Resistance to Seven Site-Specific Fungicides in Botrytis cinerea from Greenhouse-Grown Ornamentals.

The fungal pathogen Botrytis cinerea is a notorious problem on many floriculture greenhouse hosts including petunia, geranium, and poinsettia; these key crops contribute to the $6.43 billion U.S. ornamental industry. While growers use cultural strategies to reduce relative humidity and free moisture to limit Botrytis blight, fungicides remain a primary component of control programs. Isolates (n = 386) of B. cinerea sampled from symptomatic petunia, geranium, and poinsettia in Michigan greenhouses from 2018 to 2021 were screened for resistance to eight fungicides belonging to seven Fungicide Resistance Action Committee (FRAC) groups. Single-spored isolates were subjected to a germination-based assay using previously defined discriminatory doses of each fungicide. Resistance was detected to thiophanate-methyl (FRAC 1; 94%), pyraclostrobin (FRAC 11; 80%), boscalid (FRAC 7; 67%), iprodione (FRAC 2; 65%), fenhexamid (FRAC 17; 38%), cyprodinil (FRAC 9; 38%), fludioxonil (FRAC 12; 21%), and fluopyram (FRAC 7; 13%). Most isolates (63.5%) were resistant to at least four FRAC groups, with 8.7% of all isolates demonstrating resistance to all seven FRAC groups tested. Resistance frequencies for each fungicide were similar among crops, production regions, and growing cycles but varied significantly for each greenhouse. Phenotypic diversity was high, as indicated by the 48 different fungicide resistance profiles observed. High frequencies of resistance to multiple fungicides in B. cinerea populations from floriculture hosts highlight the importance of sustainable and alternative disease management practices for greenhouse growers.

A highly mutable GST is essential for bract colouration in Euphorbia pulcherrima Willd. Ex Klotsch.

BACKGROUND: Mutation breeding is an extraordinary tool in plant breeding to increase the genetic variability, where mutations in anthocyanin biosynthesis are targets to generate distinctive phenotypes in ornamental species. In poinsettia, ionizing radiation is routinely applied in breeding programs to obtaining a range of colours, with nearly all pink and white varieties being obtained after γ- or X-ray mutagenesis of red varieties. In the present study we performed a thorough characterization of a potential mutagenesis target gene as the main responsible for the 'white paradox' in poinsettia. RESULTS: We identified a GST gene in poinsettia (Bract1) as an essential factor for the expression of anthocyanin-based red colouration of bracts, which presents a high phylogenetic similarity to known anthocyanin-related GSTs. Red poinsettia varieties and white mutants generated from these varieties by X-ray were analysed for polymorphisms related to the 'white paradox' in the species. A 4 bp mutation in a short repeat within the coding region of Bract1 is most likely responsible for the appearance of white phenotypes upon irradiation treatment. The polymorphism between wild-type and mutant alleles co-segregates with the phenotype in progeny from heterozygous red and white parents. Moreover, overexpression of Bract1 wild-type allele in Arabidopsis tt19 mutants restored the anthocyanin phenotype, while the Bract1 mutated allele showed to be non-functional. CONCLUSIONS: The identified repeat seems to be highly unstable, since mutated plants can be easily detected among fewer than 200 shoots derived from 10 mutated plants. Our data indicate that particular short repeat sequences, similar to microsatellite sequences or so-called dynamic mutations, might be hot spots for genetic variability. Moreover, the identification of the Bract1 mutation fills a gap on the understanding on the molecular mechanism of colour formation in poinsettia.

Influence of temperature on the retention, absorption and translocation of fomesafen and imazamox in Euphorbia heterophylla.

Climate change will be an additional issue to the challenge to manage herbicide resistant weeds. This work investigated the impact of three temperature regimes (10/5, 20/15 and 30/25 °C) on the efficacy, foliar retention, absorption and translocation of fomesafen, protoporphyrinogen oxidase (PPO) inhibitor, and imazamox, acetolactate synthase (ALS) inhibitor, between two Euphorbia heterophylla populations, one susceptible (S) and one multiple PPO and ALS resistant (R). The R population went from 5 (fomesafen) and 12 (imazamox) times more resistant than the S population at 10/5 °C to more than 100 times to both herbicides at 20/15 and 30/25 °C. Leaf retention of fomesafen was not affected by temperature; however, imazamox retention was less at 10/5 and 20/15 °C than at 30/25 °C, and the R population always retained less imazamox than the S population. 14C-fomesafen absorption was similar between populations, but lower amounts were absorbed at 10/5 °C regardless of the evaluation time. Recovered 14C-imazamox rates decreased in both populations as the evaluation time increased, ranging from 82 to 92% at 6 h after treatment (HAT), and from 47 to 76% at 48 HAT, depending on the temperature regime. The 14C-imazamox losses were greater from 24 HAT in R plants grown at 30/25 °C and in all temperature regimes at 48 HAT. Although both populations translocated large amounts of imazamox, the S population distributed it in the rest of the plant (33%) and roots (15%), while the R population kept it mainly on the treated leaf (24%) or lost ~20% more herbicide than S population at 48 HAT, indicating the need for further studies on root exudation between these populations. Low temperatures reduced resistance levels to fomesafen and imazamox in E. heterophylla, suggesting that temperature influences the expression of the mechanisms that govern this multiple resistance.

