Precision phenotyping of imidazolinone-induced chlorosis in sunflower

Chlorosis level is a useful parameter to assess imidazolinone resistance in sunflower (Helianthus annuus L.). The aim of this study was to quantify chlorosis through two different methods in sunflower plantlets treated with imazapyr. The genotypes used in this study were two inbred lines reported to be different in their resistance to imidazolinones. Chlorosis was evaluated by spectrophotometrical quantification of photosynthetic leaf pigments and by a bioinformatics-based color analysis. A protocol for pigment extraction was presented which improved pigment stability. Chlorophyll amount decreased significantly when both genotypes were treated with 10 μM of imazapyr. Leaf color was characterized using Tomato Analyzer® color test software. A significant positive correlation between color reduction and chlorophyll concentration was found. It suggests that leaf color measurement could be an accurate method to estimate chlorosis and infer chlorophyll levels in sunflower plants. These results highlight a strong relationship between imidazolinone-induced chlorosis and variations in leaf color and in chlorophyll concentration. Both methods are quantitative, rapid, simple, and reproducible. Thus, they could be useful tools for phenotyping and screening large number of plants when breeding for imidazolinone resistance in this species.Fil: Ochogavía, Ana Claudia. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Laboratorio de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Gil, Mercedes. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Departamento de Biología. Cátedra de Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Picardi, Liliana Amelia. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Departamento de Biología. Cátedra de Genética; Argentina. Universidad Nacional de Rosario. Consejo de Investigaciones de la Universidad de Rosario; ArgentinaFil: Nestares, Graciela María. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Departamento de Biología. Cátedra de Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentin

Structure, target-specificity and expression of PN_LNC_N13, a long non-coding RNA differentially expressed in apomictic and sexual Paspalum notatum

Key message: ncRNA PN_LNC_N13 shows contrasting expression in reproductive organs of sexual and apomictic Paspalum notatum genotypes. Abstract: Apomictic plants set genetically maternal seeds whose embryos derive by parthenogenesis from unreduced egg cells, giving rise to clonal offspring. Several Paspalum notatum apomixis related genes were identified in prior work by comparative transcriptome analyses. Here, one of these candidates (namely N13) was characterized. N13 belongs to a Paspalum gene family including 30–60 members, of which at least eight are expressed at moderate levels in florets. The sequences of these genes show no functional ORFs, but include segments of different protein coding genes. Particularly, N13 shows partial identity to maize gene BT068773 (RESPONSE REGULATOR 6). Secondary structure predictions as well as mature miRNA and target cleavage detection suggested that N13 is not a miRNA precursor. Moreover, N13 family members produce abundant 24-nucleotide small RNAs along extensive parts of their sequences. Surveys in the GREENC and CANTATA databases indicated similarity with plant long non-coding RNAs (lncRNAs) involved in splicing regulation; consequently, N13 was renamed as PN_LNC_N13. The Paspalum BT068773 predicted ortholog (N13TAR) originates floral transcript variants shorter than the canonical maize isoform and with possible structural differences between the apomictic and sexual types. PN_LNC_N13 is expressed only in apomictic plants and displays quantitative representation variation across reproductive developmental stages. However, PN_LNC_N13-like homologs and/or its derived sRNAs showed overall a higher representation in ovules of sexual plants at late premeiosis. Our results suggest the existence of a whole family of N13-like lncRNAs possibly involved in splicing regulation, with some members characterized by differential activity across reproductive types.Fil: Ochogavía, Ana Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Galla, Giulio. Università di Padova; ItaliaFil: Seijo, José Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: González, Ana María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Bellucci, Michele. Consiglio Nazionale delle Ricerche; ItaliaFil: Pupilli, Fulvio. Consiglio Nazionale delle Ricerche; ItaliaFil: Barcaccia, Gianni. Università di Padova; ItaliaFil: Albertini, Emidio. Università di Perugia; ItaliaFil: Pessino, Silvina Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentin

Increased apomixis expression concurrent with genetic and epigenetic variation in a newly synthesized Eragrostis curvula polyploid

