T H E EFFECT OF COLCHICINE ON ROOT

MITOSES IN ALLIUM
BY

ALBERT LEVAN

H I L L E S H ~ G ,LANDSKRONA, SWEDEN

FTER BLAKESLEEand AVERY (1937) had made the sensational

discovery of the induction of chromosome doubling by colchicine
treatment it became a question of prime importance to investigate the
cytological mechanism causing this doubling. It has long been known
that colchicine acts disturbingly on the normal course of mitosis
(DIXON,1905). DUSTIN(1934) and LITS (1934) regarded colchicine as
a very active agent for increasing the numher of mitoses in a tissue
(a ,poison caryoclasique,). LUDFORD(1936), however, came to the
conclusion that the increase in the number of the mitoses after colchicine
treatment was due to an ,accumulation of arrested mitoses)) rather than
to a stimulation process. He attributed this effect to a ,failure of the
mitotic spindle to form and function in the normal manner)). NEBEL
and RUTTLE(1938), who first studied the effect of colchicine on plant
cells (stamen hairs of Tradescantia), arrived at a similar result.
The material of the experiments dealt with in the present paper
consists of root tips of Allium fistulosum and Cepa. Root tips of
A. fistulosum were secured from bulbils that had overwintered in the
field, while in the case of A. Cepa large bulbs were used, The fistulosum
form used had a hybrid chromosome garniture, consisting of somatically
15 medially attached chromosomes and 1 typical fistulosuni s1 (LEVAN,
1936, Fig. 1 b). The Cepa form had the chromosomes characteristic
of this species, viz. 14 medially and 2 subterminally attached chromosomes (LEVAN,1. c., Fig. 1 a ) .
The experiments were arranged in the following way. Bulbs with
rapidly growing root tips, O , . s l , o cm in length, were immersed for exposure into colchicine solutions during periods of different length. Root
tips were then fixed at certain intervals after the transfer of the bulbs
from the colchicine solutions into pure water. The fixations were made
in NAVASHINunder air evacuation. The material was embedded and
cut into transverse and longitudinal sections, the former being 20 ,u
in thickness, the latter 20-30 p. The slides were stained in gentian
violet, some also in ,lichtgriin,.

4i2

ALBERT LEVAN

I. DESCRIPTION OF THE TYPICAL COURSE OF THE

C-MITOSIS,
In the first series of experiments the conditions were the following:
Concentrations of colchicine: 0,125, O,%, 03, 1,0, 2,o %.
Exposure times: 7, 15, 30 min., 1, 2, 24, 72 hours.
Fixing took place: 0, 15, 30 min., 1, 3, 6, 12, 24, 48, 72 hours after
finishing the colchicine treatment.

16

)
I
1
P
i
I i
b

rn

Fig. 1. The first stages in the development of the c-pairs. - x 3900.

The effect of colchicine on the course of mitosis is entirely specific,

and the modification in mitotic behaviour will be abbreviated BCmitosis,. The c-mitosis can be referred to one single moment, uiz. an
inactivation of the spindle apparatus connected with a delay of the
division of the centromere. The effect thus produced may be expressed
as a completion of the chromosome mitosis without nuclear or cellular
mitosis.
The prophase stages take place normally: the chromosomes divide,
condense, and assume metaphase appearance. They are, however, not
arranged into an.equatoria1 plate. Instead they are all the time
scattered over the cell in a diakinesis-like manner. This condition lasts

473

THE EFFECT OF COLCHICINE

for a long time after-the disappearance of the nuclear membrane. The

halves of each chromosome are seen to be coiled around each other in
a relational spiral (Fig. 1 a-h). This spiral is then slowly uncoiled, and
during this process the chromosomes assume a whole series of shapes
exceedingly characteristic of the c-mitosis and never occurring norm.ally. At first loops are formed between the undivided centromeres and
the points where, on account of the spiralisation tension, the chromatids
touch each other. There usually occurs one such point on each of the
long chromosome arms (Fig. 1 i-p).
These points of contact move
slowly towards the ends, and at last the chromatids touch each other
only at the still undivided centromere and at one or both ends

6 '4 t i;
b

x
C

A'9
0

Fig. 2 . Allium fistulosum, the 16 c-pairs of one cell separately drawn.