Evaluating Fungicide Selections to Manage Pythium Root Rot on Poinsettia Cultivars with Varying Levels of Partial Resistance.

Pythium aphanidermatum is the predominant species causing Pythium root rot of commercially grown poinsettia (Euphorbia pulcherrima Willd. ex Kotzch) in North Carolina. Pythium root rot is managed primarily through a combination of sanitation practices and preventative fungicide applications of mefenoxam or etridiazole. Insensitivity to mefenoxam is common but growers continue to rely on it from lack of inexpensive and efficacious alternatives. This research was conducted to identify alternative fungicides for Pythium root rot control and to evaluate their efficacy on poinsettia cultivars with varying levels of partial resistance. Greenhouse studies were conducted to assess efficacy of fungicide treatments in seven poinsettia cultivars inoculated with a mefenoxam-sensitive isolate of P. aphanidermatum. One study examined control with a single fungicide drench made at transplant and a second study examined repeat fungicide applications made throughout the experiment. Treatments containing etridiazole, mefenoxam, fenamidone, and cyazofamid provided control of Pythium root rot across all cultivars in both experiments whereas Fosetyl-al, potassium phosphite, and Trichoderma spp. failed to offer satisfactory control. Azoxystrobin, pyraclostrobin, and propamocarb reduced disease on some cultivars but failed to control Pythium root rot on highly susceptible cultivars. Four isolates of P. aphanidermatum cultured from plants growing in commercial greenhouses were evaluated for in vitro sensitivity to fungicides labeled for Pythium root rot control at four rates. Etridiazole, fosetyl-al, and potassium phosphite completely inhibited mycelial growth, whereas isolates varied in response to mefenoxam, cyazofamid, propamocarb, fenamidone, azoxystrobin, and pyraclostrobin in vitro. Twenty-one additional isolates then were evaluated at label rates of these fungicides. Seven isolates were insensitive to label rates of all three quinone outside inhibitors and one isolate was insensitive to the quinone outside inhibitors and mefenoxam. These results provide guidelines for selecting fungicides to maximize control of Pythium root rot on poinsettia cultivars.

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.

Phytoremediation of potentially toxic elements in a polluted industrial soil using Poinsettia.

Potentially toxic elements (PTEs) pollution has become a serious environmental threat, particularly in developing countries such as China. In response, there is a growing interest in phytoremediation studies to identify plant species as designated hyperaccumulators of PTEs in polluted soils. Poinsettia was selected as a candidate species for phytoremediation of six PTEs (Zn, Pb, Hg, Cr, As, Cu) in this study. A pot cultivation experiment (randomized incomplete block experimental design with 5 treatments and 4 blocks) was conducted using contaminated soils gathered from an industrial area in southcentral China. The bioaccumulation factor (BAF), translocation factor (TF), and bioconcentration factor were analyzed to determine the phytoremediation potential of poinsettia potted in different ratios of polluted soils. One-way ANOVA with post-hoc Tukey's test showed that poinsettia had significant uptake of Zn, Pb, Cu (BAF < 1 and TF < 1, p < 0.05) and Hg (BAF < 1 and TF > 1, p < 0.05). Poinsettias can therefore effectively accumulate Zn, Pb, and Cu in their lateral roots while extracting and transferring Hg into their leaves. Moreover, poinsettia exhibited tolerance towards As and Cr. Interestingly, it was also observed that PTEs can inhibit the height of potted poinsettia at a certain concentration.

Risk of Spreading Soft Rot Through Cutting Dips Against Whiteflies in Greenhouse-Grown Poinsettia.