Eragrostis curvula includes biotypes reproducing through obligate and facultative apomixis or, rarely, full sexuality. We previously generated a ‘‘tetraploid-dihaploid-tetraploid’’ series of plants consisting of a tetraploid apomictic plant (T), a sexual dihaploid plant (D) and a tetraploid artificial colchiploid (C). Initially, plant C was nearly 100% sexual. However, its capacity to form non-reduced embryo sacs dramatically increased over a four year period (2003–2007) to reach levels of 85–90%. Here, we confirmed high rates of apomixis in plant C, and used AFLPs and MSAPs to characterize the genetic and epigenetic variation observed in this plant in 2007 as compared to 2003. Of the polymorphic sequences, some had no coding potential whereas others were homologous to retrotransposons and/or protein-coding-like sequences. Our results suggest that in this particular plant system increased apomixis expression is concurrent with genetic and epigenetic modifications, possibly involving transposable elements.Fil: Zappacosta, Diego Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida(i); Argentina. Universidad Nacional del Sur. Departamento de Agronomía; ArgentinaFil: Ochogavía, Ana Claudia. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rodrigo, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida(i); ArgentinaFil: Romero, José Rodolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida(i); ArgentinaFil: Meier, Mauro Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida(i); ArgentinaFil: Garbus, Ingrid. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida(i); ArgentinaFil: Pessino, Silvina Claudia. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias; ArgentinaFil: Echenique, Carmen Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida(i); Argentina. Universidad Nacional del Sur. Departamento de Agronomía; Argentin

Caracterización de fragmentos génicos desconocidos que fueron asociados con la apomixis y/o la poliploidía en Paspalum notatum

La apomixis es una forma de reproducción asexual vía semillas frecuentemente asociada a la poliploidía. El objetivo de este trabajo fue inferir la función de 58 transcriptos desconocidos, que participan de los procesos de apomixis y/o poliploidización en Paspalum notatum. Utilizando amplificaciones rápidas de los extremos del cDNA se obtuvieron las secuencias completas de varios candidatos, que resultaron similares a elementos repetitivos portadores de segmentos génicos transduplicados o a precursores de miRNA de plantas. Para ambos tipos de secuencias se propuso un posible rol regulatorio, y se determinaron in silico los blancos putativos. Se eligieron secuencias representativas de cada uno de estos grupos, y se realizaron estudios experimentales para lograr una anotación funcional inequívoca. Para ello se analizó el número de copias genómicas, la posición en el genoma, la expresión cuantitativa durante el desarrollo reproductivo y la localización in situ de la actividad génica. También se realizaron análisis de display diferencial específicos para aislar retrotransposones con segmentos transduplicados relacionados y estudios de clivado específico de los blancos regulatorios de los miRNAs. Los resultados presentados en esta tesis aportan las primeras evidencias acerca del rol regulatorio que los retrotransposones y los miRNAs podrían estar desempeñando durante el desarrollo apomíctico.Apomixis is an asexual mode of reproduction via seeds commonly associated with polyploidy. The objective of this research work was to ascribe a functional identity to 58 unknown transcripts related to apomixis and/or polyploidization in Paspalum notatum. We used rapid amplification of cDNA ends to obtain full sequences for several candidates, which resulted homologous to retrotransposons carrying transduplicated gene segments or plant miRNA precursors. For both sequence types a regulatory role was proposed, and the identities of the putative targets were determined in silico. Representative sequences were selected to carry out experiments in order to achieve positive functional annotation. We analyzed gene copy numbers, positions in the genome, quantitative expressions at different developmental stages, and in situ localization of gene activity. Specific differential display analysis were conducted to isolate related retroelements containing gene transduplicated segments. Specific clivage analysis of the miRNA putative targets was also performed. Results presented here provide evidence on the regulatory role that both retrotransposons and miRNAs could be performing in apomixis development.Ochogavía, Ana Claudia. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias; ArgentinaOchogavía, Ana Claudia. CONICET; Argentin

Storage-associated genes and reserves accumulation in soybean cultivars differing in physiological strategies for attaining high seed protein concentration

High seed protein concentration (HP) in soybeans [Glycine max (L.) Merr.] is attained by increases in protein content in large seed genotypes (HP large seed) or by reductions in oil and carbohydrates contents in small seed genotypes (HP small seed). We hypothesized that these alternative strategies impact seed development, component accumulation, and gene expression differently. We compared a standard protein commercial genotype with two HP genotypes having contrasting seed size. The HP large seed genotype exhibited the fastest rate and longest period of seed growth and reserves accumulation compared with the HP small genotype. Seed development of these contrasting genotypes was normalized using a moisture depletion framework. Expression levels of some of the genes involved in protein and oil synthesis were lower in the HP small seed genotype compared with the other genotypes. No difference in gene expression was observed between the commercial and the HP large seed genotypes, suggesting a role for assimilate supply controlling high protein concentration based on this strategy. Our results indicate that seed development and gene expression are not necessarily associated with high seed protein concentration per se; a better understanding of seed composition requires acknowledging the contrasting strategies, in terms of seed size, to attain high seed protein concentration.Fil: Poeta, Florencia Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Ochogavía, Ana Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Permingeat, Hugo Raúl. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Laboratorio de Biología Molecular; ArgentinaFil: Rotundo, José Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; Argentin

