Physico-Chemical Responses of Alstroemeria spp. cv. Rebecca to the presence of Salicylic Acid and Sucrose in vase solution during postharvest life.

BACKGROUND: The primary challenge in the cut flower industry, specifically in the postharvest phase, is the short vase life of flowers. This issue, along with early leaf yellowing and perianth abscission, significantly diminishes the economic value of flowers due to their accelerated senescence. To tackle this, we conducted a factorial experiment on Alstroemeria cv. Rebecca, utilizing a completely randomized design with three replications. In this experiment the effects of varying concentrations of Salicylic acid (SA) (0, 1.5, and 3 mM) and sucrose (SU) (0% and 3%) were investigated on the postharvest quality of leaves and florets, with systematic evaluations every three days throughout their vase life. RESULTS: This experiment revealed that the specific treatment combination of 1.5 mM SA + 3% SU (T5) markedly improved various parameters, such as vase life, total chlorophyll content, membrane stability index, relative fresh weight, and water uptake of cut flowers. In our analysis, we observed that this preservative solution not only extended the vase life and enhanced water uptake but also effectively preserved total chlorophyll, mitigated the loss of fresh weight, and reduced membrane deterioration in petals. Additionally, our results showed an increase in the activities of catalase (CAT) and peroxidase (POD) enzymes, as well as total protein content, alongside a decrease in malondialdehyde (MDA) and hydrogen peroxide (H2O2) levels. Moreover, this study noted a decrease in microbial populations in solutions containing different concentrations of salicylic acid. CONCLUSIONS: Our research demonstrated that alstroemeria flowers maintained in a solution with 1.5 mM SA + 3% SU exhibited a significantly prolonged vase life of up to 21 days, in contrast to the 15 days observed in control flowers kept in water. These results are highly beneficial for manufacturers in the cut flower industry, as they provide a viable method to substantially extend the vase life of cut flowers. Such an enhancement in flower longevity can lead to increased market value and customer satisfaction. Furthermore, the reduction in flower senescence and decay rates can contribute to decreased waste and greater efficiency in cut flower distribution and sales, offering a substantial advantage to manufacturers in this competitive market. The extended vase life and reduced senescence observed in alstroemeria flowers treated with 1.5 mM SA and 3% SU are attributed to SA's role in enhancing endogenous defense responses and sucrose's function as an energy source, collectively improving water uptake, and delaying the natural decay process.

Alstroemeria Micropropagation in a RITA® Temporary Immersion System.

The use of temporary immersion systems (TIS) for plant micropropagation is an efficient technique for plant production, and we have applied it for the production of alstroemerias. This method involves the cultivation of explants such as rhizomes and axillary buds in a nutrient medium to stimulate shoot growth. TIS offer advantages such as accelerated multiplication processes, uniform production, and cost reduction. This process has shown promise in meeting the growing demand for alstroemeria plants in the market. This chapter describes a specific protocol for temporary immersion bioreactor micropropagation of the "Albatroz" cultivar, with the potential for large-scale automation.

AaLaeA targets AaFla1 to mediate the production of antitumor compound in Alternaria alstroemeria.

Endophytic fungi are an important source of novel antitumor substances. Previously, we isolated an endophytic fungus, Alternaria alstroemeria, from the medicinal plant Artemisia artemisia, whose crude extracts strongly inhibited A549 tumor cells. We obtained a transformant, namely AaLaeAOE26 , which completely loses its antitumor activity due to overexpression of the global regulator AaLaeA. Re-sequencing analysis of the genome revealed that the insertion site was in the noncoding region and did not destroy any other genes. Metabolomics analysis revealed that the level of secondary antitumor metabolic substances was significantly lower in AaLaeAOE26 compared with the wild strain, in particular flavonoids were more downregulated according to the metabolomics analysis. A further comparative transcriptome analysis revealed that a gene encoding FAD-binding domain protein (Fla1) was significantly downregulated. On the other hand, overexpression of AaFla1 led to significant enhancement of antitumor activity against A549 with a sevenfold higher inhibition ratio than the wild strain. At the same time, we also found a significant increase in the accumulation of antitumor metabolites including quercetin, gitogenin, rhodioloside, liensinine, ginsenoside Rg2 and cinobufagin. Our data suggest that the global regulator AaLaeA negatively affects the production of antitumor compounds via controlling the transcription of AaFla1 in endophytic A. alstroemeria.

