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. 2023 Jul 20:14:1224268.
doi: 10.3389/fpls.2023.1224268. eCollection 2023.

A high-throughput phenotyping assay for precisely determining stalk crushing strength in large-scale sugarcane germplasm

Fumin Ma  1 , Yinjuan Shen  1   2 , De Su  1 , Muhammad Adnan  1 , Maoyao Wang  1 , Fuhong Jiang  1 , Qian Hu  1 , Xiaoru Chen  1 , Guanyong He  1 , Wei Yao  1 , Muqing Zhang  1 , Jiangfeng Huang  1
Affiliations

A high-throughput phenotyping assay for precisely determining stalk crushing strength in large-scale sugarcane germplasm

Fumin Ma et al. Front Plant Sci. .

Abstract

Sugarcane is a major industrial crop around the world. Lodging due to weak mechanical strength is one of the main problems leading to huge yield losses in sugarcane. However, due to the lack of high efficiency phenotyping methods for stalk mechanical strength characterization, genetic approaches for lodging-resistant improvement are severely restricted. This study attempted to apply near-infrared spectroscopy high-throughput assays for the first time to estimate the crushing strength of sugarcane stalks. A total of 335 sugarcane samples with huge variation in stalk crushing strength were collected for online NIRS modeling. A comprehensive analysis demonstrated that the calibration and validation sets were comparable. By applying a modified partial least squares method, we obtained high-performance equations that had large coefficients of determination (R2 > 0.80) and high ratio performance deviations (RPD > 2.4). Particularly, when the calibration and external validation sets combined for an integrative modeling, we obtained the final equation with a coefficient of determination (R2) and ratio performance deviation (RPD) above 0.9 and 3.0, respectively, demonstrating excellent prediction capacity. Additionally, the obtained model was applied for characterization of stalk crushing strength in large-scale sugarcane germplasm. In a three-year study, the genetic characteristics of stalk crushing strength were found to remain stable, and the optimal sugarcane genotypes were screened out consistently. In conclusion, this study offers a feasible option for a high-throughput analysis of sugarcane mechanical strength, which can be used for the breeding of lodging resistant sugarcane and beyond.

Keywords: NIRS; crushing strength; lodging resistance; mechanical strength; sugarcane.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Laboratory analytical method for stalk crushing strength determination in sugarcane. (A) Schematic diagram of sugarcane crushing strength determination. (B, C) Morphological changes of internode at the moment of cracks appeared (B) and complete ruptured (C), bars = 3 cm. (D) Compression force curve with multiple peaks for crushing strength determination. S1-S3: three compressive states (elasticity, yield, compaction strengthening); Red dots represent the detected peaks; X1 and X2 represent the key steps as described in B and C, respectively; (E) Comparative analysis of each detected peaks in the compressive force curves in ten representative sugarcane samples. RSD: relative standard deviation. (F) Comparative analysis of the first peak between two groups of ten representative sugarcane genotypes. Different letters indicated statistically significant differences among these genotypes via one-way ANOVA and LSD test at α ≤ 0.05 level; *** indicated statistically significant different between the two groups at p < 0.001 level. H1-H5 and L1-L5 represented five sugarcane genotypes with high (H) and low (L) mechanical strength, respectively. Each sample contained three biological replicates.
Figure 2
Figure 2
Diversity of stalk crushing strength (SCS) in collected sugarcane samples. (A) Venn diagram of sugarcane samples collected from three identical experimental field plots. (B) Heatmap and (C) violin chart displaying the stalk crushing strengths in collected sugarcane genotypes. (D) Distribution and correlation analysis of stalk crushing strength of 306 sugarcane genotypes in three planting plots. *** indicated significant correlations at p < 0.001 level. P1-P3: three planting plots.
Figure 3
Figure 3
Characterization of near-infrared spectral in 335 sugarcane samples. (A) Original spectra of sugarcane samples in three planting plots. (B-D) Principal component analysis of NIRS data. (B) Contribution of each principal component to variable explanation. (C) Cumulative contribution of principal components to variable explanation. (D) 3D score view of sugarcane samples by PCA. P1-P3: three planting plots.
Figure 4
Figure 4
Online NIRS modeling for stalk crushing strength. (A) Distribution characteristics of calibration and validation sets. (B) Online NIRS calibration and external validation. (C) Performance of the integrative final NIRS equation.
Figure 5
Figure 5
Model-based evaluation of stalk crushing strength in sugarcane germplasm. (A) Distribution of stalk crushing strength in sugarcane population. (B) Correlation analysis of stalk crushing strength between three years. *** indicated significant correlations at p < 0.001 level. (C) Comparative analysis of stalk crushing strength in the screened sugarcane germplasm. LC/HC: representing the sugarcane samples with low and high crushing strength, respectively. Different letters indicated statistically significant differences between the groups using one-way ANOVA and LSD test at α ≤ 0.05; *** indicated statistically significant different at p < 0.001 levels, respectively.
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