María Verónica Rodríguez

Dormancy in cereals (not too much, not so little): about the mechanisms behind this trait

As in other cultivated species, dormancy can be seen as a problem in cereal production, either due to its short duration or to its long persistence. Indeed, cereal crops lacking enough dormancy at harvest can be exposed to pre-harvest sprouting damage, while a long-lasting dormancy can interfere with processes that rely on rapid germination, such as malting or the emergence of a uniform crop. Because the ancestors of cereal species evolved under very diverse environments worldwide, different mechanisms have arisen as a way of sensing an appropriate germination environment (a crucial factor for winter or summer annuals such as cereals). In addition, different species (and even different varieties within the same species) display diverse grain morphology, allowing some structures to impose dormancy in some cereals but not in others. As in seeds from many other species, the antagonism between the plant hormones abscisic acid and gibberellins is instrumental in cereal grains for the inception, expression, release and re-induction of dormancy. However, the way in which this antagonism operates is different for the various species and involves different molecular steps as regulatory sites. Environmental signals (i.e. temperature, light quality and quantity, oxygen levels) can modulate this hormonal control of dormancy differently, depending on the species. The practical implications of knowledge accumulated in this field are discussed.

Expression of ABA signalling genes and ABI5 protein levels in imbibed Sorghum bicolor caryopses with contrasting dormancy and at different developmental stages.

BACKGROUND AND AIMS Pre-harvest sprouting susceptibility in grain sorghum (Sorghum bicolor) is related to low seed dormancy and reduced embryo sensitivity to inhibition of germination by abscisic acid (ABA). Intra-specific variability for pre-harvest sprouting might involve differential regulation of ABA signalling genes. METHODS Sorghum genes encoding homologues for ABA signalling components from other species (ABI5, ABI4, VP1, ABI1 and PKABA1) were studied at the transcriptional and protein level (ABI5) during grain imbibition for two sorghum lines with contrasting sprouting phenotypes and in response to hormones. KEY RESULTS Transcript levels of these genes and protein levels of ABI5 were higher in imbibed immature caryopses of the more dormant line. Dormancy loss was related to lower transcript levels of these genes and lower ABI5 protein levels in both genotypes. Exogenous ABA inhibited germination of isolated embryos but failed to prevent ABI5 rapid decrease supporting a role for the seed coat in regulating ABI5 levels. CONCLUSIONS Several genes involved in ABA signalling are regulated differently in imbibed caryopses from two sorghum lines with contrasting pre-harvest sprouting response before - but not after - physiological maturity. A role for ABI5 in the expression of dormancy during grain development is discussed.

Expression of Seed Dormancy in Grain Sorghum Lines with Contrasting Pre-Harvest Sprouting Behavior Involves Differential Regulation of Gibberellin Metabolism Genes

Grain sorghum [Sorghum bicolor (L) moench] exhibits intraspecific variability for the rate of dormancy release and pre-harvest sprouting behavior. Two inbred lines with contrasting sprouting response were compared: IS9530 (resistant) and RedlandB2 (susceptible). Precocious dormancy release in RedlandB2 is related to an early loss of embryo sensitivity to ABA and higher levels of gibberellins in imbibed grains as compared with IS9530. With the aim of identifying potential regulatory sites for gibberellin metabolism involved in the expression of dormancy in immature grains of both lines, we carried out a time course analysis of transcript levels of putative gibberellin metabolism genes and hormone content (GA(1), GA(4), GA(8) and GA(34)). A lower embryonic GA(4) level in dormant IS9530 was related to a sharp and transient induction of two SbGA2-oxidase (inactivation) genes. In contrast, these genes were not induced in less dormant RedlandB2, while expression of two SbGA20-oxidase (synthesis) genes increased together with active GA(4) levels before radicle protrusion. Embryonic levels of GA(4) and its catabolite GA(34) correlated negatively. Thus, in addition to the process of gibberellin synthesis, inactivation is also important in regulating GA(4) levels in immature grains. A negative regulation by gibberellins was observed for SbGA20ox2, SbGA2ox1 and SbGA2ox3 and also for SbGID1 encoding a gibberellin receptor. We propose that the coordinated regulation at the transcriptional level of several gibberellin metabolism genes identified in this work affects the balance between gibberellin synthesis and inactivation processes, controlling active GA(4) levels during the expression of dormancy in maturing sorghum grains.

