Ulteriori ricerche sulla germinazione di Cercis siliquastrum L. Azione dell'acido gibberellico e ruolo dell'endosperma nel fenomeno della dormienza

Abstract

Further research on the germination of Cercis siliquastrum L. Effect of gibberellic acid (GA₃) and role of endosperm in dormancy.—In order to locate the causes of the dormancy in Cercis siliquastrum L., researches have been carried out also on non-scarified seeds without integument, as well as on isolated embryos.

We have noted that gibberellic acid can substitute the cold treatment to interrupt the dormancy and we have also investigated the optimal concentrations.

The results we have obtained do not show the inhibition to be located in the integument and point out a determinant role of the endosperm.

Histological investigations are being carried out to study better the possible occurrence of a mechanical restraint imposed by the albumen on the non-dormant embryo.     

赤霉素对紫斑牡丹种子下胚轴萌发及其种皮和胚乳结构的影响

以紫斑牡丹种子为试验材料,考察不同浓度赤霉素(GA_3)处理下种子生根情况,同时对其种皮纹饰进行了扫描电镜(SEM)观察,并对胚乳结构进行了荧光倒置显微镜、透射电镜(TEM)观察,以揭示GA_3对紫斑牡丹种子下胚轴解除休眠过程中种子种皮、胚乳结构变化和生根的影响。结果表明:(1)GA_3处理能够促进紫斑牡丹种子生根,以300 mg/L GA_3处理的种子生根效果最好,与对照相比可提前14.66 d生根,生根率可达62.33%。(2)扫描电镜观察发现,紫斑牡丹种皮表面含有蜡质,种子萌发进程中蜡质明显不断增加,种皮表面网眼逐渐消失,种皮皱缩明显。(3)光学显微镜观察发现,GA_3处理的紫斑牡丹种子胚乳细胞中含有较多淀粉体和蛋白体,并随着种子萌发进程逐渐积累,可为种子萌发提供足够的养分。(4)透射电镜观察发现,GA_3处理的紫斑牡丹种子胚乳细胞内大分子营养物质不断降解,线粒体、内质网、高尔基体等细胞器相继出现,种子下胚轴休眠解除,细胞壁较薄后期消融,利于营养物质传送。研究认为,适宜浓度GA_3处理可有效加快紫斑牡丹种皮结构的改变和胚乳细胞内物质代谢进程,促进种子萌发,且300 mg/L GA_3处理紫斑牡丹种子24 h可以获得最好的生根效果。     

The Mechanism of Low Temperature Dormancy in Mature Seeds of Syringa Species

The mechanism of seed dormancy at low temperatures (15-9°C) was investigated in the seeds of Syringa josikaea, S. reflexa and S. vulgaris. Low temperature dormancy in Syringa species was mainly imposed by endosperm embedding the radicle. Different degrees of embryo dormancy may occur in S. reflexa seeds. In most cases the low temperature dormancy was broken completely by removing the endosperm around the radicle. The endosperm did not seem to contain significant quantities of germination inhibitors, and the results indicate that it prevents germination mainly due to its mechanical resistance. The mechanical resistance of endosperm did not change during chilling or during induction of dormancy by high temperature incubation. The strength of the endosperm decreased rapidly in non-dormant seeds before visible germination. Similar changes were not observed in dormant seeds. Generally, the strength of the endosperm was lower in the non- (or less) dormant species S. josikaea and S. vulgaris than in the more dormant S. reflexa seeds. The growth potential of the embryos, measured as their ability to germinate in osmotic solutions (mannitol or polyethylen glycol 4000), was increased by chilling and by GA₃-treatment. The growth potential of untreated S. josikaea and S. vulgaris embryos was generally higher than that of S. reflexa embryos. Acid ethyl acetate fractions of methanol extracts from embryos of all three species contained substances with GA³-like activity in the lettuce hypocotyl test. The activity was found at Rf 0.9–1.0 on paper chromatograms run in distilled water. No significant changes in the activity were detected during chilling or prior to visible germination.     

