Only Full Cooling of Mother Tulip Bulbs Is Necessary for Daughter

Bulb Formation from Daughter Buds

Marian Saniewski Hiroshi Okubo
Research Institute of Pomology and Floriculture Laboratory of Horticultural Science
Pomologiczna 18 Faculty of Agriculture
96-100 Skierniewice Kyushu University
Poland Fukuoka 812-8581
Japan

Keywords: tulip, Tulipa gesneriana, cooling, mother bulbs, daughter bulb formation,
daughter buds

Abstract
Mother tulip bulbs planted in the autumn require a long cold period before
satisfactory stem extension with flower development and enlargement of daughter
buds to bulbs will occur in spring. Little is known of the underlying physiological
mechanism of daughter bulb formation from daughter buds. In our studies we used
tulip bulbs cv. Apeldoorn cooled at 5C. We found that for the growth of daughter
bulbs in tulips, only full cooling of mother bulbs was necessary, not the growth of
shoot and roots when the bulbs were stored at room temperature after cooling
(without planting of bulbs). It was interesting that daughter bulbs were formed
when mother bulbs are stored for extended periods (about seven months) at 5C. In
most of the daughter bulbs that formed in these conditions, flower bud
differentiation took place and flowering occurred during the next season after
sufficient cooling. The mechanisms of transporting food reserves from the mother
bulb directly to daughter buds to grow daughter bulbs, including the role of plant
hormones and flavonoids, is discussed.

INTRODUCTION
During the development of tulip flowers, three phases can be distinguished
(Boonekamp et al., 1990): (i) the initiation and formation of a new sprout with flower (at
high temperature); (ii) the internal preparation for stem elongation (at low temperature);
and (iii) the rapid elongation of the sprout (at high temperature). Tulip bulbs, with a
terminal bud containing a complete flower, require 12-16 weeks of low temperature
treatment for floral stalk elongation. This suggests a kind of dormancy that can be released
by exposure to low temperature (Kamerbeek et al., 1972). The duration of the cold
treatment is a major factor determining stalk growth and flowering. Increasing the
duration of low temperature treatment decreases the number of days from planting to
flowering.
A low temperature treatment of tulip bulbs simultaneously induces not only a
stimulation of shoot growth but also the bulbing of the axillary buds; a faster plant
elongation is also associated with a more rapid bulbing of the axillary buds (Le Nard and
De Hertogh, 1993). Thus, extending the duration of the cold treatment results in a very
rapid expression of bulbing (Aoba, 1976; Le Nard and De Hertogh, 1993).
Daughter buds are located at the inner bases of the scales, and generally there is
one bud per scale. The transformation of the daughter buds into daughter bulbs is
especially rapid after flowering and simultaneously, the mother-bulb scales shrivel and
progressively disappear (Le Nard and De Hertogh, 1993). The innermost bud produces the
largest bulb, while the other buds produce smaller bulbs with fewer scales. At the end of
spring, daughter-bulb enlargement ceases.
The aim of the present work was to study the formation of daughter bulbs in fully
cooled mother tulip bulbs stored at room temperature without planting, including excision
of sprouts and root primordia, under light and darkness conditions.

Proc. IXth Intl. Symp. on Flower Bulbs 501

Eds.: H. Okubo, W.B. Miller and G.A. Chastagner
Acta Hort. 673, ISHS 2005
MATERIAL AND METHODS
Tulip bulbs cv. Apeldoorn, with circumference of 10-11 cm, were stored at
17-20C after lifting until October 15 and then were dry-cooled at 5C until the end of July
or until used in February-March. Other tulip bulbs were stored at the same time at 17C as
uncooled bulbs. Fully cooled and uncooled bulbs were stored at room temperature
(20-23C) without rooting as intact bulbs after excision of sprouts and roots, under light
and darkness conditions. Morphological observation of daughter bulb formation and
measurement of their weight were made in cooled and uncooled mother bulbs. A total of
20 to 25 bulbs were used per treatment and experiments were repeated during two growing
seasons.

