Rupture Test: A New Method for Evaluating Maize (Zea mays) Seed Vigour.

To explore the application of seed germination biomechanical event(s) in seed vigour tests, a new procedure for the evaluation of maize seed vigour tests based on pericarp-testa rupture (PR) and coleorhiza rupture (CR) during seed germination was developed. Twenty-four lots of hybrid maize were used to determine the feasibility of the rupture test (RT) as a seed vigour test in Zea mays. The results showed that the physiological quality pattern of 24 maize seed lots assessed through RT was similar to that obtained through analysis with other seed test methods. Correlation and regression analyses revealed that the percentage of CR and percentage of PR + CR at "15 ± 0.5 °C for 120 h ± 1 h" and "20 ± 0.5 °C for 72 h ± 15 min" exhibited positive correlations with the field seedling emergence data (p < 0.01). Hence, the proposed method (the rupture test) is cogent and effective, thus providing an important reference for more crops to select for seed germination event(s) and establishing corresponding new methods for seed vigour tests in the future.

KAR1-induced dormancy release in Avena fatua caryopses involves reduction of caryopsis sensitivity to ABA and ABA/GAs ratio in coleorhiza and radicle.

MAIN CONCLUSION: The dormancy release by KAR1 is associated with a reduction of coleorhiza and radicle sensitivity to ABA as well as with reduction the ABA/GAs ratio in the coleorhiza, by a decrease content of ABA, and in the radicle, by a decrease the ABA and an increase of the GAs contents. Both, karrikin 1 (KAR1) and gibberellin A3 (GA3), release dormancy in Avena fatua caryopses, resulting in the emergence of coleorhiza (CE) and radicle (RE). Moreover, KAR1 and GA3 stimulate CE and RE in the presence of abscisic acid (ABA), the stimulation being more effective in CE. The stimulatory effects of KAR1 and GA3 involve also the CE and RE rates. A similar effect was observed at KAR1 concentrations much lower than those of GA3. KAR1 increased the levels of bioactive GA5 and GA6 in embryos and the levels of GA1, GA5, GA3, GA6 and GA4 in radicles. The stimulatory effect of KAR1 on germination, associated with increased levels of gibberellins (GAs) and reduced levels of ABA in embryos, was counteracted by paclobutrazol (PAC), commonly regarded as a GAs biosynthesis inhibitor. Consequently, KAR1 decreased the ABA/GAs ratio, whereas PAC, used alone or in combination with KAR1, increased it. The ABA/GAs ratio was reduced by KAR1 in both coleorhiza and radicle, the effect being stronger in the latter. We present the first evidence that KAR1-induced dormancy release requires a decreased ABA/GAs ratio in coleorhiza and radicle. It is concluded that the dormancy-releasing effect of KAR1 in A. fatua caryopses includes (i) a reduction of the coleorhiza and radicle sensitivity to ABA, and (2) a reduction of the ABA/GAs ratio (i) in the coleorhiza, by decreasing the ABA content, and (ii) in the radicle, by decreasing the ABA and increasing the content GAs, particularly GA1. The results may suggest different mechanisms of dormancy release by KAR1 in monocot and dicot seeds.

Comparative transcriptome analysis of coleorhiza development in japonica and Indica rice.

BACKGROUND: Coleorhiza hairs, are sheath-like outgrowth organs in the seeds of Poaceae family that look like root hair but develop from the coleorhiza epidermal cells during seed imbibition. The major role of coleorhiza hair in seed germination involves facilitating water uptake and nutrient supply for seed germination. However, molecular basis of coleorhiza hair development and underlying genes and metabolic pathways during seed germination are largely unknown and need to be established. RESULTS: In this study, a comparative transcriptome analysis of coleorhiza hairs from japonica and indica rice suggested that DEGs in embryo samples from seeds with embryo in air (EIA) as compared to embryo from seeds completely covered by water (CBW) were enriched in water deprivation, abscisic acid (ABA) and auxin metabolism, carbohydrate catabolism and phosphorus metabolism in coleorhiza hairs in both cultivars. Up-regulation of key metabolic genes in ABA, auxin and dehydrin and aquaporin genes may help maintain the basic development of coleorhiza hair in japonica and indica in EIA samples during both early and late stages. Additionally, DEGs involved in glutathione metabolism and carbon metabolism are upregulated while DEGs involved in amino acid and nucleotide sugar metabolism are downregulated in EIA suggesting induction of oxidative stress-alleviating genes and less priority to primary metabolism. CONCLUSIONS: Taken together, results in this study could provide novel aspects about the molecular signaling that could be involved in coleorhiza hair development in different types of rice cultivars during seed germination and may give some hints for breeders to improve seed germination efficiency under moderate drought conditions.

Avena fatua caryopsis dormancy release is associated with changes in KAR1 and ABA sensitivity as well as with ABA reduction in coleorhiza and radicle.

MAIN CONCLUSION: The dormancy release in Avena fatua caryopses was associated with a reduction in the ABA content in embryos, coleorhiza and radicle. The coleorhiza proved more sensitive to KAR1 and less sensitive to ABA than the radicle. The inability of dormant caryopses and ABA-treated non-dormant caryopses to complete germination is related to inhibition and delayed of cell-cycle activation, respectively. As freshly harvested Avena fatua caryopses are dormant at 20 °C, they cannot complete germination; the radicle is not able to emerge. Both karrikin 1 (KAR1) and dry after-ripening release dormancy, enabling the emergence of, first, the coleorhiza and later the radicle. The after-ripening removes caryopse sensitivity to KAR1 and decreases the sensitivity to abscisic acid (ABA). The coleorhiza was found to be more sensitive to KAR1, and less sensitive to ABA, than radicles. Effects of KAR1 and after-ripening were associated with a reduction of the embryo's ABA content during caryopsis germination. KAR1 was found to decrease the ABA content in the coleorhiza and radicles. Germination of after-ripened caryopses was associated with the progress of cell-cycle activation before coleorhiza emergence. Inhibition of the germination completion due to dormancy or treating the non-dormant caryopses with ABA was associated with a total and partial inhibition of cell-cycle activation, respectively.

