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. 2024 Mar 5;19(3):e0299290. doi: 10.1371/journal.pone.0299290

Recommendation of Tahiti acid lime cultivars through Bayesian probability models

Renan Garcia Malikouski 1,#, Filipe Manoel Ferreira 2,#, Saulo Fabrício da Silva Chaves 3,#, Evellyn Giselly de Oliveira Couto 3,#, Kaio Olimpio das Graças Dias 1,#, Leonardo Lopes Bhering 1,*,#
Editor: Mehdi Rahimi4
PMCID: PMC10914267  PMID: 38442106

Abstract

Probabilistic models enhance breeding, especially for the Tahiti acid lime, a fruit essential to fresh markets and industry. These models identify superior and persistent individuals using probability theory, providing a measure of uncertainty that can aid the recommendation. The objective of our study was to evaluate the use of a Bayesian probabilistic model for the recommendation of superior and persistent genotypes of Tahiti acid lime evaluated in 12 harvests. Leveraging the Monte Carlo Hamiltonian sampling algorithm, we calculated the probability of superior performance (superior genotypic value), and the probability of superior stability (reduced variance of the genotype-by-harvests interaction) of each genotype. The probability of superior stability was compared to a measure of persistence estimated from genotypic values predicted using a frequentist model. Our results demonstrated the applicability and advantages of the Bayesian probabilistic model, yielding similar parameters to those of the frequentist model, while providing further information about the probabilities associated with genotype performance and stability. Genotypes G15, G4, G18, and G11 emerged as the most superior in performance, whereas G24, G7, G13, and G3 were identified as the most stable. This study highlights the usefulness of Bayesian probabilistic models in the fruit trees cultivars recommendation.

Introduction

Breeding perennial fruit crops presents a set of challenges. The 3 to 5-year juvenile phase and the variable expression of quantitative traits over time can delay and mislead the selection of superior genotypes [1]. This differential performance over time can be a reflex of the genotypes-by-harvests interaction (GHI). The GHI in perennial species refers to the variation in gene expression and, consequently, the phenotypic traits of a plant due to the different environmental conditions and agricultural practices that occur in each planting cycle [2]. Therefore, in perennial fruit breeding, repeated measures on the same plant over time are important, which increases costs and the duration of breeding cycle [3]. In the presence of complex GHI, breeders must consider the productivity and the stability of the genotypes over harvests [46].

In Tahiti acid lime (Citrus latifolia Tanaka L.), several strategies have been proposed to address the challenges related to the extended juvenile phase and the presence of GHI. Grafting, a well-established technique in citrus propagation, influences vegetative growth and shortens the juvenile phase, facilitating earlier evaluations in breeding programs [7, 8]. Furthermore, research has demonstrated that employing repeatability models allows for accurate genotype selection in Tahiti acid lime after just four measurements [9]. To address the interdependence of measurements taken on the same individual over time, approaches like random regression models using Legendre polynomials have been applied [10]. Random regression models enable the estimation of the genotypic trajectory of evaluated treatments over time.

A recent methodology proposed by Dias et al. [11] can help to optimize the Tahiti acid lime cultivars recommendation, since it uses probability concepts. This method aims to reduce the risk associated with the selection of a given genotype, which is the daily dilemma of the farmers, who seek to guide their actions to minimize the risks of low production for a given crop [12]. Furthermore, plant breeding is increasingly focused on developing genotypes capable of coping with the modifications of the current climate, as the impacts of climate change become an additional pivotal factor to consider in the agricultural sector [13]. Dias et al. [11] proposed to use Bayesian probability concepts to assist in the selection of genotypes that gather favorable alleles for performance and stability across environments and harvesters. Furthermore, it allows a straightforward recommendation based on the probability of a given genotype to be selected considering its performance and stability, and a pairwise comparison of the probabilities of the evaluated selection candidates. This methodology has been proposed for the multiple-location context. Nevertheless, we believe that the same ideas are valid for the multi-harvest. The objective of our study was to evaluate the use of a Bayesian probabilistic model for the recommendation of superior and persistent genotypes of Tahiti acid lime evaluated in 12 harvests.

Materials and methods

Trial and plant material

We evaluated 24 combinations of rootstock and scion of Tahiti acid lime for fruit yield, expressed in kg of fruit per plant (Kg/tree). The hybrids Citrumelo swingle (Citrus paradisi X Poncirus trifoliata) and Citrandarin ‘riverside’ (Citrus sunki X Poncirus trifoliata) were used as rootstock for scions of 12 clones of Tahiti acid lime (Table 1). The plant materials came from the Active Germplasm Bank of Embrapa Mandioca e Fruticultura [14]. Each combination was considered a different selection candidate. The trial was laid out in a complete randomized block design, with four replications. Each plot was composed of three plants. The inter-rows and inter-ranges spacing was 6 x 3 m, respectively. We performed the statistical analysis described in the next topic using the plots unit mean.

Table 1. Codes for 24 combinations of rootstock and scion of Tahiti acid lime.

