Auxin and carbohydrate control flower bud development in Anthurium andraeanum during early stage of sexual reproduction.

BACKGROUND: Flower buds of Anthurium andraeanum frequently cease to grow and abort during the early flowering stage, resulting in prolonged planting times and increased commercialization costs. Nevertheless, limited knowledge exists of the mechanism of flower development after initiation in A. andraeanum. RESULTS: In this study, the measurement of carbohydrate flow and intensity between leaves and flowers during different growth stages showed that tender leaves are strong sinks and their concomitant flowers are weak ones. This suggested that the tender leaves compete with their concomitant flower buds for carbohydrates during the early growth stages, potentially causing the abortion of the flower buds. The analysis of transcriptomic differentially expressed genes suggested that genes related to sucrose metabolism and auxin response play an important role during flower bud development. Particularly, co-expression network analysis found that AaSPL12 is a hub gene engaged in flower development by collaborating carbohydrate and auxin signals. Yeast Two Hybrid assays revealed that AaSPL12 can interact with AaARP, a protein that serves as an indicator of dormancy. Additionally, the application of exogenous IAA and sucrose can suppress the expression of AaARP, augment the transcriptional abundance of AaSPL12, and consequently expedite flower development in Anthurium andraeanum. CONCLUSIONS: Collectively, our findings indicated that the combination of auxin and sugar signals could potentially suppress the repression of AaARP protein to AaSPL12, thus advancing the development of flower buds in Anthurium andraeanum.

Image completion algorithm of anthurium spathes based on multi-scale feature learning.

Machine vision has been used to grade the potted anthurium plant in large-scale production recently. Images are taken to measure the number and size of anthurium spathes. However, due to the limitation of the shooting angle, the occlusion problem reduces the accuracy of measurement. It is necessary to segment the overlapping spathes and repair the incomplete ones. The traditional image completion model has good performance on missing small areas, but it is not satisfactory for missing large areas. In this article, a multi-scale fusion Recurrent Feature Reasoning (RFR) network was proposed to repair the spathe images. Unlike the traditional RFR, a multi-layer component was used in the feature reasoning module. This network can combine multi-scale features to complete the learning task and obtain more details of the spathe, which makes the network more advantageous in image completion when missing large areas of spathes. In this study, a comparison experiment between this network and the widely used image completion network was performed, and the results showed that this network performed well in all types of image completion, especially with large-area incomplete images.

A High-Resolution Linkage Map Construction and QTL Analysis for Morphological Traits in Anthurium (Anthurium andraeanum Linden).

Anthurium andraeanum Linden is a prominent ornamental plant belonging to the family Araceae and is cultivated worldwide. The morphology characteristics are crucial because they significantly impact ornamental values, commercial properties, and the efficiency of space utilization in production. However, only a few related investigations have been conducted in anthurium to date. In this study, an F1 genetic segregation population containing 160 progenies was generated through hybridization between potted and cut anthurium varieties. Fifteen morphological traits were assessed and revealed substantial levels of genetic variation and widespread positive correlation. Based on specific length amplified fragment (SLAF) sequencing technology, 8171 single nucleotide polymorphism (SNP) markers were developed, and the high-density linkage map of 2202.27 cM in length distributed on 15 linkage groups was constructed successfully, with an average distance of 0.30 cM. Using the inclusive composite interval mapping (ICIM) method, 59 QTLs related to 15 key morphological traits were successfully identified, which explained phenotypic variance (PVE) ranging from 6.21% to 17.74%. Thirty-three of those associated with 13 traits were designated as major QTLs with PVE > 10%. These findings offer valuable insights into the genetic basis of quantitative traits and are beneficial for molecular marker-assisted selection (MAS) in anthurium breeding.

Identification of key genes responsible for green and white colored spathes in Anthurium andraeanum (Hort.).

