate the
relationships b e t w e e n these features a n d n u c l e a r D N A content.
Materials and Methods

(i ) Materials
Seeds of the Briza species were either collected from plants growing in the
wild or obtained from :Botanic Gardens and Research Institutes and are listed
in Table 1.
( i i ) Methods
(a) For karyotype studies actively growing root tips were pretreated with a
saturated solution of para-dichloro-benzene for 18-20 hours at 4~ C. After fixation
in acetic-alcohol (1:3) they were treated with a 5% aqueous solution of pcctinase
(Koch-Light Laboratories Ltd.) for two hours at room temperature. Finally they
were hydrolysed in :N HC1 for 8 minutes at 60~C and stained in Feulgen for 2 hours.

Table 1
Species

Source

Origin

B. media 2x

Wild collection

Watlington Hill,
Oxfordshire, U.K.

B. media 4x

Wild collection

Zakopane, Krakow Prov.,

Poland

B. minor

Jardin Botanique,
Caen, France

--

B. maxima

Civico Orto Botanico,

Trieste, Italy

--

B. elatior

Station Nationale d'Essais

de Semances, Versailles,
France

U.S.S.R.

B. poaemorpha

Division o5 Plant Industry,

C.S.I.R.O., Australia

B. stricta

Division of Plant Industry,

C.S.I.R.O., Australia

B. triloba

Division of Plant Industry,

C.S.I.R.O., Australia

B. subaristata

Wild collection

Sierra Lihuel Calel,

La Pampa Prov., Argentina

Cytology of Briza. I

301

(h) Nuclear DNA estimation b y Feulgett photometry. The m e t h o d used in this

study was essentially t h a t of Paroda and Bees (1971). Because of the difficulty
in obtaining well stained squash preparations measurements were made on isolated
nuclei. Boot tips were placed in ice-cold M/15 phosphate buffer (ptI 7) containing
4% formaldehyde for 12 minutes. The terminal 1 m m of each root was then tapped
out on a glass slide in a small drop of the buffer. The nuclei were precipitated b y
adding one or two drops of absolute alcohol and the slides placed in ether vapour
for 30 minutes. Following hydration in distilled water the slides were hydrolysed
in N tIC1 for 12 minutes at 60 ~ C. After washing in distilled water the slides were
stained in Feulgen (pH 3.6) for two hours. All the slides in each replicate were
hydrolysed a n d stained in the same staining container to ensure t h a t hydrolysis
and staining time were the same for all species. After staining the slides were
washed in S02 water three times (10 minutes each time), dehydrated and mounted
in euparal.
Photometric measurements were t a k e n on a Barr and Stroud integrating
microdensitometer. All measurements were t a k e n on at least 25 small presumed
2C nuclei and 5 large presumed 4C nuclei. The lowest 10 values were t h e n used
to calculate the mean 2C value. A preparation of Allium cepa root tip nuclei was
included in each replicate as a standard.

Results

(i ) Chromosome Numbers and Karyotypes

All known chromosome counts for the genus Briza are given in
Table 2. The results of this investigation confirm m a n y earlier counts
and show that in the genus Briza, with the single exception of B. minor
in which x = 5, all the species have the basic number of x = 7, The
chromosome number of B. poaemorpha is recorded for the first time.
Small (2 ~m) metacentric B chromosomes have been found in B, elatior
and in both diploid and tetraploid races of B. media.
Fig. 1 and 2 show the karyotypes of some of the species. The karyotypes can be broadly arranged into three groups. The first group contains a single species, B. minor, which is unique in the genus having
only 10 chromosomes. The karyotype consists of metacentrie and submetacentric chromosomes. The second group consists of B. media,
B. elatior and B. maxima. Their karyotypes show m a n y similarities and
are made up of metacentric, sub-metacentrie and acrocentric chromosomes. Tatraploid races of B. media have karyotypes in which the
chromosomes can nsuMly be arranged into groups of four homologues.
The South American species make up the third group. With the sole
exception of Saura's report of 2 n = 14 in B. glomerata (Saura, 1947),
all these species appear to be tetraploid. The karyotypes of these species
show less symmetry than those of the other two groups and are composed mainly of sub-metaeentric and acrocentric chromosomes. I n contrast to tetraploid B. media, groups of four homologous chromosomes
are not easily distinguished in the karyotypes of these species. Only
20*