- x 3900.

(Fig. 1n, 0 ; Fig. 2 a, 1 ) . This process strikingly resembles the terminalisation of chiasmata in diakinesis bivalents. At last the ends also
slip off, and the. half-chromosomes are now held together only at the
undivided centromeres. Instances of such typical cross-shaped pairs
(called below c-pairs) are reproduced in Figs. 2 d, n and 3 b-d.
As has already been mentioned, the formation of the c-pairs is
peculiar to material treated with colchicine. Their origin is evidently
due to the delay of the division of the centromere. In the course of
normal mitoses the centromere divides at about the same time as the
chromosomes are arranged into the equatorial plate, and the orientation
towards the poles is probably simply conditioned by the division of the
centromeres- ( ,auto-orientation DARLINGTON, 1937). Anyhow, the later
stages in the uncoiling of the relational spiral normally occur within the
)),

474

ALBERT LEVAN

equatorial plate. After that the centromeres separate and proceed

towards the poles, pulling the chromosome arms behind.
The c-pairs form the configuration most commonly found the first
few hours after the colchicine treatment. This indicates that the division
of the centromere is delayed for quite a considerable time. This is,
partly at least, the cause of the apparent impression of mitotic
stimulation, which is always found after c-treatment. The prophases
arrive at metaphase and are kept at that stage for a long period until+
the centromere finally divides.

\I

,6 I I\

Fig. 3. The divisioii of the centromere within the c-pairs. .4. Ccpa: a--b, k-1;
fistulosurn: c-j, m-p. - x 3500.

During this period no normal anaphases are found in the slides. If

certain cells are at anaphase at the onset of colchicine action, they may
iorm two telophase nuclei, if the anaphase is advanced far enough, and
in certain cases a cell wall has been seen formed between the nuclei,
but in most cases the anaphase chromosomes remain in 2 groups,
whieh will later be ineluded into one nucleus.
After a few hours the division of the centromeres finally takes
place (Fig. 3 e-Z), and the two daughter chromosomes a m straightened
out and locate themselves parallelly like )pairs of skis, (NEBEL amd
RUTTLE,1. c.). The centromeres are placed opposite one another in each
pair (Fig. 3 m-p). The arrangement in pairs is of ten maintained. also

475

THE EFFECT OF COLCHICINE

through the following mitoses and is especially easy to observe in the

case of s1 chromosomes (Fig. 3 9). Incidentally this relative stability
in the chromosome arrangement through several subsequent c-mitoses
is very conspicuous and results in an accumulation of the same chromosome in one section of the nucleus. An instance of this is shown in
Fig. 4 a, wh,ere at least 8 s1 chromosomes are gathered at the same place.
It is often noticed during the c-anaphase that the division of the
centromeres does not take place quite simultaneously within one cell.
In Fig. 3, for instance, the still undivided pairs c and d are drawn from
the same cell as the two divided ones, e and f . The division of the
centromere has evidently been desyncronised as well as delayed. Now

Exposure

7 min.
15 D
30 B
1 hour
2 hours
24
72

))
))

Lp.

3
n5

A
A
A
A
A
A
A
Diagram 1.

A = the c-pairs appear.

C = normal spindle.

C
C

B
B
B

(:

B
B
B

c:
C

The rhythm of the c-mitoses.

