Seed Coat Rupture Research Articles

Diversity of Treatments in Overcoming Morphophysiological Dormancy of Paeonia peregrina Mill. Seeds.

Paeonia peregrina Mill. is a protected, herbaceous species native to Southeastern Europe and Turkey. Due to its vulnerability, it has to be protected both in its natural habitats and through cultivation. Peonies are known to have a low potential for natural regeneration due to their seed dormancy, low germination rate, and long germination period. In this study, treatments with gibberellic acid (GA3 150, 200, 250, 300, and 350 mg L-1 GA3) and warm (at 20/16 °C day/night regime) and cold stratification (at 4 °C) were used to accelerate dormancy release and increase the germination rate. The seeds of P. peregrina from four natural habitats in Serbia and the Institute's collection were collected and analyzed. They showed an underdeveloped embryo that needs to grow inside the seed before it can germinate. The application of GA3 accelerated each stage of germination (seed coat rapturing, hypocotyl dormancy release, and epicotyl dormancy release) for approximately 10 days compared to the control. It was also found that two-day imbibition with 200 mg L-1 GA3 significantly accelerated and equalized germination. Higher GA3 concentrations had a more pronounced impact on each stage but also resulted in greater seed infection after the seed coat rupture, elongated and weak seedlings, while lower concentrations did not result in obtaining uniform seedlings. There were no significant differences observed between localities. Restoring P. peregrina through seeds and nursery-produced plants is crucial for conserving the genetic diversity of the tested species.

Key Metabolite Differences Between Korean Pine (Pinus koraiensis) Seeds With Primary Physiological Dormancy and No-Dormancy.

Pinus Koraiensis seeds have physiological dormancy. Cold stratification releases seed dormancy. The changes in metabolite profiles of dormant seeds and cold stratified seeds during shorter incubation time in a favorable condition for seed germination have been studied. However, a more-long-term detection of the changes in metabolites in dormant seeds can identify the real metabolic pathways responsible for dormancy. Metabolite composition was investigated in embryo and megagametophyte of primary physiological dormant seeds (DS) of P. Koraiensis collected at 0, 1, 2, 4, and 6 weeks of incubation and of non-primary physiological dormant seeds (NDS) sampled at 0 and 1 week of incubation, seed coat rupture stage, and radicle protrusion stage. Embryos contained higher levels of most metabolites than megagametophyte. Strong accumulation of most metabolites in DS occurred at 1 and 4 weeks of incubation. A larger reduction in the relative levels of most phosphorylated sugars and amino acids in NDS was found between 1-week-incubation and seed coat rupture stage. The relative levels of metabolites involved in carbohydrate metabolism, especially the pentose phosphate pathway (PPP) and tricarboxylic acid (TCA) cycle, were higher in the embryos of 4-week-incubated DS, but the relative contents of intermediate metabolites of most amino acid metabolism were lower compared to 1-week-incubated NDS. We suggested that the disturbed carbohydrate metabolism and amino acid metabolism in the embryos of DS after 4 weeks of incubation maybe related to primary dormancy. Our study provides information for a better understanding of the mechanism of seed dormancy.

Identification of tannic cell walls at the outer surface of the endosperm upon Arabidopsis seed coat rupture.

SUMMARYThe seed coat is specialized dead tissue protecting the plant embryo from mechanical and oxidative damage. Tannins, a type of flavonoids, are antioxidants known to accumulate in the Arabidopsis seed coat and transparent testa mutant seeds, deficient in flavonoid synthesis, exhibit low viability. However, their precise contribution to seed coat architecture and biophysics remains evasive. A seed coat cuticle, covering the endosperm outer surface and arising from the seed coat inner integument 1 cell layer was, intriguingly, previously shown to be more permeable in transparent testa mutants deficient not in cuticular component synthesis, but rather in flavonoid synthesis. Investigating the role of flavonoids in cuticle permeability led us to identify periclinal inner integument 1 tannic cell walls being attached, together with the cuticle, to the endosperm surface upon seed coat rupture. Hence, inner integument 1 tannic cell walls and the cuticle form two fused layers present at the surface of the exposed endosperm upon seed coat rupture, regulating its permeability. Their potential physiological role during seed germination is discussed.