Modern anthuriums, Anthurium andraeanum (Hort.) are among the most popular flowering plants and widely used for interior decoration. Their popularity is largely attributed to the exotic spathes with different colors. Previous studies have reported color development in red spathe cultivars, but limited information is available on key genes regulating white and green colored spathes. This study analyzed anthocyanin, chlorophyll, and carotenoid contents as well as transcript differences in spathes of eight cultivars that differed in spathe colors ranging from red to white and green. Results showed that increased expression of a transcription factor AaMYB2 was associated with elevated levels of anthocyanin in spathes, but decreased expression of AaMYB2 and increased expression of AaLAR (leucoanthocyanidin reductase) and AaANR (anthocyanidin reductase) were accompanied with the accumulation of colorless proanthocyanidin, thus the white spathe. As to the green colored spathe, chlorophyll content in the green spathe cultivar was substantially higher than the other cultivars. Correspondingly, transcripts of chlorophyll biosynthesis-related genes AaHemB (porphobilinogen synthase) and AaPor (protochlorophyllide oxidoreductase) were highly upregulated but almost undetectable in white and red spathes. The increased expression of AaHemB and AaPor was correlated with the expression of transcription factor AaMYB124. Subsequently, qRT-PCR analysis confirmed their expression levels in nine additional cultivars with red, white, and green spathes. A working model for the formation of white and green spathes was proposed. White colored spathes are likely due to the decreased expression of AaMYB2 which results in increased expression of AaLAR and AaANR, and the green spathes are attributed to AaMYB124 enhanced expression of AaHemB and AaPor. Further research is warranted to test this working model.

Functional and histological effects of Anthurium schlechtendalii Kunth extracts on adenine-induced kidney damage of adult Wistar rats.

Anthurium schlechtendalii Kunth is used by the Zoque group in southeastern Mexico for kidney and urinary diseases, but its safety and effectiveness are unproven, therefore a model of adenine-induced renal failure in rats was performed. The rats were fed with solid and aqueous plant extracts for 4 weeks to study its effects on kidney histological morphology. Kidneys were examined, and statistical analysis was performed. The adenine-containing diet caused renal failure, characterized by crystal deposits, cystic dilatation of tubules, and micro-abscesses. Both extracts caused tubular damage and collagen increase without inflammation. However, when combined with adenine, the extracts showed some protective effects, although cystic dilatation and granulomatous inflammation were observed. The extracts at the tested doses resulted in glomerular and tubular damage, aggravating cystic degeneration, therefore, its indiscriminate use in Humans is not safe. Additionally, the extracts can serve as a model for studying renal damage without crystal deposits.

Grading Criteria of Anthurium DUS Quantitative Characteristics by Multiple Comparison.

The determination of the grades and interval of quantitative characteristics is an important job while we draft new distinctness, uniformity and stability (DUS) test guidelines. Grading criteria should be adjusted because of the effect of year and site; it is also a key task to establish applicable criteria in the DUS test. Excellent criteria will improve the accuracy of the DUS evaluation. In this study, we analyzed the variability and distribution patterns of nine quantitative characteristics of 251 anthurium varieties. Three methods were used to establish the grade criteria: the two standard deviation methods, the two LSD0.05 methods and the multiple comparison method. The results showed that the coefficient of variation within varieties varied from 6.96% to 10.11%. The quantitative characteristics observed in this study did not follow a normal distribution, except spadix thickness at the middle and spathe size. In most characteristics, the standard deviations and LSD0.05 were similar, except for spathe size. The state interval set by multiple comparison methods for every characteristic was variable, and its mean was about 1.25 times that of the other two methods. The process of establishing grading criteria using the multiple comparison method was simpler, and the criteria were more accurate, with a lower error rate.

The complete chloroplast genome sequence of Anthurium andraeanum Linden (Araceae; Pothoideae).

The chloroplast genome of Anthurium andraeanum Linden 1877 was assembled and analyzed in this study. The genome size is 162,560 bp, of which contains a large single-copy (LSC) region with 88,814 bp, a small single-copy (SSC) region with 22,856 bp, and two inverted repeat regions (IRA and IRB) with 25,445 bp, respectively. The plastome contains 124 genes, including 80 protein-coding genes, 37 tRNAs, six rRNAs and one pseudogene. Phylogenetic analysis indicated that A. andraeanum is a member of Pothoideae and sister to A. huixtlense.

Comparative Physiological and Transcriptome Analyses of Tolerant and Susceptible Cultivars Reveal the Molecular Mechanism of Cold Tolerance in Anthurium andraeanum.