Scion Rootstock
Citrumelo swingle Citrandarin riverside
Bello Fruit G1 G13
Eledio G2 G14
Iconha G3 G15
Itarana G4 G16
Santa Rosa G5 G17
Bearss Lime G6 G18
CNPMF01 G7 G19
CNPMF02 G8 G20
CNPMF2001 G9 G21
CNPMF5059 G10 G22
BRS Passos G11 G23
Persian 58 G12 G24

The trial was established in July 2015, in São Mateus municipality, Espírito Santo, Brazil (18°48’21"S, 39°53’30"W, 35 m of altitude). The work was conducted on a farm at Bello Fruit® company, through a partnership between the fruit production and export company and the Universidade Federal do Espírito Santo. The experimental region has a rainy season in summer and a dry season in winter, being classified as Aw, following the classification of Köppen [15]. The precipitation and temperature during the period of the experiment was illustrated in the S1 Fig of the Supplementary Material. The data was collected between July 2017 and September 2020, and consisted of 12 harvests carried out in the following days after planting: 736, 808, 861, 918, 972, 1083, 1200, 1249, 1415, 1568, 1633 and 1867.

Statistical analyses

We applied the probabilistic approach of Dias et al. [11] by fitting two Bayesian models using the the rstan package [16] and ProbBreed package [17]. The first model had a homogeneous residual variance (B-ID) and presented the following conditional normal probability:

yijkNEyijk,σ

where:

Eyijk=μ+gi+rj+hk+ghik+pij+e

in which E[yijk] is the expectation of the phenotype from the ith genotype, evaluated in the jth block, at the kth harvest; μ is the overall mean; gi is the genotypic effect; rj is the block effect; hkis the harvest effect; ghik is the genotypes-by-harvests interaction, and pij is the environmental permanent effect.

The prior probability distribution of each parameter of the model was defined as:

μ~N(0,σ[μ])
r~N(0,σ[r])
h~N(0,σ[h])
g~N(0,σ[g])
gh~N(0,σ[μ])
p~N(0,σ[p])
e~HalfCauchy(0,σ[e])
α~HalfCauchy(0,σ[α])

where N(0,σ[α]) and Half Cauchy(0,σ[α]) represent the normal and half-Cauchy distributions, respectively, with mean equal to zero and with different σ[α]2 scale parameters. The following hyperpriors were considered for the respective parameters:

σ[μ]~HalfCauchy(0,φ)
σ[r]~HalfCauchy(0,φ)
σ[h]~HalfCauchy(0,φ)
σ[g]~HalfCauchy(0,φ)
σ[gh]~HalfCauchy(0,φ)

where φ represents a predetermined global hyperparameter (φ = max(y)*10), defined in such a way that results in a weakly informative second level hyperpriors. Therefore, the data dominated the posterior distributions [18]. The half-Cauchy distribution is restricted to positive values, being often recommended as a prior distribution when modeling variance parameters [19].

The second Bayesian model had heterogeneous residual variances (B-DG). This model has the same considerations of the M-ID, except that σkHalfCauchy(0,σ[σk]). We selected the best-fitted model via the Watanabe-Akaike Information Criterion 2 (WAIC2) [18]. We used the Hamiltonian Monte Carlo algorithm in four Markov chains with 4000 samples, thin equals 1 and 50% burn-in.

Convergence diagnostics

The scale reduction factor (R^) was used to assess the effectiveness of the convergence of the Markov chain Monte Carlo (MCMC). This metric indicates whether the chains have mixed sufficiently, and the estimated model parameters have reached a stable distribution. The closer the R^to 1, the greater the quality of mixing and convergence [18]. Greater values imply that more iterations are needed.

We also conducted a graphical analysis to visually assess how well the data generated by our model aligns with the true generative process of the observed data. This involved creating samples (referred to as "ygen") from the fitted models using ancestral sampling from the conditional joint distribution and then plotting these samples against the observed data. Additionally, we employed posterior predictive p-values to gauge how closely the statistical measures (maximum, minimum, median, mean, and standard deviation) of the data generated by the fitted models resembled those of the observed data [18]. For instance, when considering the maximum statistic, we defined the Bayesian p-value as follows: Pmax = pr(T(ygen,θ) ≥ T(y,θ)|y), where T is the statistic test. The degree of similarity between the statistics derived from the generated data and those from the observed data increases as the Bayesian p-values approach 0.5.

Probability of superior performance and genotypic stability

The probability metrics for both performance and stability utilized in this study were proposed by Dias et al. [11] and implemented in the ProbBreed R package [17]. Aiming to select the top four genotypes, we sampled the posterior distribution of the marginal genotypic values given the observed phenotypic values. In each sample, we ranked these genotypes in descending order of posterior genotypic values (gis). Then, we counted the number of events where a given genotype appeared in the subset of superior genotypes (Ω), i.e., among the top four (selection proportion of 16%). The selection of 4 genotypes has been defined in a breeding program that aims to select multiple superior materials to be recommended across various regions. Moreover, diversifying varieties on a single farm is important for the sustainability of cultivating Tahiti acid lime. The probability of superior performance of a genotype is given by the number of samples it appeared in divided by the total number of samples. In summary:

PrgisϵΩsy=1ss=1STgisϵΩsy

where S(s = 1,2,…,S) is the number of samples and Igisy is an indicator variable that maps failure (0, if giS) or success (1, if gis).