Conseguenze e Significato del Prolungamento Nella Durata della Dormienza in Triticum Durum Desf

Abstract

Consequences and significance of dormancy Prolongation in «Triticum durum» Desf. — Different environment conditions have effect on the dormancy duration: this has been ascertained also for durum wheat Cappelli. Starting from this acquisition, the author does the embryo exchange between seeds of Cappelli different for dormancy degree. It is demonstrated that there is a direct relation between dormancy degree and inhibition power of endosperm.     

Nuove Prospettive Nello Studio dei Fattori che Controllano la Germinazione Dei Semi

Abstract

New perspectives in the study of factors which control seed germination. — Seedlings of Triticum durum, cv. « Cappelli », coming from unirradiated embryos grafted on to irradiated endosperms (EM(u)/EN(i) of presoaked seeds (in distilled water for 24 h. at 20[ddot]C.) (treatment: X-rays, doses, 2,4,6,8,10 and 20 Kr), grow more than seedlings of control EM(u)/EN(u) (dose 0) (fig. 1). To have this reaction, it is necessary that the used seeds be after-ripe; at the various stages of seed ripening, be ginning from the milk stage, the phenomenon is not present.

On this basis, the author has thought that a natural inhibitor occurs in the after-ripe endosperm of « Cappelli », which is neutralized or destroyed by X-rays.

As an experimental demonstration, some trials have been made of growing wheat seedlings in Petri dish, on moistened (distilled water) filter paper, together with excised embryos or isolated endosperms (fig. 2): the after-ripe endosperm is able to depress the seedling growth (fig. 3). In the same experimental conditions, X-rays, dose 6 Kr, neutralize the inhibition effect given by the endosperm. (fig. 4).

A completely different situation occurs in wheat seed, during its ripening: endosperm is inactive, embryo produces inhibition effects on the seedling growth, which, also in this case, are reduced by X-rays.

These phenomena, put in relation with dormancy in Triticum durum, cv. « Cappelli », which is a relative dormancy, having its maximum at the milk stage, have led the author to the general conclusion that, during dormancy, a germination inhibitor occurs in the embryo of wheat seed; when dormancy is finished, the inhibitor appears in the endosperm, in a situation which becomes stable and definitive.     

Potential Risk of Hybridization in Ex Situ Collections of Two Endangered Species of Sinojackia Hu （Styracaceae）

Spontaneous hybridization in ex situ facilities can undermine the genetic integrity of ex situ collections and potentially contaminate open-pollinated seeds or seedlings destined for the reintroduction of endangered plant species into the wild. In the present study, the potential risk of hybridization between two endangered Chinese endemic species, namely Sinojackia xylocarpa Hu and S. rehderiana Hu, which are naturally allopatric species but were conserved ex situ in Wuhan Botanical Garden （WBG）, Wuhan, China, were investigated over three consecutive years from 2003 to 2005. The entire overlapping flowering period of the two species was 14-20 d and the two species shared the same pollinator insects during the entire flowering season in WBG. The floral isolation between the two species was not an issue in the ex sltu collection at WBG. The results suggest an opportunity for pollen transfer between species and a potential risk of genetic Introgression and loss of genetic identity of open-pollinated seeds produced in the ex sltu Collection of these two endangered species. An artificial reciprocal cross between S xylocarpa and S. rehderlana confirmed that the two congener species could readily set seeds, indicating no post-pollination barriers to hybridization and the importance of spatial isolation as a barrier to inter-specific crossing. Therefore, to manage these crossable species with overlapping flowering times and shared pollination vectors in ex situ facilities, spatial isolation should be carefully considered to minimize the possibility of spontaneous hybridization.     