B = first indications of the spindle regeneration.

the chromosomes pass on into telophase and all of them are included
in one nucleus, which will consequently contain the double somatic
chromosome number.
After the short exposures, 7 min.-I hour, there usually occurs
only one c-mitosis, but after long exposures several c-mitoses may
follow each other in the same cell, and each of them doubles the chromosome number.
The inactivation of the spindle apparatus under the influence of
colchicine is reversible; after a period of 12-24 hours in pure water
the spindle begins to regenerate. The regeneration takes place very
characteristically and in the course of the transition to normal spindle
all kinds of abnormities are seen : multipolar spindles, asymmetrically

$76

ALBERT LEVAN

or not at all compact spindles, etc. After 36-48 hours the mitoses
again run their normal course.
In Diagr. 1 is given a survey of the rhythm of the c-mitoses,
calculated from the experiments now dealt with. From this diagram
is seen that after short exposures an interval of 30 minutes passes
before the typical c-mitoses appear. This interval is necessary for the
full development of the c-pairs, while the inactivation of the spindle
probably occurs much earlier. Diagr. 1 also indicates that long exposures require a somewhat longer time for recovery than short ones.
11. CYTOLOGICAL CONSEQUENCES

OF C-MITOSES,

About 40 hours after the c-exposure was finished, the mitoses have
reassumed their normal course and persistent changes in the root cells
produced by the colchicine treatment can be observed. After the short
exposures (7-30 min.) such changes are rarely met with, nevertheless
a few cells with 4x chromosomes may be found. The majority of cells
show the normal diploid number. After an exposure of 1-2 hours a
great percentage of 4x cells are found together with occasional 8x cells,
.and after the longest exposures (72 hours) cells with still greater chromosome numbers are seen, 32x being the upper limit in these series.
In order to get an idea of the distribution of different chromosome
numbers in the roots after different c-exposures, I examined the crosssections of the whole meristematic region of a few roots and plotted
.on diagrams all the chromosome numbers which could be determined.
Determination of greater chromosome numbers than 8x could not always be made exactly, yet there was no difficulty in deciding whether
a chromosome plate should be classified as 8x, 1Gx or 32x. The numbers were not strictly euploid, which is easy to understand in view of
the frequently occurring anaphase disturbances. The plate reproduced
in Fig. 4 d shows, for instance, 135 chromosomes, and though this
number may not be exact it is certainly greater than 128, which
corresponds to 1Gx.
The root diagrams were grouped into 5 or 6 regions from each root
and the chromosome numbers from these regions were tabulated in
'Table 1, the treatment of the roots being specified in Table 2. A
study of Table 1 will at once reveal one very important fact: there
is a decided correlation between the chromosome number of a cell
and the location of the cell in the root. A greater percentage of
changed cells occurs in older parts of the root, while close to the root

477

T H E EFFECT OF COLCHICINE

II

Slide 4693
Region

2x 3x 4x

1
2
3
4
5
6

21 - 2
232
32 5 9
16-11

Region

1
2
3
4
5
6

7 - 5
- - -

I
1 2x

I
1

Slide 4694
2x 3x

30 2
18 1
7 - 3
2 - 3
2 -

1 4
4 9
6 5
5

1
4x 8x 16x 32x I 2x
Slide 4739

I
1

Slide 4741

8
3
-

3x 4x 8x

_ _ - - - - - -

Slide 4696

2x 3x 4x

G - 1 4 3 3 - 7 - 2 5 472
5 - 3 8
2
406
3 3 1 9 1 3 2 - 1 3
- - 1 - 19 2 19
- - - - 12 - 22

- - 1 1
1 -

- - -

- 1
5 1 6 - 519 7 - 1 2 8 - - 212
3 - - 3 1 1

I 2x

4x 6x 8x

Slide 4695

4x

8x

16
39
27
11
2

5
19
15
2

16x
-

Slide 4912
2x

4x

8x

16x

1 7
12
13
10
3

- - - 9 - 15
1 18
1 7
3
1

_ - - -

TABLE 2. Particulars about the roots of Table 1

Number of
sections in

The treatment of the roots

% colcliicine

4693
4694
4695
4696
4739

10
10
10
10
5

exposure time

fixing time

72 hours

0,128

2 hours

0,250

))

))

O,5

))

))

'190
0,O

290
0,Ol

))

72 hours
))

144 hours

))

48 hours
))

72 hours

tip among the new meristematic cells normal diploid numbers prevail.
This situation is made clear in Diagr. 2, where the percentages of diploid
cells in different regions of 4 roots are graphically represented. In all
of the root tips this percentage increases towards the tip of the root.
Hereditax XXZV.

