Liliana Amelia Picardi

Reduction of hyperhydricity in sunflower tissue culture

In order to reduce the occurrence of hyperhydrated shoots, the response of three sunflower inbred lines was examined on regeneration media containing various concentrations of kinetin, silver nitrate, and casein hydrolysate, calcium nitrate and cobalt nitrate. There were differences among the inbred lines for all the parameters taken into account to outline the in vitro efficiency. Percentage of hyperhydrated primordia, average number of shoots and of primordia per total explants, and percentage of hyperhydrated shoot traits differed among all media. The genotype × culture media interaction was significant for average number of shoots and primordia per total explants, regeneration percentage and percentage of hyperhydrated primordia. Among all media tested, those containing silver nitrate significantly reduced hyperhydricity in a dose-dependent way. The addition of silver nitrate showed to be useful in improving the quality of sunflower micropropagated plants by reducing this undesirable phenomenon.

Protein profiling in F1 and F2 generations of two tomato genotypes differing in ripening time

Pericarp polypeptide profiles were analyzed at three ripening stages in the F1 hybrid and the F2 population from the cross between the accessions: LA1385 (Lycopersicon esculentum var. cerasiforme) and 804627 (L. esculentum, a homozygous genotype for the nor mutant). Six polymorphic polypeptides were observed in LA1385, while no polymorphic polypeptides among ripening stages was observed in 804627. On the other hand, some polypeptides in the F1 hybrid were not observed in the parents whereas others were present in both parental genotypes and were unnoticeable in the hybrid genotype. From a cluster analysis on the protein profiles of the F2 population, the differential expression of proteins allowed to distinguish mature green (MG) stage from the others two stages, while for breaker stage (BR) and red ripe stage, the genetic background was more important in forming groups. The differential expression of proteins could be associated with fruit morphology traits such as a 72 kDa polypeptide present in MG stage with fruit diameter, height and mass and a 47 kDa polypeptide found in BR with fruit shelf life.

An approach for understanding the heredity of two quality traits (head color and tightness) in globe artichoke (Cynara scolymus L.)

The inheritance of head color and tightness in globe artichoke was studied utilizing crosses between inbreed lines and between clones and self-pollinated clones from different genetic origins. These genetic materials were sowed in a completely randomized design with 20 plants per plot and genotype. Globe artichoke heads were classified into three colors (purple-green, purple and green) and three head tightness types (compact, fairly compact and soft) and the segregating ratios for these traits tested in each offspring using the chi-square test. Crosses between green and purple inbreed lines produced only purple-green heads but F2 generation segregated at a purple-green:purple:green ratio of 9:3:4. The self-pollinated compact head clones produced a compact head:fairly compact head:soft head ratio of 12:3:1. The remaining crosses between lines and among clones and backcrosses verified these ratios. These results suggest that two loci with a simple recessive epistasis are involved in the inheritance of head color and that two loci with simple dominant epistasis are involved in the expression of the different head tightness types. The inheritance models proposed here could be helpful in predicting the appearance of artichoke heads if breeders need to obtain hybrid seeds for a desirable phenotype.

Differential expression of acetohydroxyacid synthase genes in sunflower plantlets and its response to imazapyr herbicide.

Acetohydroxyacid synthase (AHAS) catalyzes the first reaction in branch chain amino acids biosynthesis. This enzyme is the target of several herbicides, including all members of the imidazolinone family. Little is known about the expression of the three acetohydroxyacid synthase genes (ahas1, ahas2 and ahas3) in sunflower. The aim of this work was to evaluate ahas gene expression and AHAS activity in different tissues of sunflower plantlets. Three genotypes differing in imidazolinone resistance were evaluated, two of which carry an herbicide resistant-endowing mutation known as Ahasl1-1 allele. In vivo and in vitro AHAS activity and transcript levels were higher in leaves than in roots. The ahas3 transcript was the less abundant in both tissues. No significant difference was observed between ahas1 and ahas2 transcript levels of the susceptible genotype but a higher ahas1 transcript level was observed in leaves of genotypes carrying Ahasl1-1 allele. Similar transcript levels were found for ahas1 and ahas2 in roots of genotypes carrying Ahasl1-1 allele whereas higher ahas2 abundance was found in the susceptible genotype. Herbicide treatment triggered tissue-specific, gene and genotype-dependent changes in ahas gene expression. AHAS activity was highly inhibited in the susceptible genotype. Differential responses were observed between in vitro and in vivo AHAS inhibition assays. These findings enhance our understanding of AHAS expression in sunflower genotypes differing for herbicide resistance and its response to herbicide treatment.

