Transcriptomic analysis of Verbena bonariensis roots in response to cadmium stress.

BACKGROUND: Cadmium (Cd) is a serious heavy metal (HM) soil pollutant. To alleviate or even eliminate HM pollution in soil, environmental-friendly methods are applied. One is that special plants are cultivated to absorb the HM in the contaminated soil. As an excellent economical plant with ornamental value and sound adaptability, V. bonariensis could be adapted to this very situation. In our study, the Cd tolerance in V. bonariensis was analyzed as well as an overall analysis of transcriptome. RESULTS: In this study, the tolerance of V. bonariensis to Cd stress was investigated in four aspects: germination, development, physiological changes, and molecular alterations. The results showed that as a non-hyperaccumulator, V. bonariensis did possess the Cd tolerance and the capability to concentration Cd. Under Cd stress, all 237, 866 transcripts and 191, 370 unigenes were constructed in the transcriptome data of V. bonariensis roots. The enrichment analysis of gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway revealed that differentially expressed genes (DEGs) under Cd stress were predominately related to cell structure, reactive oxygen species (ROS) scavenging system, chelating reaction and secondary metabolites, transpiration and photosynthesis. DEGs encoding lignin synthesis, chalcone synthase (CHS) and anthocyanidin synthase (ANS) were prominent in V. bonariensis under Cd stress. The expression patterns of 10 DEGs, validated by quantitative real-time polymerase chain reaction (qRT-PCR), were in highly accordance with the RNA-Sequence (RNA-Seq) results. The novel strategies brought by our study was not only benefit for further studies on the tolerance of Cd and functional genomics in V. bonariensis, but also for the improvement molecular breeding and phytoremediation.

Transcriptomic Analysis of Verbena bonariensis Leaves Under Low-Temperature Stress.

Verbena bonariensis is a valuable plant for both ornament and flower border. As a major constraint, low temperature affects the growing development and survival of V. bonariensis. However, there are few systematic studies in terms of molecular mechanism on the tolerance of low temperature in V. bonariensis. In this study, Illumina sequencing technology was applied to analyze the cold resistance mechanism of plants. Six cDNA libraries were obtained from two samples of two groups, the cold-treated group and the control group. A total of 271,920 unigenes were produced from 406,641 assembled transcripts. Among these, 19,003 differentially expressed genes (DEGs) (corrected p-value <0.01, |log2(fold change) | >3) were obtained, including 9852 upregulated and 9151 downregulated genes. The antioxidant enzyme system, photosynthesis, plant hormone signal transduction, fatty acid metabolism, starch and sucrose metabolism pathway, and transcription factors were analyzed. Based on these results, series of candidate genes related to cold stress were screened out and discussed. The physiological indexes related to response mechanism of low temperature were tested. Eleven upregulated DEGs were validated by Quantitative Real-time PCR. In this study, we provided the transcriptome sequence resource of V. bonariensis and used these data to realize its molecular mechanism under cold stress. The results contributed to valuable clues for genetic studies and helped to screen candidate genes for cold-resistance breeding.

Whole-Transcriptome Sequence Analysis of Verbena bonariensis in Response to Drought Stress.

Drought is an important abiotic factor that threatens the growth and development of plants. Verbena bonariensis is a widely used landscape plant with a very high ornamental value. We found that Verbena has drought tolerance in production practice, so in order to delve into its mechanism of drought resistance and screen out its drought-resistance genes, we used the RNA-Seq platform to perform a de novo transcriptome assembly to analyze Verbena transcription response to drought stress. By high-throughput sequencing with Illumina Hiseq Xten, a total of 44.59 Gb clean data was obtained from T01 (control group) and T02 (drought experiment group). After assembly, 111,313 unigenes were obtained, and 53,757 of them were annotated by compared databases. In this study, 4829 differentially expressed genes were obtained, of which 4165 were annotated. We performed GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analyses, and explored a lot of differently expressed genes related to plant energy production, hormone synthesis, cell signal transduction, and metabolism to understand the stress response of Verbena in drought stress. In addition, we also found that a series of TFs related to drought-resistance of Verbena and provide excellent genetic resources for improving the drought tolerance of crops.

Enhancement of phosphate absorption by garden plants by genetic engineering: a new tool for phytoremediation.

Although phosphorus is an essential factor for proper plant growth in natural environments, an excess of phosphate in water sources causes serious pollution. In this paper we describe transgenic plants which hyperaccumulate inorganic phosphate (Pi) and which may be used to reduce environmental water pollution by phytoremediation. AtPHR1, a transcription factor for a key regulator of the Pi starvation response in Arabidopsis thaliana, was overexpressed in the ornamental garden plants Torenia, Petunia, and Verbena. The transgenic plants showed hyperaccumulation of Pi in leaves and accelerated Pi absorption rates from hydroponic solutions. Large-scale hydroponic experiments indicated that the enhanced ability to absorb Pi in transgenic torenia (AtPHR1) was comparable to water hyacinth a plant that though is used for phytoremediation causes overgrowth problems.

Northwestern Argentina: a center of genetic diversity of lemon verbena (Aloysia citriodora PALÁU, Verbenaceae).

