Light-Induced Fluorescence Transient

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The Light-Induced Fluorescence Transient (LIFT) is a device to remotely measure chlorophyll fluorescence in plants in a fast and non-destructive way. By using a series of excitation light pulses, LIFT combines chlorophyll fluorescence data with spectral and RGB information to provide insights into various photosynthetic traits and vegetation indices. LIFT combines the pump-probe method with the principle of laser-induced fluorescence.

Fluorescence measurement principle

The process of photosynthesis in plants involves the absorption of light by photosynthetic pigments, transfer of energy to the reaction center, charge separation leading to oxygen evolution, and a series of electron transport processes. The efficiency of photosynthesis is determined by factors such as the rate of electron transport, carbon fixation, and energy production. The LIFT measures photosynthesis by exposing the plant to short flashes of blue light and analyzing the changes in fluorescence over time by the help of the FRR technique.

LIFT FRR technique

Light-Induced Fluorescence Transient instrument (black box) in a growth chamber measuring Arabidopsis thaliana plants.

The LIFT Fast Repetition Rate Fluorescence (FRR) technique is a method for measuring plant fluorescence. It uses a series of short bursts of blue light pulses from a LED to excite Photosystem II in the plant. When the Quinone acceptor A (Q_A) reaches its capacity for binding electrons, the system becomes saturated and consequently red fluorescence is emitted. This is regulated by a precise excitation protocol, which consists of a saturation sequence (SQA) and a relaxation sequence (RQA) with a set of short excitation flashes (1 μs).

The fluorescence can then be measured with FRR fluorometry. For that purpose, the LIFT instrument has a built-in optical interference filter to separate the red chlorophyll fluorescence from reflected light, with a wavelength of 685 ± 10 nm.

The fluorescence transient resulting from this excitation protocol shows the kinetics of the reduction of Q_A and its subsequent re-oxidation, and can be used to calculate various photosynthetic indicators. These indicators provide information on the level of photosynthetic activity, such as the efficiency of light utilization, the quantum yield of photochemical conversion, and the rate of electron transport.

LIFT-retrieved photosynthetic indicators

LIFT Transient with the saturation (SQA) and relaxation sequence (RQA) showing the initial (F'), maximum (Fm') and variable fluorescence (Fq').

The LIFT system measures chlorophyll fluorescence by stimulating the plant with excitation light, leading to an increase in fluorescence to its maximum (Fm'). The naturally occurring fluorescence (F') can also be measured without the excitation light. The variable fluorescence (Fq') can be calculated as the difference between Fm' and F'. The Photosystem II operating efficiency can be calculated using the equation:

$\mathrm{F}\mathrm{q}{\phantom{A}}^{\prime }/\mathrm{F}\mathrm{m}{\phantom{A}}^{\prime }=(\mathrm{F}\mathrm{m}{\phantom{A}}^{\prime }-\mathrm{F}{\phantom{A}}^{\prime })/\mathrm{F}\mathrm{m}{\phantom{A}}^{\prime }$

For the relaxation sequence (RQA), different relaxation parameters can be calculated according to different time sections:

• F_r1, ~ 0.625 ms after the maximum Fluorescence has occurred and
• F_r2, ~ 6.5 ms after the maximum Fluorescence has occurred.

These parameters can be used for calculating the reoxidation efficiency of Q_A i.e. the kinetics of electron transfer between QA to PQ pool to Photosystem I for light-adapted plants.

Development history and models

Light-Induced Fluorescence Transient instrument (white box) on a semi-automated phenotyping platform measuring Winter wheat plants grown under elevated CO
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The concept for an airborne LIFT instrument was developed by Dr. Zbigniew Kolber at Rutgers University in 1998. The first field test was conducted at the Biosphere 2 center in Arizona in 2002 using a stationary large LIFT setup equipped with a laser operating at a distance of up to 50 meters. The prototype instrument was later refined and improved at the Carnegie Institute, Stanford, and the Agricultural Research Center in Arizona, where the first attempt to operate it on a tractor frame was made. In 2010 several instruments were transferred from Carnegie to the Forschungszentrum Jülich where they are used for laboratory and field research in robotic positioning systems for non-invasive, high-throughput data acquisition.

Nowadays, two different kinds of LIFT models exist, terrestrial LIFT and marine LIFT.

Terrestrial LIFT

The terrestrial LIFT instrument measures photosynthetic features in higher plants up to a distance of 3 meters. The instrument is equipped with a pair of cameras, triggered by the LIFT excitation signal, to acquire information for 3-D reconstruction of the target plant. The in-line spectrometer is used to acquire reflectance spectra of the target.

Marine LIFT

A marine LIFT version can be used in an aquatic environment. It works with five excitation wavelengths (445 nm, 470 nm, 505 nm, 530 nm, and 590 nm) to distinguish between different phytoplankton groups based on the spectral composition of their light-harvesting pigments. It determines the photosynthetic performance of marine and freshwater algae and photosynthetic bacteria.

Current research

In recent research, LIFT has been used in laboratory settings to explore the kinetics of photosynthesis and was also implemented in high-throughput phenotyping platforms for early drought detection

References

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