302

B. G. Murray
Table 2. Chromosome numbers in the genus Briza

Species

Chromosome
number

Author

B. elatior Sibth. and Sm.

14+1--3B
14, 14+1,
2 and 4B

Avdulov (1928, 1931) a

Murray

B. marcowiczii Woronow

14

Sokolovskaya and Strelkova

(1948, 1962) a

B. maxima L.

14

Avdulov (1928, 1931) a

Parodi (1946) a
Delay (1947) a
Saura (1947) a
Tateoka (1953) a
Hubbard (1954) a
Larsen (1954) a
Fernandes and Quieros (1969) b
Murray

B. media L.

14

Avdulov (1928) a
Kattermann (1930, 1933) a
Tisehler (1934) a
]~ohweder (1937) a
Hubbard (1954) a
Tateoka (1955 a) a
Lepper (1968, 1970) b
Gadella and Kliphuis (1968,
1971) b
Bosemark (1957) a
Delay and Petit (1972)
Murray

Eurasian

14, 1 4 + 1 - - 2 B
14+4B
14, 1 4 + 1 - - 3
and 6B
28
28, 2 8 + 2
and 7B
B. minor L.

10

14

Cauderon and Saigne (1962)

Skalinska et al. (1971) b
Murray
Avdulov (1928, 1931) a
Parodi (1946) a
Heiser and Whitaker (1948) a
Tateoka (1953a, 1955b)
Hubbard (1954) a
Larsen (1960) a
Fernandes and Quieros (1969) b
Pohl and Davidse (1971) b
Murray
Gould (1958) b

303

Cytology of Briza. I
Table 2 (Continued)
Species

Chromosome
number

Author

B. brizoides (Lain.) Kuntze

(syn. B- elegans)

28

Bowden and Senn (1962) ~

B, erecta Lain.

28

Bowden and Selm (1962) ~

B. ]useu (Parodi) Parodi

28

Bowden and Sum~ {i962) ~

B, glomerata Hack. and Arech.

14

Saura (1947) a

B. poaemorpha (Presl) Henrard

28

Murray

B. rotundata

28

Reeder (1971) b

B. stricta (Hook. and Arn.)

Steudel

28

Saura (1947) a

South American

Bowdeu and Senn (1962) a

Murray
B. subaristata Lain.

28

Saura (1947) a
Mun'ay

B. triloba Nees

28

Bowden and Senn (1962) a

Murray

B. uniolae Nees ex Steudel

28

Bowden and Senn (1962) a

Complete reference in: Z. Bolkhovskikh et al.: Chromosome Numbers of Flowering l)lants. Leningrad: Komarov Botanical Institute 1969
b Complete reference in: Index to Plant Chromosome Numbers (g. J. Moore, ed.).
Utrecht: Oosthock's Uitgeversmaatschappji 1973.

B. stricta of t h e four species s t u d i e d has four chromosomes w i t h satellites,

in t h e o t h e r species satellites are o n l y expressed in a single pair.
( i i ) Nuclear D N A
Table 3 shows t h e n u c l e a r D N A values for six species of Briza. T h e
results show t h a t t h e r e are large differences in nuclear D N A a m o u n t
b e t w e e n some of t h e species; for e x a m p l e B. m a x i m a has a p p r o x i m a t e l y
25% m o r e D N A t h a n diploid B. media. T h e n a t u r a l l y occurring t e t r a p l o i d B, media has, as m i g h t be expected, a p p r o x i m a t e l y twice as m u c h
D N A as t h e diploid plants. H o w e v e r , t h e t h r e e t e t r a p l o i d species B.
stricta, B. 8ubaristata, a n d B. poaemorpha, all of which occur in S o u t h
A m e r i c a , h a v e c o n s i d e r a b l y less nuclear D N A t h a n t e t r a p l o i d B, media,
B. minor with o n l y 10 chromosomes contains t h e l e a s t nuclear D N A of
all t h e speciesr

IlJlJts J

IlJtll|l x(z

IIfxs l sxxsx
tg|XKKlXNx a

Fig. I a---e. Karyotypes of (a) Briza minor," (b) B. ~axima," (c) B. elatlor;
(d) B. m e d i a - diploid; (e) B. m e d i a - tetraploid. Scale -~ 10]~m
Table 3. Nuclear DNA amounts in six species of Briza, 2C values in pg. DNA using
Allium cepa as standard
Species

~. minor
B. maxima
B. media
B. media
J~. vubaristata
B. ~trieta
B. poaemorpha

2n

10
14
14
28
28
28
28

Replicate

Mean DNA
in pg.

