Alicia Carrera

Gene flow between cultivated sunflower and Helianthus petiolaris (Asteraceae)

Helianthus petiolaris (Asteraceae) native to North America has naturalized in Argentina. The extensive overlapping with sunflower crop regions, the coincidence of life cycles and the common pollinators facilitate interspecific crosses with sunflower, H. annuus var. macrocarpus. To estimate the occurrence of crop-to-wild and wild-to-crop gene flow, off-type plants in progenies of pure stands of both species flowering in coincidence and the presence of crop alleles in H. petiolaris populations were investigated in 26 wild populations and nine crop fields. Morphological traits and RAPD markers were used to attempt hybrid characterization. Off-type individuals were found in frequencies of 0.5 and 0.3% among crop progenies and wild populations, respectively. Off-type plants showed intermediate values for metric morphological traits and low fertility traits. Some off-type plants proved to carry crop alleles based on molecular analysis. The average frequency of cultivar-marker introgression across the wild populations was very low (0.02). Although observed hybridization rates seem to be low, the extension of crop-wild overlapping in Argentina make hybrid formation a noticeable process. Therefore, the new sunflower varieties and eventually GM varieties would transfer their traits through pollen flow and they would persist in H. petiolaris populations.

Isozyme variation in Helianthus petiolaris and sunflower, H. annuus

SummaryHelianthus petiolaris is a wild species used in genetic improvement of sunflower, as a donor of cytoplasmic male sterility and of genetic resistance to diseases. Isozyme variation for ADH, ACP, EST, GDH, LAP, PGI, PGD and SKDH in this species was studied using starch gel electrophoresis. The patterns thus obtained were compared with zymograms of inbred lines, hybrids and open pollinated varieties of H. annuus. The same alleles for EST and SKDH isozymes were found in both species, while ACP showed an allele that has not been found in sunflower. The rest of the isozyme systems showed both common alleles and characteristic ones for each species. ACP, GDH and PGD were monomorphic in H. petiolaris, while ADH and LAP were monomorphic in H. annuus. The isozyme markers obtained here could be useful in breeding programs involving interspecific crosses, and studies on introgression and on genetic variation in other populations of this wild species.

Variability among inbred lines and RFLP mapping of sunflower isozymes

Eight isozyme systems were used in this study: acid phosphatase (ACP), alcohol dehydrogenase (ADH), esterase (EST), glutamate dehydrogenase (GDH), malate dehydrogenase (MDH), phosphoglucoisomerase (PGI), 6-phosphogluconate dehydrogenase (PGD), and phosphoglucomutase (PGM). The polymorphism of these enzyme systems was studied in 25 elite inbred lines. A total of 19 loci were identified, but only eight of them were polymorphic in the germplasm tested. The polymorphic index for the eight informative markers ranged from 0.08 to 0.57, with a mean value of 0.36. Five isozyme loci were mapped in F2:3 populations with existing RFLP data. Est-1, Gdh-2 and Pgi-2 were mapped to linkage groups 3, 14 and 9, respectively. As in previous reports, an ACP locus and a PGD locus were found to be linked, both located in linkage group 2 of the public sunflower map.

Collection and morphological characterization of Panicum coloratum L. in Argentina

Panicum coloratum (Poaceae) is a perennial grass native to South Africa, distributed all over the world, but poorly adopted by ranchers in Argentina. Adapted to a wide range of soil and climatic conditions, it constitutes a good forage resource for zones with edaphic and climatic restrictions, where cattle rising activities are here displaced. The high polymorphism in the species has made it difficult to delineate sub specific taxonomic categories. It has been classified in botanical varieties, types and/or cultivars. In order to document the morphological variability in P. coloratum and to investigate the differences between var. makarikariense and var. coloratum, a collection was gathered in INTA Rafaela composed by fourteen accessions from different provinces of Argentina: six accessions of var. makarikariense and eight from var. coloratum. The evaluation of 152 individuals gives evidence of a wide morphological variation in the collection in traits related to forage and caryopses production; seven out of nine traits showed significant differences between var. makarikariense and var. coloratum. Based on the study of local type specimens, a morphological key to distinguish between P. coloratum varieties is provided. These results report new information about the diversity in P. coloratum in Argentina and provide valuable insights for future use of the collection as the base material of a breeding program.

