Using Cameras for Precise Measurement of Two-Dimensional Plant Features: CASS.

Images are used frequently in plant phenotyping to capture measurements. This chapter offers a repeatable method for capturing two-dimensional measurements of plant parts in field or laboratory settings using a variety of camera styles (cellular phone, DSLR), with the addition of a printed calibration pattern. The method is based on calibrating the camera using information available from the EXIF tags from the image, as well as visual information from the pattern. Code is provided to implement the method, as well as a dataset for testing. We include steps to verify protocol correctness by imaging an artifact. The use of this protocol for two-dimensional plant phenotyping will allow data capture from different cameras and environments, with comparison on the same physical scale. We abbreviate this method as CASS, CAmera aS Scanner.

Tracking the Adaptation and Compensation Processes of Patients' Brain Arterial Network to an Evolving Glioblastoma.

The brain's vascular network dynamically affects its development and core functions. It rapidly responds to abnormal conditions by adjusting properties of the network, aiding stabilization and regulation of brain activities. Tracking prominent arterial changes has clear clinical and surgical advantages. However, the arterial network functions as a system; thus, local changes may imply global compensatory effects that could impact the dynamic progression of a disease. We developed automated personalized system-level analysis methods of the compensatory arterial changes and mean blood flow behavior from a patient's clinical images. By applying our approach to data from a patient with aggressive brain cancer compared with healthy individuals, we found unique spatiotemporal patterns of the arterial network that could assist in predicting the evolution of glioblastoma over time. Our personalized approach provides a valuable analysis tool that could augment current clinical assessments of the progression of glioblastoma and other neurological disorders affecting the brain.

The Influence of Marking Methods on Mobility, Survivorship, and Field Recovery of Halyomorpha halys (Hemiptera: Pentatomidae) Adults and Nymphs.

Halyomorpha halys (Stål), the brown marmorated stink bug, is an invasive and highly polyphagous insect that has caused serious economic injury to specialty and row crops in the United States and Europe. Here, we evaluated the effects of marking adult and nymphal H. halys with four different colors of fluorescent powder (Blaze Orange, Corona Pink, Horizon Blue, and Signal Green) on mobility and survivorship in laboratory bioassays. Adults and nymphs were marked using liquified fluorescent powder solutions and allowed to dry prior to bioassay. The presence of the marking solution had no significant effects on adult or nymphal mobility, adult survivorship, nymphal development, or adult flight capacity. We also evaluated the persistence of neon marker applied to the pronotum of H. halys adults and found this technique remained detectable for 2 wk under field conditions. Although both marking techniques are inexpensive, persist for ≥1 wk, and do not affect mortality, the neon marker method is more time-consuming, taking ~12 times longer to mark 50 adult H. halys compared with the liquified fluorescent powders. Thus, we would recommend using fluorescent powders for large-scale mark-release-recapture studies.

Multi-dimensional machine learning approaches for fruit shape phenotyping in strawberry.

BACKGROUND: Shape is a critical element of the visual appeal of strawberry fruit and is influenced by both genetic and non-genetic determinants. Current fruit phenotyping approaches for external characteristics in strawberry often rely on the human eye to make categorical assessments. However, fruit shape is an inherently multi-dimensional, continuously variable trait and not adequately described by a single categorical or quantitative feature. Morphometric approaches enable the study of complex, multi-dimensional forms but are often abstract and difficult to interpret. In this study, we developed a mathematical approach for transforming fruit shape classifications from digital images onto an ordinal scale called the Principal Progression of k Clusters (PPKC). We use these human-recognizable shape categories to select quantitative features extracted from multiple morphometric analyses that are best fit for genetic dissection and analysis. RESULTS: We transformed images of strawberry fruit into human-recognizable categories using unsupervised machine learning, discovered 4 principal shape categories, and inferred progression using PPKC. We extracted 68 quantitative features from digital images of strawberries using a suite of morphometric analyses and multivariate statistical approaches. These analyses defined informative feature sets that effectively captured quantitative differences between shape classes. Classification accuracy ranged from 68% to 99% for the newly created phenotypic variables for describing a shape. CONCLUSIONS: Our results demonstrated that strawberry fruit shapes could be robustly quantified, accurately classified, and empirically ordered using image analyses, machine learning, and PPKC. We generated a dictionary of quantitative traits for studying and predicting shape classes and identifying genetic factors underlying phenotypic variability for fruit shape in strawberry. The methods and approaches that we applied in strawberry should apply to other fruits, vegetables, and specialty crops.

Alteration of TAC1 expression in Prunus species leads to pleiotropic shoot phenotypes.

Prunus persica (peach) trees carrying the "Pillar" or "Broomy" trait (br) have vertically oriented branches caused by loss-of-function mutations in a gene called TILLER ANGLE CONTROL 1 (TAC1). TAC1 encodes a protein in the IGT gene family that includes LAZY1 and DEEPER ROOTING 1 (DRO1), which regulate lateral branch and root orientations, respectively. Here we found that some of the native TAC1 alleles in the hexaploid plum species Prunus domestica, which has a naturally more upright stature, contained a variable length trinucleotide repeat within the same exon 3 region previously found to be disrupted in pillar peach trees. RNAi silencing of TAC1 in plum resulted in trees with severely vertical branch orientations similar to those in pillar peaches but with an even narrower profile. In contrast, PpeTAC1 overexpression in plum led to trees with wider branch angles and more horizontal branch orientations. Pillar peach trees and transgenic plum lines exhibited pleiotropic phenotypes, including differences in trunk and branch diameter, stem growth, and twisting branch phenotypes. Expression profiling of pillar peach trees revealed differential expression of numerous genes associated with biotic and abiotic stress, hormone responses, plastids, reactive oxygen, secondary, and cell wall metabolism. Collectively, the data provide important clues for understanding TAC1 function and show that alteration of TAC1 expression may have broad applicability to agricultural and ornamental tree industries.

Loss of a highly conserved sterile alpha motif domain gene (WEEP) results in pendulous branch growth in peach trees.

Plant shoots typically grow upward in opposition to the pull of gravity. However, exceptions exist throughout the plant kingdom. Most conspicuous are trees with weeping or pendulous branches. While such trees have long been cultivated and appreciated for their ornamental value, the molecular basis behind the weeping habit is not known. Here, we characterized a weeping tree phenotype in Prunus persica (peach) and identified the underlying genetic mutation using a genomic sequencing approach. Weeping peach tree shoots exhibited a downward elliptical growth pattern and did not exhibit an upward bending in response to 90° reorientation. The causative allele was found to be an uncharacterized gene, Ppa013325, having a 1.8-Kb deletion spanning the 5' end. This gene, dubbed WEEP, was predominantly expressed in phloem tissues and encodes a highly conserved 129-amino acid protein containing a sterile alpha motif (SAM) domain. Silencing WEEP in the related tree species Prunus domestica (plum) resulted in more outward, downward, and wandering shoot orientations compared to standard trees, supporting a role for WEEP in directing lateral shoot growth in trees. This previously unknown regulator of branch orientation, which may also be a regulator of gravity perception or response, provides insights into our understanding of how tree branches grow in opposition to gravity and could serve as a critical target for manipulating tree architecture for improved tree shape in agricultural and horticulture applications.