Low mycorrhizal diversity in the endangered and rare orchids Bipinnula volckmannii and B. apinnula of Central Chile

Several species of the family Orchidaceae have limited distribution and small populations, in which the probability of extinction is much higher than in broadly distributed species. As orchid seeds are very small and lack energy reserves, they require association with compatible mycorrhizal fungi that provide nutrients and carbon needed for germination and early development. Therefore, identification of putative orchid mycorrhizal partners is essential for the design of conservation strategies for these species. We investigated the mycorrhizal associations of two endangered rare species of orchids, Bipinnula apinnula and B. volckmannii, and compared our results with the mycorrhizal associations of two broadly distributed species of Bipinnula. We isolated mycorrhizal fungi and directly amplified DNA from colonized root segments and found an extremely low diversity of mycorrhizal fungi. The rarefaction curves of rare species populations saturated at a single OTU, showing a highly specialized association and considerably lower diversity than the other two species of Bipinnula. Nevertheless, symbiotic germination experiments with fungal strains of this specific fungal OTU only reached the stage of seed coat rupture in both orchid species, without formation of protocorms. Our results suggest that seeds may require additional treatment for successful germination and that fungal partners isolated from adult plants may not be the same that support the germination of seeds. Further studies are needed to improve germination of thiese orchid species and to contribute to their conservation.

Control of seed coat rupture by ABA-INSENSITIVE 5 inArabidopsis thaliana

AbstractIn Arabidopsis, seed germination is a biphasic process involving rupture of the seed coat followed by emergence of the radicle through the micropylar endosperm. Embryo expansion results in seed coat rupture and removal of seed coat imposed dormancy with DELLA proteins blocking embryo expansion in the absence of gibberellins. Exogenous abscisic acid (ABA) treatment does not block seed coat rupture but does block radicle emergence. We used this limited effect of exogenous ABA to further investigate the mechanism by which it blocks the onset of germination marked by seed coat rupture. We show that physical nicking of the seed coat results in exogenous ABA treatment blocking both seed coat and endosperm rupture and this block requires the transcription factors ABI3 and ABI5, but not ABI4. Furthermore, we show that the repression of expression of severalEXPANSINgenes (EXPA1,EXPA2,EXPA3,EXPA9andEXPA20) by exogenous ABA requires ABI5. We conclude that ABI5 plays an important role in the ABA-mediated repression of germination through prevention of seed coat rupture and propose that this involves EXPANSIN related control of cell wall loosening.

Dehiscence method: a seed-saving, quick and simple viability assessment in rice

BackgroundSeed viability monitoring is very important in ex situ germplasm preservation to detect germplasm deterioration. This requires seed-, time- and labor- saving methods with high precision to assess seed germination as viability. Although the current non-invasive, rapid, sensing methods (NRSs) are time- and labor-saving, they lack the precision and simplicity which are the virtues of traditional germination. Moreover, they consume a considerable amount of seeds to adjust sensed signals to germination percentage, which disregards the seed-saving objective. This becomes particularly severe for rare or endangered species whose seeds are already scarce. Here we propose a new method that is precise, low-invasive, simple, and quick, which involves analyzing the pattern of dehiscence (seed coat rupture), followed by embryonic protrusion.ResultsDehiscence proved simple to identify. After the trial of 20 treatments from 3 rice varieties, we recognized that dehiscence percentage at the 48th hour of germination (D(48)) correlates significantly with germination rate for tested seed lots. In addition, we found that the final germination percentage corresponded to D(48) plus 5. More than 70% of the seeds survived post-dehiscence desiccation for storage. Hydrogen peroxide (1 mM) as the solution for imbibition could further improve the survival. The method also worked quicker than tetrazolium which is honored as a fast, traditional method, in detecting less vigorous but viable seeds.ConclusionWe demonstrated the comprehensive virtues of dehiscence method in assessing rice seed: it is more precise and easier to use than NRSs and is faster and more seed-saving than traditional methods. We anticipate modifications including artificial intelligence to extend our method to increasingly diverse circumstances and species.

Nitrate affects sensu-stricto germination of after-ripened Sisymbrium officinale seeds by modifying expression of SoNCED5, SoCYP707A2 and SoGA3ox2 genes

The influence of nitrate upon the germination of Sisymbrium officinale seeds is not entirely controlled by after-ripening (AR), a process clearly influenced by nitrate. Recently, we have reported that nitrate affects sensu-stricto germination of non-AR (AR0) seeds by modifying the expression of crucial genes involved in the metabolism of GA and ABA. In this study, we demonstrate that nitrate affects also the germination of AR seeds because: (i) the AR negatively alters the ABA sensitivity being the seed more ABA-sensible as the AR is farthest from optimal (AR0 and AR20 versus AR7); in the presence of diniconazole (DZ), a competitive inhibitor of ABA 8′-hydroxylase, testa rupture is affected while the endosperm rupture is not. (ii) AR7 seed-coat rupture is not inhibited by paclobutrazol (PBZ) suggesting that nitrate can act by a mechanism GA-independent. (iii) The germination process is accelerated by nitrate, most probably by the increase in the expression of SoNCED5, SoCYP707A2 and SoGA3ox2 genes. Taken together, these and previous results demonstrate that nitrate promotes germination of AR and non-AR seeds through transcriptional changes of different genes involved in ABA and GA metabolism.