Dipping is a quick and cost-effective technique to reduce pest infestations on unrooted cuttings of greenhouse ornamental crops. Large numbers of cuttings are immersed in an insecticidal treatment, e.g., biopesticides and/or insecticidal soap, before they are stuck in the growing medium and rooted. This research investigated the risk of cross-contamination of poinsettia cuttings with Pectobacterium carotovorum subsp. carotovorum, a potentially devastating pathogen causing soft rot, through the dipping process. Sampling at four commercial greenhouses showed that P. carotovorum subsp. carotovorum was present in all dip suspensions during and after processing poinsettia cuttings; concentrations up to 1 × 105 CFU/ml were detected. A laboratory experiment determined that P. carotovorum subsp. carotovorum-infected cuttings could contaminate clean dip suspensions to similar levels. These results indicated that there is potential for disease transfer during dipping. The potential for cross-contamination of healthy cuttings was evaluated by immersing poinsettia cuttings in dip suspensions artificially inoculated with P. carotovorum subsp. carotovorum (from 1 × 103 to 1 × 107 CFU/ml). Disease incidence increased as P. carotovorum subsp. carotovorum concentrations in the dip suspension increased and the variety 'Prestige Red' was more susceptible than 'Freedom White.' However, even at the highest P. carotovorum subsp. carotovorum concentration of 1 × 107 CFU/ml, the proportion of diseased cuttings was low at 6% for var. 'Freedom White,' but higher at 21% for var. 'Prestige Red.' We conclude that P. carotovorum subsp. carotovorum transfer among unrooted poinsettia cuttings through the dipping process is relatively low although some varieties are sensitive to high levels of inoculum. Even so, strict sanitation practices are still important to prevent build-up of inoculum in the dip treatment.

Hybrid de novo transcriptome assembly of poinsettia (Euphorbia pulcherrima Willd. Ex Klotsch) bracts.

BACKGROUND: Poinsettia is a popular and important ornamental crop, mostly during the Christmas season. Its bract coloration ranges from pink/red to creamy/white shades. Despite its ornamental value, there is a lack of knowledge about the genetics and molecular biology of poinsettia, especially on the mechanisms of color formation. We performed an RNA-Seq analysis in order to shed light on the transcriptome of poinsettia bracts. Moreover, we analyzed the transcriptome differences of red- and white-bracted poinsettia varieties during bract development and coloration. For the assembly of a bract transcriptome, two paired-end cDNA libraries from a red and white poinsettia pair were sequenced with the Illumina technology, and one library from a red-bracted variety was used for PacBio sequencing. Both short and long reads were assembled using a hybrid de novo strategy. Samples of red- and white-bracted poinsettias were sequenced and comparatively analyzed in three color developmental stages in order to understand the mechanisms of color formation and accumulation in the species. RESULTS: The final transcriptome contains 288,524 contigs, with 33% showing confident protein annotation against the TAIR10 database. The BUSCO pipeline, which is based on near-universal orthologous gene groups, was applied to assess the transcriptome completeness. From a total of 1440 BUSCO groups searched, 77% were categorized as complete (41% as single-copy and 36% as duplicated), 10% as fragmented and 13% as missing BUSCOs. The gene expression comparison between red and white varieties of poinsettia showed a differential regulation of the flavonoid biosynthesis pathway only at particular stages of bract development. An initial impairment of the flavonoid pathway early in the color accumulation process for the white poinsettia variety was observed, but these differences were no longer present in the subsequent stages of bract development. Nonetheless, GSTF11 and UGT79B10 showed a lower expression in the last stage of bract development for the white variety and, therefore, are potential candidates for further studies on poinsettia coloration. CONCLUSIONS: In summary, this transcriptome analysis provides a valuable foundation for further studies on poinsettia, such as plant breeding and genetics, and highlights crucial information on the molecular mechanism of color formation.

The rare orange-red colored Euphorbia pulcherrima cultivar 'Harvest Orange' shows a nonsense mutation in a flavonoid 3'-hydroxylase allele expressed in the bracts.

