Phylogenetic implications of endosperm cell wall morphology in Douglasia, Androsace, and Vitaliana (Primulaceae)

We investigated differences in endosperm cell wall thickening in three genera of Primulaceae; seven species of Douglasia , 28 species of Androsace representing the four sections of the genus, and the single species of the genus Vitaliana . Endosperm cell walls have irregular wall thickenings and narrow constrictions and characterize all Douglasia species and Vitaliana , whereas evenly thickened cell walls without constrictions were found in the endosperm of Androsace and Primula . The endosperm character supports the monophyly of Douglasia and Vitaliana , and we discuss the phylogenetic implications of this conclusion.     

Complex biogeographic patterns in Androsace (Primulaceae) and related genera: evidence from phylogenetic analyses of nuclear internal transcribed spacer and plastid trnL-F sequences

We conducted phylogenetic analyses of Androsace and the closely related genera Douglasia, Pomatosace, and Vitaliana using DNA sequences of the nuclear internal transcribed spacer (ITS) and the plastid trnL-F region. Analyses using maximum parsimony and Bayesian inference yield congruent relationships among several major lineages found. These lineages largely disagree with previously recognized taxonomic groups. Most notably, (1) Androsace sect. Andraspis, comprising the short-lived taxa, is highly polyphyletic; (2) Pomatosace constitutes a separate phylogenetic lineage within Androsace; and (3) Douglasia and Vitaliana nest within Androsace sect. Aretia. Our results suggest multiple origins of the short-lived lifeform and a possible reversal from annual or biennial to perennial habit at the base of a group that now contains mostly perennial high mountain or arctic taxa. The group containing Androsace sect. Aretia, Douglasia, and Vitaliana includes predominantly high alpine and arctic taxa with an arctic-alpine distribution, but is not found in the European and northeastern American Arctic or in Central and East Asia. This group probably originated in Europe in the Pliocene, from where it reached the amphi-Beringian region in the Pleistocene or late Pliocene.     

Seed morphology and endosperm structure of selected species of Primulaceae, Myrsinaceae, and Theophrastaceae and their systematic importance

Seed size and shape, seed coat surface pattern, seed coat thickness, and endosperm structure were investigated in Androsace septentrionalis, Cortusa matthioli, Hottonia palustris, Primula elatior, Soldanella carpatica (Primulaceae), Anagallis arvensis, A. minima, Cyclamen purpurascens, Glaux maritima, Lysimachia nemorum, L. vulgaris, Trientalis europaea (Myrsinaceae), and Samolus valerandi (Theophrastaceae). Three seed size categories were distinguished on the basis of biometric measurements. Almost all seeds examined were found to be small with an angular shape classified as sectoroid or polyhedral. A new type of seed shape, suboval, was identified for H. palustris. Cyclamen purpurascens seeds differed from seeds of all other species examined because of their large size, subglobose shape, and concave hilar area. Despite the different shape types, the length/width ratio of the seeds examined was constant, while their hilum length/width ratio was a highly variable feature. Three types of seed surface patterns were observed: (1) reticulate, (2) tuberculate with secondary striations, and (3) poroid-alveolate with the presence of a spongy outer layer. For seeds of Anagallis arvensis, A. minima, Cortusa matthioli, Lysimachia nemorum, and Soldanella carpatica, secondary seed sculpture was described. The seed coats of all species examined were two-layered, and great differences in testa thickness were found (9.9?C128.6???m). In general, seeds of the Myrsinaceae species were more often characterized by thick testa. Different proportions in thickness of the inner and outer testa layers were observed. In seeds with reticulate seed patterns, the inner testa layer was twice to several times thicker than the outer layer, while the opposite proportions were observed in seeds with the tuberculate or poroid-alveolate seed sculpture pattern. In seeds of all species examined, oxalate crystals were present on the surface of the inner testa layer. Thus, the presence or absence of oxalate crystals in testa is not a feature distinguishing species with angular or subglobose seeds. Four types of endosperm structure were identified according to the thickness of the endosperm cell walls and the relief of their inner surface: (1) with evenly thickened and smooth cell walls, (2) with evenly thickened cell walls and circular or helical thickenings on their inside surfaces, (3) with very thick, but not evenly thickened cell walls with constrictions (??pitted??), and (4) with very thin papery and undulate cell walls. There is no rule concerning the seed shape, type of the seed sculpture, testa thickness, or endosperm structure that corresponds to the family affiliation of the species examined.     