Underdeveloped embryos and germination in Aristolochia galeata seeds

Aristolochiaceae have been described as having seeds with underdeveloped embryos and morphological or morphophysiological dormancy. Aristolochia galeata is a native climber found in the Cerrado biome, associated with road and gallery forest edges. The aims of this study were to investigate: embryo growth rate, morphology and seed germination parameters under different treatments. Embryos were excised to obtain embryo length at four stages: initial, seeds after coat rupture, radicle tip protrusion and cotyledon emergence from the seed coat. Germination tests were conducted at 30 °C under three nitrate concentrations (1, 10 and 20 mM), fluctuating temperature (27/20 °C) and light and dark conditions. We found that seeds have underdeveloped embryos, which take about 301 ± 178 h (±SD) to achieve seed coat rupture, another 205 ± 126 h to reach radicle protrusion and 176 ± 76 h more to the final stage of cotyledon emergence. Germinability was above 52% in all treatments, except in the dark (15%). For all treatments, average germination time was above 290 ± 123 h. Potassium nitrate increased germinability to >87%. No particular treatment was required for embryo development, but seeds in the population that continued to germinate after 1 month were probably in various states of non-deep, simple morphophysiological dormancy. Increased germinability in nitrate treatments and light requirement for germination could prevent germination under unsuitable environmental conditions and be a strategy to increase seedling establishment in the cerrado.

IN VITRO GERMINATION AND SEEDLING DEVELOPMENT OF CAPER (CAPPARIS SPINOSA L.) MATURE SEEDS

Capparis spinosa L. (Capparidaceae) is a perennial tropical and subtropical shrub plant known worldwide. Despite the increasing demand and economic importance of capers, little information is available regarding the propagation of this shrub. In fact, it is usually propagated by seed, but its percentage of germination is very low. In this report, in vitro seed germination and seedling development of Capparis spinosa L. were studied and several treatments were evaluated to determine the ability of several factors to increase the percentage of germination. High variability was observed among the germination percentages of the different treatments (3-81%) due to their different efficiency to break dormancy. Scarification through seed coat rupture has been determinant to improve germination percentage. In vitro obtained seedlings were valuable to establish a micropropagation protocol for caper.

Seed coat rupture and endosperm protrusion in developing seeds of Catharanthus roseus (L.) G. Don.

SummaryProtrusion of the endosperm due to rupture of the seed coat that occurs during seed formation in Catharanthus roseus (L.) G. Don was investigated. Rupture was not limited to a specific part of the seed coat. It occurred most frequently at the chalazal end of the seed. Seeds with protruding endosperm were formed in fruit 17 d after pollination, or later, when the endosperm filled the space enclosed by the seed coat. The frequency of seeds with protruding endosperm increased with the stage of physiological maturity. Seeds with protruding endosperm comprised up to 12% of seeds harvested at full maturity. The protruding endosperm was composed of an abnormal proliferation of endosperm cells. In seeds with protruding endosperm, the seed coat was poorly developed and the embryo protruded from a rupture in the seed coat after imbibition. The percentages of seeds that germinated normally were 61% and 15%, for normal seeds and seeds with a protruding endosperm, respectively. Protrusion of the endosperm in C. roseus seeds may have been caused by a thin seed coat and by pressure caused by the developing embryo and endosperm.

A new procedure for making observations of embryo morphology in dust-like seeds with rigid coats

AbstractObserving the embryo in dust-like seeds with a rigid coat presents a particular challenge, because it is not possible to use conventional techniques applied to normal-sized seeds. A new combination of softening and clearing techniques, which is remarkably effective for direct observations of embryos in minute seeds, is reported here. Moreover, no special seed treatment is required prior to application of this combination of techniques, and good results are obtained from living and preserved material. Dust-like seeds of Cytinus hypocistis and Cytinus ruber with a rigid coat were treated successively with Franklin's and with Jeffrey's softening fluids, and then with Herr's clearing fluid. The combined action of the two softening fluids caused sufficient seed coat rupture to allow Herr's fluid to clear seed tissues, permitting effective observation of the embryo. This technique should be applicable to a wide range of species with dust-like seeds, and its possibilities for biological studies are numerous.