32

478

ALBERT LEVAN

The absolute values are, it is true, highly different in different roots,

and they are evidently due l o fairly arbitrary conditions. However,
they afford a measure of the susceptibility of the meristematic cells to
colchicine action in the course of the exposure.
The situation just described seems to be generally valid throughout
the colchicine experiments of the present study. It is of fundamental
significance. If for some reason there occurs a population of cells with
different chromosome numbers within an embryonic tissue, a selection

146

/
I

REGION

Diagram 2.

among the cells immediately sets in. Normal 2x cells are favoured at
the expense of cells changed by colchicine. The primary cause of this
is probably the division rate, which seems to be more rapid in diploid
than in polyploid cells. Moreover, normal unchanged cells start their
first division after the c-treatment more readily than do changed cells.
At a certain moment after the transfer from the colchicine solution,
frequent diploid mitoses are seen, while highly polyploid giant nuclei
still linger in prophase stages.
As regards the roots recorded in Table 1, the diploid cells have in

THE EFFECT OF COLCHICINE

479

no case totally disappeared. Even after an exposure of 72 hours there

occur numerous diploid cells in the sections close to the tip. When
colchicine is used for the production of polyploid forms, this fact must
be kept in mind. Throughout a considerably prolongated colchicine
treatment single cells of the meristeme can keep themselves in a
condition resistant to colchicine. After the conclusion of the treatment
these cells may constitute a very trifling minority, but thanks to their
favoured situation in the competition with polyploid cells they will predominate again after some time. The tissue thus shows a tendency to
return to its original cytological state.

111. THE REITERATION OF THE C-MITOSES,

The preceding chapter leads to the question: How far can the cells
be changed by colchicine, where is the limit of the capability of the cell
to respond to an extremely prolongated c-exposure with new c-mitoses?
Even the strongest dose of colchicine used in the above experiments
( 2 %' colchicine for 72 hours) did not entirely kill the cells, After their
transfer into water the cells might revert again to the normal mitosis.
Judging from the chromosome numbers then observed there had taken
place at least 4 generations of c-mitoses in the course of the exposure.
The number 128 frequently occurred in the cells and in one case* 256
chromosomes could be counted with certainty. It is clear, however,
that in this experiment the lethality limit cannot have been far off.
In order to investigate if a longer exposure in a weaker colchicine
solution might give a still more extreme final result, the following two
experimental series were arranged. 5 bulbs were placed in 0,111% colchicine and 5 in 0,i % . They were transferred daily into new-prepared
colchicine solutions in order to diminish the risk of putrefaction. 6, 8,
10, 12, and 14 days after the beginning of the exposure one bulb from
each series was transferred into pure water, and root tips were fixed
several times from each bulb.
A poisoning effect of colchicine could actually be observed in these
experiments. The bulbs immersed in 0,i % solution manifested a much
slower growth of the leaf shoots than the bulbs in 0,oi %. On the
seventh day the former showed leaf shoots about 8 cm in length, the
latter on an average 15 cm, while an untreated control bulb had shoots
34 cm in length.
In many of these bulbs the limit of cell lethality had been passed.
C-mitoses were, it is true, going on in most of them at the first fixation

480

ALBERT LEVAN

time, but as a rule they were incapable of reverting into normal mitoses.
In these roots enormously high chromosome numbers were found. In
some cases the numbers could be fairly correctly estimated, and numbers as high as 500 and 1000 c-pairs were not rare. In c-mitoses, how-

Fig. 4. Normal mitoses after the regeneration of the spindle. A . Cepa: a-c,
fistulosum: d ; a: f.500 chromosomes, b : 16 chromosomes, c: detail of a, d : f.135
chromosomes. - a, c X 1000, b, d X 2000.