BACKGROUND: Commercially available poinsettia (Euphorbia pulcherrima) varieties prevalently accumulate cyanidin derivatives and show intense red coloration. Orange-red bract color is less common. We investigated four cultivars displaying four different red hues with respect to selected enzymes and genes of the anthocyanin pathway, putatively determining the color hue. RESULTS: Red hues correlated with anthocyanin composition and concentration and showed common dark red coloration in cultivars 'Christmas Beauty' and 'Christmas Feeling' where cyanidin derivatives were prevalent. In contrast, orange-red bract color is based on the prevalent presence of pelargonidin derivatives that comprised 85% of the total anthocyanin content in cv. 'Premium Red' and 96% in cv. 'Harvest Orange' (synonym: 'Orange Spice'). cDNA clones of flavonoid 3'-hydroxylase (F3'H) and dihydroflavonol 4-reductase (DFR) were isolated from the four varieties, and functional activity and substrate specificity of the corresponding recombinant enzymes were studied. Kinetic studies demonstrated that poinsettia DFRs prefer dihydromyricetin and dihydroquercetin over dihydrokaempferol, and thus, favor the formation of cyanidin over pelargonidin. Whereas the F3'H cDNA clones of cultivars 'Christmas Beauty', 'Christmas Feeling', and 'Premium Red' encoded functionally active enzymes, the F3'H cDNA clone of cv. 'Harvest Orange' contained an insertion of 28 bases, which is partly a duplication of 20 bases found close to the insertion site. This causes a frameshift mutation with a premature stop codon after nucleotide 132 and, therefore, a non-functional enzyme. Heterozygosity of the F3'H was demonstrated in this cultivar, but only the mutated allele was expressed in the bracts. No correlation between F3'H-expression and the color hue could be observed in the four species. CONCLUSIONS: Rare orange-red poinsettia hues caused by pelargonidin based anthocyanins can be achieved by different mechanisms. F3'H is a critical step in the establishment of orange red poinsettia color. Although poinsettia DFR shows a low substrate specificity for dihydrokaempferol, sufficient precursor for pelargonidin formation is available in planta, in the absence of F3'H activity.

Protecting Unrooted Cuttings From Bemisia tabaci (Hemiptera Aleyrodidae) During Propagation.

In North America, the sweetpotato whitefly, Bemisia tabaci Genn., is an important pest of greenhouse poinsettia. Growers have limited options to control this pest during propagation of cuttings, which are rooted under mist for several weeks. Early establishment of this pest increases the difficulty of managing the whitefly and retaining high aesthetic standard during the remaining crop production phase. We evaluated two neonicotinoids with translaminar activity, thiamethoxam (Flagship 25WG), and acetamiprid (TriStar 70 WSP), for control of B. tabaci pre-infested on unrooted cuttings propagated under mist. In an experimental greenhouse, both materials significantly reduced whitefly populations, providing an average reduction of 87.8% and 61.5% total recovered whitefly stages respectively, compared with controls. In another test, dipping cuttings in thiamethoxam (immersion treatment) did not improve control significantly, when compared with foliar sprays applied at label rate. In a commercial greenhouse operation, immersion treatments of thiamethoxam on pre-infested poinsettia cuttings maintained whiteflies at ≤ 0.02/plant, compared with up to 0.33/plant in untreated cuttings. Our data suggest that treating unrooted cuttings before or at the start of propagation can be part of an overall strategy for growers to manage whiteflies in poinsettia production.

Male Pheromones Influence the Mating Behavior of Echinothrips americanus.

Two dibasic esters, the dimethyl ester of hexanedioic acid (dimethyl adipate, DBE-6) and the dimethyl ester of pentanedioic acid (dimethyl glutarate, DBE-5) were found in head-thorax extracts of male Echinothrips americanus. DBE-5 induced abdomen wagging and raising in males and females, which is typically exhibited when encountering a male. DBE-6 was avoided by males and was detected on mated, but not on virgin, females. Both substances applied to virgin females lead to females being ignored by males. The role of both substances is discussed with regard to the male mating system.

Anthocyanin metabolic engineering of Euphorbia pulcherrima: advances and perspectives.