Arabinoxylan, β‐glucan and pectin in barley and malt endosperm cell walls: a microstructure study using CLSM and cryo‐SEM

The architecture of endosperm cell walls in Hordeum vulgare (barley) differs remarkably from that of other grass species and is affected by germination or malting. Here, the cell wall microstructure is investigated using (bio)chemical analyses, cryogenic scanning electron microscopy (cryo‐SEM) and confocal laser scanning microscopy (CLSM) as the main techniques. The relative proportions of β‐glucan, arabinoxylan and pectin in cell walls were 61, 34 and 5%, respectively. The average thickness of a single endosperm cell wall was 0.30 µm, as estimated by the cryo‐SEM analysis of barley seeds, which was reduced to 0.16 µm after malting. After fluorescent staining, 3D confocal multiphoton microscopy (multiphoton CLSM) imaging revealed the complex cell wall architecture. The endosperm cell wall is composed of a structure in which arabinoxylan and pectin are colocalized on the outside, with β‐glucan depositions on the inside. During germination, arabinoxylan and β‐glucan are hydrolysed, but unlike β‐glucan, arabinoxylan remains present in defined cell walls in malt. Integrating the results, an enhanced model for the endosperm cell walls in barley is proposed.     

The emergence of embryos from hard seeds is related to the structure of the cell walls of the micropylar endosperm, and not to endo-beta-mannanase activity

BACKGROUND AND AIMS: Seeds of carob, Chinese senna, date and fenugreek are hard due to thickened endosperm cell walls containing mannan polymers. How the radicle is able penetrate these thickened walls to complete seed germination is not clearly understood. The objective of this study was to determine if radicle emergence is related to the production of endo-beta-mannanase to weaken the mannan-rich cell walls of the surrounding endosperm region, and/or if the endosperm structure itself is such that it is weaker in the region through which the radicle must penetrate. METHODS: Activity of endo-beta-mannanase in the endosperm and embryo was measured using a gel assay during and following germination, and the structure of the endosperm in juxtaposition to the radicle, and surrounding the cotyledons was determined using fixation, sectioning and light microscopy. KEY RESULTS: The activity of endo-beta-mannanase, the major enzyme responsible for galactomannan cell wall weakening increased in activity only after emergence of the radicle from the seed. Thickened cell walls were present in the lateral endosperm in the hard-seeded species studied, but there was little to no thickening in the micropylar endosperm except in date seeds. In this species, a ring of thin cells was visible in the micropylar endosperm and surrounding an operculum which was pushed open by the expanding radicle to complete germination. CONCLUSIONS: The micropylar endosperm presents a lower physical constraint to the completion of germination than the lateral endosperm, and hence its structure is predisposed to permit radicle protrusion.     

胚乳细胞壁的建成及其与胞质微管的关系

综述了被子植物核型胚乳发育的不同阶段(合胞体时期、初始垂周壁形成期、初始平周壁形成期和胚乳细胞分裂增生期)胚乳细胞壁建成的各种观点,以及在各个阶段中微管与胚乳细胞壁建成之间关系的最新研究进展。并分别比较了胚乳细胞化过程不同阶段胚乳游离核或胚乳细胞有丝分裂与分生组织细胞有丝分裂微管周期的差异。     

Early Endosperm Development in Ovules of Soybean, Glycine max (L) Merr. (Fabaceae)

Endosperm development was studied in normally setting flowersand pods of soybean from anthesis to a pod length of 10–20mm. The free-nuclear stage following double fertilization istypified by loss of starch and increasing vacuolation. The cytoplasmprovides evidence of extensive metabolic activity. Wall ingrowths,already present at the micropylar end of the embryo sac wallprior to fertilization, develop along the lateral wall of thecentral cell as well as at the chalazal endosperm haustorium.Endosperm cellularization begins when the embryo has developeda distinct globular embryo proper and suspensor. Cellularizationstarts at the micropylar end of the embryo sac as a series ofantidinal walls projecting into the endosperm cytoplasm fromthe wall of the central cell. The free, growing ends of thesewalls are associated with vesicles, microtubules, and endoplasrnicreticulum. Pendinal walls that complete the compartmentalizalionof portions of the endosperm cytoplasm are initiated as cellplates formed during continued mitosis of the endosperm nuclei.Endosperm cell walls are traversed by plasmodesmata. This studywill provide a basis for comparison with endosperin from soybeanflowers programmed to abscise. Glycine max, soybean, endosperm, ovules     