ever, the chromosomes are equally scattered out spherically and so their
number is extremely difficult to determine. Judging from the size of
the cells and the chromosome clusters, there occurred still higher numbers than 1000 pairs. At least 6 c-mitoses, probably more, have taken

THE EFFECT OF COLCHICINE

481

place here. The fixations were excellent and the c-pairs could be closely
studied. They showed the same characteristic features as are described
in Chapter I.
In some slides fixed 2 4 4 8 hours after the exposure was finished,
normal divisions could be studied. In these slides the chromosomes
were arranged in equatorial plates and their numbers could be determined with a fairly high degree of certainty. About 500 chromosomes
constituted the upper limit so far met with in these slides; thus, greater
numbers seem to be unable to revert into normal mitosis.
The problems of cell mechanics which are elucidated in connection with the origin of these fantastically great chromosome numbers will only be touched upon on this occasion. The cells have considerably increased in volume. The macroscopically observable tumours
and AVERY, 1. c., Fig. 4 B1) are simply
(Figs. 6 and 7 ; BLAKESLEE
caused by this enormous increase in cell volume. The same cells which
at the beginning contained 16 chromosomes have been adapted to house
500 or 1000 chromosomes. The increase in volume is, however, not
sufficient to allow the mitoses with the new great chromosome numbers
to take place without being severely disturbed by crowding. Divisions
with 64 chromosomes still go on quite regularly, but plates with 128
chromosomes and more are often bent and twisted on account of lack
of space. Often the centre of a plate forms a cupola and is found in one
section while the ring-shaped periphery lies in the neighbouring section.
Part of one plate may bend into a level at right angles with the rest of
the plate or turn round 180", etc. The plate reproduced in Fig. S a
probably contains about 500 chromosomes. As is seen from the detail
drawing Fig. 4 c, the appearance of the chromosomes is quite normal,
although they are more crowded than normally. Fig. 4 a is drawn
under low magnification from 2 sections and is not intended to give
the exact number and shape of the chromosomes, but only a survey of
the whole cell.
The chromosomes are especially over-crowded in the embryonic
vascular tissue, the cells of which are extended in length. In such cells
it happens that after reiterated c-mitoses the nucleus completely fills
the cell. The nucleus thus gets lengthened and spool-shaped. Normal
mitosis can take place, however, also under such conditions. The
equatorial plate then resembles the corresponding stages in the pollen
tube mitosis.
The transition from c-mitosis to normal mitosis in these cells with
high chromosome numbers has a very characteristic course. The first

482

ALBERT LEVAN

indication of spindle regeneration is that the telophase chromosomes are

not included into a rounded nucleus. Instead they gather together into
an often large number of small groups, which are never completely
separated from each other. The nucleus thus formed gets a highly
irregular shape and is perforated by numerous holes and canals.
The next step in the spindle recovery is indicated by the plainly
multipolar spindle. This results in the gathering of the chromosomes
into some 20 clusters, which are wholly separated from each other
(Fig. 5 a), and at telophase they form as many small nuclei. Between
these nuclei there are often formed real cell walls, stained green by
Blichtgruns (Fig. 5 b). By this peculiar mechanism a giant cell is
divided into a large number of minor cells, which of course obtain
varying sets of chromosomes and will mostly be non-viable. A point

Fig. 5. The transition to nornial mitosis, a : multipolar telophase, b: a giant cell has
been divided into many minor cells. - X 750.

of considerable interest which may be noted is that as a consequence of

a spindle abnormality one way has been opened for the reduction of
the chromosome number in somatic cells. This mechanism, however,
acts very irregularly and is of quite a different nature from the case of
somatic reduction described by STEIN (1936).
When the multipolar divisions have been going on for some time
the number of poles is gradually reduced and at last all cells have
regained normal bipolar spindles.
The different stages in spindle recovery are continuously intermingled, but the different types described above are sufficiently clear to
be identified in longitudinal sections as 3 regions: in the upper region of
the root the lobed giant cells are found,then follows the region with giant
cells divided into small cells, and at the root tip normal bipolar mitoses
predominate.