RESULTS AND DISCUSSION

At planting, dormant tulip bulbs are composed of fleshy scales attached to a basal
plate that produces roots at the basal surface, differentiated apical flower bud (stem, leaves
and all parts of flower), central daughter bud, other axillary buds an H bud, and tunic
(dry outer bulb scale).
The size of flower bud and of the particular organs (leaves, perianth, stem, and
anthers, and daughter buds) in fully cooled tulip bulbs (at 5C) was a little smaller than in
uncooled bulbs continuously at 17C (Fig. 1). The growth and development of tulips after
planting of uncooled and cooled bulbs is presented in Fig. 2. Natural shoot growth and
flowering only occurred from fully cooled bulbs.
In fully cooled tulip bulbs stored at room temperature under natural light,
enlargement of daughter bulbs took place, whereas no enlargement occurred in uncooled
mother bulbs stored under the same conditions (Fig. 3, Table 1). Excision of sprouts and/or
roots primordia from fully cooled tulip mother bulbs did not prevent daughter bulb growth
at room temperature (Fig. 4, Table 1). It was interesting that daughter bulbs were also
formed when mother bulbs were continuously stored for extended periods (about seven
months) at 5C (Fig. 5). In most of the larger size daughter bulbs that formed in cooled
mother bulbs stored at room temperature, flower buds developed and flowering took place
in the field during the following season (Fig. 6).
Translocation of both carbohydrates and inorganic nutrients from mother-bulbs to
the daughter bulblets is well documented (Schmalfeld and Carolus, 1965; Aung et al.,
1973). Aung et al. (1973) found that during the ontogeny of cv. Preludium bulblets in the
field, the decrease in soluble sugars and starch contents of the mother-bulbs scales
corresponded with a concomitant increase of these carbohydrates in the bulblets. They
noted also that while all the soluble sugars in the matured bulblets were accounted for by
the initial sugars of the mother bulbs scales, only 17% of the bulblet starch could be
accounted for by the initial starch content of the bulbs, with the remaining 83% of the
starch being derived from photosynthates.
Limited information is available regarding hormonal control of daughter bulb
development in field-grown tulips. Aung and Rees (1974) determined the changes in the
endogenous gibberellins content of the developing bulblets derived from field-grown
bulbs cv. Apeldoorn. They showed that the gibberellin activity of bulblets increased
dramatically in November-March and declined sharply in April-July. The increase in
gibberellin-like substances was considered to be derived primarly by synthesis within the
bulblets with possible contributions via translocation from the mother bulb scales, roots,
and shoots.
Recently, Saniewski and Horbowicz (2004) showed the occurrence of quercetin
and kaempferol as glycosides in leaves and anthers of uncooled and cooled tulip bulbs.
During cold storage of tulip bulbs, the content of quercetin and kaempferol substantially
increased in the leaves and the level of these compounds was much lower and stable in
leaves of uncooled bulbs. In the case of anthers, the content of quercetin and apigenin
greatly increased during storage of bulbs at high temperature (17C) and was low in
cooled bulbs. The level of kaempferol in anthers was substantially higher in cooled than
uncooled bulbs.

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 These flavonoids were not detected in dry and fleshly scales, stem, perianth, and
pistil in both uncooled and cooled tulip bulbs.
The role of these changes in endogenous flavonoid levels in leaves and anthers
during cooling of tulip bulbs in dormancy release and daughter bulb formation is unknown.
Flavonoids, mainly quercetin and kaempferol, as endogenous auxin transport inhibitors
(Jacobs and Rubery, 1988; Dakora, 1995; Murphy et al., 2000), inhibitors of lipid
peroxidation (Whittern et al., 1984; Takahama, 1985; Alcaraz et al., 1986; Torel et al.,
1986), strong antioxidants (Pietta, 2000), and substrates for peroxidase (Takahama and
Oniki, 1997, 2000), may play an important role in many physiological processes,
including dormancy release and daughter-bulb enlargement in mother tulip bulbs as a
result of low temperature treatment.

ACKNOWLEDGEMENT
This work was partially supported by a Grant No 6/P06A/011/21 from State
Committee for Scientific Research (Poland).