Mannans and endo-ß-mannanases (MAN) in Brachypodium distachyon: expression profiling and possible role of the BdMAN genes during coleorhiza-limited seed germination.

Immunolocalization of mannans in the seeds of Brachypodium distachyon reveals the presence of these polysaccharides in the root embryo and in the coleorhiza in the early stages of germination (12h), decreasing thereafter to the point of being hardly detected at 27h. Concurrently, the activity of endo-ß-mannanases (MANs; EC 3.2.1.78) that catalyse the hydrolysis of ß-1,4 bonds in mannan polymers, increases as germination progresses. The MAN gene family is represented by six members in the Brachypodium genome, and their expression has been explored in different organs and especially in germinating seeds. Transcripts of BdMAN2, BdMAN4 and BdMAN6 accumulate in embryos, with a maximum at 24-30h, and are detected in the coleorhiza and in the root by in situ hybridization analyses, before root protrusion (germination sensu stricto). BdMAN4 is not only present in the embryo root and coleorhiza, but is abundant in the de-embryonated (endosperm) imbibed seeds, while BdMAN2 and BdMAN6 are faintly expressed in endosperm during post-germination (36-42h). BdMAN4 and BdMAN6 transcripts are detected in the aleurone layer. These data indicate that BdMAN2, BdMAN4 and BdMAN6 are important for germination sensu stricto and that BdMAN4 and BdMAN6 may also influence reserve mobilization. Whether the coleorhiza in monocots and the micropylar endosperm in eudicots have similar functions, is discussed.

Architectural and biochemical changes in embryonic tissues of maize under cadmium toxicity.

Heavy metals greatly alter plant morphology and architecture, however detailed mechanisms of such changes are not fully explored. Two experiments were conducted to investigate the influence of cadmium (CdCl2 · 2.5H2 O) on some germination, morphological, biochemical and histological characteristics of developing embryonic tissue of maize. In the first experiment, maize seeds were germinated in increasing levels of CdCl2 (200-2000 µm) in sand and measurements were taken of changes in germination and seedling development attributes. Based on these parameters, 1000 µM CdCl2 was chosen for detailed biochemical and histological measurements. In the second experiment, seeds were germinated in Petri dishes and supplied with 0 (control) or 1000 µM CdCl2 (Cd-treated). Radicle, plumule, coleoptile and coleorhiza were measured for biochemical and histological changes. The highest amount of Cd was in the coleorhiza and radicle. Free proline, soluble sugars, anthocyanin, soluble phenolics, ascorbic acid, H2 O2 and MDA were significantly higher in coleorhizae, followed by the coleoptile, radicle and plumule. Although the radicle and coleorhiza were relatively poor targets of Cd than the other tissues, Cd stress reduced cortical cell size and vascular tissues, and deformed xylem and phloem parenchyma in all plant parts. In conclusion, the main reason for reduced germination was the influence of Cd on architecture of the coleorhiza and coleoptile, which was the result of oxidative stress and other physiological changes taking place in these tissues.

Analogous reserve distribution and tissue characteristics in quinoa and grass seeds suggest convergent evolution.

Quinoa seeds are highly nutritious due to the quality of their proteins and lipids and the wide range of minerals and vitamins they store. Three compartments can be distinguished within the mature seed: embryo, endosperm, and perisperm. The distribution of main storage reserves is clearly different in those areas: the embryo and endosperm store proteins, lipids, and minerals, and the perisperm stores starch. Tissues equivalent (but not homologous) to those found in grasses can be identified in quinoa, suggesting the effectiveness of this seed reserve distribution strategy; as in cells of grass starchy endosperm, the cells of the quinoa perisperm endoreduplicate, increase in size, synthesize starch, and die during development. In addition, both systems present an extra-embryonic tissue that stores proteins, lipids and minerals: in gramineae, the aleurone layer(s) of the endosperm; in quinoa, the micropylar endosperm; in both cases, the tissues are living. Moreover, the quinoa micropylar endosperm and the coleorhiza in grasses play similar roles, protecting the root in the quiescent seed and controlling dormancy during germination. This investigation is just the beginning of a broader and comparative study of the development of quinoa and grass seeds. Several questions arise from this study, such as: how are synthesis and activation of seed proteins and enzymes regulated during development and germination, what are the genes involved in these processes, and lastly, what is the genetic foundation justifying the analogy to grasses.

Gene expression patterns in wheat coleorhiza under cold- and biological stratification.

This study assessed germination of wheat seeds under cold and biological stratification and determined the expression level of gibberellins (GA) and abscisic acid (ABA) genes in coleorhiza. Both cold and biological stratification significantly (P<0.05) enhanced the rate and efficacy of germination. The spatial distance between the fungal endophyte and the seed can be a determining factor of biological stratification as seeds in direct contact with fungal endophyte showed the highest rate and efficacy of germination. Consistently high expression of GA3ox2 gene was found in wheat coleorhiza throughout the tested period of germination. The expression of ABA biosynthesis gene, TaNCED, was substantially higher in cold stratification seeds, reflecting the role of abscisic acid in stress-adaptation. Overall, this study provides molecular evidence of the importance of coleorhiza in germinating wheat seeds, in addition to reporting that the spatial distance between symbiotic partners may be a critical factor driving mycovitality.