We calculated the stability across harvests based on the variance of the effect of GHI. Genotypes with lower GHI variance (var[ghik]) tend to be more stable, showing less fluctuation in their performance between harvests. One can draw a parallel of this metric and the frequentist persistence (see the next topic). Following the same idea described in the last paragraph, calculated the probability of a given candidate belonging to the subset of the top four genotypes with smaller var[ghik] (I(var[ghik]s ∈ℓV|y)). The probability of superior stability was given as follows:

PrvarghilVy=1ss=1SIvarghislVy

We used the ideas previously described for the two probabilities to perform pairwise comparisons between selection candidates. The goal is to investigate the chances of a given genotype being superior, whether in performance or stability, to its peers. The pairwise probabilities of superior performance and the pairwise probabilities of superior stability were given by, respectively:

Prgi>giy=1Ss=1SIgis>gisy

and

Prvarghi<varghiy=1ss=1SIvarghis<varghisy

where IgiS>giSyis an indicator variable mapping success if giS has higher genotypic value than giS, or failure otherwise; and I(var[ghi]s < var[ghi]s|y) is another indicator variable mapping success if ghik has lower variance than ghik, or failure otherwise.

The probability of superior performance within harvests and the pairwise probability of superior performance within harvests can be obtained by the following equations, respectively:

prgik>giky=1Ss=1SIgikS>gikSy,
prvarghik<varghiky=1ss=1sIvarghiks<varghiksy

where Igiks>giksy, is an indicator variable mapping success if giS has higher genotypic value than giS in the harvest k, or failure otherwise; and I(var[ghik]s < var[ghik]s|y) is another indicator variable mapping success if ghik has lower variance than ghik in harvest k, or failure otherwise.

Stability in the frequentist context

To compute the stability, we first fitted the following frequentist model (F-DG):

y=1μ+X1r+X2h+Z1g+Z2gh+Z3p+e

where y is the vector of phenotypic data, 1 is a vector of ones, μ is the intercept of the model, r is the vector of repetition effects (assumed to be fixed), h is the vector of harvest effects (assumed to be fixed), g is the vector of genotypic effects (assumed to be random) gN0,Iσg2, gh is the vector of the effects of the genotype-by-harvest interaction (assumed to be random) ghN0,Iσgh2, p is the vector of permanent environmental effect (assumed to be random) pN0,Iσp2, and e is the vector of residues associated with phenotypic observations (random) [eN(0,R)], where R is a residual covariance matrix. The capital letters X1 and X2 refer to the incidence matrix for the fixed effects, and Z1,Z2 and Z3 are the incidence matrix for the random effects of the respective effects.

We estimated the variance components and predicted the genetic values using the residual maximum likelihood–REML [20], and the best linear unbiased predictor–BLUP [21], respectively. The significance tests of the random effects were verified via likelihood ratio test (LRT) [22].

A concept analogous to stability called “persistence” is used by breeders of perennial forages and refers to the ability to survive and keep producing dry matter for long periods [23, 24]. In perennial fruit plants, this concept can be readjusted as the ability to maintain a high fruit yield for several years [2]. Therefore, persistence is analogous to an ecological stability of the genotypes (Pi) based on the distance between each genotype in relation to the ideotype. The ideotype (gmax) was defined as the maximum genotypic value estimated on each harvest. We used the following expression to estimate the persistence [24]:

Pi=1k=112gigmax2i=1241k=112gigmax2

We compared the genotype ranking of the Bayesian probabilistic model and persistence via the frequentist model using Spearman correlation, following the expression below [25]:

ρ=16i=1nd2nn21

where ρ is the Spearman correlation, d is the difference between the rank positions of the genotypes in each methodology, and n is the number of genotypes.

We performed the analysis using R software environment, version 4.2.1 [26]. The Bayesian models were fitted using the probabilistic programming language Stan [27] from the rstan package [16], and the ProbBreed R package [17]. We fitted the linear mixed models using ASReml-R (version 4.1) [28].

Results

Probability of superior performance of genotypes

The Bayesian models (B-ID and B-DG) displayed a mean value of the statistic R^ close to 1, indicating strong convergence of the model parameters (Table 2). Notably, among these models, the B-DG model exhibited the best fit, as evidenced by its lower WAIC2 value (Table 2). Note how the density of the data generated by B-DG model follows the same trend as the density of the real data. This indicates the model’s effectiveness in replicating the distribution of observed data through the generated data, highlighting its reliability in capturing the underlying patterns (Fig 1A). Considering the findings from the B-DG, the posterior distribution of genotypic values of the 24 genotypes exhibited a variable overlapping pattern among their highest posterior density intervals (Fig 1B).

Table 2. Comparative statistics of Bayesian models.

Predictive a posteriori verification statistic Homogeneous residual variance model Heterogeneous (diagonal) residual variance model
WAIC2 8420.87 6899.61
R^ 1.00 0.99

Where WAIC2 is the Watanabe–Akaike information criterion, and R^ is the potential scale reduction factor.

Fig 1.

Fig 1

Bayesian distribution of the observed and generated data of the Tahiti acid lime dataset (A). Caterpillar plot of the genotypic posterior effects (and their 95% and 97.5% HPDs, represented by the thick and thin lines, respectively) of 24 genotypes of posterior effects (B). Marginal probability of superior performance of the 24 genotypes (C). Probability of superior stability of the 24 genotypes (D).

Genotypes G15, G4, G18 and G11 presented the highest posterior genotypic values (Fig 1B) and the highest probability of superior performance (Fig 1C). Genotypes G15, G4, G18, G11, G3, G23, G22, G1, G19, G14, G2, G13 and G16 were selected at least in a few samples, while the remaining 11 genotypes did not appear among the selected in any sample (Fig 1C). Genotypes G15 and G4 offers low risk of bad performance if selected (probability of superior performance equal to or higher than 75%). Genotypes G24 and G7 had the highest probability of superior stability, meaning that they have the less variable performance across harvests (Fig 1D).