Multi-walled carbon nanotubes wrapped with polyvinylpyrrolidone can control the leaf yellowing of Alstroemeria cut flowers.

The rapid yellowing of the leaves on cut flowers with leafy stems severely limits their vase life and commercial value. In this study, the effect of a composite of multi-walled carbon nanotubes (MWCNTs) and polyvinyl pyrrolidone (PVP) on the longevity of cut Alstroemeria flowers (Alstroemeria hybrida) was investigated to obtain a solution to this problem. A range of MWCNTs/PVP composite concentrations (0, 3, 6, and 9 mg L-1) was applied in a vase solution (for 24 h) as pulse treatments. Our findings indicate that the composite of MWCNTs and PVP exhibits excellent dispersibility in a vase solution. The results demonstrate that a 3 mg L-1 MWCNTs/PVP concentration was the most effective, extending the vase life of cut Alstroemeria flowers by up to 27 days. Pulsing with MWCNTs/PVP delayed the onset of floret abscission and leaf yellowing by 5 and 18 days, respectively. Additionally, when MWCNTs/PVP solution was applied to cut stems, water uptake remained consistently greater than that of the control. Additionally, MWCNTs/PVP increased the total chlorophyll content, soluble protein content, and POX enzyme activity of leaves while decreasing the malondialdehyde (MDA) content. The results indicate that this composite exhibited antimicrobial activity against gram-positive and -negative bacteria, particularly at a concentration of 3 mg L-1. This study demonstrated that adding MWCNTs/PVP to a vase solution of Alstroemeria cut flowers increased their longevity with minimal leaf yellowing symptoms compared to untreated cut stems. As a result, this nanocomposite can be used safely and effectively in vase solutions and in combination with other preservatives.

Global regulatory factor AaLaeA upregulates the production of antitumor substances in the endophytic fungus Alternaria alstroemeria.

The global regulatory factor LaeA has been shown to be involved in the biosynthesis of secondary metabolites in various fungi. In a previous work, we isolated an endophytic fungus from Artemisia annua, and its extract had a significant inhibitory effect on the A549 cancer cell line. Phylogenetic analysis further identified the strain as Alternaria alstroemeria. Overexpression of AalaeA gene resulted in significantly increased antitumor activity of this strain's extract. The 3-(4, 5- dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay results showed that the inhibition rate of the AalaeAOE29 mutant extract on A549 cancer cells was significantly higher than that of the WT extract, as the IC50 decreased from 195.0 to 107.4 µg/ml, and the total apoptosis rate was enhanced. Overexpression of the AalaeA gene significantly increased the contents of myricetin, geraniol, ergosterol, and 18 other antitumor compounds as determined by metabolomic analysis. Transcriptomic analysis revealed significant changes in 95 genes in the mutant strain, including polyketide synthases, nonribosomal peptide synthases, cytochrome P450s, glycosyltransferases, acetyl-CoA acetyltransferases, and others. These results suggested that AaLaeA mediated the antitumor activity of the metabolites in A. alstroemeria by regulating multiple metabolic pathways.

Decellularized Alstroemeria flower stem modified with chitosan for tissue engineering purposes: A cellulose/chitosan scaffold.

Utilizing plant-based scaffolds has pulled in the consideration of tissue engineers. Plant tissues own different structures with particular porosity and structure. In this study, the stem of the Alstroemeria flower was designated for decellularization to fabricate a new scaffold. The stems were decellularized and called AFSP and then modified by chitosan and named AFSPC. Osteoblast precursor cell line was employed to assess the biological potential of the final scaffolds. The results uncovered that AFSP owns linear microchannels with a smooth surface. AFSPC delineated uniform chitosan coating on the walls with appropriate roughness. AFSPC showed higher potential in swelling, degradation, diffusion, and having a porous structure than AFSP. Modification with chitosan improved mechanical behavior. Biological assays depicted no cytotoxicity for AFSP and AFSPC. AFSPC showed good cell attachment, proliferation, and migration. In conclusion, modified tissue plants can be a good candidate for tissue engineering of both soft and hard tissues.