THE EFFECT OF COLCHICINE

IV.

483

THE LOWER THRESHOLD VALUE OF COLCHICINE

ACTION.

In order to determine the lowest concentration necessary for the

induction of c-mitoses, two series of experiments were performed. The
irst of them included the following concentrations: 0,0001, 0,0005, 0,001,
0,005, O,oi, (405, 0,i %. The roots were exposed in these solutions for
4 hours, and were then fixed 0, 8, 24, and 48 hours after the transfer
to pure water.
% ) did not produce cThe lowest concentrations (0,0001-0,005
mitosis, while Prom 0,oi % and upwards the c-mitoses were conspicuous. Also on the second occasion of fixing (after 8 hours) the
c-mitoses were taking place in all the concentrations except 0,oi % , where
the new meristematic cells showed normal mitosis. Ih older parts of
the 0,oi % roots, however, the colchicine-induced disturbances earlier
observed could be traced. The recovery of the spindle is evidently more
rapid after exposures in low concentrations. At first, however, the
c-mitoses were as conspicuous in 0,oi % as in the stronger solutions.
The second experimental series was intended especially to elucidate
the threshold region limited by the former series to 0,005-0,oi %. It
contained the following 11 concentrations: 0,0050, O,oos, O,OOCO,0,0065,
0,0070, 0,0075, 0,u080, 0,0085, 0,0090, 0,0095, 0,oioo %.
Also this time the exposures were made for 4 hours and fixing was made after 0, 8 and
24 hours. The same variety of A. Cepa was used in both series.
Even the concentration 0,005s caused disturbances of the spindle
function. Of great interest is the observation of a fractionating of the
disturbances. In treatment with increasing concentrations the exterior
spindle is first damaged (in 0,0055%), while the centromeres are
normally divided still in a concentration of 0,0070 5%. Above this concentration the first traces of c-pair formation appear. The c-pairs are,
however, not of the characteristic appearance until a concentration close
to 0,oi % is reached. After 8 hours all new mitoses were normal throughout the whole series, and the spindle apparatus had completely recovered.
In this connection reference should be made to the observations of
DARLINGTON
and THOMAS(1937) as to the individuality of the two cooperating factors of normal spindle function, the exterior or centrosomic
factor and the interior or centromeric factor.
The threshold value of root mitoses of A. Cepa has thus been fixed
to an exposure of 4 hours in 0,005-0,oi % colchicine solution. As a comparison it may be mentioned that NEBEL and RUTTLE (1. c.) found the

484

ALBERT LEVAN

threshold value of Tradescantia to be just above 0,004 %. LUDFORD

(1. c.), working with animal material, found a very much higher susceptibility to colchicine: the action arresting mitosis could be observed as
far down as in a 0,ooooi % solution.

THE PRACTICAL SIGNIFICANCE OF COLCHICINE.

Although the studies on the action of colchicine dealt with on the
present occasion are of a preliminary nature, based as they are on only
1 month of experimenting, they nevertheless furnish, in the writers
a

Fig. 6.
The formation of colchicine tumours.
a : control bulb, b-e: c-treated bulbs.

opinion, many facts of interest. The expectations of BLAKESLEE

and
AVERY as to the value for practical breeding work of the colchicine
method have been fully confirmed in the present study. A great many
facts go to prove that colchicine will constitute the agent long sought
for, which, without detrimental secondary effects and with full certainty,
induces polyploidy. From a practical point of view colchicine has many
advantages over the methods hitherto prevalent for increasing the chromosome number. Of importance is, for instance, that the colchicine
action is specific and total. No other disturbances besides the inactivation of the spindle apparatus are observed, at least if the c-exposure is not prolonged too much and the concentrations employed are
not too strong. The completeness of the action of colchicine is remarkable. All the mitoses taking place within the exposed tissue turn into