南京老山秤锤树空间分布格局及种间关联性

以南京老山1 hm 2样地秤锤树(Sinojackia xylocarpa)天然种群为研究对象,运用成对g(r)函数,选择完全随机模型、异质泊松模型与先决条件零模型,分析秤锤树种群结构和空间分布格局及其空间关联性,从空间格局角度来深入认识其种群结构和分布格局及形成该格局可能存在的机制并提出保护建议。结果表明:(1)秤锤树天然种群中小径个体数量占优,属于增长型种群。(2)种内空间分布研究中,基于完全随机模型分析,秤锤树种群在尺度0~26 m时为聚集分布,尺度29~30 m时为均匀分布;基于异质泊松模型分析,秤锤树种群在0~23 m时为聚集分布,尺度27~30 m时为均匀分布。秤锤树空间分布表现为由聚集分布向均匀分布变化。(3)主要种间关联性研究中,秤锤树与朴树(Celtis sinensis)的种间关联性表现为小尺度下负关联,随着空间尺度的增加变为正关联。秤锤树与黄连木(Pistacia chinensis)和秤锤树与三角槭(Acer buergerianum)的种间关联性大致相同,基本为大尺度下正关联,偶尔出现负关联和无关联。上述结果表明,秤锤树种群更新状况良好,种群空间分布以聚集分布为主,其主要受种间竞争、扩散限制与密度制约的影响。基于种群现状开展就地保护与适当干扰其生存群落,是濒危物种秤锤树的科学有效的保护措施。     

Changes in the Endosperm Cell Walls of Two Datura Species before Radicle Protrusion

The possibility of an association between changes in cell walls of the micropylar portion of the endosperm and the induction of germination was explored in seeds of Datura ferox and Datura stramonium. The structure of the inner surface of the endosperm was studied by scanning electron microscopy and the composition of cell wall polysaccharides analyzed by gas chromatography and gas chromatography-mass spectrometry. Both scanning electron microscope images and chemical analysis showed changes in the micropylar portion of the endosperm in induced seeds before radicle protrusion. The inner surface of the endosperm appeared eroded, and in some areas, wall material seemed to be missing. The content of the main component of the cell wall polysaccharides, containing predominantly 4-linked mannose, decreased well before the emergence of the radicle through the endosperm. We propose that the degradation of a mannan type polysaccharide is an important factor in the reduction in mechanical strength of the endosperm, thus facilitating germination.     

Le Basi Anatomiche Dei Rapporti Nutritivi Tra Pianta Madre,Seme Ed Embrione

Abstract

The anatomical basis of the nutritive relationships between mother plant, seed, and embryo. — The morphology and anatomy of the fruits and seeds of the Angiosperms show a great variety of structures and adaptments, even within the same family, and one must be cautious in drawing generalized conclusions.

If we first examine the ovary we see that the single carpel receives three vascular traces from which the three main bundles originate, a dorsal and two ventral ones, all more or less reduced. Except in the case of laminar placentation the ovule traces are connected to the ventral vascular system, but often the entire vascular system of the ovary is anastomosed and therefore reticular. However especially when the placentae are at the centre or at the basis of the ovary, it is possible to detect a tendency towards a separation between the vascular system of the ovarian wall and that of the placentae.

The ovular bundle runs through the funicle reaching the chalaza, where it can either end or continue towards the micropyle with a single bundle or with a few branches or even forming a complete reticular envelope surrounding the ovule. The ovular vascular bundles are normally found in the outer integument.

The ovule is made of an inner part (nucellus), and an outer one (integuments). The integuments play a very important role in the processes of seed maturation, dormancy, and germination. They are isolated from the interior of the seed by a cuticle which is a common production of the inner integumentary epidermis, and of the nucellus. The cuticle is not present in the chalaza and can be dissolved in the micropylar region: through these two apertures nutrients can penetrate into the seed or haustoria can grow out of it. During the course of maturation these openings become closed by various means, often through the formation of a new cuticle or of a suberised chalazal plate.