15.49
23.11
17.64
-25.09
----

14.96
21.67
15.09
-22.19
24.6i
21.01

13.38
20.00
18.03
31.96
21.38
22.97
20.65

14.61
21.59
16.92
31.96
22.89
23.34
20.83

varl OI = ~

"~)~.*-~ "~ (P) .'~9~/a~o~og "fir (~) .'~qol*,~ "g (q) /,)~).t~z~)qn~ ~.~,~fir (~) to sod~2~o2.~ N "p--~eg '~!~I

~uo~m ttc ~ s t ~ u t tt n J t fcl~ ~Y.t I~J II tl d

306

B.G. Murray

Discussion
Karyotypic changes are frequently associated with or involved in
the process of speciation, but the direction and mechanism of the change
is not always easily perceived. The problem of how the karyotype of
B. minor evolved remains to be confirmed and work is in progress to
produce hybrids of this species with the other European species of the
genus to study pairing behaviour at meiosis. One likely explanation is
that Robertsonian translocations have occurred between acrocentric
chromosomes bringing about a reduction in chromosome number as a
result of the loss of the small centric fragment. Examples of this sort
of fusion are not unknown in plants and have been reported to have
occurred in the genus Crepis (Stebbins, 1950), Haplopappus gracilis
(Jackson, 1965) and Gibasis scheideana (Jones, 1974). In B. minor, as
in Gibasis, a change in breeding system provides evidence to support
the derived nature of the reduced chromosome number. B. minor is
a self-compatible inbreeding species whereas B. media, the species to
which it shows the greatest morphological and chemical similarity
(Williams and Murray, 1972) is a self-incompatible outbreeder (Murray,
1974). There is also a complete absence of acrocentric chromosomes in
B. minor.
The nuclear DNA values for the different species may also shed
some light on the problem of karyotype evolution. A comparison of the
values for B. minor and B. media shows that the former has only 2.32 pg
of DNA less than the latter. This would support the idea that only a
small amount of nuclear DNA has been lost during the evolution of the
karyotype of B. minor. The nuclear DNA values together with the
karyotypic differences would also appear to show that there are two
different kinds of polyploids in the genus. Though there arc reports of
diminishing nuclear DNA content in a polyploid series of birches (Grant,
1969; Taper and Grant, 1973) in Briza the tetraploid race of B. media
has approximately twice the nuclear DNA content of the diploids. This,
coupled with the occurrence of many groups of four chromosomes in
the karyotype and the formation of numerous quadrivalents at meiosis
(Cauderon and Saigne, 1962; Murray, uupub.) confirms the autopolyploid
origin of the tetraploid races of B. media. Karyotype studies give little
indication of an autopolyploid origin of the South American species.
The variation in nuclear DNA content between the species is not
easily correlated with differences in habit or breeding system. Of the
two annual self-compatible species, B. minor has less and B. maxima
more nuclear DNA than the perennial self-incompatible species B. media.
The South American species B. poaemorpha, B. stricta and B. subaristata are all self-compatible perennials but have nuclear DNA values
similar to that of B. maxima. These species are, however, facultative

Cytology of Briza. I

307

p e r e n n i a l s able to complete t h e i r life cycle w i t h i n 6-8 m o n t h s . T h u s

these results are in a g r e e m e n t with those p u b l i s h e d b y B e n n e t t (1972)
as he could f i n d no significant difference in nuclear D N A c o n t e n t b e t w e e n
a s a m p l e of a n n u a l a n d f a c u l t a t i v e perenniM species.
I t is now a p p a r e n t t h a t changes in nuclear D N A c o n t e n t h a v e little
p r e d i c t i v e value in e v o l u t i o n a r y studies in higher plants. U n d o u b t e d l y
w i t h i n a genus correlations can be f o u n d b e t w e e n different levels of
nuclear D N A a n d e v o l u t i o n a r y a d v a n c e m e n t or differences in b r e e d i n g
system, b u t no such correlations exist when comparisons are m a d e
b e t w e e n different genera. I n one genus t h e o u t b r e e d i n g species m a y
have m o r e nuclear D N A t h a n t h e inbreeding species whereas in a n o t h e r
genus t h e reverse in t h e case (l~ees a n d Jones, 1972). I t would a p p e a r
therefore t h a t e v o l u t i o n a r y change can be i n d e p e n d e n t of changes in
nuclear D N A content. H o w e v e r studies ol n u c l e a r D N A c o n t e n t can be
of considerable use in helping to u n d e r s t a n d t h e processes i n v o l v e d in
chromosome evolution.

