Effects of phosphorus stress on root

development in Arabidopsis thaliana

Charlie Page, Haseeb Shah, and Allie Shoffner

BIOL 407
4/11/2012
Introduction
Phosphorus is a critical plant nutrient
Required for phosphate groups in DNA, RNA,
phospholipids, proteins, and more
Often supplemented with classic NPK fertilizer
Problem: phosphorus is practically immobile
in the soil.
To combat this plants respond to low phosphorus
levels with adaptations such as increased lateral
root and root hair growth (Lopez-Bucio et al.
2002)
Root Hairs
Plants absorb much of their phosphorus
through root hairs
Root hairs provide increased surface area and
contact with fresh soil that has not been depleted
of phosphorus
Root hairs alone can provide as much as 60% of a
plants phosphorus intake (Gahoonia and Nielsen,
1998)
Unsurprisingly, root hair length and density
increases under nutrient stress (Gilroy and
Jones 2000)
Lateral Roots
Lateral roots also grow faster in
response to phosphorus stress
(Lopez-Bucio et al. 2002)
This serves to increase root contact
with fresh soil to acquire more
phosphorus
This lateral growth may come at
the expense of primary root
growth (Gahoonia and Nielson,
1998)
Hypothesis
Based off of the findings of other researchers,
we predict that Arabidopsis thaliana grown in
a low-phosphorus medium will exhibit the
following traits relative to plants grown in high
phosphorus medium:
Increased root hair density
Increased lateral root density
Potentially decreased main root length
Materials and methods
Nutrients
Stock Concentration (M) Volume Added (ml or g)
ZnSO4(7H20) 0.001 1
H3BO3 0.0125 1
(NH4)6Mo8O24(4H2O) 0.0005 .1
KNO3 1 3
Ca(NO3)2(4H20) 1 2
MgSO4(7H20) 0.5 1
(NH4)2SO4 0.5 1
KH2PO4 (low P - 1 uM) 1 0.001
KH2PO4 (high P - 50 uM) 1 0.05
Micronutrient Solution 0.5
MES (solid) .1
sucrose (solid) 0.5
myo-inositol (solid) 10
Materials and Methods
Created 2 nutrient solutions
Nutrients: Ca, Mg, etc.

High Phosphorus (HP) and Low Phosphorus (LP)
HP-50 uM and LP-1 uM
Created 12 gel plates
6 for HP and 6 for LP
30 ml of nutrient solution in each plate
Planted 3 Arabidopsis Thaliana seeds in each plate

Materials and Methods
Took photos of each plate on the 5th, 8
th
,
12
th
, and 15
th
day after planting to
observe root hairs
Obtained with Jenoptik ProgRes C14plus camera
and stereomicroscope (model Olympus SZX16) at
3.2x magnification
Images calibrated with 1mm reticule
Scanned plates on 8
th
, 12
th
, and 15
th
days
to observe lateral roots and total root
length
Images obtained from scanner at 600 px/in
resolution
Measured total root length, root hair
density, and lateral root density in ImageJ,
and analyzed in Minitab
Results
Total root length
Day 15 Low P: 76.65 mm; High P: 73.30 mm
t-test: P = 0.633
Lateral roots
Lateral roots
emerged
between day 6
and day 10
Lateral root
density on day
15:
t-test: P = 0.175



Results
Root hair density
No clear pattern
across days
Root hair density
decreases over
time for both low
and high P plants
Discussion
Expected results Main root growth and
lateral root growth
Main root growth should be unaffected by
phosphorus (Lynch and Brown, 2000)
Lateral root branching was higher in HP
Unexpected results of root hair density
No overall difference. Should have had higher
density in LP
Maybe P was depleted in the gel media after a
certain time (Raghothoma, 2009)
Root tip length should have been higher in LP
(Bates and Lynch, 2000)
Discussion
Sample size may have been to small for root
tip among other variables
Possible error includes the positioning error
for some plates and then repositioning
May have tampered with lateral root branching
and main root growth
Future experimentation growing plants in
buffer systems to measure nutrient
concentrations and make consistent diffusion
gradient of gel media over time (Gourley,
1993)
References
Allan, D. L., Gourley, C. J. ., Bloom, P. R., & Russelle, M. P. (1993). (1993)
Evaluation and Improvements of a Sand-Alumina Culture Technique to
Screen Plants for Low Phosphorus Tolerance. Soil Sci. Soc. Am. J., 57, 103-
110.
Bates, T. R., & Lynch, J. P. (2000). The efficiency of Arabidopsis thaliana
(Brassicaceae) root hairs in phosphorus acquisition. American journal of
botany, 87(7), 964-70. Retrieved from
http://www.ncbi.nlm.nih.gov/pubmed/10898773
Gahoonia TS, Nielsen NE (1998) Direct evidence on participation of root
hairs in phosphorus (
32
P) uptake from soil. Plant and Soil 198:147152.
Lopez-Bucio J, Hernandez-Abreu E, Sanchez-Calderon L, Nieto-Jacobo MF,
Simpson J, and Herrera-Estrella, L (2002) Phosphate availability alters
architecture and causes changes in hormone sensitivity in the Arabidopsis
root system. Plant Physiology 129:244-256.
Lynch, Jonathan P, & Brown, K. M. (2005). Whole plant adaptations to low
phosphorus availability. Plant-Environment Interactions, 3, 814-863.