Plant Growth Analysis & Time-Lapse Characterization

of Arabidopsis Starch Metabolism Mutants

Zachary J. Jarou*, Sean E. Weise & Thomas D. Sharkey
Department of Biochemistry & Molecular Biology, Michigan State University
Columbia
sex1

Introduction Measuring Leaf Area with Photoshop

During the early exponential growth phase of plants, the majority of
photoassimilates are reinvested into new leaf material. Once the pho-
tosynthetic capacity to reach sufficient carbon mass has been
achieved, carbon metabolism can be redirected to cause the accumu-
lation of end products, such as starch and oil. To optimize the timing of
this redirection, we have developed growth analysis methods to mini-
mize yield penalty.
Classical Growth Analysis
Relative growth rate (RGR) is defined as the net dry biomass increase
per unit dry mass per day and can be further factorized into three
components, specific leaf area (SLA), leaf mass fraction (LMF), and unit
leaf rate (ULR).
The irregular shape of leaves can make measurement by traditional
means difficult, especially for large numbers of plants. By taking a digi-
tal picture of the leaves to be measured, along with an object of
known size, the pixel area of the leaves can be can quantified into phy-
sicial units. This is accomplished using the color range and histogram
tools in Adobe Photoshop. A screen capture demonstration of this
process is available on YouTube
(http://www.youtube.com/watch?v=E3O-V6WLw0g).

ln(M2) - ln(M1)
RGR = = SLA x LMF x ULR
t2 - t1
SLA is calculated by dividing the leaf area by the leaf mass. LMF is de-
Time-Lapse Characterization
Leaf area, one of many growth parameters, has been examined by
fined as the leaf mass divided by the total plant mass. ULR is the in-
time-lapse photography in Arabidopsis starch metabolism mutants.
crease of biomass per unit leaf area per day. It can be calculated from
Weekly time-course images of Columbia and severe starch excess
plant mass and leaf area on two time points or by gas exchange.
plants (sex1) are shown (far above). Measurement of rosette areas
(below) shows that mild starch accumulation (sex4) imposed very
Arabidopsis Hydroponics Systems little yield penalty. Videos of these and additional mutants are avail-
Growth analysis requires that the masses of each plant component be able on YouTube (http://www.youtube.com/watch?v=b-XSnuuXFE4).
determined each harvest, including the roots. Because of the difficul-
ties associated with root washing, hydrponic cultivation is ideal as the
roots are easily accessible. Seeds are germinated in small, rockwool-
filled microfuge tubes with the bottoms cut off. Once the roots have
Rosette area (cm^2)

protruded out of the bottom, the tubes are transferred into the pre-
drilled lids of 50mL centrifuge tubes, which also have their bottoms re-
moved. This method prevents tangling of roots while growing in aer-
ated nutrient solution.

(images from Grant Cramer, University of Nevada)

Weeks of Development