Abstract

Micropropagation techniques offer opportunity to proliferate, maintain, and study dynamic plant responses in highly controlled environments without confounding external influences, forming the basis for many biotechnological applications. With medicinal and recreational interests for Cannabis sativa L. growing, research related to the optimization of in vitro practices is needed to improve current methods while boosting our understanding of the underlying physiological processes. Unfortunately, due to the exorbitantly large array of factors influencing tissue culture, existing approaches to optimize in vitro methods are tedious and time-consuming. Therefore, there is great potential to use new computational methodologies for analyzing data to develop improved protocols more efficiently. Here, we first tested the effects of light qualities using assorted combinations of Red, Blue, Far Red, and White spanning 0–100 μmol/m2/s in combination with sucrose concentrations ranging from 1 to 6% (w/v), totaling 66 treatments, on in vitro shoot growth, root development, number of nodes, shoot emergence, and canopy surface area. Collected data were then assessed using multilayer perceptron (MLP), generalized regression neural network (GRNN), and adaptive neuro-fuzzy inference system (ANFIS) to model and predict in vitro Cannabis growth and development. Based on the results, GRNN had better performance than MLP or ANFIS and was consequently selected to link different optimization algorithms [genetic algorithm (GA), biogeography-based optimization (BBO), interior search algorithm (ISA), and symbiotic organisms search (SOS)] for prediction of optimal light levels (quality/intensity) and sucrose concentration for various applications. Predictions of in vitro conditions to refine growth responses were subsequently tested in a validation experiment and data showed no significant differences between predicted optimized values and observed data. Thus, this study demonstrates the potential of machine learning and optimization algorithms to predict the most favorable light combinations and sucrose levels to elicit specific developmental responses. Based on these, recommendations of light and carbohydrate levels to promote specific developmental outcomes for in vitro Cannabis are suggested. Ultimately, this work showcases the importance of light quality and carbohydrate supply in directing plant development as well as the power of machine learning approaches to investigate complex interactions in plant tissue culture.

Highlights

  • The multifaceted value of Cannabis sativa L. as a quality fiber, seed oil, and therapeutic crop have been recognized for millennia (Sandler et al, 2019; Hesami et al, 2020)

  • To evaluate the efficiency and reliability of the hybrid GRNNevolutionary optimization algorithms, the predicted-optimized treatments obtained from evolutionary optimization algorithms (GA, interior search algorithm (ISA), symbiotic organisms search (SOS), and biogeography-based optimization (BBO)) were separately evaluated in the lab as Effects of Light and Carbohydrate Sources on Cannabis Shoot Growth and Development

  • The greatest shoot length was acquired from 25 μmol/m2/s W + 25 μmol/m2/s far-red light (Fr) + 3% Sucrose (154.68 ± 51.228 mm), while shoot length was most stunted when grown with 100 μmol/m2/s

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Summary

Introduction

The multifaceted value of Cannabis sativa L. (cannabis) as a quality fiber, seed oil, and therapeutic crop have been recognized for millennia (Sandler et al, 2019; Hesami et al, 2020). Over the past two decades, interest relating to its medicinal applications have largely been emphasized due to the discovery of over 500 unique secondary metabolites (ElSohly and Gul, 2014) Of these compounds, there are more than 100 cannabinoids that contribute to cannabis’ pharmacological properties (Fathordoobady et al, 2019). Tissue culture techniques can be applied to examine essential plant responses to external stimuli in highly controlled environments under axenic conditions for biotechnological (Shukla et al, 2017), conservation (Ayuso et al, 2019), and various –omics related technologies (Andre et al, 2016) These approaches can be re-applied to suit the needs of the emerging cannabis industry

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