Parthenocarpy allows fruit set independently of fertilization. In parthenocarpic-prone tomato genotypes, fruit set can be achieved under pollen-limiting environmental conditions and in sterile mutants. Parthenocarpy is also regarded as a quality-related trait, when seedlessness is associated with positive fruit quality aspects. Among the different sources of genetic parthenocarpy described in tomato, the parthenocarpic fruit (pat) mutation is of particular interest because of its strong expressivity, high fruit set, and enhanced fruit quality. The complexity of the pat “syndrome” associates a strong competence for parthenocarpy with a complex floral phenotype involving stamen and ovule developmental aberrations. To understand the genetic basis of the phenotype, we mapped the pat locus within a 0.19-cM window of Chr3, comprising nine coding loci. A non-tolerated missense mutation found in the 14th exon of Solyc03g120910, the tomato ortholog of the Arabidopsis HD-Zip III transcription factor HB15 (SlHB15), cosegregated with the pat phenotype. The role of SlHB15 in tomato reproductive development was supported by its expression in developing ovules. The link between pat and SlHB15 was validated by complementation and knock out experiments by co-suppression and CRISPR/Cas9 approaches. Comparing the phenotypes of pat and those of Arabidopsis HB15 mutants, we argued that the gene plays similar functions in species with fleshy and dry fruits, supporting a conserved mechanism of fruit set regulation in plants.
Cadmium (Cd) contamination is a severe problem in the environment and produces detrimental effects on crop productivity and quality. Characterization of crop performance at different Cd concentrations is crucial to identify pollution-safe cultivars with low translocation efficiency to aboveground organs to be used for food safety. Here, we estimated germination, survival, growth, photosynthetic pigments, Cd bioaccumulation, among-organs translocation, and ionic balance in six Solanum melongena L. (eggplant) cultivars from the central Mediterranean basin. On two cultivars, we also analyzed expression of genes involved in Cd uptake, i.e., heavy metal ATPases (HMAs) and metal tolerance proteins (MTPs). We found that Cd has a negative effect on all the investigated parameters but with relevant among-cultivar differences. Cd-treated plants showed a decrease in germination rate and survival. Photosynthetic pigments showed opposite trends, i.e., with increasing Cd contents, we observed a decrease in chlorophylls and an increase in carotenoids. The investigated cultivars showed high ability of sequestrating Cd in roots but a low translocation efficiency to the aboveground organs, suggesting a good potential for food safety. The response of plants to Cd was mediated by a different expression of the MTP and HMA gene families. Our study represents the first comprehensive investigation of Cd tolerance in eggplant varieties from the Mediterranean basin and highlights the importance of comparative studies to identify Cd-tolerant cultivars.
The tomato parthenocarpic fruit (pat) mutation associates a strong competence for parthenocarpy with homeotic transformation of anthers and aberrancy of ovules. To dissect this complex floral phenotype, genes involved in the pollination-independent fruit set of the pat mutant were investigated by microarray analysis using wild-type and mutant ovaries. Normalized expression data were subjected to one-way ANOVA and 2499 differentially expressed genes (DEGs) displaying a >1.5 log-fold change in at least one of the pairwise comparisons analyzed were detected. DEGs were categorized into 20 clusters and clusters classified into five groups representing transcripts with similar expression dynamics. The “regulatory function” group (685 DEGs) contained putative negative or positive fruit set regulators, “pollination-dependent” (411 DEGs) included genes activated by pollination, “fruit growth-related” (815 DEGs) genes activated at early fruit growth. The last groups listed genes with different or similar expression pattern at all stages in the two genotypes. qRT-PCR validation of 20 DEGs plus other four selected genes assessed the high reliability of microarray expression data; the average correlation coefficient for the 20 DEGs was 0.90. In all the groups were evidenced relevant transcription factors encoding proteins regulating meristem differentiation and floral organ development, genes involved in metabolism, transport and response of hormones, genes involved in cell division and in primary and secondary metabolism. Among pathways related to secondary metabolites emerged genes related to the synthesis of flavonoids, supporting the recent evidence that these compounds are important at the fruit set phase. Selected genes showing a de-regulated expression pattern in pat were studied in other four parthenocarpic genotypes either genetically anonymous or carrying lesions in known gene sequences. This comparative approach offered novel insights for improving the present molecular understanding of fruit set and parthenocarpy in tomato.