花生胚乳细胞化的超微结构观察

花生（ＡｒａｃｈｉｓｈｙｐｏｇｅａｅＬ．）心形胚期的胚乳游离核多瓣裂,或具长尾状结构。胚乳细胞质内有大量线粒体、质体、高尔基体、小泡及少量内质网。中央细胞壁有壁内突。球胚及心形胚期常见胚乳瘤。心形胚晚期,胚乳开始细胞化,胚乳细胞壁形成有３种方式,分别存在于不同的胚珠中：（１）从胚囊壁产生自由生长壁形成初始垂周壁,具有明显的电子密度深的中层,其生长主要靠末端的高尔基体小泡及内质网囊泡的融合。两相邻的自由生长壁末端或其分枝末端相连形成胚乳细胞。（２）核有丝分裂后产生细胞板,细胞板向外扩展并可分枝。间期的非姊妹核间也观察到形成了细胞板。小泡与微管参与细胞板的扩展,高尔基体和内质网是小泡的主要来源。细胞板的扩展末端相互连接,形成胚乳细胞的前身。小泡继续加入细胞板的组成,以后形成胚乳细胞壁。（３）胚乳细胞质中,出现一些比较大的不规则形的片段性泡状结构,它们可能来源于高尔基体小泡,这些片段性泡状结构随机相连形成细胞壁,未见微管参与。胚乳细胞外切向壁及经向壁上有壁内突。     

Endo-[beta]-Mannanase Activity Present in Cell Wall Extracts of Lettuce Endosperm prior to Radicle Emergence

Lettuce (Lactuca sativa L.) endosperm cell walls isolated prior to radicle emergence underwent autohydrolysis, the rate of which was correlated with whether radicle emergence would subsequently occur. Extraction of endosperm cell walls with 6 M LiCl suppressed autohydrolysis, and the desalted extract possessed activity that was capable of hydrolyzing purified locust bean galactomannan but not arabinogalactan, carboxymethylcellulose, glucomannan, polygalacturonic acid, tomato galactomannan, or native lettuce endosperm cell walls. Some hydrolytic activity was detected on endosperm cell walls if they were modified by partial trifluoroacetic acid hydrolysis or pretreatment with guanidinium thiocyanate. In extended incubations the cell wall enzyme extract released only large molecular mass fragments from locust bean galactomannan, indicating primarily endo-activity. Galactomannan-hydrolyzing activity in the cell wall extract increased as a function of imbibition time and was greatest just prior to radicle emergence. Thermoinhibition (imbibition at 32[deg]C) or treatment with abscisic acid at a temperature optimal for germination (25[deg]C) suppressed both germination and endosperm cell wall mannanase activity, whereas alleviation of thermoinhibition with gibberellic acid was accompanied by significant enhancement of mannanase activity. We conclude that a cell wall-bound endo-[beta]-mannanase is expressed in lettuce endosperm prior to radicle emergence and is regulated by the same conditions that govern germination.     

Studies on the Development of the Embryo and Endosperm of Nitraria sibirica Pall

The pollen tube enters the embryo sac through the crassinucellus at the micropylar end, and brings about the porogamy. The embryogeny corresponds to the Solanad type. The defference of the suspensor structure is notable by comparing it with the other genera of Zygophyllaceae that have been studied. The endosperm is of the Nuclear type. Mitosis is the main form of the free endosperm nuclei proliferation. No cell plates develop in the early free nuclear division, however, they appear in late development, without developing into the cell wall and disappear ultimately. At the late globular embryo stage, cell formation in endosperm starts first from the micropylar end. The first anticlinal walls develop from the cell plate that is initiated from tile phragmoplast as a result of normal cytokinesis. Follwing this a wall begins to grow from the base of the cell plates, the outer growing margin soon fuses with the wall of the central cell, and the inner growing margin continues to grow towards the central vacuole. The growing walls branch and eventually fuse on the side nearest the central vacuole. Thus, the first periclinal walls are initiated, and a complete endosperm cell is formed. Along with the development of embryo, cell is gradually formed in the endosperm from the micropylar end towards the chalazal end, but the chalazal endosperm is still coenocytic until the endosperm disintegrate completely. The mature seed has no endosperm.     

Microtubular configurations during the cellularization of coenocytic endosperm in Ranunculus sceleratus L.

Summary Endosperm cellularization in Ranunculus sceleratus was studied in terms of the initiation of cell-wall formation in the coenocytic endosperm. The first endosperm cell walls were in an anticlinal position relative to the cell wall of the embryo sac and originated from the cell plates and not from wall ingrowths from the embryo-sac wall itself. Alveolar endosperm was formed 3 days after pollination. Microtubules were associated with the freely growing wall ends of the anticlinal walls and were observed in various orientations that generally ranged from angles of 45 ° to 90 ° to the plane of the wall. They were absent in the regions where vesicles had already fused. These microtubules may function in maintaining the growth and the direction of growth of the anticlinal wall until cellularization is completed. At the site where three neighbouring alveoli share their freely growing wall ends, remarkable configurations of microtubules were observed: in each alveolus, microtubules ran predominantly parallel to the bisector of the angle formed by the common walls. These microtubules may form a physically stable framework and maintain the direction of growth of the wall edges. It is concluded that the growing edge of the anticlinal endosperm wall and its associated microtubules are a special continuum of the original phragmoplast that gave rise to the anticlinal wall.     