Identification of suitable reference genes by quantitative real-time PCR for gene expression normalization in sunflower

Quantitative real-time PCR (qPCR) is currently the most accurate method for detecting differential gene expression, but depends greatly on normalization to stably expressed housekeeping genes. Transcriptomics analyses and experimental validation in different plant species have shown that the reliability of these endogenous controls can be influenced by the plant species, growth conditions and organs/tissues examined. Thus, reliable validation of reference genes is required to ensure proper normalization. This paper presents a systematic comparison of ten potential reference genes in sunflower: five commonly used genes (Actin, Elongation Factor1, Plastid-encode RNA polymerase, Tubulin, and Ubiquitin, as ACT, EF1, PEP, TUB, and UBQ respectively), as well as five new candidates (Translation initiation factor, MicroRNA precursors 171 and 156, Ask-interacting protein, and Protein of unknown function, as ETIF5, MIR171, MIR156, SKIP, and UNK2 respectively). Reference gene expression stability was examined by qPCR across 20 biological samples, representing different tissues at various developmental stages. Expression of all 10 genes was variable to some extent, but that of ACT, UNK2, and EF1 was overall the most stable. A combination of ETIF5/UNK2/EF1 would be appropriate to use as a reference panel for normalizing gene expression data among vegetative tissues, whereas the combination of ACT/MIR156/UNK2 is most suitable for reproductive tissues. Reference genes selected in this study were further validated by examining relative expression of ahas1, one of three acetohydroxyacid synthase genes of sunflower. Our identification and validation of suitable normalizer genes will be of use to ensure accurate results in future transcriptomics studies in this crop.Fil: Ochogavía, Ana Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Novello, Maria Angelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Picardi, Liliana Amelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; Argentina. Consejo de Investigaciones de la Universidad Nacional de Rosario; ArgentinaFil: Nestares, Graciela María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; Argentin

Precision phenotyping of imidazolinone-induced chlorosis in sunflower

Chlorosis level is a useful parameter to assess imidazolinone resistance in sunflower (Helianthus annuus L.). The aim of this study was to quantify chlorosis through two different methods in sunflower plantlets treated with imazapyr. The genotypes used in this study were two inbred lines reported to be different in their resistance to imidazolinones. Chlorosis was evaluated by spectrophotometrical quantification of photosynthetic leaf pigments and by a bioinformatics-based color analysis. A protocol for pigment extraction was presented which improved pigment stability. Chlorophyll amount decreased significantly when both genotypes were treated with 10 μM of imazapyr. Leaf color was characterized using Tomato Analyzer(®) color test software. A significant positive correlation between color reduction and chlorophyll concentration was found. It suggests that leaf color measurement could be an accurate method to estimate chlorosis and infer chlorophyll levels in sunflower plants. These results highlight a strong relationship between imidazolinone-induced chlorosis and variations in leaf color and in chlorophyll concentration. Both methods are quantitative, rapid, simple, and reproducible. Thus, they could be useful tools for phenotyping and screening large number of plants when breeding for imidazolinone resistance in this species

An integrated approach for the characterization of one‐ and two‐gene imazamox‐resistant wheat lines

