Green plastids, maximal PSII photochemical efficiency and starch content of inner stem tissues of three Mediterranean woody species during the year

Wood and pith of 1–2-year-old stems of three woody species with different life strategies common in the Mediterranean basin were studied during the year regarding (i) the occurrence of green plastids, (ii) their maximal photosystem II photochemical efficiency (F_v/F_m) and (iii) their starch content. Green plastids were identified from the red chlorophyll auto-fluorescence under epi-fluorescence microscope, F_v/F_m was estimated using imaging-PAM fluorometry and starch content was recorded under bright field microscope after iodine staining. The evergreen sclerophyll Nerium oleander, the summer deciduous Euphorbia acanthothamnos and the winter deciduous species Platanus orientalis were selected for the study.

Epi-fluorescence microscopy revealed that (i) all species possess abundant green plastids in their wood ray and pith cells throughout the year. In the winter deciduous species chlorophyll fluorescence was found to be strong during the leafless period. By contrast, in the evergreen and the summer deciduous species chlorophyll fluorescence was found uniformly bright during the year; (ii) F_v/F_m value variation during the year seems to be species-specific: in the wood of N. oleander it remains unchanged whereas in the pith it is low during spring–summer; in both tissues of E. acanthothamnos F_v/F_m value reaches maximal value during spring and in P. orientalis during autumn; (iii) in N. oleander and E. acanthothamnos starch is accumulated during spring, whereas in P. orientalis starch content is high during winter.

The scanning electron microscopy (SEM) investigation revealed that the stem epidermis of all three species lacks stomata and formation of lenticels is delayed. Provided that gas exchange is therefore minimized and that PSII photochemical efficiency of inner stem tissues is relatively high, it is further supported that green plastids of wood ray and pith cells may help toward the re-fixation of the internally respired CO₂.     

Distribution of leaf assimilates in the stem of Picea abies L.

Summary Autoradiographic and microautoradiographic studies of 2-year-old Picea abies plants show that in summer leaf assimilates from the second-year shoot are translocated basipetally. Leaf assimilates are first transported to the stem via leaf trace phloem, then to the base of the stem in the sieve cells of the latest increment of secondary phloem. On the way down leaf assimilates move radially from sieve cells into cells of the phloem parenchyma, the vascular cambium, the rays, the inner periderm and certain cells of pith and cortex, including the epithelial cells surrounding the resin ducts. Other cells of pith and cortex remain nearly free of label, despite the long translocation time (20 h). With the exception of the vascular cambial cells, the stem cells that gain leaf assimilates by radial distribution coincide with those that contain chlorophyll and starch.     

Seasonal variation of storage substances in stem cells of Diapensia lapponica. A light microscopic study

Owerwintering strategy of the sub-arctic evergreen Diapensia lapponica has been studied by examining storage substances in the stem cells throughout different seasons. Stem segments were taken from 5 and 20 mm behind the shoot tip, fixed in glutaraldehyde/formaldehyde and embedded in Epon. Semithin sections were stained with Toluidine blue for general observation, Sudan black for lipids, Amido black for proteins and periodic acid-Schiff s reagent (PAS) for carbohydrates. Lipids occurred abundantly throughout the year and seem to be the main storage substances. They were particularly common in mid-winter in the lower part of the shoot. The highest lipid content was found in the pith cells and parenchyma cells of the conductive tissues. Large amyloplasts appeared in mid-summer in higher amounts in the cortex than in the pith of the upper shoot, but vice versa in the lower stem. Starch was almost entirely lacking from November to April. Amido black staining did not indicate presence of protein bodies at any season.
The occurrence of lipids and starch grains in the stem of Diapensia is in general similar to the occurrence of these substances in the leaves throughout the annual cycle.     

