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 198:147–152.
• 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.