The nutrients which pass through the chalaza penetrate into the nucellus where in some cases one can find some structures which facilitate the communications between the chalaza and embryo sac. The endosperm feeds at the expense of the nucellus but often it can establish a direct contact with the chalaza or the integuments or even the placentae. This occurs often thanks to haustoria.

The embryo is normally surrounded at first by a more or less liquid endosperm: in a second stage the endosperm becomes cellular and the embryo grows at its expense through the digestive activity of the cotyledonar epidermis.

From an anatomo-physiological point of view the following points seem of particular interest:

(I) The endosperm and the embryo show a remarkable autonomy in respect of the mother plant: from an anatomical point of view this is shown by the isolation of the endosperm and embryo by means of a cuticular covering or substitutive structures and by the interposition of nutritive tissues between the vascular system of the mother plant and the endosperm.

(II) Given the importance of the inner cuticle its presence and its constitution should be ascertained in the various species having also in mind the properties of selective permeability shown by the testa.

(III) Two nutritive mechanisms exist: translocation of nutrients via the vascular system and the nutritive tissues, and digestion of surrounding cells. In the digestive phenomena it is important to explain the mechanisms by which only the right cells are digested and not the others.

(IV) The embryo very frequently is immersed at first in a more or less liquid endosperm and is later surrounded by a compact tissue; the nutritive mechanisms are probably different in the two cases.

(V) Two endospermic zones are often distinguishable: one having an haustorial or at least a digestive or elaborative function, and being typically non cellular; another zone, typically cellular, forms a tissue which is sooner or later absorbed by the embryo. The cellularization of this zone seems to coincide with the establishment of polarity and with the beginning of maximum growth of the embryo.

(VI) The relationships between the inner seed and the integuments is complex and there is a correlation between the histoanatomical and biochemical changes of these two parts during seed development. The modifications undergone by the integuments are important steps also towards the preparation of the seed to the processes of dispersal, dormancy, and germination.     

Sull'evoluzione Dei Mitocondri Nell'endosperma di Semi di Ricino in Germinazione

Abstract

On the behavior of mitochondria in the castor bean seed endosperm during the early phases of germination. — In the endosperm of the castor bean seed the oxidative activity and the protein nitrogen contents of the mitochondrial fraction markedly increase during the first period of germination (Beevers and coworkers). The activation of the mitochondrial system is paralleled by a similar increase of the activity of several soluble enzymes; the latter process is severely depressed by protein synthesis inhibitors (Cornaggia, Aberghina).

The present research is aimed to understand at what extent phenomena of activation and/or, respectively, of « ex novo » synthesis are responsible of the increase of mitochondrial activity. The following aspects of the mitochondrial behavior during the early period of germination were investigated:

a) Changes in the activity of cytochrome oxydase, malate dehydrogenase and of the succinate-citochrome reductase system.

b) Changes in the morphology of mitochondria and other particulated cell structures, as revealed by electron microscopy.

In the mitochondrial preparation all of the three enzymatic activities investigated were found to increase rapidly during the first days of germination. The increase during the first 24 hours was almost as large when measured as specific activity (activity per mg protein in the mitochondrial fraction) than when measured on an absolute (i.e. per seed) basis; moreover, it was not significantly inhibited by puromycin or by actinomycin. The increase of the three activities during the following period of germination (second-third day) was accompanied by an increase of the protein nitrogen (per seed) in the mitochondrial fraction, and was consistently depressed by the protein synthesis inhibitors.

In the mitochondrial preparation all of the three enzymatic activities investigated were found to increase rapidly during the first days of germination. The increase during the first 24 hours was almost as large when measured as specific activity (activity per mg protein in the mitochondrial fraction) than when measured on an absolute (i.e. per seed) basis; moreover, it was not significantly inhibited by puromycin or by actinomycin. The increase of the three activities during the following period of germination (second-third day) was accompanied by an increase of the protein nitrogen (per seed) in the mitochondrial fraction, and was consistently depressed by the protein synthesis inhibitors.