The range of floral colors is determined by the type of plant pigment accumulated by the plant. Anthocyanins are the most common flavonoid pigments in angiosperms; they provide a wide range of visible colors from red-magenta to blue-purple, products of cyanidin and delphinidin biosynthesis, respectively. For the floriculture industry, floral color is one of the most important ornamental characteristics for the development of new commercial varieties; however, most plant species are restricted to a certain color spectrum, limited by their own genetics. In fact, many ornamental crops lack bluish varieties due to the lack of activity of essential biosynthetic enzymes for the accumulation of delphinidin. An example is the poinsettia (Euphorbia pulcherrima Willd. ex Klotzsch), the ornamental plant symbol of Christmas and native to Mexico. Its popularity is the result of the variety of colors displayed by its bracts, a kind of modified leaves that accumulate reddish pigments based mainly on cyanidin and, to a lesser extent, on pelargonidin. The commercial success of this plant lies in the development of new varieties and, although consumers like the typical red color, they are also looking for poinsettias with new and innovative colors. Previous research has demonstrated the possibility of manipulating flower color through metabolic engineering of the anthocyanin biosynthesis pathway and plant tissue culture in different ornamental plant species. For example, transgenic cultivars of flowers such as roses, carnations or chrysanthemums owe their attractive bluish colors to a high and exclusive accumulation of delphinidin. Here, we discuss the possibilities of genetic engineering of the anthocyanin biosynthetic pathway in E. pulcherrima through the introduction of one or more foreign delphinidin biosynthetic genes under the transcriptional control of a pathway-specific promoter, and the genome editing possibilities as an alternative tool to modify the color of the bracts. In addition, some other approaches such as the appropriate selection of the cultivars that presented the most suitable intracellular conditions to accumulate delphinidin, as well as the incorporation of genes encoding anthocyanin-modifying enzymes or transcription factors to favor the bluish pigmentation of the flowers are also revised.

Population changes of Bemisia tabaci (Hemiptera: Aleyrodidae) on different colored poinsettia leaves with different trichome densities and chemical compositions.

The whitefly, Bemisia tabaci, is a destructive and invasive pest of many horticultural plants including poinsettia (Euphorbia pulcherrima). Outbreaks of B. tabaci cause serious damage by direct feeding on phloem sap, and spreading 100+ plant viruses to crops. Bemisia tabaci were observed more frequently on green than red poinsettia leaves, and the factors responsible for this are unknown. Here, we investigated the development rate, survivorship, fecundity of B. tabaci feeding on green versus red leaves, as well as the leaves' volatiles, trichome density, anthocyanin content, soluble sugars, and free amino acids. Compared to red leaves, B. tabaci on green leaves showed increased fecundity, a higher female sex ratio, and survival rate. The green color alone was more attractive to B. tabaci than red. Red leaves of poinsettia contained more phenol, and panaginsene in their volatiles. Alpha-copaene and caryophyllene were more abundant in the volatiles of poinsettia green leaves. Leaf trichome density, soluble sugars and free amino acids were higher in green than red leaves of poinsettia, anthocyanin was lower in green than red leaves. Overall, green leaves of poinsettia were more susceptible and attractive to B. tabaci. The morphological and chemical variation between red and green leaves also differed; further investigation may reveal how these traits affect B. tabaci's responses.

2,5-Diketopiperazines via Intramolecular N-Alkylation of Ugi Adducts: A Contribution to the Synthesis, Density Functional Theory Study, X-ray Characterization, and Potential Herbicide Application.

To investigate the herbicidal potential of 2,5-diketopiperazines (2,5-DKPs), we applied a known protocol to produce a series of 2,5-DKPs through intramolecular N-alkylation of Ugi adducts. However, the method was not successful for the cyclization of adducts presenting aromatic rings with some substituents at the ortho position. Results from DFT calculations showed that the presence of voluminous groups at the ortho position of a benzene ring results in destabilization of the transition structure. Lower activation enthalpies for the SN2-type cyclization of Ugi adducts were obtained when bromine, instead of a chlorine anion, is the leaving group, indicating that the activation enthalpy for the cyclization step controls the formation of the 2,5-DKP. Some Ugi adducts and 2,5-DKPs formed crystals with suitable qualities for single-crystal X-ray diffraction data collection. Phytotoxic damage of some 2,5-DKPs on leaves of the weed Euphorbia heterophylla did not differ from those caused by the commercial herbicide diquat.

Synthesis of Tyrosol 1,2,3-Triazole Derivatives and Their Phytotoxic Activity against Euphorbia heterophylla.

The synthesis and phytotoxic activity of a series of tyrosol 1,2,3-triazole derivatives are reported herein. Target compounds were synthesized through the copper(I)-catalyzed azide-alkyne cycloaddition reaction (CuAAC), known as click reaction, and these were tested for phytotoxic activity on leaves of wild poinsettia (Euphorbia heterophylla), fleabane (Conyza sumatrensis), and tropical spiderwort (Commelina benghalensis). These are three highly noxious agricultural weeds that challenge available weed control methods, including the use of chemical herbicides. Twenty-five compounds were synthesized and tested. None of the compounds showed phytotoxic activity against C. benghalensis and C. sumatrensis, but almost all of them produced yellowing, bleaching, and necrosis on leaves of E. heterophylla. Two of the tyrosol 1,2,3-triazole derivatives produced more extensive lesions than those produced by the commercial herbicide diquat, used as a positive control (p ≤ 0.05). When applied on leaves of E. heterophylla, these compounds interfered with the stomatal conductance, net photosynthesis, internal carbon concentration, transpiration rate, water-use efficiency, and chlorophyll A and B contents. The interference of such compounds on such photosynthesis-related variables indicates that tyrosol 1,2,3-triazole derivatives may be capable of lowering the competitiveness of E. heterophylla and acting as additional tools for managing this competitive weed in agricultural lands.