NEW HARVESTING DEVICE FOR SOYBEANS

This study is part of a major project to develop a new and simplified concept of a soybean harvester. The prototype threshes soybeans from standing, uncut plants using impact energy provided by plastic fingers mounted on opposite rotating shafts, and uses a pneumatic system for capturing, separating, cleaning, and transporting the beans. Compared to a conventional combine, the prototype harvested significantly better seeds with respect to their physical characteristics such as broken/split beans, material other than grain, discardable small seeds, and good seeds. Physiological characteristics such as seeds vigor, germination, and seed coat rupture were also significantly better in the prototype samples, while invisible mechanical damages were not statistically different between the prototype and conventionally combined samples. The overall results raised the level of enthusiasm for further development of the new and simplified soybean harvester.

The effect of micronization of full-fat canola seed on digestion in the rumen and total tract of dairy cows

The effect of micronization of full-fat canola seed on dry matter disappearance (DMD) and total nitrogen disappearance (TND) in the rumen and intestine were investigated. Full-fat canola seed was left untreated (C) or micronized for 1.5 min (M), and left whole (CW, MW) or ground to pass through a 1.25-mm sieve (CG, MG). In vitro gas production from CW and MW were determined at 0, 1, 2, 3, 8, 12, 20, 24, 32, 48, 52, 68 and 72 h. For subsequent studies, CW was hand-cracked (CH) for comparison against MW. In situ DMD and TND from CH, MW, CG and MG were determined after 0, 2, 4, 8, 16, 24, 48 and 96 h of ruminal incubation. The mobile nylon bag technique was used to estimate intestinal digestion (DM and N disappearance) of samples incubated in the rumen for 16 h and in acid-pepsin for 1 h. Scanning electron microscopy revealed the pattern and extent of seed coat rupture from hand-cracking and from micronization to be similar, but micronization-mediated changes to the internal structures were evident in ground samples. Micronization increased (P < 0.05) in vitro gas production from whole canola seed (CW vs. MW), but relative to CH, DMD and TND from MW were lower (P < 0.001) in situ and in the whole digestive tract (P < 0.001). Estimated intestinal DMD and TND did not differ (P > 0.05) between CH and MW. Ruminal DMD and TND from ground seed were reduced (P < 0.05) by micronization (CG vs. MG) in the rumen, but intestinal DMD and TND were increased (P < 0.05 and P < 0.01, respectively), thus in the whole digestive tract, these values did not differ (P > 0.05) between treatments. Micronization reduced potential ruminal protein degradability (P < 0.05) and effective ruminal protein degradability (P < 0.01) of full-fat canola, and in combination with grinding, increased the proportion of protein digestion in the intestine. Key words: Micronization, full-fat canola seed, digestibility, rumen, dairy cow, in situ

Effects of gibberellins, darkness and osmotica on endosperm rupture and class I β-1,3-glucanase induction in tobacco seed germination

The “Havana 425” cultivar of Nicotiana tabacum L. is photodormant. Gibberellins (e.g. 10−5 M GA4 or GA7) can substitute for light in releasing dormancy. Measurements of β-1,3-glucanase activity, mRNA accumulation and the activity of the class I β-1,3-glucanase B promoter indicated that class I β-1,3-glucanases are induced by GA4 in the dark in association with germination. As in the light, this induction occurred prior to endosperm rupture and was localized exclusively in the micropylar region of the endosperm where the radicle will penetrate. Abscisic acid (ABA, 10−5 M) did not appreciably affect GA-induced release of photodormancy or seed-coat rupture, but it delayed endosperm rupture and inhibited the rate of class I β-1,3-glucanase accumulation. Seeds imbibed in the light in the presence of osmotica, e.g. 0.04 M polyethylene glycol 6000, showed delayed seed-coat and endosperm rupture, delayed onset of β-1,3-glucanase induction, and decreased rates of β-1,3-glucanase accumulation. These delays were shortened by GA4 treatment. Our results suggest that GAs and ABA act at two distinct sites during germination and that expansive growth of the embryo acts in two ways by triggering β-1,3-glucanase induction and by providing force for endosperm penetration. This provides further support for our working hypothesis that class I β-1,3-glucanases promote endosperm weakening and facilitate radicle penetration.