These results, integrated with those of other investigations on the same material are in agreement with the hypothesis that the activation of metabolism in the endosperm during germination depends in a very early phase mainly on the transition of enzyme systems from an inactive to an active state; while in a second phase synthesis « ex novo » of enzymes and cell structures predominates.     

Achene germination of the spring ephemeroid species Carex physodes in the Gurbantunggut Desert

Much of the seed germination research on Carex has focused on wetland species, and little is known about the species of arid habitats. Here, we investigated seed dormancy and germination of Carex physodes, which is an important component of the plant community of the Gurbantunggut Desert of the Junggar Basin in Xinjiang, China. Our studies included the effects of mechanical and chemical scarification, dry storage, treatment with GA₃, wet‐cold stratification, and burial in the field. No freshly matured achenes germinated over a range of temperature regimes after treatment with GA₃, 6 months of dry storage or removal of part of the endosperm. The mechanical scarification resulted in < 5% achene germination, however, higher percentage of achene germination occurred after removal of the pericarp (60%), H₂SO₄ scarification (30%) or scarification in 10% NaOH (85%). Six and nine months of wet‐cold stratification promoted < 40% achene germination. The optimal germination temperatures ranged from 25/10°C to 35/20°C. Maximum germination after 9 months of burial at a depth of 3 cm in the field was 36%. These results indicate that the seeds have non‐deep physiological dormancy (PD) and that the pericarp contains germination inhibitors and has strong mechanical resistance to germination.     

A comparison of the seed dormancy characteristics of Spartina patens and Spartina alterniflora (Poaceae)

Seed dormancy characteristics of Spartina alterniflora were delineated previously by Plyler and Carrick (American Journal of Botany, vol. 80, pp. 752–756, 1993). This study was undertaken in order to determine whether or not the dormancy characteristics of S. patens are similar. As in the previous study, the site of a dormancy mechanism was determined by assessing the germinability of surgically altered dormant seeds. Likewise, the effects of three growth-regulating substances (abscisic acid, fusicoccin, and gibberellic acid), prechilling, and light were studied by assessing the germinability of appropriately treated dormant seeds. Surgical injury to the endosperm, and to a lesser extent the scutellum, produced significant germination in S. patens, whereas only injury to the scutellum produced germination in S. alterniflora. Exogenously applied abscisic acid was highly effective in maintaining dormancy in injured seeds of both species. Gibberellic acid and fusicoccin were ineffective in promoting germination in S. patens, but fusicoccin was highly effective in breaking dormancy in 5. alterniflora. Prechilling was effective in breaking dormancy in S. patens but not in S. alterniflora. In S. patens, treatments that broke dormancy were only successful when seeds were exposed to light during the germination period. It is concluded that the dissimilarities in the dormancy mechanisms may be manifestations of the different environmental adaptations these closely related species display.     

Floral ontogeny of Sinojackia xylocarpa (Styracaceae), with special reference to the development of the androecium

Using scanning electron microscopy, we studied the floral ontogeny of Sinojackia xylocarpa. There are 6–7 (–8) sepals. Sepal initiation is staggered; adaxial sepals arise later than abaxial and lateral ones. There are (5–) 6–7 (–8) petals, initiated simultaneously. Petals alternate with the sepals, and occasionally there are two petals instead of one between two sepals. The (10–) 12–14 (–16) stamens are initiated centripetally in two sets (whorls). These floral organ numbers deviate from those of the otherwise mostly pentamerous family Styracaceae. The ovary consists of three (rarely four) locules. In each of the locules, two rows of ovules are differentiated basipetally. Placentation is axile with (5–) 6 (–7) ovules in each locule. Ovules are unitegmic and are ascending with the micropyle directed downwards. Intra‐ovarian trichomes are present as in other representatives of Styracaceae and seem to be an apomorphic character of the family as they are absent in the closely related Symplocaceae and Diapensiaceae. Various levels of organ union occur in anthetic S. xylocarpa. The calyx is synsepalous and the ovary syncarpous. Possibly, the basal connation of petals and stamens is postgenital (and not congenital), but this needs further study. The outward curvature of the young anthers of the inner stamen whorl superficially simulates an obdiplostemonous androecium. However, the sequence of stamen initiation shows a diplostemonous pattern.     