Acknowledgments. Part of this work was carried out while the author was the
holder of a University of Reading Postgraduate Studentship in the Department
of Botany. This studentship is gratefully acknowledged. Thanks are also due to
Dr. Leslie Crowe, University of Reading, for her help and supervision and to Dr.
Keith Jones and Mr. Simon Owens of the Jodrell Laboratory, Royal Botanic
Gardens, Kew, for the use of their mierodensitometer and many helpful discussions.

References
Bennett, M. D. : Nuclear DNA content and minimum generation time in herbaceous plants Proc. roy Soc. London, B 181, 109-135 (1972)
Caunderon, Y., Saigne, B.: Sur la presence et l'6tude cytog~ndtique de Brizes
diploid et tetraploid dans le Massif Central. Rev. Cytol. Biol. v6g. 25, 461-464
(1962)
Delay, J., Petit, D. : Chromosomes surnum4raires chez les Gramindes. Inf. Ann.
Cytosyst~matique Cytog~n6tique 6, 1 4 (1972)
Grant, W . F . : Decreased DNA content of birch (Betula) chromosomes at high
ploidy as determined by cytophotometry. Chromosoma (Berl.) 26, 326-336
(1969)
Jackson, R. C. : A cytogenetie study of a three-paired race of Haplopappus gracilis.
Amer. J. Bet. 52, 946-953 (1965)
Jones, K.: Chromosome evolution by Robertsonian transloeation in Gibasis
(Commelinaceae). Chromosoma (Berl.) 45,353-368 (1974)
Jones, 1%.N., Rees, H. : Nuclear DNA variation in Allium. Heredity 23, 591-605
(1968)
Kadir, Z. B.A.: DNA values in the genus Phalaris (Gramineae). Chromosoma
(Berl.) 45, 376-386 (1974)
Kattermann, G.: Chromosomenuntersuchungen bei Graminecn. Planta (Berl.) 12,
19-37 (1930)

308

B.G. Murray

Kattermann, G. : Weitere zytologischc Untersuchungen an Briza media mit besonderer Berficksichtigung der durch Verb/~nde aus vier Chromosomen ausgezeichneten Pflanzen. Jb. wiss. Bot. 78, 43-91 (1933)
Kattermann, G.: Zur Kenntnis der strukturellen 14ybriden yon Briza media.
PIanta (Berl.) 27, 669-673 (1938a)
Kabtermann, G. : Krenzungsversuche bei Briza media zur Kl~rung der strukturellen
14ybriditgt. III. Planta (Berl.) 27, 674-679 (1938b)
Murray, B. G. : Breeding system and floral biology in the genus Briza L. (Gramineae). Heredity 33, 285-292 (1974)
Paroda, R. S., Rees, H.: Nuclear DNA variation in eu-Sorghums. Chromosoma
(Berl.) 32, 353-363 (1971)
Rees, H., 14azarika, M.H.: Chromosome evolution in Lathyrus. Chromosomes
today 2, 158-165 (1969)
Rees, H., Jones, G. I4. : Chromosome evolution in Lolium. 14eredity 22, 1-18 (1967)
Rees, 14., Jones, R. N. : The origin of the wide species variation in nuclear DNA
content. Int. Rev. Cytol. 32, 53-92 (1972)
Saura, F. : Complemento diploide en algunas especes de Briza. Rev. Fac. Agron.
Vet. (Buenos Aires) l l , 330-333 (1947)
Stebbins, G. L. : Variation and evolution in plants. New York: Columbia University Press 1950
Taper, L. J., Grant, W. F. : The relationship between DNA content and chromosome size in birch (Betula) species. Caryologia (Firenze) 26, 263-273 (1973)
Tateoka, T. : Karyotype analysis of Briza minor L. La Kromosomo (Tokyo) 22-24,
786-787 (1955)
Williams, C.A., Murray, B. G. : Flavonoid variation in the genus Briza. Phytochemistry 11, 2507-2512 (1972)
Received November 15, 1974 / Accepted by H. Bauer
Ready for press November 17, 1974
Dr. B. G. Murray
Department of Botany & Biochemistry
Westfield College
(University of London)
London NW3 7St
England