The pairwise comparison graph presents two symmetrical sides, which indicate the probability of success (lower diagonal) and failure (upper diagonal) of the genotypes in the x-axis being superior to the ones of the y-axis (Fig 2A). G15, for example, has a high probability of beating all genotypes. On the other hand, G9 is beaten by all its peers, except for G21, which wins three-thirds of the time. The greenish color indicates genotypes that tie in their performance (probability close to 50%), like G18 and G11, G2 and G14, and G10 and G24 (Fig 2A).

Fig 2.

Fig 2

Pairwise probability of superior performance among genotypes (A). Probabilities of superior performance within environments (B).

The probabilities of a given genotype belonging to the group of selected ones in each harvest varies for high performance genotypes (Fig 2B). Genotypes G15, G4, G18, and G11 consistently displayed probabilities exceeding 50% across nearly all harvests. Conversely, genotypes G9 and G21 exhibited nil probabilities throughout all harvests, implying that they are not recommended due to their consistently poor performance (Fig 2B). According to the probability of superior performance within the harvests, high production by certain genotype in one harvest did not guarantee the same level of performance in subsequent harvests (Supplementary material–S2 Fig).

Probability of genotypic stability

The probability of encountering a Tahiti acid lime genotype with minimal variation for stability was generally low. Only G24 exhibited values exceeding 0.3, indicating it to be the most stable genotype among those under evaluation (Fig 1D). The frequentist model (F-DG) demonstrated the significance of GHI according to the LRT. Also, the variance components exhibited similar magnitudes for both models (Table 3). The σg2overshadowed the σgh2 and σp2, with values of 2.21 for the F-DG model and 2.44 for the B-DG. The residual variance displayed varying values across different harvests, ranging from 0.88 to 412.17 in the F-DG and from 0,63 to 384.17 in the B-DG (Table 3).

Table 3. Variance components estimates for a frequentist heterogeneous (diagonal) residual variance model (F-DG) and a Bayesian heterogeneous (diagonal) residual variance model (B-DG).

For the Bayesian model, it includes the corresponding lower (L) and upper (U) highest posterior density (HPD), considering a confidence level α = 0.05.

Parameter F-DG B-DG
Component L-HPD Component U-HPD
σg2 2.21 1.31 2.44 4.22
σgh2 0.81 0.44 0.76 1.09
σp2 0.24 0.11 0.26 0.42
σe2 H1 1.19 0.89 1.24 1.67
H2 0.88 0.63 0.91 1.29
H3 0.86 0.70 0.97 1.33
H4 5.60 4.48 5.88 7.69
H5 3.07 2.72 3.60 4.72
H6 4.22 3.37 4.52 5.99
H7 10.26 8.53 11.23 14.78
H8 20.17 20.57 26.71 34.32
H9 412.17 328.66 418.67 531.69
H10 136.74 113.61 146.63 186.56
H11 404.45 324.73 413.08 521.54
H12 473.61 384.17 492.71 621.24

σg2: genotypic variance, σgh2: genotypes-by-harvesters interaction, σp2: variance of the environmental permanent effect, and σe2: residual variance for the 12 harvesters (H1 to H12).

Persistence by the frequentist sense presented different results from the probability of superior stability of Bayesian models. However, in both contexts, the values were low. Genotypic persistence in the frequentist context ranged from 5.7 to 3.4 (Fig 3). Except for G3, which presented the highest value and well above the others, the other genotype values were in a range of 1.1, showing the low ability of the F-DG to distinguish the persistence of these genotypes. The correlation between the persistence rankings in the Bayesian context (Fig 1D) and those in the frequentist context (Fig 3) exhibited a coefficient of 0.69. This suggests a statistically significant correlation between the classifications provided by the two methods, considering a confidence level of α = 0.05.

Fig 3. Persistence of 24 genotypes of Tahiti acid lime via the frequentist model with diagonal residual variance.

Fig 3

Discussion

The consideration of the GHI is very important in the genetic evaluation of perennial species. This is because gene expression varies in response to environmental factors across different harvests [29]. The model selection based on WAIC2, coupled with the observed increase in residual variance over successive harvests, provides robust evidence in favor of the suitability of the heterogeneous residuals model for data fitting over the homoscedastic model. These findings underscore the critical importance of appropriate modeling and accounting for diverse sources of variation in repeated measures datasets.

Comparing the genotypic values associated with a posteriori probability ensures greater confidence in the analysis of the performance of the Tahiti acid lime genotypes. Bayesian probability measures offer breeders the opportunity to delve into the probability of a particular genotype surpassing others, including scenarios where a candidate genotype may outperform a widely adopted cultivar [11]. This probability-based approach aids decision-making, especially when the difference between genotypes’ performance is small. These probabilities are dependent on the selection intensity, a value that is often predefined in breeding programs, depending on the stage [30]. Thence, probabilities are an intuitive metric and offer an enhanced reliability for recommendations, since it provides information about the risks. Its application simplifies decision-making processes and opens avenues for application in various domains beyond plant breeding [11]. Defining a probability value as a threshold would be very useful for practical purposes, as it would make it easy to classify comparisons as significative or non-significative. However, due to the different selective intensities that can be employed, and the diversity of selection candidates, the threshold depends on each reality and dataset the analysis is adjusted for.