侧金盏胚和胚乳发育的细胞胚胎学研究

利用石蜡切片技术对毛茛科植物侧金盏胚及胚乳发育进行了研究,以明确其胚胎发育的特征,为毛茛科植物的系统研究提供资料。结果表明:(1)侧金盏胚的发育属于柳叶菜型,胚乳发育为核型;初生胚乳核的分裂早于合子的第一次分裂。(2)种子成熟时,种胚尚未分化完全,尚处于球形胚后期或心形胚早期阶段,整个胚发育大约需要50~60d。(3)侧金盏种子存在明显的形态生理休眠现象,经后熟作用逐渐完成种胚的分化与生长,形成子叶形胚;侧金盏种子在相同处理条件下胚分化和发育的速度存在差异。     

Molecular mechanisms and hormonal regulation underpinning morphological dormancy: a case study using Apium graveolens (Apiaceae)

Underdeveloped (small) embryos embedded in abundant endosperm tissue, and thus having morphological dormancy (MD) or morphophysiological dormancy (MPD), are considered to be the ancestral state in seed dormancy evolution. This trait is retained in the Apiaceae family, which provides excellent model systems for investigating the underpinning mechanisms. We investigated Apium graveolens (celery) MD by combined innovative imaging and embryo growth assays with the quantification of hormone metabolism, as well as the analysis of hormone and cell-wall related gene expression. The integrated experimental results demonstrated that embryo growth occurred inside imbibed celery fruits in association with endosperm degradation, and that a critical embryo size was required for radicle emergence. The regulation of these processes depends on gene expression leading to gibberellin and indole-3-acetic acid (IAA) production by the embryo and on crosstalk between the fruit compartments. ABA degradation associated with distinct spatiotemporal patterns in ABA sensitivity control embryo growth, endosperm breakdown and radicle emergence. This complex interaction between gibberellins, IAA and ABA metabolism, and changes in the tissue-specific sensitivities to these hormones is distinct from non-MD seeds. We conclude that the embryo growth to reach the critical size and the associated endosperm breakdown inside MD fruits constitute a unique germination programme.     

Caryopsis germination and seedling emergence in an inland dune dominant grass Leymus secalinus

Leymus secalinus (Georg.) Tzvel. (Poaceae) is a dominant sand dune grass inhabiting the Mu-Us Sandland, semiarid China. Freshly harvested caryopses (seeds) are in non-deep physiological dormancy (non-deep PD) because of low percentage and slow rate of germination. Experiments were conducted to examine the effects of temperature, cold stratification, caryopsis coat scarification or partial removal of endosperm and sand burial on caryopsis dormancy, germination and seedling emergence. Caryopsis germination was significantly influenced by duration of cold stratification, temperature and their interactions. After 8 weeks of cold stratification, caryopsis germination percentage at 30 °C reached to 90%, equally in light or darkness. Rate and percentages of germination were also hastened and increased by scarifying the caryopsis coat or by artificial removal of different proportions of the endosperm. However, seedling developmental characteristics were significantly influenced by the proportion of the endosperm that remained in the caryopses. Seedling emergence, caryopsis germination and enforced dormancy in sand were significantly affected by sand burial depth. As sand burial depth increased, caryopsis germination and seedling emergence decreased whereas caryopsis enforced dormancy increased. 1–2 cm was the optimal depths for caryopses germination and seedling emergence. Although there were still 30% caryopses germinated at 8 cm, the maximal burial depth for seedling emergence was only 4 cm. The partial germination strategy regulated by non-deep PD, temperature and sand burial ensures that only a few caryopses germinated each time and may reduce risks for seedling survival.