In vitro binding of Sorghum bicolor transcription factors ABI4 and ABI5 to a conserved region of a GA 2-OXIDASE promoter: possible role of this interaction in the expression of seed dormancy

The precise adjustment of the timing of dormancy release according to final grain usage is still a challenge for many cereal crops. Grain sorghum [Sorghum bicolor (L.) Moench] shows wide intraspecific variability in dormancy level and susceptibility to pre-harvest sprouting (PHS). Both embryo sensitivity to abscisic acid (ABA) and gibberellin (GA) metabolism play an important role in the expression of dormancy of the developing sorghum grain. In previous works, it was shown that, simultaneously with a greater embryo sensitivity to ABA and higher expression of SbABA-INSENSITIVE 4 (SbABI4) and SbABA-INSENSITIVE 5 (SbABI5), dormant grains accumulate less active GA4 due to a more active GA catabolism. In this work, it is demonstrated that the ABA signalling components SbABI4 and SbABI5 interact in vitro with a fragment of the SbGA 2-OXIDASE 3 (SbGA2ox3) promoter containing an ABA-responsive complex (ABRC). Both transcription factors were able to bind the promoter, although not simultaneously, suggesting that they might compete for the same cis-acting regulatory sequences. A biological role for these interactions in the expression of dormancy of sorghum grains is proposed: either SbABI4 and/or SbABI5 activate transcription of the SbGA2ox3 gene in vivo and promote SbGA2ox3 protein accumulation; this would result in active degradation of GA4, thus preventing germination of dormant grains. A comparative analysis of the 5′-regulatory region of GA2oxs from both monocots and dicots is also presented; conservation of the ABRC in closely related GA2oxs from Brachypodium distachyon and rice suggest that these species might share the same regulatory mechanism as proposed for grain sorghum.

Reduced embryo sensitivity to abscisic acid in a sprouting-susceptible sorghum ( Sorghum bicolor ) variety is associated with altered ABA signalling

In the work reported in this paper, we attempted to elucidate the nature of the different abscisic acid (ABA) sensitivities presented by developing embryos from sorghum [Sorghum bicolor (L.) Moench] lines with contrasting pre-harvest sprouting (PHS) behaviour (Redland B2, susceptible; IS 9530, resistant). We explored two different hypotheses for a possible mechanism: (1) a different functionality of the ABA signalling pathway, and (2) a different rate of ABA degradation/conjugation in the apoplast of embryos from these genotypes. To assess the first possibility, we used an ABA-responsive gene (Rab17 )a s a reporter of changes in endogenous ABA content, which were artificially induced in embryos from both genotypes by means of fluridone application immediately after anthesis, to reduce ABA content, and embryo incubation in the presence of ABA. A defect in ABA signalling should be seen as a level of Rab17 expression that is independent of endogenous ABA content. For testing the second possibility at two stages of development, embryos from both lines were isolated and incubated in water for different periods. ABA concentrations in embryos and the incubation media were quantified through radioimmunoassay. In contrast to our findings for the resistant IS 9530 line, Rab17 expression did not respond to changes in ABA levels in sensitive Redland B2 embryos. The ABA degradation/conjugation rates in embryos and incubation media did not show clear differences between sorghum lines for any of the developmental stages analysed. These results suggest that a disruption in the ABA signal transduction pathway in Redland B2 underlies the low ABA sensitivity shown by embryos from this line.

Parámetros bioquímico-endocrinos de utilidad en la etapa del crecimiento y desarrollo del Ovejero Alemán, Doberman y Gran Danés

En el control bioquimico del crecimiento se deben tener en cuenta los aspectos metabolico-fisiologicos que en el intervienen y que variables son utiles para evaluarlos. Debido a la falta de datos por edad, sexo y raza en la bibliografia se propuso determinar valores de FAS, Ca, Pi, Mg, proteinas totales, albumina, CPK, T4, T3 y T4 libre. Se trabajo con 800 cachorros (350 ovejeros alemanes, 270 doberman y 180 gran danes) divididos por edad, sexo y raza. El estadistico utilizado fue el analisis de varianza, comparacion de medias por Test de Tukey y Bonferroni y calculo de percentiles. No hubo diferencias significativas (p0.05) por sexo y raza, quedando agrupados solo por edad (diferencias significativas p<0.05) desde 1 mes a 8 meses. Segun los resultados obtenidos los parametros de utilidad son: FAS, Pi, albumina, T4 y T4L; siendo las proteinas totales T3 y Ca complementarias a los mencionados anteriormente. En los primeros cinco meses de vida los requerimientos metabolicos son maximos, siendo importante el control en este momento para realizar el diagnostico precoz como tambien una adecuada profilaxis

Pre-harvest Sprouting and Grain Dormancy in Sorghum bicolor: What Have We Learned?