Noteworthy idioblastic sclereids in the stems of Eulychnia (Cactaceae)

Stems of Eulychnia (a genus of six to nine species of candelabriform or arborescent cacti) have a parenchymatic cortex with two distinct regions. The outer chlorenchymatic layer is characterized by a conspicuous parallel striping, whereas the inner cortex region devoid of chlorophyll has a coarsely granular aspect. Stem samples from nine accessions, collected in the field or taken from cultivation, were studied from resin-embedded microtome sections and maceration. Two different forms of lignified sclereids were found dispersed in the cortex and the pith. The sclereids of the outer palisade-like cortex layer are distinctly elongated and strictly oriented at right angle to the stem surface, whereas those of the inner cortex and pith are globular or subglobular and conspicuously enlarged compared with the surrounding parenchyma cells. The ontogeny of the sclereids was studied from stem samples of different ages. Formation of the secondary cell walls starts only after cell growth is completed. A screening of numerous South American cacti for the presence of idioblastic sclereids showed that these structures are unique for the genus Eulychnia. Finally, functional aspects of the sclereids are shortly discussed. It is assumed that the sclereids contribute to the mechanical support and reinforcement of the plants.     

Sugar concentrations along and across the Ricinus communis L. hypocotyl measured by single cell sampling analysis

Single cell sap sampling and analysis were used to measure the longitudinal and radial distribution of sucrose, glucose and fructose in the apical cell division zone and in the basal, elongated zone of the Ricinus hypocotyl. Sucrose and hexose increased in concentration from the apex to the base of the seedling axis. In the cell division zone low hexose and sucrose concentrations prevailed in cortex and pith, with a slightly higher hexose concentration in pith cells. The sucrose concentrations in sieve tubes and in phloem were much higher than in the cortex and pith cells. In the basal zone of the hypocotyl high levels of sucrose in phloem, cortex and pith were found, therefore radial, diffusional sucrose flow away from the phloem was considered unlikely. It is proposed that radial flow of growth-water to the hypocotyl periphery together with the down-regulation of a sucrose transporter at the phloem leads to a preferential sucrose flow to the expanding cortex. The pith cells, which do not experience flow of growth-water, are probably insufficiently supplied with sucrose from the phloem resulting eventually in cell death as the plant grows. Shortage of sucrose supply, experimentally achieved by removal of the endosperm, led to sucrose hydrolysis in the pith. The sucrose levels in the other tissues decreased less. It appears that the hydrolysis to hexose was initiated to maintain the osmotic value in the pith cell sap. It is speculated that high hexose levels in the cells are indicative of insufficient sucrose supply via the phloem and that the pith cells are confronted with that situation during early seedling development.     

Determination of the Cellular Mechanisms Regulating Thermo-Induced Stem Growth in Thlaspi arvense L

Field pennycress (Thlaspi arvense L.) is a species with a cold requirement for the initiation of reproductive development (thermoinduction). Work in this laboratory has been focused on elucidating the biochemical and molecular mechanisms underlying the bolting or rapid stem elongation response that is an intricate part of reproductive development in this species. In the present paper the cellular basis for thermo-induced stem growth was determined. Evidence is presented indicating that bolting results from the production of new cells that elongate to their original length before thermoinduction. This increase in cell division occurs in the pith and cortex approximately 0.5 to 5.0 millimeters below the stem apex. For at least the early stages of thermo-induced stem growth, enhanced cell elongation does not appear to be a factor because average lengths of pith cells from stems of thermo-induced plants were similar or less than noninduced controls. In addition, both the amount of increase in the production of new pith cells and stem growth were positively correlated with the length of the cold treatment. Two other lines of evidence are presented corroborating previous assertions (JD Metzger [1985] Plant Physiol 78: 8-13) that gibberellins mediate thermo-induced stem growth in field pennycress. First, treatment of noninduced plants with gibberellin A₃ completely mimicked the effects of a 4-week cold treatment on mitotic activity in the pith and cortex. Second, very little increase in the production of new cells was observed in the pith and cortex of thermo-induced plants of a gibberellin-deficient dwarf mutant of field pennycress. It is also shown that the influence of photoperiod on stem growth is mediated by an effect on the final length that cells ultimately attain.     

掌叶大黄茎和叶多糖的贮藏分布特征

采用组织化学方法和苯酚硫酸比色法研究了掌叶大黄茎和叶中大黄多糖的分布特征和含量变化规律.结果表明:大黄多糖在掌叶大黄茎和叶内的分布各有特点,在茎中大黄多糖分布于靠近维管束外方的皮层薄壁细胞、维管束的韧皮薄壁细胞、靠近皮层的髓射线细胞和少数髓细胞,皮层和髓射线是茎中大黄多糖贮藏较集中的部位,且随着茎的成熟其含量有一定程度的增加;在叶的表皮、叶肉和叶脉中不同程度地分布着较少量的大黄多糖;大黄多糖在叶柄维管束外围的基本组织细胞中不同程度地分布较多,而维管束中分布较少.茎和叶中大黄多糖的含量每年在植物生长的前期逐渐增高,后期略有下降.     