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.

Adding a Natural Enemy to Respond to Pest Immigration and Delayed Natural Enemy Releases in Augmentative Biological Control.

Whether increased natural enemy density or adding a second natural enemy species will provide superior pest suppression in greenhouse augmentative biological control is unknown for many commercially available natural enemy species. In this study, we use sweetpotato whiteflies, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), on poinsettias, Euphorbia pulcherrima Willd. ex Klotzsch (Malpighiales: Euphorbiaceae), to determine whether adding Amblyseius swirskii (Athias-Henriot) (Acari: Phytoseiidae) to Eretmocerus eremicus Rose and Zolnerowich (Hymenoptera: Aphelinidae) is better for B. tabaci suppression compared with either natural enemy alone, both with and without challenges with whitefly immigration or delayed natural enemy releases. The number of whiteflies on caged poinsettias treated with different natural enemy release rates (single or double rate), natural enemy species (one or two species), natural enemy delayed release (weeks 4 and 8), and whitefly immigration treatments (introduced at week 4 or week 8) was censused biweekly for 16 wk. Both species used in combination provided similar or better suppression of whiteflies compared with either natural enemy alone. Both species combined also provided superior suppression of whiteflies when challenged with whitefly immigration or delays in natural enemy releases compared with E. eremicus alone. Whitefly immigration or delays in E. eremicus releases did not increase whitefly populations, suggesting that suppression of whiteflies by E. eremicus alone is relatively robust. This study found no evidence for negative interactions between E. eremicus and A. swirskii for suppressing B. tabaci.

Allelopathic activity and chemical constituents of extracts from roots of Euphorbia heterophylla L.

Euphorbia heterophylla L. is regarded as a major weed worldwide. Its high aggressiveness in agricultural environment prompted us to investigate the allelopatic activity and chemical constitution of extracts from roots of this plant. Hexane extract showed low phytotoxic activity. Methanol extract at 2.0 mg mL-1 inhibited 100% of germination, root and shoot growth of the indicator plants Sorghum bicolor and Lactuca sativa. ß-sitosterol, stigmasterol, and esters of lupeol, germanicol, taraxasterol, pseudotaraxasterol, α-amyrin and ß-amyrin were isolated from the hexane extract and their structures elucidated on the basis of MS and 1H, 13C and DEPT-135 NMR data. GC-MS analysis of the derivatized methanol extract allowed for identifying a series of allelopathic organic acids potentially involved in allelopathic interactions of E. heterophylla. This is the first study on the allelopathic activity of extracts and identification of metabolites from roots of E. heterophylla.

Silicon Affects Root Development, Tissue Mineral Content, and Expression of Silicon Transporter Genes in Poinsettia (Euphorbia pulcherrima Willd.) Cultivars.

The effects of silicon (Si) on root development, mineral content, and expression of Si transporter genes in Euphorbia pulcherrima Willd. 'Flame', 'Mable Bell', 'Green Star', 'Pink Bell', and 'Peach Bowl' cultivars were investigated in this study. Stem cuttings in a propagation bench were drenched regularly with a solution containing either 0 (control) or 50 ppm of silicon (Si treatment) from potassium silicate (K2SiO3), with a 25 °C mean air temperature and 80% relative humidity (RH) under 70% shading. The results showed that the 'Flame' treated with Si had a significantly higher survival ratio as compared with that of the control (P ≤ 0.05) and that the Si treatment improved number of roots, length of longest root, fresh root weight, and dry root weight in all cultivars except 'Mable Bell'. Supplementary Si increased the content of magnesium (Mg) and decreased the content of boron (B) and zinc (Zn) in the roots. The content of sulfur (S) in the shoots was increased by supplementary Si. The relative expression of Lsi1 and Lsi2 was higher in 'Peach Bowl', while it was lower in 'Mable Bell' and 'Green Star', which may be caused by the differing accumulation of Si in the shoot. Overall, supplementary Si had beneficial effects during cutting propagation of poinsettia cultivars, although these effects were cultivar-dependent.