Non-destructive evaluation of in vitro-stored plants: A comparison of visual and image analysis

Aynalem, Hailu M.; Righetti, Timothy L.; Reed, Barbara M.

doi:10.1079/ivp2006816

Cited by 12 publications

(5 citation statements)

References 24 publications

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“…Since the visual evaluation was done on the whole plant and the aim of this project was to compare the visual evaluation with the image analysis, a consistent value was needed for the comparison. This result differs from our earlier study of pear where the lower node was more indicative of change during storage (Aynalem et al, 2006). In neither case was the mean of the two nodes as useful as the individual node.…”

Section: Image Analysis Of In Vitro Stored Hopscontrasting

confidence: 99%

“…The goal of this study was to determine if a newly developed imageanalysis procedure developed to detect deterioration for pear shoot cultures (Aynalem et al, 2006) could be applied to cultures of Humulus germplasm and if it could be applied to experimental evaluations. Image analysis was compared to visual observations in an experiment designed to determine if the improved vigor of growthroom plantlets on medium with sequestrene iron would be maintained during cold storage and whether the mean storage duration would change with changes in the iron content of the medium.…”

Section: Introductionmentioning

confidence: 99%

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Iron formulation affects in vitro storage of hops: An image analysis

Aynalem

Righetti

Reed

2006

In Vitro Cell. Dev. Biol. - Plant

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SummaryIn vitro-stored plant germplasm is usually evaluated by visual analysis of the plant based on subjective characters. To reduce the variability in these evaluations, we developed a digital-image evaluation system for in vitro-stored plantlets. This study compares the standard visual evaluation system with a digital analysis system to determine if digital analysis can effectively quantify the health of diverse Humulus germplasm. Eight cultivars of Humulus lupulus L. were stored on standard Murashige and Skoog (MS) medium with iron alone (EDTA chelated) and on MS iron with 100 or 200 mg l 21 sequestrene 138 iron (EDDHA chelated). Digital images of the upper two nodes of each plantlet were evaluated for red, green, blue, green/red ratio, and modified normalized difference vegetation index (MNDVI ¼ R 2 G/R þ G). Evaluation of each plantlet for MNDVI values showed consistent significant differences for all treatments only at the upper node. Significant differences for visual and the MNDVI values among the three iron treatments were observed at the upper node of most of the eight hop cultivars. Regression analysis of the upper node MNDVI values vs. whole-plant visual ratings showed positive correlations for most cultivars. Effects of iron treatments on storage duration were also analyzed for both visual and digital systems. There were significant differences among MNDVI values for plantlets stored on medium with standard MS iron alone (EDTA chelated) and with the addition of sequestrene 138 iron. In general, the MNDVI value of the upper node correlated well with visual ratings and could be used to determine the health of in vitro stored hops.

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Section: Image Analysis Of In Vitro Stored Hopscontrasting

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Iron formulation affects in vitro storage of hops: An image analysis

Aynalem

Righetti

Reed

2006

In Vitro Cell. Dev. Biol. - Plant

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“…Although plant in vitro culture is the basis of most biotechnological methods for breeding and propagation of disease-free plants, very limited research using automated sensors in plant tissue culture has been reported, mainly using “plant to sensor” approaches [ 2 - 7 ] and thus involved a significant degree of invasiveness. So far, only few sensor technologies were used, including monochromatic imaging sensors [ 2 ], RGB cameras [ 3 , 5 - 7 ], modified RGB camera setups with a near infrared (NIR) channel [ 4 , 8 ] and thermal imaging sensors [ 9 ]. Therefore, most studies (reviewed by Gupta and Karmakar [ 5 ]) focused on image analysis to estimate parameters like biomass of callus [ 10 ], classification of somatic embryos and regenerated shoots [ 11 , 12 ], as well as chlorophyll determination [ 13 ] and growth of embryogenic suspension cultures [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

Low-cost and automated phenotyping system “Phenomenon” for multi-sensor in situ monitoring in plant in vitro culture

et al. 2023

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Background The current development of sensor technologies towards ever more cost-effective and powerful systems is steadily increasing the application of low-cost sensors in different horticultural sectors. In plant in vitro culture, as a fundamental technique for plant breeding and plant propagation, the majority of evaluation methods to describe the performance of these cultures are based on destructive approaches, limiting data to unique endpoint measurements. Therefore, a non-destructive phenotyping system capable of automated, continuous and objective quantification of in vitro plant traits is desirable. Results An automated low-cost multi-sensor system acquiring phenotypic data of plant in vitro cultures was developed and evaluated. Unique hardware and software components were selected to construct a xyz-scanning system with an adequate accuracy for consistent data acquisition. Relevant plant growth predictors, such as projected area of explants and average canopy height were determined employing multi-sensory imaging and various developmental processes could be monitored and documented. The validation of the RGB image segmentation pipeline using a random forest classifier revealed very strong correlation with manual pixel annotation. Depth imaging by a laser distance sensor of plant in vitro cultures enabled the description of the dynamic behavior of the average canopy height, the maximum plant height, but also the culture media height and volume. Projected plant area in depth data by RANSAC (random sample consensus) segmentation approach well matched the projected plant area by RGB image processing pipeline. In addition, a successful proof of concept for in situ spectral fluorescence monitoring was achieved and challenges of thermal imaging were documented. Potential use cases for the digital quantification of key performance parameters in research and commercial application are discussed. Conclusion The technical realization of “Phenomenon” allows phenotyping of plant in vitro cultures under highly challenging conditions and enables multi-sensory monitoring through closed vessels, ensuring the aseptic status of the cultures. Automated sensor application in plant tissue culture promises great potential for a non-destructive growth analysis enhancing commercial propagation as well as enabling research with novel digital parameters recorded over time.

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“…Correlating projected plant area to plant height and fresh mass [7] Monitoring plant quality of in vitro conserved plants [8] Ibaraki & Gupta 2011…”

Section: Shoot Culturementioning

confidence: 99%

Towards automated phenotyping in plant tissue culture: in situ fluorescence monitoring

Bethge,

Tapia León,

Rüter

et al. 2024

Photonic Technologies in Plant and Agricultural Science

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Plant in vitro culture techniques are fundamental for research, propagation, and breeding. Automated phenotyping of in vitro cultures can revolutionize trait evaluation by transitioning to continuous and objective quantification, as well as by enhancing accuracy, speed, and efficiency. Limited research exists on automated sensor usage in plant tissue culture, mainly focusing on "plant-to-sensor" approaches. While reflection-based imaging techniques have dominated research to date, fluorescence-based imaging could offer advantages for the application of phenotyping in commercial in vitro propagation and plant research. We developed a new detector head for our "Phenomenon" plant phenotyping system to investigate the potential of fluorescence-based in situ monitoring of plant in vitro culture. In this study, we demonstrate the acquisition of fluorescence image data from plant in vitro cultures as an advanced imaging technique for phenotyping approaches. Over time and qualitatively, we were able to document the development of hairy roots in N. tabacum after transformation with Rhizobium rhizogenes carrying the recently developed reporter gene eYGFPuv.

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Non-destructive evaluation of in vitro-stored plants: A comparison of visual and image analysis

Cited by 12 publications

References 24 publications

Iron formulation affects in vitro storage of hops: An image analysis

Iron formulation affects in vitro storage of hops: An image analysis

Low-cost and automated phenotyping system “Phenomenon” for multi-sensor in situ monitoring in plant in vitro culture

Towards automated phenotyping in plant tissue culture: in situ fluorescence monitoring

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