Anthurium andraeanum is a tropical ornamental flower. The cost of Anthurium production is higher under low temperature (non-freezing) conditions; therefore, it is important to increase its cold tolerance. However, the molecular mechanisms underlying the response of Anthurium to cold stress remain elusive. In this study, comparative physiological and transcriptome sequencing analyses of two cultivars with contrasting cold tolerances were conducted to evaluate the cold stress response at the flowering stage. The activities of superoxide dismutase and peroxidase and the contents of proline, soluble sugar, and malondialdehyde increased under cold stress in the leaves of the cold tolerant cultivar Elegang (E) and cold susceptible cultivar Menghuang (MH), while the soluble protein content decreased in MH and increased in E. Using RNA sequencing, 24,695 differentially expressed genes (DEGs) were identified from comparisons between cultivars under the same conditions or between the treatment and control groups of a single cultivar, 9132 of which were common cold-responsive DEGs. Heat-shock proteins and pectinesterases were upregulated in E and downregulated in MH, indicating that these proteins are essential for Anthurium cold tolerance. Furthermore, four modules related to cold treatment were obtained by weighted gene co-expression network analysis. The expression of the top 20 hub genes in these modules was induced by cold stress in E or MH, suggesting they might be crucial contributors to cold tolerance. DEGs were significantly enriched in plant hormone signal transduction pathways, trehalose metabolism, and ribosomal proteins, suggesting these processes play important roles in Anthurium's cold stress response. This study provides a basis for elucidating the mechanism of cold tolerance in A. andraeanum and potential targets for molecular breeding.

Identification of microRNA158 from Anthurium andraeanum and Its Function in Cold Stress Tolerance.

Anthurium andraeanum is a tropical flower with high ornamental and economic value. Cold stress is one of the major abiotic stresses affecting the quality and value of A. andraeanum; thus, improving the cold tolerance of this species is an important breeding objective. MicroRNAs (miRNAs) have a critical role in plant abiotic stress responses, but their specific molecular regulatory mechanisms are largely unknown, including those related to the cold stress response in A. andraeanum. Here, we identified and cloned the precursor of miR158 from A. andraeanum (Aa-miR158). Both Aa-miR158 and its target gene (c48247) had higher expression levels in strong leaves than in other tissues or organs. Further study revealed that the transcript level of Aa-miR158 was increased by cold stress. Heterologous overexpression of Aa-miR158 improved cold stress tolerance in Arabidopsis, which was associated with decreases in the malondialdehyde (MDA) concentration and relative electrical conductivity (REC) as well as increases in peroxidase (POD) and catalase (CAT) activity. Moreover, overexpressing Aa-miR158 significantly increased the expression of endogenous genes related to cold stress tolerance and reactive oxygen species (ROS) levels in transgenic Arabidopsis under cold stress. Overall, our results demonstrate that Aa-miR158 is significantly involved in the cold stress response and provide a new strategy for cold tolerance breeding of A. andraeanum.

Somatic Embryogenesis of Anthurium andraeanum Linden., -A Tropical Florists' Plant.

The global floriculture market is expected to reach US$41.1 billion by 2027 at a CAGR of 5% over the analysis period 2020-2027; on the year 2020, the recorded market value in this trade was US$29.2 billion. The florists mainly use Anthurium andraeanum flowers in fashionable bouquets and floral arrangements because of their beautiful, attractive bright colored eye-catching spathe, candle-like spadix, prolonged vase life, etc. The cut flower industry always seeks elite cultivars and new hybrids of A. andraeanum, that in turn depend on the availability of large numbers of clonal planting propagules. In vitro somatic embryogenesis is an important technique for large-scale clonal propagation, development of transgenic plants, creation of new variety by somaclonal variation, mutagenesis on in vitro plants, and germplasm preservation for future use. Here, we describe the protocol of somatic embryogenesis of Anthurium andraeanum cv. Cancan, an important commercial cultivated variety. The protocol has been optimized by using 4 different types of culture media which are used during embryogenic callus induction, multiplication of callus, induction of somatic embryogenesis, and maturation plus conversion of embryos into plantlets. The protocol outlines the detailed methods from mother plant procurement to hardening of micro plants that is ready to transfer in the field and it can be used for large-scale commercial propagation.

Integrated small RNA profiling and degradome analysis of Anthurium andraeanum cultivars with different-colored spathes.