Genotypes with the highest probability of superior performance, namely G15 (Iconha x Citrandarin riverside), G4 (Itarana x Citrumelo swingle), G18 (Bearss Lime x Citrandarin riverside), and G11 (BRS Passos x Citrumelo swingle), emerged as strong candidates for recommendation. These genotypes, as determined through probabilistic methods, possess alleles that impart adaptation to the changes in environmental conditions encountered through the harvests, maintaining consistently good performance. Indeed, the probability of superior performance is a measure of stability in an agronomic sense. The presence of GHI imposes changes for the selection of superior genotypes based on a single or a few harvests. Therefore, considering the genotype`s performance across multiple harvests is advisable for making well-informed decisions when selecting genetically superior candidates.

Initially employed in forage species to assess the maintenance of productivity levels through multiple cuts [24], the concept of persistence is also relevant in the context of perennial fruit crops [31], given the perennial behavior of both. We can make a parallel between persistence and the probability of superior stability, as both represent ecological stability, i.e., invariance of performance. In the Bayesian framework, the probability of being selected among the four most stable genotypes was, in general, low (values below 0.4). This metric had a 50% agreement rate in identifying the top four most persistent genotypes with the frequentist persistence. Both approaches selected G24 and G3 among the four most persistent genotypes.

Certain advantages of the Bayesian model deserve attention. The incorporation of priors enhances confidence in selecting materials with varying levels of persistence [11, 32]. Furthermore, Bayesian models offer the advantage of obtaining variance components with associated high probability density intervals. These credibility intervals provide a more intuitive means of quantifying component uncertainty. Also, from an asymptotic perspective, Bayesian credibility intervals outperform frequentist confidence intervals [18], since frequentist confidence intervals may prove inaccurate for small or moderate sample sizes and may, in certain instances, fail to converge to the true parameter value as the sample size increases [33]. Likewise in mixed model, Bayesian models work well in common situation of plant breeding, such as unbalanced data, heterogeneous residual variance [6, 34, 35].

Conclusion

By applying probabilistic Bayesian models in Tahiti acid lime in the genetic evaluation, we estimated the probability of superior performance of a genotype and the pairwise probabilities of superior performance between genotypes for both across and within harvests. Genotypes G15, G4, G18 and G11 were considered superior, and genotypes G24, G7, G13 and G3 were considered the most stable ones. Therefore, we believe that Bayesian probabilistic models can assist to more accurate recommendation in perennial fruit crops evaluated along many harvests, since it allows a more direct and precise interpretation of the performance and persistence of the candidate’s genotypes.

Supporting information

S1 Fig. Climatic data of precipitation (mm) and temperature (°C) from July 2017 to July 2020 in the field trial location.

(TIF)

pone.0299290.s001.tif (407.4KB, tif)
S2 Fig. Pairwise probability of superior performance among genotypes within harvest.

(TIF)

pone.0299290.s002.tif (8.3MB, tif)

Acknowledgments

We appreciate the Federal University of Espírito Santo, the Brazilian Agricultural Research Corporation, and the Bello Fruit Company for making possible the execution of these experiments. Also, we thank the Federal University of Viçosa, which provided infrastructure and human training for data analysis.

Data Availability

The data relevant to this study are available from GitHub at: https://github.com/malikouskirg/PONE-D-23-38484.

Funding Statement

This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) - Finance Code 001, Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), Fundação de Amparo à Pesquisa do Estado do Espírito Santo (FAPES), and Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP). FMF was supported by FAPESP (São Paulo Research Foundation, Grant 2023/04881-3), and LLB was supported by CNPq (Research Productivity Fellowship, Grant 310610/2021-4).

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Decision Letter 0

Mehdi Rahimi

26 Dec 2023

PONE-D-23-38484Application of Bayesian probabilistic models for recommendation of ‘Tahiti’ acid lime using longitudinal dataPLOS ONE

Dear Dr. Bhering,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

**********

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Reviewer #1: No

Reviewer #2: Yes

**********

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Reviewer #2: Yes

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Reviewer #1: The authors aimed to use probability concepts to recommend Tahiti acid lime genotypes. To do this, they evaluated, in a field trial, 24 combinations of rootstock and scion of Tahiti acid lime regarding fruit yield, over 12 harvests. They concluded that the Bayesian analysis outperformed the frequentist analysis. This work appears to be the first to use probability concepts to recommend cultivars of fruit trees and has merit for publication in PLOS ONE.

Random regression models are the state-of-the-art for the analysis of repeated measures. They are parsimonious and deal with genotype-harvest interactions. In this work, Bayesian and frequentist random regression models could provide better results?

See more comments in the attached file.

Reviewer #2: In my point of view, the article is comprehensive in terms of statistical analyses and arguments that support its hypotheses. The writing is well-structured, lacking only some additional information about the cultivation of the 'Tahiti' acid lime, which would further reinforce the importance of the article. Therefore, my recommendation is to accept it for publication with minor revisions.

**********

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Reviewer #1: No

Reviewer #2: No

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Attachment

Submitted filename: PONE-D-23-38484-R0.pdf

pone.0299290.s003.pdf (1.7MB, pdf)
Attachment

Submitted filename: PONE-D-23-38484_reviewed.pdf

pone.0299290.s004.pdf (1.7MB, pdf)
PLoS One. 2024 Mar 5;19(3):e0299290. doi: 10.1371/journal.pone.0299290.r002

Author response to Decision Letter 0


27 Jan 2024

To the Editorial Office of PLOS ONE

Dear Academic Editor Mehdi Rahimi, Ph.D.

We are sending the revised version of the manuscript PONE-D-23-38484. The changes based on the reviewers’ comments, were highlighted in blue in the manuscript.