Satellite DNA probes of Alstroemeria longistaminea (Alstroemeriaceae) paint the heterochromatin and the B chromosome, reveal a G-like banding pattern, and point to a strong structural karyotype conservation.

Alstroemeria species present a well-conserved and asymmetric karyotype. The genus is divided into a Chilean clade, rich in heterochromatin, and a Brazilian clade, poor in heterochromatin. We investigated the distribution of the main repetitive sequences in the chromosomes of the Brazilian species A. longistaminea (2n = 16 + 0-6B) aiming to evaluate the role played by these sequences on the structural organization of the karyotype. In situ hybridization of the three most abundant retrotransposons, corresponding to ~ 45% of the genome, was uniformly distributed. Three satellite DNA sequences, representing near half of the whole satellite fraction (1.93% of the genome), were mainly concentrated on the heterochromatin and one of them painted the whole B chromosome. Noteworthy, some satellites were located on euchromatin, either dispersed or concentrated in clusters along the chromosomes, revealing a G-band-like pattern. The two satellites that presented more C-band- and G-band-like labeling were also hybridized in situ in two other Alstroemeria species. They revealed astonishing similar patterns of distribution, indicating an unusually structural karyotype conservation among Brazilian species.

Origin and evolution of highly polymorphic rDNA sites in Alstroemeria longistaminea (Alstroemeriaceae) and related species.

Alstroemeria (Alstroemeriaceae) displays a conserved and highly asymmetric karyotype, where most rDNA sites can be properly recognized by the size and morphology of the chromosomes. We analyzed the intraspecific variation of rDNA sites in A. longistaminea and compared with their distribution in other species (A. caryophyllaea and A. piauhyensis) and a representative of a sister genus, Bomarea edulis. All three species of Alstroemeria presented 2n = 16, and one to six B chromosomes were found in some individuals of A. longistaminea. There was a set of 12 conserved rDNA sites (four 5S and eight 35S) and up to 11 variable sites. B chromosomes were almost entirely covered by 35S signals, coupled with tiny 5S sites. Noteworthy, most rDNA sites found in A. caryophyllaea and A. piauhyensis were localized in chromosome positions similar to those in A. longistaminea, suggesting the existence of conserved hotspots for rDNA accumulation. Some of these hotspots were absent in Chilean Alstromeria as well in B. edulis. We propose that insertions of rDNA sequences on chromosomes do not occur randomly but rather on preferential sites or hotspots for insertions. The maintenance of these arrays, however, may be favored/constrained by different factors, resulting in stable or polymorphic sites.

Molecular characterization and complete genome of alstroemeria mosaic virus (AlMV).

Even though alstroemeria mosaic virus (AlMV) is one of the most important viruses affecting alstroemeria plants, its genome is only partially available in public sequence databases. High throughput sequencing (HTS) of RNA from alstroemeria plants with symptoms of mosaic and streaking, collected in Lasso-Ecuador, indicated the presence of AlMV and lily symptomless virus. In this study, we aimed to assemble and characterize the complete genome sequence of AlMV. Reads from Illumina sequencing of ribosomal RNA-depleted total RNA were assembled into contigs that were mapped to the sunflower chlorotic mottle virus genome, revealing the 9774 [corrected] bp complete genome sequence of AlMV. Multiple sequence alignment of the AlMV polyprotein with close homologs allowed the identification of ten mature proteins P1, HC-Pro, P3, 6K1, CI, 6K2, NIa-VPg, NIa-Pro, NIb and CP. Furthermore, several potyvirus motifs were identified in the AlMV polyprotein including those related to potyvirus aphid transmission 334KMTC337, 592PTK594 and 2800DAG2802. Phylogenetic analysis based in the polyprotein showed that AlMV belongs to the potato virus Y clade and its closest relative is sunflower ring blotch virus. This study describes the first complete genome of AlMV and its placement within the genus Potyvirus, providing valuable information for future studies on this economically important virus.