Internal axial light conduction in the stems and roots of herbaceous plants

In order to reveal any roles played by stems and roots of herbaceous plants in responding to the surrounding light environment, the optical properties of the stem and root tissues of 18 herbaceous species were investigated. It was found that light was able to penetrate through to the interior of the stem and was then conducted towards the roots. Light conduction was carried out within the internodes and across the nodes of the stem, and then in the roots from the tap root to lateral roots. Light conduction in both the stem and root occurred in the vascular tissue, usually with fibres and vessels serving as the most efficient axial light conductors. The pith and cortex in many cases were also involved in axial light conduction. Investigation of the spectral properties of the conducted light made it clear that only the spectral region between 710 nm and 940 nm (i.e. far-red and near infra-red light) was the most efficiently conducted in both the stem and the root. It was also found that there were light gradients in the axial direction of the stem or root, and the light intensity generally exhibited a linear attenuation in accord with the distance of conduction. These results revealed that tissues of the stem and root are bathed in an internal light environment enriched in far-red light, which may be involved in phytochrome-mediated metabolic activities. Thus, it appears that light signals from above-ground directly contribute to the regulation of the growth and development of underground roots via an internal light-conducting system from the stem to the roots.     

接骨草的显微结构及其绿原酸组织化学定位研究

运用石蜡切片法和荧光显微镜观察法研究了3个不同接骨草(Sambucus chinensis Lindl.)居群营养器官的显微结构及其绿原酸的分布规律。结果表明:(1)接骨草地上茎厚角组织明显,髓部由大小不等的两类薄壁细胞组成,且有单宁细胞分布;地下根状茎厚角组织细胞小,髓部薄壁细胞大小差异不明显,皮层及髓中有油细胞分布。(2)叶片为异面叶,栅栏组织细胞为短柱状,油细胞不明显。(3)绿原酸分布在根状茎皮层部分细胞、茎厚角组织部分细胞及叶片的海绵组织中,以海绵组织中含量最高。研究认为,髓部薄壁细胞大小的差异可作为接骨草的一个鉴别特征;荧光显微镜观察法可迅速准确显示绿原酸的分布;在所研究的3个接骨草居群中,怀化居群的绿原酸含量最高,若以绿原酸为有效成分来采收接骨草,可以只采收叶。     

A comparative karyological and cytophotometric study of normal and wounded stem tissues ofLathyrus odoratus L.

Wounding of stems ofLathyrus odoratus induced increased mitoses with polyploid in vascular and pith tissues, but not in cortex. Cell division area extended 200 μm from the wound edge. These facts confirm the previous observations inPisum sativum. DNA content of normal stem nuclei was high in vascular and pith tissues, but not in cortex.     

Microtubule reorientation in shoots precedes bending during the gravitropic response of cut snapdragon spikes

The microtubule reorientation during the gravitropic bending of cut snapdragon (Antirrhinum majus L.) spikes was investigated. Using indirect immunofluorescence methods, we examined changes in microtubule orientation in the cortex, endodermis and pith tissues of the shoot bending zone, in response to gravistimulation. Our results show that dense microtubule arrays were visible throughout the cortical, endodermal and pith shoot tissues, and that the transverse orientation of the microtubules (perpendicular to the growth axis) was specifically associated with the shoot growing bending zone. Microtubules showed gravity-induced kinetics of changes in their orientation, which occurred only in the upper stem flank and preceded shoot bending. While this observation, that the gravity-induced microtubule orientation precedes bending, was previously reported only in special above-ground organs such as coleoptiles and hypocotyls, our present study is the first to show that such patterns of change occur in mature flowering shoots. These changes were exhibited first in the upper flank of the cortex and then in the upper flank of the endodermis. No changes in microtubule orientation were observed in the cortex or endodermis tissues of the lower flanks or in the pith, suggesting that these tissues continue to grow during shoot gravistimulation. Our results imply that microtubules may be involved in growth cessation of the upper shoot flank occurring during the gravitropic bending of snapdragon cut spikes.     