The possibility of obtaining sorghum grains with quality to match the standards for a diversity of end-uses is frequently hampered by the susceptibility to pre-harvest sprouting (PHS) displayed by many elite genotypes. For these reasons, obtaining resistance to PHS is considered in sorghum breeding programs, particularly when the crop is expected to approach harvest maturity under rainy or damp conditions prevalence. As in other cereals, the primary cause for sprouting susceptibility is a low dormancy prior to crop harvest; in consequence, most research has focused in understanding the mechanisms through which the duration of dormancy is differentially controlled in genotypes with contrasting sprouting behavior. With this aim two tannin-less, red-grained inbred lines were used as a model system: IS9530 (sprouting resistant) and Redland B2 (sprouting susceptible). Redland B2 grains are able to germinate well before reaching physiological maturity (PM) while IS9530 ones can start to germinate at 40–45 days after pollination, well after PM. Results show that the anticipated dormancy loss displayed by Redland B2 grains is related reduced embryo sensitivity to abscisic acid (ABA) and increased levels of GA upon imbibition. In turn, transcriptional data showed that ABA signal transduction is impaired in Redland B2, which appears to have an impact on GA catabolism, thus affecting the overall GA/ABA balance that regulates germination. QTL analyses were conducted to test whether previous candidate genes were located in a dormancy QTL, but also to identify new genes involved in dormancy. These analyses yielded several dormancy QTL and one of them located in chromosome 9 (qGI-9) was consistently detected even across environments. Fine mapping is already in progress to narrow down the number of candidate genes in qGI-9.

Interspecific Potato Breeding Lines Display Differential Colonization Patterns and Induced Defense Responses after Ralstonia solanacearum Infection

Potato (Solanum tuberosum L.) is one of the main hosts of Ralstonia solanacearum, the causative agent of bacterial wilt. This plant pathogen bacteria produce asymptomatic latent infections that promote its global spread, hindering disease control. A potato breeding program is conducted in Uruguay based on the introgression of resistance from the wild native species S. commersonii Dun. Currently, several backcrosses were generated exploiting the high genetic variability of this wild species resulting in advanced interspecific breeding lines with different levels of bacterial wilt resistance. The overall aim of this work was to characterize the interaction of the improved potato germplasm with R. solanacearum. Potato clones with different responses to R. solanacearum were selected, and colonization, dissemination and multiplication patterns after infection were evaluated. A R. solanacearum strain belonging to the phylotype IIB-sequevar 1, with high aggressiveness on potato was genetically modified to constitutively generate fluorescence and luminescence from either the green fluorescence protein gene or lux operon. These reporter strains were used to allow a direct and precise visualization of fluorescent and luminescent cells in plant tissues by confocal microscopy and luminometry. Based on wilting scoring and detection of latent infections, the selected clones were classified as susceptible or tolerant, while no immune-like resistance response was identified. Typical wilting symptoms in susceptible plants were correlated with high concentrations of bacteria in roots and along the stems. Tolerant clones showed a colonization pattern restricted to roots and a limited number of xylem vessels only in the stem base. Results indicate that resistance in potato is achieved through restriction of bacterial invasion and multiplication inside plant tissues, particularly in stems. Tolerant plants were also characterized by induction of anatomical and biochemical changes after R. solanacearum infection, including hyperplasic activity of conductor tissue, tylose production, callose and lignin deposition, and accumulation of reactive oxygen species. This study highlights the potential of the identified tolerant interspecific potato clones as valuable genetic resources for potato-breeding programs and leads to a better understanding of resistance against R. solanacearum in potato.

Seed Dormancy: Preharvest Sprouting

This article is a revision of the previous edition article by R.L. Benech-Arnold, R.A. Sanchez, volume 3, pp. 1333–1339,