Abstract
BackgroundRecently emerging approaches to high-throughput plant phenotyping have discovered their importance as tools in unravelling the complex questions of plant growth, development and response to the environment, both in basic and applied science. High-throughput methods have been also used to study plant responses to various types of biotic and abiotic stresses (drought, heat, salinity, nutrient-starving, UV light) but only rarely to cold tolerance.ResultsWe present here an experimental procedure of integrative high-throughput in-house phenotyping of plant shoots employing automated simultaneous analyses of shoot biomass and photosystem II efficiency to study the cold tolerance of pea (Pisum sativum L.). For this purpose, we developed new software for automatic RGB image analysis, evaluated various parameters of chlorophyll fluorescence obtained from kinetic chlorophyll fluorescence imaging, and performed an experiment in which the growth and photosynthetic activity of two different pea cultivars were followed during cold acclimation. The data obtained from the automated RGB imaging were validated through correlation of pixel based shoot area with measurement of the shoot fresh weight. Further, data obtained from automated chlorophyll fluorescence imaging analysis were compared with chlorophyll fluorescence parameters measured by a non-imaging chlorophyll fluorometer. In both cases, high correlation was obtained, confirming the reliability of the procedure described.ConclusionsThis study of the response of two pea cultivars to cold stress confirmed that our procedure may have important application, not only for selection of cold-sensitive/tolerant varieties of pea, but also for studies of plant cold-response strategies in general. The approach, provides a very broad tool for the morphological and physiological selection of parameters which correspond to shoot growth and the efficiency of photosystem II, and is thus applicable in studies of various plant species and crops.
Highlights
In plants, acclimation to cold, causes reduced growth, increase in antioxidant content, reduced water content, and changes in gene regulation, hormone balance, membrane composition, osmotic regulation, and photosynthetic function [1]
Visible imaging used for shoot growth To compare the influence of cold acclimation on biomass production, two putative cold-resistant cultivars of pea Terno and Enduro were selected
The green area of each individual seedling was extracted from particular projections (Figure 1) and combined to account for the overall shoot biomass
Summary
Acclimation to cold, causes reduced growth, increase in antioxidant content, reduced water content, and changes in gene regulation, hormone balance, membrane composition, osmotic regulation, and photosynthetic function [1]. Humplík et al Plant Methods (2015) 11:20 have developed so-called sustained non-photochemical quenching (reviewed, e.g., by Verhoeven [4]) as a protection mechanism against absorbed light which is in excess with respect to the capacity of the carbon photosynthetic reactions and which is decreased during winter. These plants sense the upcoming cold period through the perception of environmental impulses, mainly temperature and day length. High-throughput methods have been used to study plant responses to various types of biotic and abiotic stresses (drought, heat, salinity, nutrient-starving, UV light) but only rarely to cold tolerance
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