A transcriptome-based association study of growth, wood quality, and oleoresin traits in a slash pine breeding population.

Slash pine (Pinus elliottii Engelm.) is an important timber and resin species in the United States, China, Brazil and other countries. Understanding the genetic basis of these traits will accelerate its breeding progress. We carried out a genome-wide association study (GWAS), transcriptome-wide association study (TWAS) and weighted gene co-expression network analysis (WGCNA) for growth, wood quality, and oleoresin traits using 240 unrelated individuals from a Chinese slash pine breeding population. We developed high quality 53,229 single nucleotide polymorphisms (SNPs). Our analysis reveals three main results: (1) the Chinese breeding population can be divided into three genetic groups with a mean inbreeding coefficient of 0.137; (2) 32 SNPs significantly were associated with growth and oleoresin traits, accounting for the phenotypic variance ranging from 12.3% to 21.8% and from 10.6% to 16.7%, respectively; and (3) six genes encoding PeTLP, PeAP2/ERF, PePUP9, PeSLP, PeHSP, and PeOCT1 proteins were identified and validated by quantitative real time polymerase chain reaction for their association with growth and oleoresin traits. These results could be useful for tree breeding and functional studies in advanced slash pine breeding program.

Genetic analysis and elite tree selection of the main resin components of slash pine.

Pine resin, as a natural material, has been widely used in food, pharmaceutical, and chemical industries. Slash pine (Pinus elliottii Engelm var. elliottii) is the primary tree species for resin tapping due to its high resin yield, low resin crystallization rate, and high turpentine content. Current researches focuse on the targeted improvement of several significant components to meet industrial needs rather than just resin yield. The objective of this study was to examine the genetic variation and correlation of genetic and phenotype for four main resin components (α pinene, ß pinene, abietic acid, and levoprimaric acid) of 219 half-sib progenies from 59 families. The results showed that the levopimaric acid had the largest content (mean value = 21.63%), while the ß pinene content had the largest variation coefficient (CV = 0.42). The α pinene content has the highest heritability (h2 = 0.67), while levopimaric acid has the lowest heritability (h2 = 0.51). There was a significant negative correlation between α pinene and the other three components and a significant positive correlation between ß pinene and the two diterpenes. The family ranking and genetic gain suggested that it is possible to improve the contents of main resin components of slash pine through genetic breeding selection.

Phenomic selection in slash pine multi-temporally using UAV-multispectral imagery.

Genomic selection (GS) is an option for plant domestication that offers high efficiency in improving genetics. However, GS is often not feasible for long-lived tree species with large and complex genomes. In this paper, we investigated UAV multispectral imagery in time series to evaluate genetic variation in tree growth and developed a new predictive approach that is independent of sequencing or pedigrees based on multispectral imagery plus vegetation indices (VIs) for slash pine. Results show that temporal factors have a strong influence on the h2 of tree growth traits. High genetic correlations were found in most months, and genetic gain also showed a slight influence on the time series. Using a consistent ranking of family breeding values, optimal slash pine families were selected, obtaining a promising and reliable predictive ability based on multispectral+VIs (MV) alone or on the combination of pedigree and MV. The highest predictive value, ranging from 0.52 to 0.56, was found in July. The methods described in this paper provide new approaches for phenotypic selection (PS) using high-throughput multispectral unmanned aerial vehicle (UAV) technology, which could potentially be used to reduce the generation time for conifer species and increase the genetic granularity independent of sequencing or pedigrees.

Prediction of Needle Physiological Traits Using UAV Imagery for Breeding Selection of Slash Pine.

Leaf nitrogen (N) content and nonstructural carbohydrate (NSC) content are 2 important physiological indicators that reflect the growth state of trees. Rapid and accurate measurement of these 2 traits multitemporally enables dynamic monitoring of tree growth and efficient tree breeding selection. Traditional methods to monitor N and NSC are time-consuming, are mostly used on a small scale, and are nonrepeatable. In this paper, the performance of unmanned aerial vehicle multispectral imaging was evaluated over 11 months of 2021 on the estimation of canopy N and NSC contents from 383 slash pine trees. Four machine learning methods were compared to generate the optimal model for N and NSC prediction. In addition, the temporal scale of heritable variation for N and NSC was evaluated. The results show that the gradient boosting machine model yields the best prediction results on N and NSC, with R 2 values of 0.60 and 0.65 on the validation set (20%), respectively. The heritability (h 2) of all traits in 11 months ranged from 0 to 0.49, with the highest h 2 for N and NSC found in July and March (0.26 and 0.49, respectively). Finally, 5 families with high N and NSC breeding values were selected. To the best of our knowledge, this is the first study to predict N and NSC contents in trees using time-series unmanned aerial vehicle multispectral imaging and estimating the genetic variation of N and NSC along a temporal scale, which provides more reliable information about the overall performance of families in a breeding program.

CountShoots: Automatic Detection and Counting of Slash Pine New Shoots Using UAV Imagery.