Class I [beta]-1,3-Glucanases in the Endosperm of Tobacco during Germination.

Rupture of the seed coat and rupture of the endosperm are separate events in the germination of Nicotiana tabacum L. cv Havana 425 seeds. Treatment with 10-5 M abscisic acid (ABA) did not appreciably affect seed-coat rupture but greatly delayed subsequent endosperm rupture by more than 100 h and resulted in the formation of a novel structure consisting of the enlarging radicle with a sheath of greatly elongated endosperm tissue. Therefore, ABA appears to act primarily by delaying endosperm rupture and radicle emergence. Measurements of [beta]-1,3-glucanase activity, antigen content, and mRNA accumulation together with reporter gene experiments showed that induction of class I [beta]-1,3-glucanase genes begins just prior to the onset of endosperm rupture but after the completion of seed-coat rupture. This induction was localized exclusively in the micropylar region of the endosperm, where the radicle will penetrate. ABA treatment markedly inhibited the rate of [beta]-1,3-glucanase accumulation but did not delay the onset of induction. Independent of the ABA concentration used, onset of endosperm rupture was correlated with the same [beta]-1,3-glucanase content/seed. These results suggest that ABA-sensitive class I [beta]-1,3-glucanases promote radicle penetration of the endosperm, which is a key limiting step in tobacco seed germination.

A Possible Mechanism for Seed Coat Rupture by the Embryo in Germinating Wheat

The process of seed-coat splitting, terminating germination, in red wheats is described. We show that sequential expansion of the coleorhiza and coleoptile is required for germinative growth to be normal. Normal emergence requires the seed-coats to be split by the expanding coleorhiza. We here describe a group of cells associated with the coleorhizal tip forming a papilla-like projection which could aid this process.

Allelopathic effects of Engelmann spruce bark stilbenes and tannin–stilbene combinations on seed germination and seedling growth of selected conifers

The allelopathic effects of Engelmann spruce bark and bark extacts on seed germination and seedling growth of several conifer species were examined. Extracts were hydroxystilbenes (isorhapontin and astringin) and condensed tannins. Experiments included the following: (i) seed germination in petri dishes with various concentrations of stilbenes and tannin–stilbenes; (ii) seed germination in petri dishes containing Engelmann spruce bark fragments; (iii) seedling growth in varying mixtures of peat/Engelmann spruce bark, the latter added either before or after seedling establishment; (iv) seedling growth in samples of Engelmann spruce forest soil horizons. In all experiments and in all species there was an allelopathic response to bark extracts. This response was expressed through inhibition of seed germination (seed-coat rupture and radicle emergence), necrotic discoloration and protoplasmic disruption of root-tip cells, suppression of root hairs, and repressed growth in general. Among the conifers tested Pinus ponderosa and Tsuga mertensiana appeared to be the most tolerant to bark extracts, Pinus contorta the most sensitive. Picea engelmannii was moderately sensitive.

Fracture Resistance of Soybeans to Compressive Loading

ABSTRACT THE fracture resistance of whole soybeans was measured in terms of average compressive force, deformation, and toughness at seedcoat rupture. Amsoy-71, Corsoy, and Williams varieties at moisture contents of 8, 11, 14, and 17 percent w.b. were quasi-statically loaded in a horizontal hilum and vertical hilum position. Force required to initiate seedcoat rupture decreased as soybean moisture content in-creased from 8 to 17 percent. Soybeans loaded in the vertical hilum position consistently developed seedcoat cracks at lower levels of force than those in the horizontal hilum position. Maximum toughness occurred in the 11 to 14 percent moisture range, indicating an optimum moisture range for absorbing compressive energy. Soybeans loaded in the vertical hilum position required less energy for seedcoat rupture than those in the horizontal hilum position. Maximum allowable com-pressive energy levels often fell below the impact energy levels that would occur during normal dropping and other handling operations.

The inheritance of seed coat rupture in snap beans Phaseolus vulgaris L.

Seed coat rupture known also as ‘fish face’, is a common seed problem in snap beans. Lines form the cross Tendercrop × Streamliner were classified as high (H), medium (M), and low (L) in their susceptibility to ruptured seed coat and were used as parents in an inheritance study. Crosses of H × L gave an F2 segregation pattern of 1:2:1 for H:M:L classes and this was confirmed by F3 data. The results suggest that seed coat rupture is due to a single incompletely dominant gene (Tr) with 25–50% penetrance. Tr is not linked with seed color or maternal factors.