Alstroemeria yellow spot virus (AYSV): a new orthotospovirus species within a growing Eurasian clade.

An orthotospovirus distinct from all other orthotospoviruses was isolated from naturally infected alstroemeria plants. Disease symptoms caused by this virus mainly consisted of yellow spots on the leaves based on which the name alstroemeria yellow spot virus (AYSV) was coined. A host range analysis was performed and a polyclonal antiserum was produced against purified AYSV ribonucleoproteins which only reacted with the homologous antigen and not with any other (established or tentative) orthotospovirus from a selection of American and Asian species. Upon thrips transmission assays the virus was successfully transmitted by a population of Thrips tabaci. The entire nucleotide sequence of the M and S RNA segments was elucidated by a conventional cloning and sequencing strategy, and contained 4797 respectively 2734 nucleotides (nt). Simultaneously, a next generation sequencing (NGS) approach (RNAseq) was employed and generated contigs covering the entire viral tripartite RNA genome. In addition to the M and S RNA nucleotide sequences, the L RNA (8865 nt) was obtained. The nucleocapsid (N) gene encoded by the S RNA of this virus consisted of 819 nucleotides with a deduced N protein of 272 amino acids and by comparative sequence alignments to other established orthotospovirus species showed highest homology (69.5% identity) to the N protein of polygonum ringspot virus. The data altogether support the proposal of AYSV as a new orthotospovirus species within a growing clade of orthotospoviruses that seem to share the Middle East basin as a region of origin.

Changes in postharvest physio-biochemical characteristics and antioxidant enzymes activity of cut alsteroemeria aurantiaca flower as affected by cycloheximide, coconut water and 6-benzyladenine / Alterações nas características físico-bioquímicas pós-colheita e atividades de enzimas antioxidantes da flor de alstroemeria aurantiaca cortada como afetada pela cicloheximida, água de coco e 6-benziladenina

Early leaf yellowing in cut alstroemeria (Alstroemeria aurantiaca) flowers before flower development and petal abscission is an important limiting postharvest quality and vase life factors. Early leaf senescence reduces postharvest longevity of cut flowers and promotes petal's wilting. A study was made to evaluate the response of cut alstroemeria flowers at varying concentrations of cycloheximide (CHI) (50, 100 and 200 mg l-1), coconut water (5, 10 and 20%) and 6-benzyladenine (BA) (50, 100 and 200 mg l-1). CHI, coconut water and BA extended the vase life at all concentrations compared to the control, but coconut water at 5% concentration (with 17.39 days) was the most effective treatment. Control cut flowers showed the least vase life (10.76 days). Ethylene production in cut flowers promoted flower senescence. All concentrations of CHI, coconut water and BA delayed ethylene production compared to the control. Treatment of cut flowers with coconut water at concentration of 5% maintained the highest fresh weight of flowers and increased the content of water uptake. The chlorophyll degradation was significantly reduced by the application of CHI, coconut water and BA. The maximum content of membrane's lipid peroxidation and antioxidant enzymes activity (super oxide dismutase and peroxidase) was obtained in control cut flowers. Thus, 5% fresh coconut water has the potential to be applied as vase solution (preservative medium) due to prolongs of cut alstroemeria flowers.

O amarelecimento precoce das folhas em flores de alstroemeria (Alstroemeria aurantiaca) cortadas antes do desenvolvimento floral e da abscisão de pétalas é um importante limitante da qualidade pós-colheita e dos fatores de vida do vaso. A senescência precoce da folha reduz a longevidade pós-colheita das flores cortadas e promove o murchamento da pétala. Um estudo foi realizado para avaliar a resposta de flores de alstroemeria cortadas em diferentes concentrações de cicloheximida (CHI) (50, 100 e 200 mg l-1), água de coco (5, 10 e 20%) e 6-benziladenina (BA) 50, 100 e 200 mg l-1). CHI, água de coco e BA prolongou a vida do vaso em todas as concentrações em comparação com o controle, mas a água de coco a 5% de concentração (com 17,39 dias) foi o tratamento mais eficaz. As flores cortadas de controlo mostraram a menor vida útil do vaso (10,76 dias). A produção de etileno em flores cortadas promoveu a senescência da flor. Todas as concentrações de CHI, água de coco e BA atrasaram a produção de etileno em comparação com o controle. O tratamento de flores cortadas com água de coco a uma concentração de 5% manteve o maior peso fresco de flores e aumentou o conteúdo de absorção de água. A degradação da clorofila foi significativamente reduzida pela aplicação de CHI, água de coco e BA. O teor máximo de atividade de enzimas antioxidantes e de peroxidação lipídica da membrana (super óxido dismutase e peroxidase) foi obtido em flores cortadas de controle. Assim, 5% de água de coco fresca tem potencial para ser aplicada como solução de vaso (meio de conservação) devido a prolongamentos das flores de alstroemeria cortadas.

