Recent substantial advances in high-throughput field phenotyping have provided plant breeders with affordable and efficient tools for evaluating a large number of genotypes for important agronomic traits at early growth stages. Nevertheless, the implementation of large datasets generated by high-throughput phenotyping tools such as hyperspectral reflectance in cultivar development programs is still challenging due to the essential need for intensive knowledge in computational and statistical analyses. In this study, the robustness of three common machine learning (ML) algorithms, multilayer perceptron (MLP), support vector machine (SVM), and random forest (RF), were evaluated for predicting soybean (Glycine max) seed yield using hyperspectral reflectance. For this aim, the hyperspectral reflectance data for the whole spectra ranged from 395 to 1005 nm, which were collected at the R4 and R5 growth stages on 250 soybean genotypes grown in four environments. The recursive feature elimination (RFE) approach was performed to reduce the dimensionality of the hyperspectral reflectance data and select variables with the largest importance values. The results indicated that R5 is more informative stage for measuring hyperspectral reflectance to predict seed yields. The 395 nm reflectance band was also identified as the high ranked band in predicting the soybean seed yield. By considering either full or selected variables as the input variables, the ML algorithms were evaluated individually and combined-version using the ensemble–stacking (E–S) method to predict the soybean yield. The RF algorithm had the highest performance with a value of 84% yield classification accuracy among all the individual tested algorithms. Therefore, by selecting RF as the metaClassifier for E–S method, the prediction accuracy increased to 0.93, using all variables, and 0.87, using selected variables showing the success of using E–S as one of the ensemble techniques. This study demonstrated that soybean breeders could implement E–S algorithm using either the full or selected spectra reflectance to select the high-yielding soybean genotypes, among a large number of genotypes, at early growth stages.
A hybrid artificial intelligence model and optimization algorithm could be a powerful approach for modeling and optimizing plant tissue culture procedures. The aim of this study was introducing an Adaptive Neuro-Fuzzy Inference System- Non-dominated Sorting Genetic Algorithm-II (ANFIS-NSGAII) as a powerful computational methodology for somatic embryogenesis of chrysanthemum, as a case study. ANFIS was used for modeling three outputs including callogenesis frequency (CF), embryogenesis frequency (EF), and the number of somatic embryo (NSE) based on different variables including 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BAP), sucrose, glucose, fructose, and light quality. Subsequently, models were linked to NSGAII for optimizing the process, and the importance of each input was evaluated by sensitivity analysis. Results showed that all of the R 2 of training and testing sets were over 92%, indicating the efficiency and accuracy of ANFIS on the modeling of the embryogenesis. Also, according to ANFIS-NSGAII, optimal EF (99.1%), and NSE (13.1) can be obtained from a medium containing 1.53 mg/L 2,4-D, 1.67 mg/L BAP, 13.74 g/L sucrose, 57.20 g/L glucose, and 0.39 g/L fructose under red light. The results of the sensitivity analysis showed that embryogenesis was more sensitive to 2,4-D, and less sensitive to fructose. Generally, the hybrid ANFIS-NSGAII can be recognized as a powerful computational tool for modeling and optimizing in plant tissue culture.
Background: Optimizing the somatic embryogenesis protocol can be considered as the first and foremost step in successful gene transformation studies. However, it is usually difficult to achieve an optimized embryogenesis protocol due to the cost and time-consuming as well as the complexity of this process. Therefore, it is necessary to use a novel computational approach, such as machine learning algorithms for this aim. In the present study, two machine learning algorithms, including Multilayer Perceptron (MLP) as an artificial neural network (ANN) and support vector regression (SVR), were employed to model somatic embryogenesis of chrysanthemum, as a case study, and compare their prediction accuracy. Results: The results showed that SVR (R 2 > 0.92) had better performance accuracy than MLP (R 2 > 0.82). Moreover, the Non-dominated Sorting Genetic Algorithm-II (NSGA-II) was also applied for the optimization of the somatic embryogenesis and the results showed that the highest embryogenesis rate (99.09%) and the maximum number of somatic embryos per explant (56.24) can be obtained from a medium containing 9.10 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 4.70 μM kinetin (KIN), and 18.73 μM sodium nitroprusside (SNP). According to our results, SVR-NSGA-II was able to optimize the chrysanthemum's somatic embryogenesis accurately. Conclusions: SVR-NSGA-II can be employed as a reliable and applicable computational methodology in future plant tissue culture studies.
