ROLE OF AUXINS IN

PLANT TISSUE
CULTURE

Ruchika Bajaj
A0901209001
INTRODUCTION
 Auxins:
 class of plant growth substance and
morphogens
 essential role in coordination of many growth
and behavioral processes in the plant life
cycle
 first revealed by the Dutch scientist Frits
Went
 Kenneth V. Thimann isolated and determined
its chemical structure.
 regulates the amount, type, and direction of
plant growth
 abundantly produced in growth areas, e.g.,
root and shoot tips, but are also produced
elsewhere
 play a role in cell division and
differentiation, fruit development, the
formation of roots from cuttings, the
inhibition of lateral branching and leaf fall
 Naturally occurring: indole-3-acetic acid(IAA)
FUNCTIONS OF AUXIN
 Stimulates cell elongation
 Stimulates differentiation of phloem and
xylem
 Stimulates root initiation on stem cuttings
and lateral root development in tissue
culture
 Can inhibit or promote (via ethylene
stimulation) leaf and fruit abscission
 Can induce fruit setting and growth in some
plants
 Delays fruit ripening
 Stimulates growth of flower parts
IAA STIMULATES POLLEN TUBE
GROWTH AND MEDIATES THE
MODIFICATION OF ITS WALL
COMPOSITION AND STRUCTURE IN
TORENIA FOURNIERI
Juan-Zi Wu,1 Yi Lin,2 Xue-Lian Zhang,1 Dai-Wen
Pang,2 and Jie Zhao1
 1Key Laboratory of the Ministry of Education for
Plant Developmental Biology, College of Life
Sciences, Wuhan University, Wuhan, 430072, China
 2College of Chemistry and Molecular Sciences, and
State Key Laboratory of Virology, Wuhan University,
Wuhan 430072, China
INTRODUCTION
 IAA has important roles in plant sexual
reproduction: controlling the development of
stamens, gynoecia, and ovary, promoting the
maturation of egg cells, inducing the axial
polarity and polar development of the
embryo (Nemhauser et al., 2000; Mol et al.,
2004; Aloni et al., 2006)
  IAA probably promotes pollen tube growth in
the pistils (Kovaleva and Zakharova,
2003; Aloni et al., 2006)
MATERIALS AND METHODS
 Plant materials: Pollens of Torenia fournieri
 Pollen tube growth:
 Pollens were grown with different hormones like IAA,
zeatin, gibberellins etc
 Mixed hormone culture also used
 Growth rate was studied
 FM 4-64 labelling:
 After culture for 2 h, pollen tubes were stained with 3
μM FM4-64 for 12 min
 washed three times with basal medium
 observed under a microscope using green light
excitation
 Experiments were repeated three times.
 SVs observed by TEM
 Immunofluorescent labelling of PM H+-
ATPases in pollen tubes
 Immunofluorescent labelling of pectins
 Detection of cellulose
 FTIR analysis:
 Thespectrum between 800 and 1800 cm −1, which
contains information of polysaccharides, was
selected in order to monitor cell wall
modifications
 AFM imaging
RESULTS
 Effects of various exogenous hormones on
pollen tube growth
 IAA was the most effective hormone stimulating tube
growth
 IAA interacting with ZT and/or GA3 stimulated tube
growth much more than IAA used alone
 Control tube had a rougher surface compared with
the IAA-treated tube
 Shank of the IAA-treated tubes was narrower and
straighter than that of control tubes, but ZT and
GA3 had no effect on tube shape
 These results suggest that IAA might be a primary
hormonal promoter stimulating pollen tube growth.
 FM4-64 staining:
 The FM4-64 labelling pattern was similar in
control and IAA-treated tubes, i.e. the
fluorescence staining focused in the tube apex
and extended to the subapical region with a
gradually lower intensity
 However, the labelled region, which was deemed
to be filled with SVs, was longer and the
intensity of staining was stronger in IAA-treated
tubes than in control tubes
 Ultrastructure of pollen tubes observed by
TEM
 Polarity with four distinct zones along the tubes,
i.e. clear zone, organelle zone, nuclear zone, and
vacuolar zone.
 TEM observation showed that the clear zone was
abundant in SVs.
 In the organelle zone (10–150 μm from tube tip),
more SVs and organelles, such as mitochondria,
occurred in the IAA-treated tubes than in the
control tubes.
 In both the nuclear and the organelle zone, there
were bigger vacuoles in IAA-treated tubes
 PM H+-ATPase distribution in pollen tubes
 After plasmolysis, the immunosignal of PM H +-
ATPase appeared in the plasma membrane, not
in the cell wall of the pollen tubes.
 Enzyme was concentrated in the apical region of
control tubes, while it was distributed
throughout the tubes and was focused at the
apical region after IAA treatment.
 Pectin distribution in pollen tubes
 The distribution pattern was similar in IAA-
treated and control tubes, but there was
stronger labelling of both pectins in the former
 Cellulose distribution in pollen tubes
 As tubes grew, the average cellulose fluorescence intensity
significantly declined
 IAA lowered the average cellulose fluorescence intensity in
tubes, especially in the apical region, though the
distribution pattern of cellulose was alike in IAA-treated and
control tubes
 Changes in chemical components of the cell wall of
pollen tubes
 IAA induced displacements of the peaks and changes in
absorbance.
 The difference spectra generated by digital subtraction of
control spectra from IAA-treated spectra showed that
protein and pectin increased while cellulose decreased in
tubes after IAA treatment
 Ultrastructure of pollen tubes observed by AFM
 The apical region of the control tube surface was rich
in disorganized CMFs and block-like structures, while
the IAA-treated tube surface was covered with both
block-like structures approximately parallel to each
other and a few CMFs
 CMFs formed sinuous networks in the subapical region
of control tube surface while in IAA-treated tubes
were parallel to each other and showed a
predominant orientation
 For IAA-treated tubes, the density of CMFs in the
apical region was lower than that in the subapical
region
 CONCLUSION
 IAA stimulated the growth
of T. fournieri pollen
tubes by increasing the
secretion of SVs and PM
H+-ATPase, and modifying
the tube wall composition
and structure, especially
CMF orientation
 This study highlighted the
role of the cell wall, esp.
CMF orientation, in pollen
tube growth and direction
REFERENCES
 http:/www.plant-hormones.info/auxins.html

 http://users.rcn.com/jkimball.ma.ultranet/Biol
ogyPages/A/Auxin.html.

 Juan-Zi Wu, Yi Lin, Xue-Lian Zhang, Dai-Wen

Pang and Jie Zhao “IAA stimulates pollen tube
growth and mediates the modification of its
wall composition and structure inTorenia
fournieri”,
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC
2423660/