Heterotrimeric G proteins composed of a, b, and g subunits are central signal transducers mediating the cellular response to multiple stimuli in most eukaryotes. Gg subunits provide proper cellular localization and functional specificity to the heterotrimer complex. Plant Gg subunits, divided into three structurally distinct types, are more diverse than their animal counterparts. Type B Gg subunits, lacking a carboxyl-terminal isoprenylation motif, are found only in flowering plants. We present the functional characterization of type B Gg subunit (SlGGB1) in tomato (Solanum lycopersicum). We show that SlGGB1 is the most abundant Gg subunit in tomato and strongly interacts with the Gb subunit. Importantly, the green fluorescent proteinSlGGB1 fusion protein as well as the carboxyl-terminal yellow fluorescent protein-SlGGB1/amino-terminal yellow fluorescent protein-Gb heterodimer were localized in the plasma membrane, nucleus, and cytoplasm. RNA interference-mediated silencing of SlGGB1 resulted in smaller seeds, higher number of lateral roots, and pointy fruits. The silenced lines were hypersensitive to exogenous auxin, while levels of endogenous auxins were lower or similar to those of the wild type. SlGGB1-silenced plants also showed strong hyposensitivity to abscisic acid (ABA) during seed germination but not in other related assays. Transcriptome analysis of the transgenic seeds revealed abnormal expression of genes involved in ABA sensing, signaling, and response. We conclude that the type B Gg subunit SlGGB1 mediates auxin and ABA signaling in tomato.
Polyploid plants have been induced in differentAsparagus officinalis L. breeding programs in order to obtain plants with improved agronomical traits, such as large spear diameter or segregation ratios with a higher number of males. The polyploidization methods can produce somaclonal variation in the polyploid plants obtained and, therefore, unwanted changes in the agronomical traits of the initial elite plants. We used two different polyploidization methods to induce polyploid plants from diploid genotypes of commercial varieties and tetraploid genotypes of the Spanish landrace ''Morado de Hue ´tor''. The first method was the culture of rhizome buds in the medium ARBM-3 (Asparagus Rhizome Bud Medium), supplemented with different concentrations of colchicine (0.1-0.75 g l -1 ) for 10 and 20 days. The best polyploidization rate obtained was 25 % (0.5 g l -1 colchicine for 10 days). The second method was the regeneration of polyploid plants from callus culture, resulting in a polyploidization rate of 40 and 12.5 % for the diploid genotype CM077 and the tetraploid genotype HT156, respectively. Additionally, we have developed a new protocol to separate the mixoploids generated into their different genetic components, obtaining plants with a unique ploidy level. EST-SSRs markers were employed to analyze the genetic stability of polyploidy plants. Somaclonal variation was not detected for polyploidy plants obtained through the culture of rhizome bud explants. Therefore, these polyploid plants should maintain the agronomical traits of the initial elite plants. However, somaclonal variation was detected in the polyploid plants regenerated from callus culture.
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