Improving genetic yield potential in major food grade crops such as soybean (Glycine max L.) is the most sustainable way to address the growing global food demand and its security concerns. Yield is a complex trait and reliant on various related variables called yield components. In this study, the five most important yield component traits in soybean were measured using a panel of 250 genotypes grown in four environments. These traits were the number of nodes per plant (NP), number of non-reproductive nodes per plant (NRNP), number of reproductive nodes per plant (RNP), number of pods per plant (PP), and the ratio of number of pods to number of nodes per plant (P/N). These data were used for predicting the total soybean seed yield using the Multilayer Perceptron (MLP), Radial Basis Function (RBF), and Random Forest (RF), machine learning (ML) algorithms, individually and collectively through an ensemble method based on bagging strategy (E-B). The RBF algorithm with highest Coefficient of Determination (R2) value of 0.81 and the lowest Mean Absolute Errors (MAE) and Root Mean Square Error (RMSE) values of 148.61 kg.ha-1, and 185.31 kg.ha-1, respectively, was the most accurate algorithm and, therefore, selected as the metaClassifier for the E-B algorithm. Using the E-B algorithm, we were able to increase the prediction accuracy by improving the values of R2, MAE, and RMSE by 0.1, 0.24 kg.ha-1, and 0.96 kg.ha-1, respectively. Furthermore, for the first time in this study, we allied the E-B with the genetic algorithm (GA) to model the optimum values of yield components in an ideotype genotype in which the yield is maximized. The results revealed a better understanding of the relationships between soybean yield and its components, which can be used for selecting parental lines and designing promising crosses for developing cultivars with improved genetic yield potential.
Recent advanced high-throughput field phenotyping combined with sophisticated big data analysis methods have provided plant breeders with unprecedented tools for a better prediction of important agronomic traits, such as yield and fresh biomass (FBIO), at early growth stages. This study aimed to demonstrate the potential use of 35 selected hyperspectral vegetation indices (HVI), collected at the R5 growth stage, for predicting soybean seed yield and FBIO. Two artificial intelligence algorithms, ensemble-bagging (EB) and deep neural network (DNN), were used to predict soybean seed yield and FBIO using HVI. Considering HVI as input variables, the coefficients of determination (R2) of 0.76 and 0.77 for yield and 0.91 and 0.89 for FBIO were obtained using DNN and EB, respectively. In this study, we also used hybrid DNN-SPEA2 to estimate the optimum HVI values in soybeans with maximized yield and FBIO productions. In addition, to identify the most informative HVI in predicting yield and FBIO, the feature recursive elimination wrapper method was used and the top ranking HVI were determined to be associated with red, 670 nm and near-infrared, 800 nm, regions. Overall, this study introduced hybrid DNN-SPEA2 as a robust mathematical tool for optimizing and using informative HVI for estimating soybean seed yield and FBIO at early growth stages, which can be employed by soybean breeders for discriminating superior genotypes in large breeding populations.