The density of new shoots on pine trees is an important indicator of their growth and photosynthetic capacity. However, traditional methods to monitor new shoot density rely on manual and destructive measurements, which are labor-intensive and have led to fewer studies on new shoot density. Therefore, in this study, we present user-friendly software called CountShoots, which extracts new shoot density in an easy and convenient way using unmanned aerial vehicles based on the YOLOX and Slash Pine Shoot Counting Network (SPSC-net) models. This software mainly consists of 2 steps. Firstly, we deployed a modified YOLOX model to identify the tree species and location from complex RGB background images, which yielded a high recognition accuracy of 99.15% and 95.47%. These results showed that our model produced higher detection accuracy compared to YOLOv5, Efficientnet, and Faster-RCNN models. Secondly, we constructed an SPSC-net. This methodology is based on the CCTrans network, which outperformed DM-Count, CSR-net, and MCNN models, with the lowest mean squared error and mean absolute error results among other models (i.e., 2.18 and 1.47, respectively). To our best knowledge, our work is the first research contribution to identify tree crowns and count new shoots automatically in slash pine. Our research outcome provides a highly efficient and rapid user-interactive pine tree new shoot detection and counting system for tree breeding and genetic use purposes.

Spectrometric Prediction of Nitrogen Content in Different Tissues of Slash Pine Trees.

The internal cycling of nitrogen (N) storage and consumption in trees is an important physiological mechanism associated with tree growth. Here, we examined the capability of near-infrared spectroscopy (NIR) to quantify the N concentration across tissue types (needle, trunk, branch, and root) without time and cost-consuming. The NIR spectral data of different tissues from slash pine trees were collected, and the N concentration in each tissue was determined using standard analytical method in laboratory. Partial least squares regression (PLSR) models were performed on a set of training data randomly selected. The full-length spectra and the significant multivariate correlation (sMC) variable selected spectra were used for model calibration. Branch, needle, and trunk PLSR models performed well for the N concentration using both full length and sMC selected NIR spectra. The generic model preformatted a reliable accuracy with R2 C and R2 CV of 0.62 and 0.66 using the full-length spectra, and 0.61 and 0.65 using sMC-selected spectra, respectively. Individual tissue models did not perform well when being used in other tissues. Five significantly important regions, i.e., 1480, 1650, 1744, 2170, and 2390 nm, were found highly related to the N content in plant tissues. This study evaluates a rapid and efficient method for the estimation of N content in different tissues that can help to serve as a tool for tree N storage and recompilation study.

Prediction and Comparisons of Turpentine Content in Slash Pine at Different Slope Positions Using Near-Infrared Spectroscopy.

Pine resin is one of the best known and most exploited non-wood products. Resin is a complex mixture of terpenes produced by specialized cells that are dedicated to tree defense. Chemical defenses are plastic properties, and concentrations of chemical defenses can be adjusted based on environmental factors, such as resource availability. The slope orientation (south/sunny or north/shady) and the altitude of the plantation site have significant effects on the soil nutrient and the plant performance, whereas little is known about how the slope affects the pine resin yield and its components. In total, 1180 slash pines in 18 plots at different slope positions were established to determine the effects on the α- and ß-pinene content and resin production of the slash pine. The near-infrared spectroscopy (NIR) technique was developed to rapidly and economically predict the turpentine content for each sample. The results showed that the best partial least squares regression (PLS) models for α- and ß-pinene content prediction were established via the combined treatment of multiplicative scatter correction-significant multivariate correlation (MSC-sMC). The prediction models based on sMC spectra for α- and ß-pinene content have an R2 of 0.82 and 0.85 and an RMSE of 0.96 and 0.82, respectively, and they were successfully implemented in turpentine prediction in this research. The results also showed that a barren slope position (especially mid-slope) could improve the α-pinene and ß-pinene content and resin productivity of slash pine, and the ß-pinene content in the resin had more variances in this research.

Prediction and Utilization of Malondialdehyde in Exotic Pine Under Drought Stress Using Near-Infrared Spectroscopy.

Drought is a major abiotic stress that adversely affects the growth and productivity of plants. Malondialdehyde (MDA), a substance produced by membrane lipids in response to reactive oxygen species (ROS), can be used as a drought indicator to evaluate the degree of plasma membrane damage and the ability of plants to drought stress tolerance. Still measuring MDA is usually a labor- and time-consuming task. In this study, near-infrared (NIR) spectroscopy combined with partial least squares (PLS) was used to obtain rapid and high-throughput measurements of MDA, and the application of this technique to plant drought stress experiments was also investigated. Two exotic conifer tree species, namely, slash pine (Pinus elliottii) and loblolly pine (Pinus taeda), were used as plant material exposed to drought stress; different types of spectral preprocessing methods and important feature-selection algorithms were applied to the PLS model to calibrate it and obtain the best MDA-predicting model. The results show that the best PLS model is established via the combined treatment of detrended variable-significant multivariate correlation algorithm (DET-sMC), where latent variables (LVs) were 6. This model has a respectable predictive capability, with a correlation coefficient (R 2) of 0.66, a root mean square error (RMSE) of 2.28%, and a residual prediction deviation (RPD) of 1.51, and it was successfully implemented in drought stress experiments as a reliable and non-destructive method to detect the MDA content in real time.