Parthenocarpy is a desired trait in fruit crops; it enables fruit set under environmental conditions suboptimal for pollination, and seedless fruits represent a valuable consumer product. We employed TILLING-based screening of a mutant tomato population to find genetic lesions in Aux/IAA9, a negative regulator of the auxin response involved in the control of fruit set. We identified three mutations located in the coding region of this gene, including two single-base substitutions and one single-base deletion, which leads to a frame shift and premature stop codon. The transcription of IAA9 was strongly reduced in the frame-shift mutant, and partial loss of mutated protein activity was evidenced by an in vitro transactivation assay. Whereas missense mutations were predicted to be tolerated and did not cause mutant phenotypes, the frame-shift mutation-induced phenotypes expected for a loss of IAA9 function, including altered axillary shoot growth, reduced leaf compoundness and a strong tendency to produce parthenocarpic fruits. Mutant flowers showed pleiotropic anther cone defects, a phenotype frequently associated with parthenocarpy in tomato and other species. Mutant fruits were larger than those of the seeded control, with higher brix values and similar firmness. Fruit set was higher in the mutant than in wild type in the greenhouse, but lower in the open field. Facultative expression of parthenocarpy indicated that the mutant is suitable for hybrid seed production and for increasing seeds of parental lines. The results highlight the utility of this novel IAA9 allele for exploiting parthenocarpy by breeding tomato adapted to pollination-limiting growth conditions.
The production of anthocyanins in the tomato (Solanum lycopersicum L.) fruit is normally absent or poor, but a number of mutants or introgression lines are known to increase anthocyanin levels in vegetative and reproductive tissues. Through conventional breeding, a genetic combination was obtained with the remarkable phenotype of a deep purple fruit pigmentation, due to an accumulation of anthocyanins on the peel. Such a genotype was named Sun Black (SB) as a consequence of its sensitivity to light induction. When characterized for morpho-agronomic traits, SB plants showed increased fertility. Purple fruits displayed an arrangement of the epicarp cells different from normal tomatoes, a feature that could account for different mechanical properties and shelf-life potential. The SB genotype and, to a lesser extent, its single mutant parents showed the capacity to accumulate anthocyanins in the seedling root when grown under light. This phenotype, which was greatly improved by the addition of sucrose to the germination medium, proved to be useful as selection index and gave new insights for in vitro production of anthocyanin extracts. To assess the nutraceutical potential of purple tomatoes, we tested the activity of SB skin extracts on the proliferation of two human cancer cells lines. Cell proliferation was significantly inhibited by SB extract in a dose-dependent manner. When the bioactivity of SB extracts was compared with that of other anthocyanin-containing fruits or vegetables, a significant "Extract*Line" interaction was evidenced, suggesting a crucial role for the extract composition in terms of anthocyanidins and other eventual cell growth-inhibiting compounds.
Silver nitrate has been found to enhance plant regeneration frequency probably due to its anti-ethylene activity. It has also been found to be a potential inhibitor of Agrobacterium tumefaciens growth after co-cultivation due to its bactericidal properties. A. tumefaciens strain GV2260, harboring a plasmid with the marker gene neomycin phosphotransferase II (nptII) and the chimeric gene Def H9-iaaM was used to assess the extent of susceptibility to AgNO3. Bacterial growth curves were generated using turbidimetric measurements of bacterial density to evaluate bacterial response in the presence of different AgNO3 concentrations. At low initial cell densities (OD600 = 0.01 and 0.03) bacteria completely stopped growing at 8 and 10 mg l of AgNO3, respectively while at the initial cell density of OD600 = 0.1, silver ions strongly retarded cell growth at 25 mg l; however, after 48 hrs they continued to grow, although at very slow rates. Also, bacterial fitness was assessed in the presence of kanamycin (Kan) after treatment with cefotaxime (0, 50, 100, 200 mg l) (Cef) in combination with AgNO3 to evaluate the frequency of plasmid loss. The presence of AgNO3 was correlated with a decrease in plasmid stability in the presence of Cef (50 mg l). Colonies that developed on non-Kan plates were analyzed by PCR to determine if functional copies of the nptII gene were still present after treatment with AgNO3 and Cef. The effect on the growth and survival of bacterial cells investigated here may provide a useful clue to the interaction between plants and Agrobacterium during and after co-cultivation in the presence of AgNO3, taking advantage that this chemical also stimulates plant regeneration. _____________________________________________________________________________________________________________
Dikegulac, a growth regulator, was shown to stimulate in vitro shoot multiplication of olive cultivars Canino, Frantoio, Moraiolo, but not cultivars Rosciola and Piantone di Moiano. The Rugini Olive Medium including zeatin (4.5 μM) was used supplemented with a range of dikegulac concentrations (0–133.4 μM). An optimal result in number of shoots and nodes was obtained on cultivars Canino, Frantoio and Moraiolo at 66.7 μM dikegulac. Higher concentrations did not stimulate additional shoot and node formation and resulted in a drastic reduction in height of shoots. Elongated shoots were rooted and acclimatised and showed normal development compared to control plants.