Ficus religiosa is known as a long-lived multipurpose forest tree. The tree plays an important role for religious, medicinal, and ornamental purposes. However, the propagation rate of Ficus religiosa is low in natural habitat so the plant tissue culture techniques are an applicable method for multiplication of this valuable medicinal plants. Thus, the aim of this study is to understand the effect of different auxin/cytokinin ratios on indirect shoot organogenesis of this plant. According to our results, the maximum callus induction frequency (100%) was obtained on Murashige and Skoog (MS) medium supplemented with 0.5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) plus 0.05 mg/l 6-benzylaminopurine (BAP) from petiole segments. For shoot induction purpose, the yellow-brownish, friable, organogenic calli were inoculated on shoot induction medium. On MS medium supplemented with 1.5 mg/l BAP and 0.15 mg/l Indole-3-butyric acid (IBA), 96.66% of the petiole-derived calli responded with an average number of 3.56 shoots per culture. The highest root formation frequency (96.66%), root number (5.5), and root length (4.83 cm) were achieved on MS medium containing 2.0 mg/l IBA plus 0.1 mg/l Naphthaleneacetic acid (NAA). The rooted shoots were successfully transferred to field condition and the substrate with the mixture of cocopeat and perlite (1:1) had the highest survival rate (96.66%). This is the first report of an effective in vitro organogenesis protocol for F. religiosa by indirect shoot organogenesis through axenic seedling derived petiole explants, which can be efficiently employed for conservation of this important medicinal plant species as well as the utilization of active biomolecules.
Chenopodium quinoa Willd. is known as a multipurpose crop that could be employed as an alternative on unsuitable lands with limited water resources such as arid and semi-arid regions. However, the biggest problem in cultivating this plant is contamination by viruses that are carried by quinoa seeds and cause diseases. Therefore, in vitro propagation methods could be applied to produce virus-free plants. The capacity of sodium hypochlorite to sterilize seeds and the regeneration potential of cotyledonary node segments were studied on Murashige and Skoog (MS) medium consisting of various concentrations of cytokinins and auxins either alone or in combinations. Although no contamination was observed after sterilization with 20% sodium hypochlorite and 10 and 15 min of immersion; with 25% sodium hypochlorite and 5, 10, and 15 min of immersion, the highest seed germination (100%) was obtained after sterilization with 20% sodium hypochlorite for 5 minutes. MS medium supplemented with 2.0 mg/l 6-benzylaminopurine (BAP) had the highest shoot induction frequency (93.33%) with an average of 4.96 shoots per node explant. This study is the first report of an appropriate and effective in vitro regeneration protocol for Chenopodium quinoa via direct organogenesis through cotyledonary node explants derived from axenic-seedlings, which can be effectively employed for the genetic manipulations and exploitation of active biomolecules of this valuable plant. This protocol can also increase the speed of quinoa breeding programs.
In conjunction with big data analysis methods, plant omics technologies have provided scientists with cost-effective and promising tools for discovering genetic architectures of complex agronomic traits using large breeding populations. In recent years, there has been significant progress in plant phenomics and genomics approaches for generating reliable large datasets. However, selecting an appropriate data integration and analysis method to improve the efficiency of phenome-phenome and phenome-genome association studies is still a bottleneck. This study proposes a hyperspectral wide association study (HypWAS) approach as a phenome-phenome association analysis through a hierarchical data integration strategy to estimate the prediction power of hyperspectral reflectance bands in predicting soybean seed yield. Using HypWAS, five important hyperspectral reflectance bands in visible, red-edge, and near-infrared regions were identified significantly associated with seed yield. The phenome-genome association analysis of each tested hyperspectral reflectance band was performed using two conventional genome-wide association studies (GWAS) methods and a machine learning mediated GWAS based on the support vector regression (SVR) method. Using SVR-mediated GWAS, more relevant QTL with the physiological background of the tested hyperspectral reflectance bands were detected, supported by the functional annotation of candidate gene analyses. The results of this study have indicated the advantages of using hierarchical data integration strategy and advanced mathematical methods coupled with phenome-phenome and phenome-genome association analyses for a better understanding of the biology and genetic backgrounds of hyperspectral reflectance bands affecting soybean yield formation. The identified yield-related hyperspectral reflectance bands using HypWAS can be used as indirect selection criteria for selecting superior genotypes with improved yield genetic gains in large breeding populations.
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