In wheat (Triticum aestivum L.), cultivars resistant to imidazolinone herbicides arean important tool for weed management. The target site of imidazolinones is acetohydroxyacid synthase (AHAS), an enzyme coded by three homeologous genes (ahasLA1, ahasL-B1, and ahasL-D1). This study aimed to evaluate the effect of the imidazolinone herbicide imazamox on a single-gene (AhasL-D1) resistant line (R1) and atwo-gene (AhasL-B1 and AhasL-D1) resistant line (R2) at multiple biological levels.These lines showed variation in the transcriptional levels of ahasL homeologs, butno differences in total AHAS activity. The relative contribution of resistant alleles tothe ahasL transcript pool accounted for 47% in the R1 line and 77% in the R2 line.The in vitro AHAS inhibition of the R1 line was intermediate between the R2 andthe susceptible lines. At an anatomical level, high imazamox concentrations inducedalterations in the root tip features of R1 plants but did not affect R2 plants, whereas atin vivo AHAS activity and whole-plant levels, both resistant lines showed high resistance. The herbicide dose that reduced shoot biomass of resistant lines by 50% was35-fold higher than that of the susceptible line. We conclude that homeolog variationallows a higher contribution of resistance genes to the ahasL transcript pool in theR1 and R2 lines. Although both resistant lines showed high whole-plant resistance,the differential root damage induced by imazamox between wheat lines suggests thattwo-gene resistant cultivars could be an effective option to avoid the negative impactscaused by herbicide residues in the soil.Fil: Anastasini, Valentin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Depetris, Mara Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Ochogavía, Ana Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Nestares, Graciela María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Breccia, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; Argentin

Transcript profiling of non-target-site imidazolinone resistance in imisun sunflower

Imidazolinone resistance found in a wild sunflower (Helianthus annuus L.) population was successfully transferred to a cultivated inbred line developing ‘Imisun’ sunflowers. Genetic regulation of this trait has been reported to involve two genes: Imr1, an allelic variant of ahasl1 locus that codes for acetohydroxyacid synthase catalytic subunit, and the modifier Imr2, whose identity remains unknown, but it could be related to non-target-site resistance such as xenobiotic metabolism. The aim of the present study was to characterize the gene expression of resistant and susceptible sunflower lines in response to imazethapyr herbicide by complementary DNA amplified fragment-length polymorphism (cDNA-AFLP). Three assays were performed to determine (i) optimal herbicide treatment concentration, (ii) duration of herbicide treatment, and (iii) in vitro acetohydroxyacid synthase activity to assess enzyme inhibition levels. An important number of genes related to metabolism of xeno-biotics and stress was found: cytochrome P450 monooxygenases, UDP-glucuronosyl/UDP-glucosyltransferases, glycosyltransferases, and ATP-binding cassette transporters, among others. These results suggest that non-target-site resistance mechanisms may contribute to herbicide resistance in Imisun sunflower and could be related to the modifier gene Imr2. Using cDNA-AFLP, we were able to detect candidate detoxification-related genes potentially involved in imidazolinone resistance in sunflower.Fil: Gil, Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Ochogavía, Ana Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Vega, Tatiana Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Felitti, Silvina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Nestares, Graciela María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; Argentin

Partial male sterility in imisun sunflower: Imazapyr treatment in advanced vegetative stages decreases pollen yield and alters ahas gene expression

Imidazolinones are powerful herbicides that inhibit branched-chain amino acid biosynthesis by targeting the catalytic subunit of acetohydroxyacid synthase (AHAS). Imidazolinone application in the advanced vegetative or early reproductive developmental stages is associated with male sterility in resistant sunflower (Helianthus annuus L.); however, the underlying mechanism of this sterility remains unknown. This study describes the morphological, cytoem-bryological, and molecular alterations induced by imazapyr (IM) treatment on reproductive tissues at different developmental stages in two sunflower genotypes, resistant and intermediate resistant, respectively. Pollen and seed physiological variables were compared between the treated and control plants. The number of pollen grains per flower and viable seeds were negatively affected by IM treatment in the intermediate-resistant genotype, and the biometric traits of early developed disc flower were also significantly different in this genotype. Differential interference contrast microscopy revealed that IM treatment slightly accelerates megagamethophyte development. Anther observations at microsporogenesis using confocal microscopy show that the sporogenous tissue was damaged. Furthermore, the expression profiles of the sunflower AHAS paralogs (ahas1, ahas2, and ahas3) were measured by quantitative polymerase chain reaction in the anthers and pistils of two developmental stages in treated and control plants. Imazapyr treatment in early reproductive growth stages clearly induces divergent expression patterns in the ahas gene family. These findings provide new insight into a novel chemical method for inducing male sterility in sunflowers and enhance our understanding of the effects of AHAS-inhibitor herbicides in reproductive tissues.Fil: Ochogavía, Ana Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Novello, María Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Bianchi, Marta Beatriz. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias; ArgentinaFil: Picardi, Liliana Amelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; ArgentinaFil: Nestares, Graciela María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Investigaciones en Ciencias Agrarias de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Instituto de Investigaciones en Ciencias Agrarias de Rosario; Argentin