木姜子油细胞的发育解剖学研究

利用薄切片法对木姜子茎叶油细胞的发育以及油细胞分布的研究结果表明：油细胞最早发生于第一叶原基以及茎端皮层和髓的基本分生组织中,在未出现油细胞以痛,上述器官的基本分生组织和原分生组织,难以区分油细胞的原始细胞与周围细胞。当油细胞原始细胞呈现出体积较大,液泡化程度较低,细胞核大而明显的特征才明显可辩,以后经过液泡融合,油细胞成熟和油细胞细胞质解体阶段而成为一贮油的囊,油细胞中未出现杯形构造。叶和茎中,     

白花前胡营养器官结构及其分泌道分布规律的研究

运用石蜡切片方法,观察白花前胡营养器官的显微结构及其分泌道的分布特征,以明确营养器官内分泌结构的分布规律,为揭示白花前胡次生代谢产物的积累提供解剖学依据。结果表明,(1)白花前胡成长根从外到内由周皮、中柱鞘薄壁组织和次生维管组织组成,而且中柱鞘薄壁组织不同于一般双子叶植物根的结构;茎从外到内由表皮、皮层和维管柱组成;叶为异面叶结构。(2)白花前胡根、茎及叶中均有分泌道存在,分泌道在根中分布于中柱鞘薄壁组织和次生韧皮部中,茎中分布于皮层和髓中,叶中分布于维管束上下两侧的薄壁组织中。     

Increasing plant diversity of experimental grasslands alters the age and growth of Plantago lanceolata from younger and faster to older and slower

The persistence of plant populations depends on the ability of individuals to cope with the conditions provided by the community. So far, it is not known whether differences in the diversity and composition of plant communities affect the age structure of plant populations or the expression of stem anatomical traits reflecting investment into plant growth and storage. We analyzed annual growth rings in the secondary xylem and measured stem anatomical traits in individuals from 18 populations of Plantago lanceolata growing in a 12‐year old grassland biodiversity experiment (Jena Experiment). Plant individuals of P. lanceolata were on average older and reproduced later with increasing species richness. Individuals of P. lanceolata were slightly younger and the age distribution within populations skewed to younger individuals in the presence of grasses. The presence of legumes did not affect mean age, but led to a more even age distribution within populations. The width of growth‐related tissues (xylem, phloem, phellem) decreased with increasing species richness. Plant diversity‐effects on storage‐related tissues (pith, cortex) were less consistent, as pith showed increasing width with species richness, while cortex did not change with plant diversity. Our results imply that plant diversity effects on population age structure and the expression of stem anatomical traits of P. lanceolata reflect a tradeoff: growth and turnover is fast at low diversity (younger age, higher allocation to growth‐related tissue, faster generative reproduction), while it is slow at high diversity (older age, higher allocation to storage‐related tissue, later generative reproduction).     

Anomalous Growth in the Stem of Ipomoea batatas (L.) Lam. Infested with Megastes Larvae

The stem of Ipomoea batatas (L.) Lam. is characterized by thepossession of a ring of bicollateral, leaftrace bundles. Lacticifersoccur in the pith, in the parenchyma between neighbouring islandsof medullary phloem, and in the cortex. The xylem groups become united by the activity of the inter-fascicularcambium. The production of a certain amount of secondary xylemtakes place before the production of secondary phloem begins.The former is produced more extensively in some areas than inothers, so that the original symmetry of the vascular cylinderis lost. The phellogen originates in the cells of the epidermis. When the stem is attacked by the larvae of Megastes grandalisGuen., which remove most of the internal tissues, anomalousgrowth takes place as a result of the activity of accessorycambia, which develop in the primary cortex, the secondary phloem,and the phelloderm. Residual parenchyma of the pith and/or xylemundergoes hyperplasia to produce a callus tissue which linesthe cavity made by the larvae.     