Each observation from the reviewers and the editor has been addressed in the file "Response to Reviewers."

Below, we paste the content present in the "Response to Reviewers" file.

To the Editorial Office of PLOS ONE

Dear Academic Editor Mehdi Rahimi, Ph.D.

We are sending the revised version of the manuscript PONE-D-23-38484. The changes, based on the reviewers’ comments, were highlighted in the reviewed manuscript file in blue color. We would like to thank the reviewers for the excellent contributions to the improvement of this manuscript. Below we responded individually to each comment made in the decision letter and reviewed manuscript. We are available for any further questions.

Academic Editor

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming.

The authors are grateful for this observation from the Editor. A second check on the provided links was conducted, and a few changes were made, e.g., titles of captions and figures were bolded, and minor alterations were made to the acknowledgments. Other than that, everything is in accordance with the established norms. Changes are highlighted in blue in the Manuscript.

2. In your Methods section, please provide additional information regarding the permits you obtained for the work. Please ensure you have included the full name of the authority that approved the field site access and, if no permits were required, a brief statement explaining why.

Thank you for this observation. We have added more information about the location of the experiment. This partnership does not present any conflict of interest, and there is no formal documentation regarding it. The company provides the site for carrying out the experiment and labor for managing the plants. In return, the university can generate fruit cultivation information for all the farmers in the region, without any right to exclusivity, and the transference of information takes place through the execution of field days held on the Bello Fruit farm.

3. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match.

When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section.

Thank you and sorry for this misunderstanding. We have revised the Acknowledgments section, removing any issues of funding. However, we reiterate the need to disclose this information as funding agencies commonly require acknowledgment of their contributions in papers and abstracts. We added the information previously contained in the Acknowledgments to the cover letter (file named “cover_letter_financial_disclosure_changes.pdf”) so that it may be placed in the appropriate section on the published paper.

4. Thank you for stating the following in the Acknowledgments Section of your manuscript: "The authors are grateful to the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Capes, Code 001), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (Fapemig) and Fundação de Amparo à Pesquisa do Estado do Espírito Santo (Fapes). Filipe Manoel Ferreira was supported by FAPESP (São Paulo Research Foundation, Grant 2023/04881-3),"

We note that you have provided funding information that is not currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form.

Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: "“The authors received no specific funding for this work.”

Please include your amended statements within your cover letter; we will change the online submission form on your behalf.

Thank you for the observation. As this point is related to the previous matter, it has already been addressed with the modifications mentioned in topic 3. We have modified the Acknowledgments section and have moved the paragraph mentioning the Funding information to the cover letter (file named “cover_letter_financial_disclosure_changes.pdf”) so that it may be placed in the appropriate section on the published paper.

5. When completing the data availability statement of the submission form, you indicated that you will make your data available on acceptance. We strongly recommend all authors decide on a data sharing plan before acceptance, as the process can be lengthy and hold up publication timelines. Please note that, though access restrictions are acceptable now, your entire data will need to be made freely accessible if your manuscript is accepted for publication. This policy applies to all data except where public deposition would breach compliance with the protocol approved by your research ethics board. If you are unable to adhere to our open data policy, please kindly revise your statement to explain your reasoning and we will seek the editor's input on an exemption. Please be assured that, once you have provided your new statement, the assessment of your exemption will not hold up the peer review process.

We appreciate the comment and have now added the complete dataset that was used for the analyses to GitHub repository with public access. Consequently, we have inserted the link in the Supporting Information section. All authors of the manuscript have agreed to publish the full dataset to the scientific community. The link to access the dataset is:

https://github.com/malikouskirg/PONE-D-23-38484

6. Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

Thank you for the information; we have reviewed the reference list, and everything is in accordance with the standards.

Reviewer #1

• Optimization of Tahiti Acid Lime Cultivars Recommendation Through Probability Concepts

Thank you for your observation. We believe that the name suggestion given by the reviewer is very relevant. We like the idea of adding the term "Bayesian" term to the title. We made a small adjustment to the reviewer’s idea. Thus, the new title is " Recommendation of ‘Tahiti’ acid lime cultivars through Bayesian probability models".

• Tahiti Acid Lime Breeding

Thank you for the suggestion. We adopted this short title.

• any breeding program

We made the recommend. Thank you.

• a

Thank you, we add the letter “a” in the sentence “The objective of our study was to evaluate the use of a Bayesian probabilistic model”.

• The objective of our study was to evaluate the use of Bayesian probabilistic models for the recommendation of superior and persistent genotypes of ‘Tahiti’ acid lime evaluated in 12 harvests.

Thank you for your suggestion. We have removed the "s" from the word "models." However, we believe that keeping the word "persistent" would be important, as in addition to performance, the objective of this work was also focused on persistence.

• Genotypes G15, G4, G18, and G11 emerged as the most superior in performance, whereas G24, G7, G13, and G3 were identified as the most stable.

Thank you for your suggestion. However, we believe that this information is very important in practical terms, as it concerns the recommendation of genotypes. We have provided details about the coding in the Methods section, and therefore, the authors feel it is better to retain this information.

• Usefulness

Thank you, we changed for the suggested word.

• the fruit trees cultivars recommendation

We made the recommend change. Thank you.

• In the presence of complex GHI, breeders must consider the productivity and the stability of the genotypes over harvests [3–5].

We made the recommend change. Thank you.

• To address the common issue of non-orthogonal longitudinal data, resulting from the interdependence…. Despite considerable progress in 'Tahiti' acid

lime breeding, straightforward methodologies for exploring GHI and identifying high performance and stable genotypes are yet to be established.