Comparative genome analysis and phylogenetic relationship of order Liliales insight from the complete plastid genome sequences of two Lilies (Lilium longiflorum and Alstroemeria aurea).

Monocots are one of the most diverse, successful and economically important clades of angiosperms. We attempt to analyse the complete plastid genome sequences of two lilies and their lengths were 152,793bp in Lilium longiflorum (Liliaceae) and 155,510bp in Alstroemeria aurea (Alstroemeriaceae). Phylogenetic analyses were performed for 28 taxa including major lineages of monocots using the sequences of 79 plastid genes for clarifying the phylogenetic relationship of the order Liliales. The sister relationship of Liliales and Asparagales-commelinids was improved with high resolution. Comparative analyses of inter-familial and inter-specific sequence variation were also carried out among three families of Liliaceae, Smilacaceae, and Alstroemeriaceae, and between two Lilium species of L. longflorum and L. superbum. Gene content and order were conserved in the order Liliales except infA loss in Smilax and Alstroemeria. IR boundaries were similar in IRa, however, IRb showed different extension patterns as JLB of Smilax and JSB in Alstroemeria. Ka/Ks ratio was high in matK among the pair-wise comparison of three families and the most variable genes were psaJ, ycf1, rpl32, rpl22, matK, and ccsA among the three families and rps15, rpoA, matK, and ndhF between Lilium.

Ribosomal DNA distribution and a genus-wide phylogeny reveal patterns of chromosomal evolution in Alstroemeria (Alstroemeriaceae).

PREMISE OF THE STUDY: Understanding the flexibility of monocot genomes requires a phylogenetic framework, which so far is available for few of the ca. 2800 genera. Here we use a molecular tree for the South American genus Alstroemeria to place karyological information, including fluorescent in situ hybridization (FISH) signals, in an explicit evolutionary context. METHODS: From a phylogeny based on plastid, nuclear, and mitochondrial sequences for most species of Alstroemeria, we selected early-branching (Chilean) and derived (Brazilian) species for which we obtained 18S-25S and 5S rDNA FISH signals; we also analyzed chromosome numbers, 1C-values, and telomere FISH signals (in two species). KEY RESULTS: Chromosome counts for Alstroemeria cf. rupestris and A. pulchella confirm 2n = 16 as typical of the genus, which now has chromosomes counted for 29 of its 78 species. The rDNA sites are polymorphic both among and within species, and interstitial telomeric sites in Alstroemeria cf. rupestris suggest chromosome fusion. CONCLUSIONS: In spite of a constant chromosome number, closely related species of Alstroemeria differ drastically in their rDNA, indicating rapid increase, decrease, or translocations of these genes. Previously proposed Brazilian and Chilean karyotype groups are not natural, and the n = 8 chromosomes in Alstroemeria compared to n = 9 in its sister genus Bomarea may result from a Robertsonian fusion.

Reduced transcription of a LEAFY-like gene in Alstroemeria sp. cultivar Green Coral that cannot develop floral meristems.