MicroRNAs (miRNAs) are known to play vital roles in coloration of leaves, flowers, and fruits in plants. However, their functions in spathe coloration are poorly known. Anthurium andraeanum is a popular ornamental plant with various spathe colors. In this study, small RNA and degradome libraries from three A. andraeanum cultivars with different-colored spathes were constructed and sequenced. Illumina sequencing resulted in 94 conserved miRNAs, and 34 novel miRNAs in total were then identified based on precursor sequences and hairpin structures. Differential expression analysis showed that 52, 51, and 49 miRNAs were differentially expressed in comparisons of orange- versus white-colored spathe, purple- versus white-colored spathe, and purple- versus orange-colored spathe, respectively. The expression patterns of miRNAs and their corresponding targets involved in spathe coloration were further analyzed, and displayed that miR156b and miR529 were highly abundant in the spathes with higher anthocyanin content. These two miRNAs co-targeted a gene encoding SPL17, which may function as a negative regulator in anthocyanin accumulation. In addition, miR408 was also abundantly expressed in purple- and orange-colored spathes, and its typical targets were also identified. This comprehensive integrated analysis provides insight into the miRNA-mediated genetic regulation in spathe coloration of A. andraeanum.

Profiling of Volatile Compounds and Associated Gene Expression in Two Anthurium Cultivars and Their F1 Hybrid Progenies.

Anthurium is an important ornamental crop in the world market and its floral scent can enhance its ornamental value. To date, studies of the components and formation mechanism of the floral scent of Anthurium are relatively few. In this study, the scent profiles of two Anthurium varieties were measured by gas chromatograph-mass spectrometer (GC-MS). There were 32 volatile organic compounds (VOCs) identified in Anthurium 'Mystral', and the most abundant compound was eucalyptol (57.5%). Extremely small amounts of VOCs were detected in Anthurium 'Alabama'. Compared with A. 'Alabama', most genes related to floral scent synthesis exhibited a higher expression in A.'Mystral', including AaDXS, AaDXR, AaMDS, AaHDS, AaTPS, AaDAHPS, AaADT2, AaPAL1, and AaPAL2. In order to produce new varieties of Anthurium with fragrance, 454 progenies of two crossbred combinations of A. 'Mystral' and A. 'Alabama' were obtained. Four F1 generation plants with different floral scent intensities were selected for further study. The major components of floral scent in the progenies were similar to that of the parental A.'Mystral' plant. The expression patterns of genes related to floral scent synthesis were consistent with the relative contents of different types of VOCs. This study revealed the profiles of volatile compounds and associated gene expression in two Anthurium cultivars and their F1 hybrids, which provided a basis for the floral scent inheritance of Anthurium andraeanum.

Typifications of some species names in Anthurium section Pachyneurium (Araceae).

During a taxonomic study of Anthurium sect. Pachyneurium, it was found that the names of four species required typification. Verification of the protologues and cited collections is discussed and typifications are proposed as follows: the illustration Schott Icones Aroideae No. 465 is designated as the neotype of A. affine Schott. A lectotype is designated for A. bonplandii G.S.Bunting since the holotype, cited in the protologue at MY, was not found there. An epitype is selected for A. solitarium Schott because the lectotype illustration of J.M.C. Vellozo (Flora Fluminensis t. 123) lacks sufficient detail to determine it unambiguously to species in A. sect. Pachyneurium. A lectotype is selected for A. glaziovii Hook.f., a synonym of A. solitarium.

Understanding the molecular basis on the biological suppression of bacterial leaf blight of anthurium exerted by Bacillus subtilis (BIO3) through proteomic approach.

We attempted to study the antibacterial activity of rhizospheric Bacillus spp., to curb the bacterial blight of anthurium caused by Xanthomonas axonopodis pv. dieffenbachiae (Xad). Twenty-eight bacterial isolates from rhizospheric regions were identified as different Bacillus spp. and Ochrobactrum sp. using 16S rRNA gene sequencing. B. subtilis BIO3 effectively inhibited the growth of Xad up to 1450.7 mm2, and extracted volatile organic metabolites from the isolate BIO3 inhibited the growth of Xad up to 1024 mm2. Tritrophic interaction of anthurium leaves bacterized with B. subtilis BIO3 and challenged with Xad resulted in the expression of 12 unique proteins compared to untreated control. Mascot Peptide Mass Fingerprint-based identification indicated that one was glutathione peroxidase, involved in defence mechanism, other six proteins were identified as leghemoglobin II, CTP synthase-like, predicted protein (Physcomitrella patens), centromere-associated protein E, grain size protein, and five proteins were hypothetical proteins. Foliar application with 1% liquid formulations (108 CFU/ml) of B. subtilis BIO3 significantly suppressed the bacterial leaf blight of anthurium up to 78% over untreated control and also increased the stem length and flower yield.