白刺胚乳早期发育的超微结构研究

白刺(Nitraria sibirica)胚乳发育经历游离核阶段、细胞化阶段和被吸收解体阶段。游离核胚乳沿胚囊壁均匀排列为一层,胞质浓厚,其中有丰富的质体、线粒体、高尔基体、内质网和各种小泡等细胞器。珠孔区域的胚囊壁具发达的分枝状壁内突,而周缘区域的胚囊壁具间隔的钉状内突,内突周围的细胞质中具多数线粒体和小泡。胚乳细胞化时,初始垂周壁源于核有丝分裂产生的细胞板。在细胞板两端开始壁的游离生长,一端与胚囊壁相连接,另一端向心自由延伸。壁的游离生长依赖于小泡的融合。早期胚乳细胞具大液泡,具核或无核,细胞质中有大量的线粒体,质体缺乏,其壁仍由多层膜结构组成。     

Ultrastructural study of change from free-nuclear to cellular endosperm in Ipomoea purpurea Roth and Cytinus hypocistis L.

Abstract

The process of cellularization of the endosperm was studied in Ipomoea purpurea Roth and Cytinus hypocistis L. In both the examined species normal cytochinesis, involving the formation of the phragmoplast, characterizes the change from the nuclear to cellular condition. Nevertheless, some ultrastructural aspects of the cell wall initiation seem to indicate that the modality of freely-growing walls cannot be excluded. The hypothesis of a unique method of wall initiation for the endosperm of the nuclear type, formulated by some Authors, is discussed.     

Changes in cell wall polysaccharides of germinating barley grains

Decorticated barley grains were germinated at 25° for 6 days, until the endosperm reserves were nearly exhausted. The neutral monosaccharide components of the hydrolysates of the cell walls and gums from the embryo, aleurone layer and starchy endosperm and the endospermic starch were determined at daily intervals. The amount of embryo cell wall polysaccharide increased 40 times and glucose became the major component, followed in abundance by xylose and arabinose. The cell wall and gum polysaccharides of the aleurone layer (plus testa) and the starchy endosperm declined during germination and their compositions altered. The endospermic starch also decreased. In the early stages of germination the apparent composition of the cell walls of the aleurone layer and starchy endosperm depended upon how they had been prepared. After 6 days the cell walls and gums had provided a significant carbohydrate supply to the living tissues, equivalent to 18.5% of the endospermic polysaccharide degraded during growth, starch having provided the remaining 81.5%.     

Fermentation characteristics of cell-wall sugars from soya bean meal,and from separated endosperm and hulls of soya beans

Two experiments were conducted to investigate the degradation of cell-wall sugars from soya bean meal (in situ), and soya bean endosperm and hulls (in vitro). Soya bean meal, soya bean endosperm, and soya bean hulls were extracted with different chemicals to obtain the cell-wall fraction. Soya bean meal cell walls were incubated in the rumen of a fistulated cow. The individual cell-wall sugars were degraded at different rates: galactose (13.6% h⁻¹), arabinose (7.8% h⁻¹), uronic acids (5.1% h⁻¹), xylose (3.5% h⁻¹) and glucose (3.2% h⁻¹). Microscopic evaluation of the cell walls and degraded material revealed the presence of two cell wall types, with distinctly different degradation characteristics: one originating from the hull (thick, slowly degraded) and one from the endosperm (thin, rapidly degraded). Furthermore, the cell-wall sugar composition of endosperm and hull cell walls was different, most markedly for galactose (281 vs. 12 g kg⁻¹) and glucose (132 vs. 508 g kg⁻¹). The degradation of endosperm and hull cell walls was measured in vitro by use of in vitro cumulative gas production. Degradation rates of the individual cell-wall sugars for hull cell walls were similar (ranging from 2.4% to 4.6% h⁻¹). For endosperm cell walls, the degradation rates of the individual sugars were different but with the same ranking as in the in situ experiment (ranging from 20.9% to 7.0% h⁻¹). It was concluded that for soya bean meal cell walls, the cell-wall sugar degradation pattern is influenced by the presence of two cell-wall types (hull and endosperm cell-wall), which differ in their rate of degradation and sugar composition. The difference in cell-wall sugar degradation pattern between hull and endosperm cell walls is likely to be caused by a combined effect of particle size and cell-wall thickness.     