Thank you for the suggestion. We removed these parts of the text.

• optimize the Tahiti acid lime cultivars recommendation, since it uses probability concepts.

We made the recommend change. Thank you.

• Dias et al. [10] proposed to use probability concepts from Bayesian models to assist in the selection of genotypes that gather favorable alleles for performance and stability across environments.

We appreciate your suggestion and have adopted it in part, as retaining the information on alleles favorable for performance and stability is relevant. This is the genetic definition of the GHI interaction; thus, the text has been revised accordingly.

“Dias et al. [11] proposed to use Bayesian probability concepts to assist in the selection of genotypes that gather favorable alleles for performance and stability across environments and harvesters.”

• This methodology has been proposed for the multiple location context. Nevertheless, we believe that the same ideas are valid for the multi-harvest scenario

Thank you for the suggestion. We removed these parts of the text.

• The objective of our study was to evaluate the use of a Bayesian probabilistic model for the recommendation of superior and persistent genotypes of

Tahiti acid lime evaluated in 12 harvests.

Thank you. We have adopted your suggested objective.

• statistical

Thank you. We added this part to the manuscript.

• was

We made the recommend change. Thank you.

• Codes

We made the recommend change. Thank you.

• method

We made the recommend change. Thank you.

• How the permanent environmental effect was accounted here? Based on the results presented in Table 3, it seems that this effect was fitted, however it was not described here.

We appreciate the comment and apologize for the oversight. Indeed, there is a permanent environmental effect, which we have now correctly included in the model. With this adjustment, we identified values in the table that were switched.

• N(0, σ[α])? N(0, σ[.]), where [.] can be μ, r, ..., e.

We revised the notations.

• We used a Monte Carlo Hamiltonian algorithm (the Hamiltonian Monte Carlo algorithm) in four Markov chains with 4000 samples (thin?) and 50% burn-in.

Thank you for the observation; we have revised the text to "the Hamiltonian Monte Carlo algorithm" and have added the thinning information of the model, which in this case was the default of the "rstan" package, 1.

• Package

Thank you. We changed the word “software” to “package”.

• Proportion

Thank you. We changed the word “intensity” to “proportion”.

• Effects

Thank you. We changed the word “effect” to “effects”.

• Was.. Were

Thank you. We changed the words “is/are” to “was/were”.

• Comparative statistics of Bayesian models using the 'Tahiti' acid file dataset.

Thank you for the suggestion; we have removed the term "using the Tahiti acid file dataset" from all captions, table titles, and figures where this text appeared.

• ‘Tahiti’

Thank you for your observation. In scientific articles, the word "Tahiti" is found both with and without quotation marks. Following the reviewer's suggestion, and after grammatical research, we decided to adopt the reviewer's advice to remove the quotation marks from the name Tahiti, resulting in 'Tahiti lime'. We have carried out the removal throughout the text.

• is the genotypes-by-harvesters interaction… is the variance of the environmental permanent effect

Thank you for the suggestion. We have changed the caption of Table 3 to the form proposed by the reviewer.

• by

Thank you. We changed the word “in” to “by”.

• important

Thank you. We changed the word “informative” to “important”.

• genetic evaluation

Thank you. We changed the word “assessment” to “genetic evaluation”.

• with heterogeneous residuals

Thank you. We changed the word “heterogeneous model” to “heterogeneous residuals model”.

• repeated measures

Thank you. We changed the word “longitudinal” to “repeated measures”.

• [https://doi.org/10.1007/s11295-023-01596-9]

Thank you. We have added this citation in the mentioned part of the text, as well as in the references.

• By applying probabilistic Bayesian models in longitudinal `Tahiti` acid lime dataset

Thank you. We have removed the terms "longitudinal" and “dataset” from the conclusion, as well as the previously mentioned quotation marks from 'Tahiti' throughout the text.

• This information is irrelevant to the reader.

As previously mentioned, the authors believe that maintaining the information about the recommended genotypes regarding performance and persistence is important in practical terms, since these materials are publicly available to farmers. This work, in addition to its statistical nature, is also aimed at assisting in the recommendation for planting of the Tahiti acid lime.

Reviewer #2

• I suggest adding brief information about the importance of the Tahitian lime culture, emphasizing the significance of work.

Thank you for your suggestion. Indeed, despite the statistical nature of the work, the authors believe that the importance of the crop should be highlighted in the introductory parts of the article, therefore, we have emphasized the importance of the culture in the initial sentences of the Abstract section.

• Highlight information about the juvenile phase of the culture. For example, how long does it last?

Thank you for the suggestion. The juvenile phase of the Tahiti acid lime can take from 3 to 5 years; we have added this information to the aforementioned sentence, highlighted in blue in the reviewed manuscript.

• Highlight to a non-expert what GHI means.

The authors thank you for your suggestion. We have added an introductory sentence that briefly defines the GHI interaction, with citation.

“The GHI in perennial species refers to the variation in gene expression and, consequently, the phenotypic traits of a plant due to the different environmental conditions and agricultural practices that occur in each planting cycle [2]”

• I wouldn't say they are mandatory, just that they are important for the recommendation and selection of superior individuals.

Thank you. We changed the word “mandatory” to “important”.

• I believe that studies have already been conducted exploring the performance and stability of genotypes. In this case, what is not yet present in the literature are works related to providing probabilities associated with these metrics. I suggest modifying this part.