Literature Cited
Alcaraz, M.J., Ferrandiz, M.L. and Villar, A. 1986. Flavonoid inhibition of soybean
lipoxygenase. Pharmazie 41:299.
Aoba, T. 1976. Effects of temperature on bulb and tuber formation in bulbous- and
tuberous-crop. (in Japanese with English Summary). IX. On the bulb formation in tulip.
Bull. Yamagata Univ., Agri. Sci. 7:387-399.
Aung, L.H. and Rees, A.R. 1974. Changes in endogenous gibberellin levels in Tulipa
bulblets during ontogeny. J. Exp. Bot. 25:745-751.
Aung, L.H., Tognoni, F. and De Hertogh, A.A. 1973. Changes in the carbohydrates of tulip
bulbs during development. HortScience 8:207-208.
Boonekamp, P.M., Beijersbergen, J.C.M. and Fanssen, J.M. 1990. The development of
flowering assays for cold-treated tulip bulbs. Acta Hort. 266:177-181.
Dakora, F.D. 1995. Plant flavonoids: Biological molecules for useful exploitation. Austr. J.
Plant Physiol. 22:87-99.
Jacobs, M. and Rubery, P.H. 1988. Naturally occurring auxin transport regulators. Science
241:346-349.
Kamerbeek, G.A., Beijersbergen, J.C.M. and Schenk, P.K. 1972. Dormancy in bulbs and
corms. In: N. Goren and K. Mehdel (eds.), Proc. XVIIIth Intl. Hort. Congr., Tel Aviv,
vol. V, p.233-239.
Le Nard, M. and De Hertogh, A.A. 1993. Tulipa. p.617-682. In: A. De Hertogh and M. Le
Nard (eds.), The Physiology of Flower Bulbs, Elsevier, Amsterdam, London, New
York, Tokyo.
Murphy, A., Peer, W.A. and Tavi, L. 2000. Regulation of auxin transport by
aminopeptidases and endogenous flavonoids. Planta 211:315-324.
Pietta, P.-G. 2000. Flavonoids as antioxidants. J. Nat. Prod. 63:1035-1042.
Saniewski, M. and Horbowicz, M. 2004. Changes in endogenous flavonoids level during
cold storage of tulip bulbs. Acta Hort. 669:245-252.
Schmalfeld, H.W. and Carolus, R.L. 1965. Nutrient redistribution in the tulip. Proc. Amer.
Soc. Hort. Sci. 86:701-707.
Takahama, U. 1985. Inhibition of lipoxygenase-dependent lipid peroxidation by quercetin:
mechanism of antioxidative function. Phytochemistry 24:1443-1446.
Takahama, U. and Oniki, T. 1997. A peroxidase/phenolics/ascorbate system can scavange
hydrogen peroxidase in plant cells. Physiol. Plant. 101:845-952.
Takahama, U. and Oniki, T. 2000. Flavonoids of some other phenolics as substrates of
peroxidase: physiological significance of the redox reactions. J. Plant Res.
113:301-309.
Torel, J., Cillard, J. and Cillard, P. 1986. Antioxidant activity of flavonoids and reactivity
with peroxy radical. Phytochemistry 25:383-385.
Whittern, C.C., Miller, E. and Pratt, D. 1984. Cotton seed flavonoids as lipid antioxidants.

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 J. Am. Oil Chem. Soc. 61:1975-1978.

Tables

Table 1. The effect of removal of sprouts and root primordia in fully cooled mother tulip
bulbs and their storage in light or darkness at room temperature on daughter bulb
weight.

Treatments Storage Weight of daughter bulbs (g)

conditions: Outermost Inner daughter bulbs
darkness (D) or axillary bud Innermost
light (L) H A B C
Intact bulbs L 0.7 7.9 4.2 1.5
D 0.7 7.6 3.5 1.6
Excised sprouts L 0.9 5.8 2.6 0.9
D 0.9 6.2 3.4 0.8
Excised sprouts and L 0.7 5.1 2.3 0.6
root primordia D 0.6 4.1 2.1 0.6

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Figures

Fig. 1. Tulip bulbs without cooling stored at 17C (on left) and cooled stored at 5C
(on right); photographed on March 15. A intact bulbs, B isolated flower bud and
daughter buds.

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Fig. 2. Growth and development of tulips after planting of uncooled (on left) and fully
cooled bulbs (on right) (see Fig. 1.)

Fig. 3. Uncooled and cooled tulip bulbs (see Fig. 1) were kept for one month at room
temperature (20 to 23C) in natural light conditions. A intact bulbs, uncooled and
cooled, respectively, B daughter buds and daughter bulbs, isolated from uncooled
and cooled mother bulbs, respectively.

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Fig. 4. Cooled tulip bulbs (see Fig. 1) were kept for one month at room temperature (20 to
23C) in natural light conditions after the following treatments: On left intact
mother bulb, in middle removed upper part of sprout, on right removed upper
part of sprouts and root primordial. In all cases, outermost axillary bulb and inner
daughter bulbs are formed.

Fig. 5. Tulip bulbs were stored at 5C from October 15 until the end of August; long
storage of mother bulbs at low temperature causes formation of outermost axillary
bulb and inner daughter bulbs.

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Fig. 6. Daughter bulbs formed in fully cooled mother bulbs after storage for one month at
room temperature (A) and flowering of these daughter bulbs under field conditions
(B).

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