Alstroemeria sp. cv. Green Coral has numerous bracts instead of flowers, and its cyme structures are repeated eternally. Observations of the development and morphology of inflorescence in cv. Green Coral revealed that transition from inflorescence to floral meristem was restricted. We isolated and characterized floral meristem identity genes LEAFY-like (AlsLFY) and SQUAMOSA-like (AlsSQa and AlsSQb) genes from Alstroemeria ligtu. In situ hybridization results indicated that AlsSQa and AlsSQb were expressed in the dome-shaped floral meristems and all floral organ primordia in A. ligtu. Transcripts of AlsLFY accumulated early in the dome-shaped floral meristems; the signals were restricted later to the outer region of the floral meristem. These results indicate that AlsLFY, AlsSQa, and AlsSQb function as floral meristem identity genes. Expression profiles of AlsLFY, AlsSQa, AlsSQb, and other MADS-box genes were compared between A. ligtu and cv. Green Coral. AlsLFY, AlsDEFa, and AlsAGL6 transcripts were not detected at the shoot apices of cv. Green Coral but were detected in A. ligtu. The early induction and accumulation of AlsLFY transcripts in the inflorescence meristem of A. ligtu prior to development of the floral meristem suggest that downregulation of AlsLFY is likely to restrict the inflorescence-to-floral meristem transition in cv. Green Coral.

Volatile emissions of scented Alstroemeria genotypes are dominated by terpenes, and a myrcene synthase gene is highly expressed in scented Alstroemeria flowers.

Native to South America, Alstroemeria flowers are known for their colourful tepals, and Alstroemeria hybrids are an important cut flower. However, in common with many commercial cut flowers, virtually all the commercial Alstroemeria hybrids are not scented. The cultivar 'Sweet Laura' is one of very few scented commercial Alstroemeria hybrids. Characterization of the volatile emission profile of these cut flowers revealed three major terpene compounds: (E)-caryophyllene, humulene (also known as α-caryophyllene), an ocimene-like compound, and several minor peaks, one of which was identified as myrcene. The profile is completely different from that of the parental scented species A. caryophyllaea. Volatile emission peaked at anthesis in both scented genotypes, coincident in cv. 'Sweet Laura' with the maximal expression of a putative terpene synthase gene AlstroTPS. This gene was preferentially expressed in floral tissues of both cv. 'Sweet Laura' and A. caryophyllaea. Characterization of the AlstroTPS gene structure from cv. 'Sweet Laura' placed it as a member of the class III terpene synthases, and the predicted 567 amino acid sequence placed it into the subfamily TPS-b. The conserved sequences R(28)(R)X(8)W and D(321)DXXD are the putative Mg(2+)-binding sites, and in vitro assay of AlstroTPS expressed in Escherichia coli revealed that the encoded enzyme possesses myrcene synthase activity, consistent with a role for AlstroTPS in scent production in Alstroemeria cv. 'Sweet Laura' flowers.

Deletions and recombinations with the RNA1 3' ends of different tobraviruses have created a multitude of tobacco rattle virus TCM-related RNA2 species in Alstroemeria and tulip.

In vegetatively propagated Alstroemeria plants that showed pronounced stunting and necrotic leaf spots, a tobravirus infection was diagnosed in which one tobacco rattle virus (TRV, strain AL) RNA1 species was associated with seven different RNA2 species. The latter differed considerably in size and in the types of their 3' RNA1-related sequences. The 5' RNA2-specific part of all these RNA2 molecules showed almost 100% sequence identity with that of RNA2 of the TRV isolate TCM from tulip, but in some of these RNA2 molecules it was shorter than in the TCM isolate, whereas in others it was longer. One of the TRV AL RNA2 molecules, i.e. TC3'PE-a, contained the full set of three full-length RNA2-specific ORFs (ORF2a, -2b and -2c), whereas the previously analysed TCM sequence contained only ORF2a and -2b. In four of these TRV AL RNA2 molecules, i.e. those that had a relatively short RNA2-specific part, the 3' end was identical to that of the cognate TRV AL RNA1, but in the other three, which had a long RNA2-specific part, it was closely related to that of pea early browning virus (PEBV) RNA1, which was not detected in the infected plants. A comparison with previously described TRV/PEBV RNA2 recombinants suggested that the various TRV AL RNA2 molecules may represent various steps and side steps in an evolutionary process, which is apt to open the wide host range of TRV also to PEBV-derived RNA2 species.

A specific group of genes respond to cold dehydration stress in cut Alstroemeria flowers whereas ambient dehydration stress accelerates developmental senescence expression patterns.