The aerial parts of lemon verbena (Aloysia citriodora PALÁU) are worldwide used due to their medicinal and aromatic properties. The essential-oil and acteoside contents have been proposed as the main quality markers for their pharmacological and organoleptic features. The northwestern region of Argentina has been repeatedly proposed as the place of origin for this species. For this reason, the essential-oil yields and chemical compositions of leaves of 25 populations of lemon verbena from both wild collections and experimental crops from this region were studied. Plants from six different collections were subsequently grown on the same experimental parcel located at Cerrillos, Salta province, during more than seven years. In addition, the acteoside contents determined in all the samples collected in 2010 showed significant variations (from 0.5 to 4.0%). Large differences were observed in the essential-oil composition and yields, which ranged from 0.4 to 2.1% (v/w). Nevertheless, most of the samples complied with the European Pharmacopoeia specifications. A remarkable chemical diversity with at least four clearly defined chemotypes was detected in this region. Therefore, it would be urgent to encourage actions to protect these genotypes of lemon verbena in the northwestern Argentina.

[Verbena canadensis Britt mutants with altered development of flowers and inflorescences generated by chemical mutagenesis].

Rose vervain is a little-known ornamental annual plant. The adaptive properties (drought and cold resistance) and long period of flowering make this species promising for growing in flower gardens and containers. Chemical mutagenesis widely used for various plant species was applied to induce character variation in Rose vervain. The properties of development of flowers and inflorescences in lines descending from the M3--M5 mutants generated by the seed exposure to chemical mutagens diethyl sulfate and nitrosomethylurea were considered.

A species-level phylogenetic study of the Verbena complex (Verbenaceae) indicates two independent intergeneric chloroplast transfers.

Two major impediments to infer plant phylogenies at inter- or intra- species level include the lack of appropriate molecular markers and the gene tree/species tree discordance. Both of these problems require more extensive investigations. One of the foci of this study is examining the phylogenetic utility of a combined chloroplast DNA dataset (>5.0kb) of seven non-coding regions, in comparison with that of a large fragment (ca. 3.0kb) of a low-copy nuclear gene (waxy), in a recent, rapidly diversifying group, the Verbena complex. The complex includes three very closely related genera, Verbena (base chromosome number x=7), Glandularia (x=5), and Junellia (x=10), comprising some 150 species distributed predominantly in South and North America. Our results confirm the inadequacy of non-coding cpDNA in resolving relationships among closely related species due to lack of variation, and the great potential of low-copy nuclear gene as source of variation. However, this study suggests that when both cpDNA and nuclear DNA are employed in low-level phylogenetic studies, cpDNA might be very useful to infer organelle evolutionary history (e.g., chloroplast transfer) and more comprehensively understand the evolutionary history of organisms. The phylogenetic framework of the Verbena complex resulted from this study suggests that Junellia is paraphyletic and most ancestral among the three genera; both Glandularia and Verbena are monophyletic and have been derived from within Junellia. Implications of this phylogenetic framework to understand chromosome number evolution and biogeography are discussed. Most interestingly, the comparison of the cpDNA and nuclear DNA phylogenies indicates two independent intergeneric chloroplast transfers, both from Verbena to Glandularia. One is from a diploid North American Verbena species to a polyploid North American Glandularia species. The other is more ancient, from the South American Verbena group to the common ancestor of a major Glandularia lineage, which has radiated subsequently in both South and North America. The commonly assumed introgressive hybridization may not explain the chloroplast transfers reported here. The underlying mechanism remains uncertain.

A MADS-box gene of Populus deltoides expressed during flower development and in vegetative organs.

A MADS-box gene (PdPI) was isolated from a cDNA library constructed from male flower buds of Populus deltoides Bartr. ex Marsh. According to an analysis of genomic DNA structure and putative protein structure, and a phylogenetic study, PdPI is an ortholog of the Arabidopsis PI gene. Relative-quantitative real-time polymerase chain reaction analysis showed that PdPI has a broader expression pattern than PI in Arabidopsis. PdPI was strongly expressed in floral buds and roots and weakly expressed in immature xylem, leaves and apical buds of the male P. deltoides tree. In male inflorescences, PdPI expression was abundant in the perianth and anther, and weak in the peduncle and mature pollen. The large differences in PdPI expression at various phases of male floral bud development were closely related to the development of flower organs (perianth and stamen) and pollen. PdPI was also expressed in female inflorescences. Our results suggest that PdPI has multiple functions in the development of P. deltoides.

Identification of the genotype from the content and composition of the essential oil of lemon verbena (Aloysia citriodora Palau).

Aloysia citriodora accessions from cultivated material, botanical collections, and wild populations were studied by means of their biomass and essential oil production and composition to assist the selection of the most promising genotype. The study was carried out through both field experiments during two year's time and laboratory processes. Data were evaluated by means of univariate and multivariate techniques. Aloysia citriodora intraspecific variation was accounted for by differences in both yield and chemical profiles of the essential oils, but no differences were found in the biomass production. Three chemotypes were identified according to qualitative and quantitative differences of the essential oils. For the higher contents of neral and geranial, Mendoza accession was the most promising to be encouraged for future crops.

Regeneration of transformed verbena (Verbena x hybrida) by Agrobacterium tumefaciens.

Verbena (Verbena x hybrida), an important floricultural species, was successfully regenerated from stem segments on Murashige and Skoog's basal medium supplemented with thidiazuron and indole-3-acetic acid. A transformation system was developed using cvs. Temari Scarlet, Temari Sakura, Tapien Rose and TP-P2. Agrobacterium tumefaciens strain Agl0 harboring the sGFP gene was infected into stem segments. Transformation efficiency was improved by evaluating and manipulating the age of the plant material, the concentration of kanamycin in the medium during selection, and the length of the culture period in the dark. After 2-3 months of culture on the selection medium, GFP-positive shoots were obtained in all four of the cultivars tested. These shoots were successfully acclimated and set flowers within 2-3 months in a greenhouse. GFP was expressed in all of the organs including the floral parts. Stable genomic transformation was confirmed by Southern blot analysis. No morphological differences were observed between the transformed plants and their host plants.