University of the Philippines Los Baños

College of Agriculture and Food Science

Institute of Crop Science

FLOWER INDUCTION THROUGH LOW TEMPERATURE

HASMAYE MARIE M. PINTO
HORT 132

1
A scientific paper submitted in partial fulfillment of the requirements in Horticulture 132 under
Ms. Hazel Makahiya, 1st semester, 2020-2021
 INTRODUCTION
Temperature plays a dominant role in controlling proper growth and flowering in many
plant species (Khodorova & Boitel-Conti, 2013). According to Hoenicka et al. (2016), plant
morphogenesis is dependent on intricate interactions between environmental signals and
genetically encoded developmental programs. Among several naturally occurring environmental
factors, promotion of flowering in response to prolonged exposure to cold temperatures is a useful
adaptation and growth control for plant species. Vernalization, which is the cold requirement for
flower induction, ensures that flowering can occurs in appropriate season of the year. Since
flowering is a vital stage in the life history of a plant, the time for a plant to flower must coincide
with favorable conditions so that viable seeds can be produced, ensuring the continued survival of
the species in subsequent generations (Dennis & Peacock, 2009). Many plant species, including
both dicots and the monocots, require vernalization to stimulate their flowering (Dennis &
Peacock, 2009).

The time required for complete vernalization varies with species and optimum
vernalization temperatures range from 5-8 °C with few exceptions of some plant species (Kaymak
& Guvenc, 2010). Vernalization response can be facultative or obligate. Facultative vernalization
response is observed in winter annuals where cold exposure is not required for flowering, but cold
treatment allows flowering to occur more rapidly after. Obligate vernalization, on the other hand,
is exhibited by biennials where cold treatment is needed for flowering and that without prior cold
exposure, the plant species cannot flower (Michaels & Amasino, 2000). There have been many
studies through the years that evaluates vernalization and photoperiodism (Salisbury, 1986; Sasani
et al., 2009). In many plant species, esearch have speculated flowering as a response to low
temperatures or to the combined effects of low temperatures and photoperiodism and studied the
effects of low temperature and daylength cues on expression of flowering-time genes during
vernalization-induced flowering. This exercise aims to examine the effect of photoperiodism to
the optimum period of cold treatment to induce early flowering in radish.

METHODOLOGY
The experiment conducted using radish seeds yielded no conclusive data using the
experimental set-ups made. A hypothetical data from Kaymak & Guvenc (2010) was used for the
results and discussion instead. For the failed experiment, the treatments used were T1 – control or
no cold treatment and T2 – refrigerated/cold environment treatment. The seeds were planted in
medium sized pots with soil. After the radicle of the seeds broke out, three sets were refrigerated.
For the refrigerated treatment, one set of seeds was taken out after 2 days, 4 days and 8 days. The
seedlings were transplanted into pots and grown until flowering. The data on the age at flowering,
number of leaves at flowering and total number of flowers per plant (or intensity of flowering) was
supposed to be collected but no plants grew until flowering.

RESULTS AND DISCUSSION

The vernalization period of radish lasts at least 10 days depending on the cultivar. Radish
makes an ideal candidate for vernalization studies because the maximum effect of vernalization
time is obtained at 20-30 days period. In the results of Kaymay and Guvenc (2010), the day length
and the changes in vernalization times were provided by different sowing dates of radish cultivars.
For one cultivar (Siyah raddish), it was determined that flowering did not occur when the
vernalization time was less than 15 days during both the long days (LD) and short days (SD) for
Siyah. Furthermore, except for Siyah, other cultivars could not flower without being vernalized
during SD. According to the results, under the field conditions, different sowing dates had an
impact on flowering rates. In Table 1, the highest day length mean has the least vernalization time.
Vernalization as a process has several unique properties. The initial perception and response to the
period of cold occurs in dividing cells (germinating seedlings). Its molecular aspect, as described
by Dennis & Peacock (2009), is due to the regulation of key genes by epigenetic modification
(modifications to chromatin that do not alter the DNA sequence itself.

Table 1. Effect of vernalization time and day length on anthesis of five radish cultivars
(Kaymak & Guvenc, 2010)

Day Length
Day
Vernalization Long Short
Length
time Day Day
Mean
(16h) (8h)

0 day 59.73 - 29.9

5 days 44.67 75.53 60.1
10 days 38.13 61.27 49.7
15 days 39.94 54.61 47.3
20 days 36.83 45.61 41.2
 The key protein accompanying vernalization, FLOWERING LOCUS C (FLC), was first
worked out in Arabidopsis. FLC codes for a repressor of flowering and ensures vegetative growth
of plant species. Vernalization results in histone modifications that repress FLC. The absence of
the repressor protein FLC following vernalization then allows the expression of two other
genes, FLOWERING LOCUS T (FT) and SUPPRESSOR OF OVER EXPRESSION OF
CONSTANS (SOC1) and the activity of their gene products triggers the genes that control flower
development. FT encodes a protein that acts as a mobile flowering signal, or 'florigen', traveling
via the phloem from the leaf to the apex to cause flower formation. For cereals, one of the
genetically defined genes, VRN1, was shown to be the key response gene in the vernalization
process. However, in this case the cold treatment induces gene activity, rather than repressing it as
in FLC in Arabidopsis. VRN1 is a promoter of the transition from the vegetative to reproductive
state of the growing shoot apex. According to Dennis & Peacock (2009), “induction of VRN1 is
accompanied by the repression of another genetically defined gene, VERNALISATION 2 (VRN2),
which, when active, prevents transcriptional activity of the FT gene and production of the mobile
flowering signal.” Although many studies on vernalization have been reported, the molecular
mechanism of vernalization is still largely unknown in radish. However, illumina sequencing and
mapping against radish reference genomes by Liu et al. (2017) found that up-regulated genes were
more than the down-regulated genes during radish vernalization. Additionally, they also identified
thioglucoside glucohydrolase gene (TGG1) on their analysis implying it may play an important
role in vernalization in addition to defenses against insects and disease.

Figure 1. Diagram showing the key genes controlling vernalization in Arabidopsis and cereals.
(Dennis & Peacock, 2009)
 CONCLUSIONS
Among several naturally occurring environmental factors, promotion of flowering in
response to prolonged exposure to cold temperatures is a useful adaptation and growth control for
plant species. This exercise aims to examine the effect of photoperiodism to the optimum period
of cold treatment to induce early flowering in radish. The experiment conducted using radish seeds
yielded no conclusive data using the experimental set-ups made. A hypothetical data from Kaymak
& Guvenc (2010) was used for the results and discussion instead. Results showed that under the
field conditions, different sowing dates (day length) had an impact on flowering rates and
vernalization period. Furthermore, vernalization is due to the regulation of key genes by epigenetic
modification.
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