Degree of pectin methyl esterification in endosperm cell walls is involved in embryo bending in Arabidopsis thaliana

The endosperm is a transitory structure involved in proper embryo elongation. The cell walls of mature seed endosperm are generally composed of a uniform distribution of cellulose, unesterified homogalacturonans, and arabinans. Recent studies suggest that changes in cell wall properties during endosperm development could be related to embryo growth. The degree of methyl esterification of homogalacturonans may be involved in this endosperm tissue remodelling. The relevance of the degree of homogalacturonan methyl esterification during seed development was determined by immunohistochemical analyses using a panel of probes with specificity for homogalaturonans with different degrees of methyl esterification. Low-esterified and un-esterified homogalacturonans were abundant in endosperm cells during embryo bending and were also detected in mature embryos. BIDXII (BDX) could be involved in seed development, because bdx-1 mutants had misshapen embryos. The methyl esterification pattern described for WT seeds was different during bdx-1 seed development; un-esterified homogalacturonans were scarcely present in the cell walls of endosperm in bending embryos and mature seeds. Our results suggested that the degree of methyl esterification of homogalacturonans in the endosperm cell wall may be involved in proper embryo development.     

Seed structure and germination in Cuscuta pedicellata with some notes on C. campestris

Dry seeds of Cuscuta pedicellata have a deeply pitted surface due to invaginated epidermal cell walls. After water uptake these walls bulge outwards and the seed surface becomes papillose. The seed coat consists of an epidermis, two palissade cell layers, and a multiple layer of parenchyma cells. The epidermis contains starch and mucilage, the parenchyma cells are compressed but some contain starch. The endosperm consists of starch–filled cells, but has a peripheral aleuron layer. The endosperm cell walls are gelatinous. The variable structure of the seed coat epidermis is believed to function in wind dispersal and rapid water uptake. Seed dormancy is common in the genus, but does apparently not occur in C. pedicellata.     

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.     

Change in wall composition of transfer and aleurone cells during wheat grain development

In addition to the starchy endosperm, a specialized tissue accumulating storage material, the endosperm of wheat grain, comprises the aleurone layer and the transfer cells next to the crease. The transfer cells, located at the ventral region of the grain, are involved in nutrient transfer from the maternal tissues to the developing endosperm. Immunolabeling techniques, Raman spectroscopy, and synchrotron infrared micro-spectroscopy were used to study the chemistry of the transfer cell walls during wheat grain development. The kinetic depositions of the main cell wall polysaccharides of wheat grain endosperm, arabinoxylan, and (1–3)(1–4)-β-glucan in transfer cell walls were different from kinetics previously observed in the aleurone cell walls. While (1–3)(1–4)-β-glucan appeared first in the aleurone cell walls at 90°D, arabinoxylan predominated in the transfer cell walls from 90 to 445°D. Both aleurone and transfer cell walls were enriched in (1–3)(1–4)-β-glucan at the mature stage of wheat grain development. Arabinoxylan was more substituted in the transfer cell walls than in the aleurone walls. However, arabinoxylan was more feruloylated in the aleurone than in the transfer cell walls, whatever the stage of grain development. In the transfer cells, the ferulic acid was less abundant in the outer periclinal walls while para-coumarate was absent. Possible implications of such differences are discussed.     

西瓜胚乳衰退过程中的超微结构变化及其对胚发育的作用

西瓜胚乳细胞衰退过程中 ,质膜、液泡膜突起、形成体积较大的囊泡 ,内质网断裂形成体积较小的囊泡 ;细胞质和细胞核降解形成电子致密的碎片沿细胞壁分布 ;细胞壁在衰退过程逐渐变薄 ,由于部分区域分解而使整个壁呈波浪型 ,细胞降解后的物质可直接穿越薄壁处或通过宽约 5 0 nm的胞间连丝向近胚端的胚乳细胞转移。胚乳与珠心组织分界壁 -胚囊壁上有发达的壁内突 ,有利于珠心组织内的物质向胚乳内转运 ;胚乳发育早期与胚共有的壁上内外两侧均有胼胝质沉积 ,壁上无外连丝型的胞间连丝存在 ,胚乳发育后期共有壁上的胼胝质消失 ,胚乳细胞降解物可穿越共有壁进入胚细胞内。实验结果表明西瓜胚乳在发育后期对胚的发育具有重要的作用。