THE EFFECT OF COLCHICINE

485

c-mitoses and bring about chromosome doubling. Of importance is

also the reversibility of the inactivation process of the spindle. After
the conclusion of the exposure the spindle apparatus recovers and is able
to function in the normal manner. Cells with increased chromosome
number are thus brought back to normal mitoses after their period
of rest.
The results of the present study refer exclusively to the root
meristeme, where the chromosome conditions are easy to investigate,
but probably there is no essential difference in the c-effect in other
a

Fig. 7. The formation of colchicine tumours. a : bulb treated with 0,i % colchicine
for 8 days; the calyptra is clearly visible; b: colchicine treatment alternating with
periods in pure water; the tumours take the shape of strings of pearls.

embryonic tissues. The root mosaic is characterised by the different

elements being mixed at random, and no tendency could be found to
the formation of whole sectors with changed chromosome number.
The possibility of. macroscopically diagnosing colchicine-induced
polyploidy may be of some practical use. As is seen from Figs. 6 and 7,
tumours are formed by the root meristeme,while the length growth ceases
altogether [compare control Fig. 6 a with c-treated bulbs (with 0,i %
colchicine for.5 days) Fig. G b-el.
As has already been emphasized,
this phenomenon is caused by the increase in volume of the meristematic
cells, while the formation of new cells is completely suppressed. An
idea of the sensitivity of this macroscopic reaction may be had from

406

ALBERT LEVAN

Fig. 7 b. This bulb has been treated with 0,5 % colchicine solution
alternating with periods in pure water. The colchicine tumour then
takes the shape of a string of pearls, each period in water being
distinguishable as a constriction of the tumour. In Fig. 7 a the calyptra
is very conspicuous; its meristeme contains fewer cells than the root
meristeme and consequently it cannot swell to the same degree as the
rest of the root tip.
Considering all the obvious advantages of colchicine for producing
varieties with increased chromosome number, it seems very likely that
colchicine, on account of its specific and infallible action, will be of
great practical use and possibly take the place of the more whimsically
acting agents for the production of polyploids hitherto in vogue.

LITERATURE CITED.
1. BLAKESLEE,
A. F. and AVERY,A. G.

2.
3.
4.
5.

6.

7.
8.

1937. Methods of inducing doubling of

a
chromosomes in plants. - Journ. of Hered. 28: 393-411.
DARLINGTON,
C. D. 1937. Recent advances in cytology. Second ed. - London.
C. D. and THOMAS,P. T. 1937. The breakdown of cell division in
DARLINGTON,
a Festucn-Lolium derivative. - Ann. of Bot., N. S. 1:717-761.
DIXON,W. E. 1905. A manual of pharmacology. - London.
DUSTIN,A. P. 1934. Action de la colchicine sup le sarcome greffb, type Crocker,
de la Souris. - Bull. Acad. R. Med. Belg. 14:487-?105.
LEVAN,A. 1936. Die Zytologie von Allium Cepa X fistulosum. - Hereditas
XXI : 195-214.
LITS, F. J. 1934. Contributions B l'8tude des reactions cellulaires provoquees
par la colchicine. - C. R. SOC.Biol. 115: 1421-1423.
LUDFORD,R. J. 1936. The action of toxic substances upon the division of
normal and malignant cells in vitro and in vivo. - Arch. f. exp. Zellf.
18: 411-441.

NEBEL,B. R. and RUTTLE,M. L. 1938. The cytological and genetical significance of colchicine. - Journ. of Hered. 29:s-9.
10. STEIN,EMMY. 1936. Die Doppelchromosomen im Bliitenbezirk der durch Kadiumbestrahlung erzeugten Mutanle wancroidea, von Antirrhinum majus.
(Somatische Chromosomen-Reduktion). - Zschr. f. ind. Abst.- u. Vererbungslehre 72: 267-266.
9.