Sumarlina (16/401975/PBI/01424) 1

Wardha Ayu A. (16/401938/PBI/01434

Hiqma Widya I. D.
(16/401953/PBI/01402)

The Plant Journal (2011) 68, 147-158

JOURNAL RESUME
TCP14 AND TCP15 Affect Internode Length and Leaf Shape in Arabidopsis
Author: Martin Kieffer, Vera Master, Richard Waites, and Brendan Davies

The TCP family of plant-specific, non-canonical bHLH transcription factors has been
specifically linked to regulation of cell proliferation during development. Understanding of
this family is essential for establishment of a genetic model to describe the interaction
between cell proliferation and spatial development. Representatives of the two distinct TCP
sub-families found in Arabidopsis and known as class I (TCPp) and class II (TCPc). These
two sub families are also found in algae Cosmarium spp. There has been many research about
the TCP family for both class and how they work together in plant. According to those
researches, TCP factors have been suggested to act antagonistically on plant cell division and
growth, with class I promoting growth, and class II factors inhibiting it. This journal describe
the developmental roles of two closely related Arabidopsis class I TCP factors, TCP14
(At3g47620) and TCP15 (At1g69690). The research shows that these factors act redundantly
to regulate plant stature by promoting cell proliferation in young internodes.
The plant material is consist of tcp 15-3, tcp15-1, tcp 14-6, tcp 14-4, tcp 14-5, and tcp
15-2 isolated by screening T-DNA alpha population of Arabidopsis. Before double mutant
construction, tcp 14-4 was backcrossed four times. All mutants were genotyped by PCR using
specific primer for each sample. Plant were grown at 20 oC under long-day conditions with
60% humidity. Theres a stage for vector construction where the mutant carrying the gene
reporter (using GUS assay). The RNA is analyzed by Real-time PCR machine using specific
primers. Leaves were collected from WT and mutants and scanned at resolution of 300 dpi
then processed using LEAFANALYSER software. Scanning electron microscopy samples
were also analyzed in this research.
The results show that TCP14 and TCP15 redundantly regulate internode elongation.
Three independent TCP14 T-DNA insertion lines were studied in detail and the Northern blot
analysis showed that the T-DNA insertions in the three mutant lines caused a reduction in
transcript level and resulted in production of truncated transcripts in all cases. While for the
 Sumarlina (16/401975/PBI/01424) 2
Wardha Ayu A. (16/401938/PBI/01434
Hiqma Widya I. D.
(16/401953/PBI/01402)
TCP15, northern blot analysis shows that it was disrupted. Biometric analysis of
inflorescence stems revealed that internode elongation was significantly reduced in the
double mutants. The scanning micrographs of shoot apices also showed that internode
elongation is reduced in the double mutant from early stages. The expression of five effectors
of cell division analyzed by quantitative real-time PCR also showed that the five gene
expressing in cell cycle was reduced in mutant apices relative to the wild type. These mutants
also showed leaf curling in mature leaves of plants grown under short-day condition. The
double mutant leaves were upwardly curled at their margins but only found in mature leaves.
These suggested that TCP14 and TCP15 also involved in regulating cell proliferation within
the leaf.
The results also showed that TCP14 and TCP15 are dynamically expressed in young
proliferating tissues. As expected from the mutant analysis, both genes are expressed in all
internodes of the young inflorescence stems and in young flower pedicles, and their
expression gradually ceases as the tissues mature. There is a spatial regulation for these two
genes along the leaves maturation. These genes are also expressed in flower, particularly in
sepals. In older flowers, they are expressed in young stamens and carpel valves. As the
gynecium matures, expression in the valves is reduced.
The quantitative imaging shows that TCP14 and TCP15 modulate leaf blade expansion.
According to the result of LEAFANALYSER software, it consistent with their dynamic
expression patterns during leaf development, TCP14 and TCP15 modulate overall leaf shape.
These findings suggest that organ shape is defined by rigorous control of cell proliferation in
specific cells and tissues, mediated by expression of a combination of TCP factors. It can be
also concluded that TCP14/15 genes repress cell proliferation in leaf and floral tissues. It can
be observed from the double mutant phenotype where the plants showing reduced cell
proliferation in internodes and enhanced cell proliferation in leaves and floral tissues. So that
the influence of TCP factors on cell proliferation is context dependent.