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Abstract BackgroundStalk lodging (the premature breaking of plant stalks or stems prior to harvest) is a persistent agricultural problem that causes billions of dollars in lost yield every year. Three-point bending tests, and rind puncture tests are common biomechanical measurements utilized to investigate crops susceptibility to lodging. However, the effect of testing rate on these biomechanical measurements is not well understood. In general, biological specimens (including plant stems) are well known to exhibit viscoelastic mechanical properties, thus their mechanical response is dependent upon the rate at which they are deflected. However, there is very little information in the literature regarding the effect of testing rate (aka displacement rate) on flexural stiffness, bending strength and rind puncture measurements of plant stems. ResultsFully mature and senesced maize stems and wheat stems were tested in three-point bending at various rates. Maize stems were also subjected to rind penetration tests at various rates. Testing rate had a small effect on flexural stiffness and bending strength calculations obtained from three-point bending tests. Rind puncture measurements exhibited strong rate dependent effects. As puncture rate increased, puncture force decreased. This was unexpected as viscoelastic materials typically show an increase in resistive force when rate is increased. ConclusionsTesting rate influenced three-point bending test results and rind puncture measurements of fully mature and dry plant stems. In green stems these effects are expected to be even larger. When conducting biomechanical tests of plant stems it is important to utilize consistent span lengths and displacement rates within a study. Ideally samples should be tested at a rate similar to what they would experience in-vivo. 

Stalk lodging in the monocot Zea mays is an important agricultural issue that requires the development of a genome-to-phenome framework, mechanistically linking intermediate and high-level phenotypes. As part of that effort, tools are needed to enable better mechanistic understanding of the microstructure in herbaceous plants. A method was therefore developed to create finite element models using CT scan data for Zea mays. This method represents a pipeline for processing the image stacks and developing the finite element models. 2-dimensional finite element models, 3-dimensional watertight models, and 3-dimensional voxel-based finite element models were developed. The finite element models contain both the cell and cell wall structures that can be tested in silico for phenotypes such as structural stiffness and predicted tissue strength. This approach was shown to be successful, and a number of example analyses were presented to demonstrate its usefulness and versatility. This pipeline is important for two reasons: (1) it helps inform which microstructure phenotypes should be investigated to breed for more lodging-resistant stalks, and (2) represents an essential step in the development of a mechanistic hierarchical framework for the genome-to-phenome modeling of herbaceous plant stalk lodging. 

This study sought to better understand how time of day (ToD) or turgor pressure might affect the flexural stiffness of sweet sorghum stalks and potentially regulate stalk lodging resistance. Stalk flexural stiffness measured across a 48 h period in 2019 showed a significant diurnal association with leaf water potential and stalk flexural stiffness. While the correlation between stalk flexural stiffness and this proxy for internal turgor status was statistically significant, it only accounted for roughly 2% of the overall variance in stiffness. Given that turgor status is a dynamic rather than fixed physiological variable like the cellular structure, these data suggest that internal turgor plays a small yet significant role in influencing the flexural stiffness of fully mature stalks prior to a stalk lodging event. The association was assessed at earlier developmental stages across three distinct cultivars and found not to be significant. Panicle weight and stalk basal weight, but not stalk Brix or water content, were found to be better predictors of stalk flexural stiffness than either ToD or turgor status. Observation across three cultivars and four distinct developmental stages ranging from the vegetative to the hard-dough stages suggests that stalk flexural stiffness changes significantly as a function of time. However, neither ToD nor turgor status appear to meaningfully contribute to observed variations in stalk flexural stiffness in either individual stalks or across larger populations. As turgor status was not found to meaningfully influence stalk strength or flexural stiffness at any developmental time point examined in any of the three sweet sorghum cultivars under study, turgor pressure likely offers only inconsequential contributions to the biomechanics underlying sweet sorghum stalk lodging resistance. 