Physiological and transcriptome changes induced by exogenous putrescine in anthurium under chilling stress.

BACKGROUND: Chilling stress is the major factor limiting plant productivity and quality in most regions of the world. In the present study, we aimed to evaluate the effects of putrescine (Put) and polyamine inhibitor D-arginine (D-arg) on the chilling tolerance of anthurium (Anthurium andraeanum). RESULTS: Anthurium seedlings were pretreated with five different concentrations of Put solution or D-arg solution. Subsequently, the seedlings were subjected to chilling stress at 6 °C for 3 days, followed by a recovery at 25 °C for 1 day. Relative permeability of the plasma membrane, as well as physiological and morphologic parameters was assessed during the experiments. Additionally, transcriptome sequencing and patterns of differential gene expression related to chilling response were analyzed by qRT-PCR in 1.0 mM Put-treated and untreated anthurium seedlings. Results indicated that the supplementation of exogenous Put decreased the extent of membrane lipid peroxidation and the accumulation of malondialdehyde (MDA), promoted the antioxidant activities and proline content and maintained the morphologic performances compared with the control group. This finding indicated that the application of exogenous Put could effectively decrease the injury and maintain the quality of anthurium under chilling conditions. In contrast, the treatment of D-arg exhibited the opposite effects, which confirmed the effects of Put. CONCLUSIONS: This research provided a possible approach to enhance the chilling tolerance of anthurium and reduce the energy consumption used in anthurium production.

Complete Chloroplast Genomes of Anthurium huixtlense and Pothos scandens (Pothoideae, Araceae): Unique Inverted Repeat Expansion and Contraction Affect Rate of Evolution.

The subfamily Pothoideae belongs to the ecologically important plant family Araceae. Here, we report the chloroplast genomes of two species of the subfamily Pothoideae: Anthurium huixtlense (size: 163,116 bp) and Pothos scandens (size: 164,719 bp). The chloroplast genome of P. scandens showed unique contraction and expansion of inverted repeats (IRs), thereby increasing the size of the large single-copy region (LSC: 102,956 bp) and decreasing the size of the small single-copy region (SSC: 6779 bp). This led to duplication of many single-copy genes due to transfer to IR regions from the small single-copy (SSC) region, whereas some duplicate genes became single copy due to transfer to large single-copy regions. The rate of evolution of protein-coding genes was affected by the contraction and expansion of IRs; we found higher mutation rates for genes that exist in single-copy regions as compared to those in IRs. We found a 2.3-fold increase of oligonucleotide repeats in P. scandens when compared with A. huixtlense, whereas amino acid frequency and codon usage revealed similarities. The ratio of transition to transversion mutations was 2.26 in P. scandens and 2.12 in A. huixtlense. Transversion mutations mostly translated in non-synonymous substitutions. The phylogenetic inference of the limited species showed the monophyly of the Araceae subfamilies. Our study provides insight into the molecular evolution of chloroplast genomes in the subfamily Pothoideae and family Araceae.

Postharvest Spectral Light Composition Affects Chilling Injury in Anthurium Cut Flowers.

The effect of the lighting environment during postharvest storage of ornamentals has largely been neglected in previous research. Anthurium is a cold-sensitive species originating from tropical climates and is widely cultivated all around the world for its colorful spathes. To investigate the effects of light spectrum on the performance of Anthurium cut flowers under cold storage, two cultivars [Calore (red spathe) and Angel (withe spathe)] were placed at low temperature (4°C), either in darkness (D) or under different light spectra [red (R), blue (B), 70:30% red:blue (RB), and white (W)] at an intensity of 125 µmol.m-2.s-1. In both cultivars, the longest and shortest vase lives were observed in spathes exposed to the R and B spectra, respectively. In both cultivars, electrolyte leakage (EL) of spathe was highest under the B and W spectra and lowest under the R spectrum. The highest rate of flower water loss from the spathes was observed under the B-containing light spectra, whereas the lowest rate of water loss was observed in D and under the R spectrum. Negative correlations were observed between EL and vase life and between anthocyanin concentration and EL for both Anthurium cultivars. A positive correlation was found between anthocyanin concentration and vase life. For both Anthurium cultivars, spectral light composition with higher percentage of B resulted in higher EL and as a result shorter vase life in cut flowers under cold storage condition. The negative effect of the B light spectrum on vase life of Anthurium can be explained through its effect on water loss and on oxidative stress and membrane integrity. The quality of Anthurium cut flowers should benefit from environments with restricted B light spectrum during postharvest handling.

