Expression Level of a Phenylalanine Ammonia-Lyase Gene in Poinsettia Is Negatively Correlated with Poinsettia Branch-Inducing Phytoplasma Titer.

Poinsettia is an important ornamental cultivated worldwide. Commercial poinsettias are almost universally infected with a pathogen known as the poinsettia branch-inducing phytoplasma (PoiBI), which can increase the level of branching in host plants and make the plants more desirable to consumers. Despite PoiBI's crucial role in poinsettia production, little is known about PoiBI-poinsettia interactions in regard to the pathogen's in planta population dynamics. The expression profiles of a phenylalanine ammonia-lyase gene (Euphorbia pulcherrima PAL [EpPAL]) and the PoiBI titers in poinsettia tissues were investigated. Differential gene expression analyses using quantitative PCR (qPCR) showed that EpPAL expression levels differed significantly across tissue types. The highest expression levels were detected in stems, followed by root. Lower EpPAL expression levels were detected in leaf tissues, particularly in source leaves closer to the base; the average expression level in these leaves was only one-seventh of that detected in stems. Phytoplasma concentrations in source leaves close to the base were significantly greater than the other tissue types; the average value was 7.6-fold of that detected in stem tissues, which had the lowest phytoplasma titers. A negative correlation between EpPAL expression level and PoiBI load was detected, suggesting that the products of EpPAL-associated pathways or other genes indirectly associated with EpPAL may interfere with PoiBI's growth. While additional studies are needed to validate these interpretations, the results from this work provide new insights into PoiBI-poinsettia interaction and showed that correlations between pathogen load and defense-related genes could be detected in phytoplasma-associated pathosystems. IMPORTANCE Phytoplasma-plant interactions are interesting subjects for fundamental and applicative research. Although many studies have characterized molecular interplays between these pathogens and hosts, knowledge on relationships between phytoplasmas' in planta population dynamics and host gene expression remains scarce. By using the poinsettia branch-inducing phytoplasma (PoiBI) and poinsettia as a model system, a negative correlation was observed between the expression level of a plant defense-related gene and the pathogen's titer. The findings provide potential explanations to PoiBI's distribution patterns in the plant and highlight the importance of studying phytoplasma-plant interactions in regard to the pathogen's population dynamics in other pathosystems.

Variability of Phytoplasma Infection Density in Poinsettia and Evaluation of its Association with the Level of Branching in Host Plants.

It has long been established that phytoplasma infection is the cause of the free-branching phenotype in poinsettia. However, relatively little is known about the ecology of the pathogen in planta. The present study evaluated the infection pattern of poinsettia branch-inducing phytoplasma (PoiBI) and its association with the poinsettia phenotype during cutting propagation. The presence of this pathogen in the poinsettia variety Luv U Pink was determined using PCR and sequence analysis. The infection density of PoiBI in distinct tissue types of different plant segments were then determined using quantitative PCR coupled with plasmid-based standard curves. Both vegetative-stage and flowering-stage plants were tested. The results showed that, despite being considerably variable among plants, the infection densities of PoiBI tend to be higher in source leaves located in the lower parts of the plant. The densities were consistently lower in tissues located at the top of the plants, regardless of the tissue type. Analysis of the infection densities among samples collected from six stock plants used in commercial production also revealed significantly different levels of PoiBI load. An association between PoiBI infection density in the stock plants and the level of branching in cutting-propagated plants (derived from the stock plants) was also observed; stock plants with low infection densities tended to produce smaller proportions of plants exhibiting higher degrees of branching both before and after pinching. These data suggest that uneven distribution of PoiBI within and among stock plants may lead to the production of cuttings with variable phytoplasma densities which may, in turn, affect the phenotypic uniformity of the plants produced. Overall, findings from the present work add to the understanding of PoiBI's ecology and could provide implications to commercial poinsettia production.