RNA interference (RNAi) can be used for the protection against agricultural pests through the silencing of genes required for pest fitness. To assess the potential of RNAi approaches in the two-spotted spider mite, Tetranychus urticae, we compared 5 methods for the delivery of double-stranded RNA (dsRNA). These methods include mite feeding on either (i) leaves floating on a dsRNA solution, (ii) dsRNA-expressing plants, (iii) artificial diet supplemented with dsRNA, or (iv) dsRNA-coated leaves, and (v) mite soaking in a dsRNA solution. In all cases, the gene targeted for method validation was the Vacuolar-type H+-ATPase (TuVATPase), encoding a constitutively expressed ATP-driven proton pump located in the membrane. Down-regulation of TuVATPase increased mortality and/or reduced fecundity in all methods, but with variable efficiency. The most efficient methods for dsRNA delivery were direct soaking of mites in the dsRNA solution and mite feeding on dsRNA-coated leaves that mimics dsRNA application as a sprayable pesticide. Both resulted in a dark-body phenotype not observed in mites treated with a control dsRNA. Although with lower efficiency, dsRNA designed for TuVATPase silencing and expressed in transgenic Arabidopsis plants impacted the fitness of mites feeding on these plants. RNAi may thus be a valuable strategy to control spider mite populations, either as a sprayable pesticide or through transgenic crops. This comparative methodological study focusing on the induction of RNAi-based gene silencing in T. urticae paves the way for reverse genetics approaches in this model chelicerate system and prepares large-scale systematic RNAi screens as a first step towards the development of specific RNA-based pesticides. Such alternative molecules may help control spider mites that cause significant damages to crops and ornamental plant species, as well as other chelicerates detrimental to agriculture and health.

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Pollen morphology of 19 species of Allium representing six subgenera and 12 sections, were studied. The following characters were recognized as important for separating taxa at different taxonomic ranks: the sulcus, presence or absence of perforations on the pollen surface, density of perforations, size of perforations and size of the pollen grains. Rugulate, rugulate–striate, and simple–perforate exine ornamentation were distinguished. The type of sulcus is very characteristic in A. subgen. Allium sect. Allium. The extended sulcus was not observed in any species the other sections of subgen. Allium studied here, such as sect. Caerulea (A.capitellatum), sect. Codonprasum (A.lenkoranicum and A.stamineum) and sect. Avulsea (A.rubellum and A.umblicatum). Imperforated pollen grains were observed in representatives of A. subgen. Reticulatobulbosa and subgen. Polyprason.

Saffron, made from the dried stigmas of Crocus sativus L., contains pigments and valuable aromatic compounds, and can be used in medicine and as a spice. Nowadays its production is lower than demand. Tissue culture presents an alternative biochemical tool which can be used to produce stigma-like structure (SLS) in vitro. In this study, the origin and induction of SLS formation was investigated in ovary and style explants of floral buds on MS medium supplemented with 1-naphthalene acetic acid (NAA) and 6-benzlaminopurine (BAP). SLS were directly originated through meristematic cells or indirectly in the form of colorless globular structures from parenchyma tissue. The colorless globular structures initially were conical and pale yellow color at the sharp ends; subsequently they matured into trumpet-like red stigmas with or without finger-like papillae at the margins. Light and electron microscopic observations of ultra-and semithin sections of different developmental stages of SLS showed that these structures possess two kinds of cells: (1) small cells close to parenchyma tissues and (2) large cells oriented towards the peripheral area and apparently originated from the small ones. Our results suggest that the SLS originated from internal parenchyma tissues.K Ke ey y w wo or rd ds s: : Crocus sativus L., meristematic cells, tissue culture, stigma-like structures, parenchyma tissues.

Pollen morphology of 19 species of Allium representing six subgenera and 12 sections, were studied. The following characters were recognized as important for separating taxa at different taxonomic ranks: the sulcus, presence or absence of perforations on the pollen surface, density of perforations, size of perforations and size of the pollen grains. Rugulate, rugulateÁstriate, and simpleÁperforate exine ornamentation were distinguished. The type of sulcus is very characteristic in A. subgen. Allium sect. Allium. The extended sulcus was not observed in any species the other sections of subgen. Allium studied here, such as sect. Caerulea (A. capitellatum), sect. Codonprasum (A. lenkoranicum and A. stamineum) and sect. Avulsea (A. rubellum and A. umblicatum). Imperforated pollen grains were observed in representatives of A. subgen. Reticulatobulbosa and subgen. Polyprason.