Stalk lodging (structural failure crops prior to harvest) significantly reduces annual yields of vital grain crops. The lack of standardized, high throughput phenotyping methods capable of quantifying biomechanical plant traits prevents comprehensive understanding of the genetic architecture of stalk lodging resistance. A phenotyping pipeline developed to enable higher throughput biomechanical measurements of plant traits related to stalk lodging is presented. The methods were developed using principles from the fields of engineering mechanics and metrology and they enable retention of plant-specific data instead of averaging data across plots as is typical in most phenotyping studies. This pipeline was specifically designed to be implemented in large experimental studies and has been used to phenotype over 40,000 maize stalks. The pipeline includes both lab- and field-based phenotyping methodologies and enables the collection of metadata. Best practices learned by implementing this pipeline over the past three years are presented. The specific instruments (including model numbers and manufacturers) that work well for these methods are presented, however comparable instruments may be used in conjunction with these methods as seen fit. • Efficient methods to measure biomechanical traits and record metadata related to stalk lodging. • Can be used in studies with large sample sizes (i.e., > 1,000). 

Abstract This study presents a methodology for a high-throughput digitization and quantification process of plant cell walls characterization, including the automated development of two-dimensional finite element models. Custom algorithms based on machine learning can also analyze the cellular microstructure for phenotypes such as cell size, cell wall curvature, and cell wall orientation. To demonstrate the utility of these models, a series of compound microscope images of both herbaceous and woody representatives were observed and processed. In addition, parametric analyses were performed on the resulting finite element models. Sensitivity analyses of the structural stiffness of the resulting tissue based on the cell wall elastic modulus and the cell wall thickness; demonstrated that the cell wall thickness has a three-fold larger impact of tissue stiffness than cell wall elastic modulus. 

Context: Stalk lodging causes up to 43 % of yield losses in maize (Zea mays L.) worldwide, significantly worsening food and feed shortages. Stalk lodging resistance is a complex trait specified by several structural, material, and geometric phenotypes. However, the identity, relative contribution, and genetic tractability of these intermediate phenotypes remain unknown. Objective: The study is designed to identify and evaluate plant-, organ-, and tissue-level intermediate phenotypes associated with stalk lodging resistance following standardized phenotyping protocols and to understand the variation and genetic tractability of these intermediate phenotypes. Methods: We examined 16 diverse maize hybrids in two environments to identify and evaluate intermediate phenotypes associated with stalk flexural stiffness, a reliable indicator of stalk lodging resistance, at physiological maturity. Engineering-informed and machine learning models were employed to understand relationships among intermediate phenotypes and stalk flexural stiffness. Results: Stalk flexural stiffness showed significant genetic variation and high heritability (0.64) in the evaluated hybrids. Significant genetic variation and comparable heritability for the cross-sectional moment of inertia and Young’s modulus indicated that geometric and material properties are under tight genetic control and play a combinatorial role in determining stalk lodging resistance. Among the twelve internode-level traits measured on the bottom and the ear internode, most traits exhibited significant genetic variation among hybrids, moderate to high heritability, and considerable effect of genotype × environment interaction. The marginal statistical model based on structural engineering beam theory revealed that 74–80 % of the phenotypic variation for flexural stiffness was explained by accounting for the major diameter, minor diameter, and rind thickness of the stalks. The machine learning model explained a relatively modest proportion (58–62 %) of the variation for flexural stiffness. 

Abstract Background Measurements of rind and culm thickness and stem radius/diameter are important to biomechanical, ecological and physiological plant studies. However, many methods of measuring rind thickness and diameter are labor intensive and induce plant fatality. A novel rind puncture methodology for obtaining measurements of rind thickness and diameter has been developed. The suitability of the new method for implementation in plant studies is presented. Results The novel rind puncture technique was used to obtain measurements of rind thickness and diameter for samples of Poison Hemlock ( Conium maculatum ). The rind puncture measurements were strongly correlated with caliper measurements (R 2  > 0.97) and photographic image analysis measurements (R 2  > 0.84). The capacity for high throughput measurements using the rind puncture technique was determined to exceed that of caliper measurements and image analysis techniques. Conclusions The rind puncture technique shows promise as a high throughput method for determining rind thickness and diameter as it is cost effective and non-lethal. The authors are currently working to develop a custom handheld apparatus to allow the novel rind puncture method to be used in field work. High throughput field-based measurements of rind thickness and diameter are needed to help address the problem of stalk lodging (failure of grain crops to remain upright until harvest).