Bioreduction: the biological activity, characterization, and synthesis of silver.

Today, nanoparticles are effectively used in different areas. Initially, physical and chemical methods were used in the synthesis of nanoparticles. Biosynthesis (green synthesis) has emerged as an alternative to overcome the toxic effects of chemically synthesized nanoparticles. In this study, green synthesis of silver nanoparticles (AgNPs) with the leaf extract of Anthurium andraeanum was performed. UV-Vis spectrophotometry, scanning transmission electron microscopy, and XRD were applied to characterize the biosynthesized nanoparticles. As a result of the characterization, the spectra showed that a spectrum at a wavelength of about 419 nm and a spherical size of 38 nm nanoparticles was formed. Antibacterial and biofilm inhibition activities of AgNPs against gram-positive and gram-negative bacteria were determined. AgNPs at a concentration of 1 mg/mL showed antibacterial activity against all of the bacterial strains. In the antibiofilm activity study, the highest inhibition percentage was obtained against the P. fluorescens strain at 87.1%, at a concentration of 0.5 mg/mL.

The AabHLH35 Transcription Factor Identified from Anthurium andraeanum is Involved in Cold and Drought Tolerance.

Anthurium andraeanum Lind. is a popular potted and cut-flower plant with an attractive spathe and foliage. It is native to tropical rainforest areas and is able to blossom throughout the year under suitable conditions. However, various abiotic stresses seriously restrict the ornamental value of A. andraeanum and increase the costs of cultivation. A dark green (dg) leaf color mutant of A. andraeanum 'Sonate', which accumulates high levels of anthocyanin, has shown increased vigor and tolerance to stresses during cultivation and is, thus, an ideal germplasm for studying stress tolerance in this species. Here, we show that the anthocyanin content in dg mutant plants at different stages of leaf development was higher than in wild-type (WT) plants, and the ability to tolerate under low-temperature (LT, 14 °C) stress was stronger in dg than in WT plants. RNA-Seq of cDNA libraries from young leaves of dg and WT identified AabHLH35 as a differentially expressed gene (DEG) that was significantly up-regulated in dg. Furthermore, heterologous expression of AabHLH35 improved tolerance to cold and drought stresses in Arabidopsis. These results have built an important molecular foundation for further study of stress tolerance in A. andraeanum.

A duplex quantitative real-time PCR assay for the detection and quantification of Xanthomonas phaseoli pv. dieffenbachiae from diseased and latently infected anthurium tissue.

Anthurium bacterial blight caused by Xanthomonas phaseoli pv. dieffenbachiae (formerly Xanthomonas axonopodis pv. dieffenbachiae) is the major phytosanitary threat in many anthurium growing areas worldwide. Reliable and sensitive diagnostic tools are required for surveillance and certification programs. A duplex real-time quantitative PCR assay was developed for the detection and quantification of X. phaseoli pv. dieffenbachiae from anthurium tissue. This PCR assay targeted a X. phaseoli pv. dieffenbachiae-specific gene encoding an ABC transporter and an internal control encoding for chalcone synthase in Anthurium andreanum. A cycle threshold (Ct), using a receiver-operating characteristic approach (ROC), was implemented to ensure that the declaration of a positive sample was reliable. The duplex real-time assay displayed very high performance with regards to analytical specificity (100% inclusivity, 98.9% exclusivity), analytical sensitivity (LOD95% = 894 bacteria/ml corresponding to 18 bacteria per reaction) and repeatability. We demonstrated the pertinence of this real-time quantitative PCR assay for detecting X. phaseoli pv. dieffenbachiae from diseased leaf tissue (collected from outbreaks on anthurium) and from asymptomatic, latently infected anthurium plants. This assay could be useful for surveillance, as well as for indexing propagative plant material for the presence of X. phaseoli pv. dieffenbachiae.