We appreciate your suggestion. As a suggestion from the previous reviewer, we have removed that part of the manuscript.

• In a specific period of time or under certain adverse conditions... I suggest adding the part about climate challenges. Given the current variations in temperature and precipitation, it is necessary to select genotypes that can cope well with these uncertainties.

Thank you. Indeed, adding that part about climate change and its influence on breeding is very enriching for the work, thus we have included it in the introduction with a citation.

• and harvesters (measurements)

Thank you. We added this part in the text.

• plot unit mean

Thank you. We added this part in the text.

• ranges.

Thank you. We changed the word “columns” to “ranges”.

• Explain in more detail the reasons for having four selected genotypes

Thank you for your comment. The choice of 4 genotypes as selected was determined by the authors due to the fact that in a breeding program, there are typically several materials recommended, and not just one. Moreover, on the same farm, it's sometimes important to have a diversity of cultivars due to the susceptibility of some to biotic or abiotic factors. We have briefly added this explanation in the methods section.

• I believe that adding the calculation of the ratio between events with lower GHI and the total number of events would be interesting, as mentioned for performance.

We appreciate the suggestion. We have modified the text to make it similar to the definition of probability for performance. Below is the part added to the revised manuscript.

“Following the same idea described in the last paragraph, calculated the probability of a given candidate belonging to the subset of the top four genotypes with smaller ...”

• Explain in more detail the reason why knowing the genotypes in each harvest is important.

The authors appreciate your suggestion. However, we were unable to include that part in the materials and methods, as we believe that throughout the manuscript we aim to show that the objective of the work is performance and stability. In this case, evaluating performance at each separate harvest is a form of stability assessment, which is highlighted throughout the manuscript.

• Describe the Spearman correlation model as well as the citation.

We appreciate the suggestion. We have added the Spearman correlation estimator, as well as the original citation of the work.

• Indicate what is the probability value that this range varied (i.e., present the minimum and maximum values).

Thank you for your suggestion. However, since the figure does not show the exact frequentist persistence value, it is not possible to extract the minimum and maximum information from it. Nonetheless, the information on the range gives an idea of magnitude, and it is only from this that one can see that the variation was not so pronounced for the mentioned genotypes.

• In the materials and methods section, describe the correlation model and also cite relevant literature.

Thank you. As mentioned in a previous topic, we have added the estimator, as well as the reference for the Spearman correlation.

• Additionally, state that the variation in environmental conditions throughout different harvests creates diverse environments.

Thank you for this comment.

• It would be interesting to indicate in this part of the text the correlation of this case with the existence of GHI.

Thank you for the observation, the correlation part was conducted between frequentist and Bayesian approaches; in this case, the authors believe that its linkage should be made further down in the discussion, and it was done, as you can see in the text below.

“In the Bayesian framework, the probability of being selected among the four most stable genotypes was, in general, low (values below 0.4). This metric had a 50% agreement rate in identifying the top four most persistent genotypes with the frequentist persistence. Both approaches selected G24 and G3 among the four most persistent genotypes.”

• It would be beneficial to indicate a probability value as thresholds. However, since it is still a new methodology, there may not be a well-defined value yet.

Thank you for your suggestion. Although it depends on various factors, having a threshold would be interesting. We have added the comment below in the revised manuscript.

“Defining a probability value as a threshold would be very useful for practical purposes, as it would make it easy to classify comparisons as significative or non-significative. However, due to the different selective intensities that can be employed, and the diversity of selection candidates, the threshold depends on each reality and dataset the analysis is adjusted for.”

• Certainly! It would be helpful to include the names of the genotypes here for the convenience of the readers. The format could be "GENOTYPE NAME (ENCODING)".

Thank you for your suggestion. As we defined the names of the genotypes by encoding throughout the manuscript, we changed to the suggested order, we put ENCODING (scion x rootstock).

• Because, in this case, both forage crops and fruit trees are perennials.

We appreciate your comment. We added the term "given the perennial behavior of both" to the mentioned sentence in the text.

Attachment

Submitted filename: Response to Reviewers.pdf

pone.0299290.s005.pdf (185.8KB, pdf)

Decision Letter 1

Mehdi Rahimi

8 Feb 2024

Recommendation of Tahiti acid lime cultivars through Bayesian probability models

PONE-D-23-38484R1

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Acceptance letter

Mehdi Rahimi

22 Feb 2024

PONE-D-23-38484R1

PLOS ONE

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on behalf of

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Associated Data

    This section collects any data citations, data availability statements, or supplementary materials included in this article.

    Supplementary Materials

    S1 Fig. Climatic data of precipitation (mm) and temperature (°C) from July 2017 to July 2020 in the field trial location.

    (TIF)

    pone.0299290.s001.tif (407.4KB, tif)
    S2 Fig. Pairwise probability of superior performance among genotypes within harvest.

    (TIF)

    pone.0299290.s002.tif (8.3MB, tif)
    Attachment

    Submitted filename: PONE-D-23-38484-R0.pdf

    pone.0299290.s003.pdf (1.7MB, pdf)
    Attachment

    Submitted filename: PONE-D-23-38484_reviewed.pdf

    pone.0299290.s004.pdf (1.7MB, pdf)
    Attachment

    Submitted filename: Response to Reviewers.pdf

    pone.0299290.s005.pdf (185.8KB, pdf)

    Data Availability Statement

    The data relevant to this study are available from GitHub at: https://github.com/malikouskirg/PONE-D-23-38484.


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