Petal development and senescence entails a normally irreversible process. It starts with petal expansion and pigment production, and ends with nutrient remobilization and ultimately cell death. In many species this is accompanied by petal abscission. Post-harvest stress is an important factor in limiting petal longevity in cut flowers and accelerates some of the processes of senescence such as petal wilting and abscission. However, some of the effects of moderate stress in young flowers are reversible with appropriate treatments. Transcriptomic studies have shown that distinct gene sets are expressed during petal development and senescence. Despite this, the overlap in gene expression between developmental and stress-induced senescence in petals has not been fully investigated in any species. Here a custom-made cDNA microarray from Alstroemeria petals was used to investigate the overlap in gene expression between developmental changes (bud to first sign of senescence) and typical post-harvest stress treatments. Young flowers were stressed by cold or ambient temperatures without water followed by a recovery and rehydration period. Stressed flowers were still at the bud stage after stress treatments. Microarray analysis showed that ambient dehydration stress accelerates many of the changes in gene expression patterns that would normally occur during developmental senescence. However, a higher proportion of gene expression changes in response to cold stress were specific to this stimulus and not senescence related. The expression of 21 transcription factors was characterized, showing that overlapping sets of regulatory genes are activated during developmental senescence and by different stresses.

A distinct tospovirus causing necrotic streak on Alstroemeria sp. in Colombia.

A tospovirus causing necrotic streaks on leaves was isolated from Alstroemeria sp. in Colombia. Infected samples reacted positively with tomato spotted wilt virus (TSWV) antiserum during preliminary serological tests. Further analysis revealed a close serological relationship to tomato chlorotic spot virus (TCSV) and groundnut ringspot virus (GRSV). A major part of the S-RNA segment, encompassing the nucleocapsid (N) protein gene, the 5' untranslated region and a part of the intergenic region 3' of the N gene, was cloned and sequenced. The deduced N protein sequence showed highest amino acid identity (82%) to that of TCSV, indicating that the virus represents a new tospovirus species, for which the name Alstroemeria necrotic streak virus (ANSV) is coined. Phylogenetic analysis based on the N protein sequence revealed that this Alstroemeria-infecting tospovirus clustered with tospoviruses from the American continent. Frankliniella occidentalis was identified as potential vector species for ANSV.

Isolation and characterization of the plant glsA promoter from Alstroemeria.

The differentiation of a vegetative cell and a generative cell is a critical event during pollen development. The Lilium GlsA is known to localize in pollen and is considered to be involved in development of the generative cell. Here, we cloned a glsA ortholog from Alstroemeria, a commercially important cut flower. The expression of AaglsA (Alstroemeria aurea glsA) transcripts increased gradually after pollen mitosis I (PMI) and reached a significant level when the generative cell started to elongate. Analysis of the promoter of AaglsA suggests that AaglsA expression is controlled by several cis-regulatory elements during pollen development. This is the first investigation of reproductive factors regulating male gametogenesis in Alstroemeria.

Detection of changes in the nuclear phase and evaluation of male germ units by flow cytometry during in vitro pollen tube growth in Alstroemeria aurea.

This study aimed to analyze male gamete behavior from mature pollen to pollen tube growth in the bicellular pollen species Alstroemeria aurea. For mature pollen, pollen protoplasts were examined using flow cytometry. The protoplasts showed two peaks of DNA content at 1C and 1.90C. Flow cytometry at different developmental stages of pollen tubes cultured in vitro revealed changes in the nuclear phase at 9 and 18 h after culture. Sperm cell formation occurred at 6-9 h after culture, indicating that the first change was due to the division of the generative cells into sperm cells. After sperm cell formation, the number of vegetative nucleus associations with sperm cells showed a tendency to increase. This association was suggested as the male germ unit (MGU). When sperm cells, vegetative nuclei, and partial MGUs were collected separately from pollen tubes cultured for 18 h and analyzed using a flow cytometer, the sperm cells and vegetative nuclei contained 1C DNA, while the DNA content of partial MGUs was counted as 2C. Therefore, the second change in the nuclear phase, which results in an increase in 2C nuclei, is possibly related to the formation of MGUs.