Phenotypic and molecular characterization of selected tomato recombinant inbred lines derived from the cross Solanum lycopersicum × S. pimpinellifolium

An important trait defining fresh tomato marketability is fruit shelf life. Exotic germplasm of Solanum pimpinellifolium is able to prolong shelf life. Sixteen recombinant inbred lines with differing values of shelf life and fruit weight were derived by antagonistic-divergent selection from an interspecific cross involving Solanum pimpinellifolium. The objective of this study was to evaluate these recombinant inbred lines for many fruit quality traits such as diameter, height, size, acidity, colour, firmness, shelf life and weight, and to characterize them by amplified fragment length polymorphism markers. For most traits, a wide range of genetic variability was found and a wide range of molecular variation was also detected. Both sets of data allowed the identification of recombinant inbred lines by means of cluster analysis and principal component analysis. Genetic association among some amplified fragment length polymorphism markers and fruit quality traits, suggested by the principal component analysis, could be identified by single point analysis. Potential molecular markers underlying agronomical traits were detected in these recombinant inbred lines.

Soil-less bioassays for early screening for resistance to imazapyr in sunflower (Helianthus annuus L.)

BACKGROUND Rapid and efficient diagnostic tests for early screening of herbicide resistance are convenient alternatives to field screening methods. There is a need for a quick, reliable and cost-effective method for rapid diagnosis of imidazolinone resistance in sunflower (Helianthus annuus L.). RESULTS Two seed germination bioassays were developed. Seeds from three sunflower inbred lines differing in resistance to imidazolinones were germinated either on solid culture medium or placed in plastic pots filled with commercial perlite. After 8 days incubation under controlled conditions, both assays successfully distinguished susceptible genotype from the resistant and intermediate ones. The susceptible genotype showed arrested root growth at all herbicide treatments (root length < 1 cm). The resistant genotype developed a complete root system even when exposed to the highest dose of herbicide. However, no definite differences were observed for the intermediate and resistant genotypes with respect to root growth under the different herbicide treatments. CONCLUSION The simple and rapid screening assays described in the present study were useful in discriminating imidazolinone resistance at the seedling stage. Therefore, these bioassays could be potential tools for early screening of imidazolinone resistance genes from large sunflower populations.

Responsive regions for direct organogenesis in sunflower cotyledons

Regeneration efficiency from three different regions of cotyledonary explants was examined in six sunflower inbred lines. Proximal, middle and distal regions from seedling cotyledons were cultured on regeneration medium supplemented with growth regulators. Plant regeneration by direct organogenesis was observed after four weeks. Significant differences among inbred lines were found for regeneration percentage and average number of shoots per total explants. Also a decreasing regeneration capacity was observed from proximal to distal sections for all inbred lines. Regeneration ability from cotyledonary explants in this species is strongly influenced by the genotype and by the region from which the explant was obtained. The distance to the cotyledonary node plays a preponderant role in the expression of shoot forming capacity. Shoot differentiation via seedling cotyledons depends upon the presence of the proximal region of cotyledon regardless of the genotype.

Characterization of the segregating generation of a tomato hybrid carrying nor and exotic genes

The objective of this work was to study recombination of the productive and fruit quality traits in the segregating generation of the hybrid between a cultivated variety of Lycopersicon esculentum homozygous for nor and the accession LA1385 of L. esculentum var. cerasiforme using multivariate statistical analysis. F 1 and F2 generations and the parents were evaluated for vegetative and productive traits (internode length, stem perimeter at the basal, middle and apical parts, number of flowers per cluster, number of clusters per plant and days to harvest) and quality fruit traits (weight, shape, soluble solids content, acidity, firmness, color and shelf life). A canonical correlation between vegetative and productive traits and those of fruit quality and clustering for fruit traits performed with the F 1 and F2 and the parents were used. The productive and fruit quality traits showed recombination in the segregating generation. Shelf life was the most outstanding fruit trait to discriminate groups in the F2. For three cluster levels each group of F 2 individuals behaved each one of the parents and the F 1.

Transgressive segregation for fruit quality traits in a cross between exotic and mutant genotypes of Lycopersicon

Abstract A cross between a Lycopersicon esculentum accession carrying the mutant nor and an accession of L. esculentum var. cerasiforme was made to analyse the joint genetic effect upon the phenotypic segregation of some fruit quality traits in the F2 generation. Fruit from the F1, F2, and the parents (as testers) were evaluated for shelf life and another quality traits. Because of the presence of individuals falling beyond their parental phenotypes in the F2, a transgressive inheritance was estimated by a chi‐square test. Transgressive inheritance was present for relevant traits such as shape, colour, firmness, and shelf life. The broad sense heritability values were highly significant for all traits and the shelf life was not associated with any other fruit quality trait. The use of this wild accession as a parent in tomato breeding programmes becomes an alternative for increasing fruit quality and especially prolonging the fruit shelf life.

Variability for the in vitro culture response in tomato recombinant inbred lines

The aim of this work was to estimate genetic variability for in vitro culture response of recombinant inbred lines (RILs) of the genus Lycopersicon. The callus percentage (C), the regeneration percentage (R) and the productivity rate (PR) were evaluated 45 d after culture initiation in a set of 16 elite tomato RILs and their parents. The narrow sense heritability (h2) values were 0.38 ± 0.04 for C, 0.46 ± 0.04 for R, and 0.28 ± 0.03 for R, while the genetic correlation (rg) values were −0.96 ± 0.07 between C and R, 0.81 ± 0.14 between PR and R, and −0.79 ± 0.16 between PR and C. Three AFLP markers associated to the in vitro traits were identified.