Identification of genes induced by Fusarium graminearum inoculation in the resistant durum wheat line Langdon(Dic-3A)10 and the susceptible parental line Langdon.

The wheat recombinant chromosome inbred line LDN(Dic-3A)10, obtained through introgression of a Triticum dicoccoides disomic chromosome 3A fragment into Triticum turgidum spp. durum var. Langdon, is resistant to fusarium head blight (FHB) caused by Fusarium graminearum. To identify genes involved in FHB resistance, we used a cDNA-AFLP approach to compare gene expression between LDN(Dic-3A)10 and the susceptible parental line LDN at different time points post-inoculation. In total, 85 out of the ∼ 500 transcript-derived fragments (TDFs) were found to be differentially expressed: 36 and 19% were upregulated in LDN(Dic-3A)10 and LDN, respectively, whereas 45% were induced in both genotypes. Several of the cloned TDFs showed similarity to proteins involved in specific recognition of plant pathogens or associated with early responses to infection. Some TDFs specific to the inoculation response did not show similarity to characterized proteins. The availability of T. aestivum genome sequences allowed the in silico mapping of 28 TDFs and the acquirement of the corresponding gene sequences and, in some cases, their regulatory regions. Analysis of promoter regions revealed the potential existence of shared transcription regulation mechanisms. For instance, three TDF-associated genes contained binding sites for WRKY transcription factors, which have been implicated in the regulation of genes associated with pathogen defense, and three for abscisic acid-responsive element (ABRE). Collectively, our results revealed specific pathogen recognition in the interactions of LDN and LDN(Dic-3A)10 with F. graminearum. Such recognition leads to changes in the expression of several transcripts, attributable to the presence of the wheat QTL Qfhs.ndsu-3AS.

In Silico Analysis of Fatty Acid Desaturase Genes and Proteins in Grasses

Fatty acid desaturases (FADs) catalyze the introduction of a double bond into acyl chains. Two FAD groups have been identified in plants: acyl-acyl carrier proteins (ACPs) and acyl-lipid or membrane-bound FAD. The former catalyze the conversion of 18:0 to 18:1 and to date have only been identified in plants. The latter are found in eukaryotes and bacteria and are responsible for multiple desaturations. In this study, we identified 82 desaturase gene and protein sequences from 10 grass species deposited in GenBank that were analyzed using bioinformatic approaches. Subcellular localization predictions of desaturase family revealed their localization in plasma membranes, chloroplasts, endoplasmic reticula, and mitochondria. The in silico mapping showed multiple chromosomal locations in most species. Furthermore, the presence of the characteristic histidine domains, the predicted motifs, and the finding of transmembrane regions strongly support the protein functionality. The identification of putative regulatory sites in the promotor and the expression profiles revealed the wide range of pathways in which fatty acid desaturases are involved. This study is an updated survey on desaturases of grasses that provides a comprehensive insight into diversity and evolution. This characterization is a necessary first step before considering these genes as candidates for new biotechnological approaches.

A valuable QTL for Fusarium head blight resistance from Triticum turgidum L. ssp. dicoccoides has a stable expression in durum wheat cultivars

Langdon(Dic-3A)-10 line carrying the QTL Qfhs.ndsu-3AS from T. turgidum ssp. dicoccoides that confers Type II resistance to Fusarium head blight (FHB) was crossed with Argentinean durum wheat cultivars. F4 progeny were screened with the microsatellite locus Xgwm2, tightly linked to the Qfhs.ndsu-3A region. Reaction of these plants and parents to FHB was evaluated at 7, 14 and 21 days post-inoculation (dpi) with F. graminearum; severity (% symptomatic spikelets/spike) and AUDPC (area under disease progress curve) were calculated. F4 progeny carrying the resistance allele in heterozygous or in homozygous condition showed significantly lower scab damage at 21 dpi and slower progress of disease than cultivated parents. Our results indicate that the resistance Qfhs.ndsu-3AS has a stable dominance expression in genetic backgrounds of durum cultivars and demonstrate that the linked microsatellite is an effective molecular tool for resistance screening. This work offers valuable information for Qfhs.ndsu-3AS utilization in wheat breeding programs.