落葵粘液细胞分布及发育的解剖学研究

采用石蜡切片法对落葵粘液细胞的分布及发育构造进行观察研究.结果表明:(1)除花药、子房及种子外,粘液细胞普遍存在于落葵植株的地上部分内.茎中的粘液细胞多单个散生分布于皮层、髓及髓射线;叶内的粘液细胞主要分布于海绵组织,栅栏组织中则很少见;叶柄中的粘液细胞主要沿叶柄"U"型皮层的两边分布;发育后期作为果实的花萼片中粘液细胞则散生分布很多.(2)根据发育过程的不同形态,可将粘液细胞的发育分为4个阶段:原始细胞阶段、液泡化阶段、成熟阶段和细胞质解体阶段;粘液细胞最早可见于第三叶原基,并且粘液细胞的发育与植株器官的发育不同步.     

Tissue specific protochlorophyll(ide) forms in dark-forced shoots of grapevine (<Emphasis Type="Italic">Vitis vinifera</Emphasis>L.)

Cuttings of grapevine (Vitis vinifera L. cv. Chardonnay) were dark-forced at least three weeks. Pigment contents, 77 K fluorescence emission, excitation spectra of the leaves, petioles, stems, transmission electron micrographs of the etioplasts from leaves, the chlorenchyma tissues of the stems were analysed. The dark-grown leaves, stems contained 8 to 10, 3 to 5 μg/g fresh weight protochlorophyllide, its esters, respectively. HPLC analysis showed that the molar ratio of the unesterified, esterified pigments was 7:3 in the shoot developed in darkness. The dark-forced leaves contained carotenoids identified as: neoxanthin, violaxanthin, antheraxanthin, lutein, β-carotene. Detailed analyses of the fluorescence spectra proved that all tissues of the dark-forced shoots had protochlorophyllide or protochlorophyll forms with emission maxima at 628, 636, 644, 655, 669 nm. The 628, 636 nm emitting forms were present in all parts of the dark-forced shoot, but dominated in the stems, which may indicate an organ specificity of the etioplast development. Variations in the distribution of the pigment forms were even found in the different tissues of the stem. The subepidermal layers were more abundant in the 655 nm form than the parenchyma cells of the inner part of the cortex, the pith. In the latter cells, the plastid differentiation stopped in intermediary stages between proplastids, etioplasts. The plastids in the subepidermal layers had developed prolamellar body structures, which were similar to those of etiolated leaves. The results highlight the importance of organ-, tissue specificity of plastid differentiation for chlorophyll biosynthesis, greening of different plant organs. This revised version was published online in June 2006 with corrections to the Cover Date.     

Auxin Transport in Secondary Tissues1

Auxin transport was investigated in excised stem segments ofNicotiana tabacum L. by the agar block technique using [1-¹⁴C]indol-3yl-acetic acid (IAA). The ability of the stems to transportauxin basipetally increased as secondary development proceeded;by contrast the ability of the pith to transport auxin declinedwith age. By separation of the stem tissues it was shown thatthe great majority of auxin transport took place in cells associatedwith the internal phloem and in cells close to the cambium;in both cases similar velocities of transport were found (c.5.0 mm h^–1 at 22°C). The effects of osmotic gradientson auxin transport through the internal phloem were investigated.IAA was found by chromatography to account for practically allthe radioactivity in receiver blocks and other extracts of stemsegments. The significance of these results is discussed.     

Effects of shoot inversion on stem structure in Pharbitis nil

The effects of shoot inversion on stem structure over 72 hr were investigated in Pharbitis nil by analyzing cell number, cell length, and the cross sectional areas of cells, tissues, and regions. An increase in stem diameter can be attributed to an increase in both cell number and cross sectional area of pith (primarily) and vascular tissue (secondarily). Qualitative observations of cell wall thickness in the light microscope did not reveal any significant effects of shoot inversion on this parameter. The inhibition of shoot elongation was accompanied by a significant decrease in cell length in the pith. The results are generally consistent with an ethylene effect on cell dimensions, especially in the pith.     

Appearance of a State 1 – State 2 transition during chloroplast development in the wheat leaf: Energetic and structural considerations

The ability of developing chloroplasts to dynamically regulate the distribution of excitation energy between photosystem 1 and photosystem 2, and thus perform a State 1 – State 2 transition, was examined from analyses of chlorophyll fluorescence kinetics in 4- and 8-day-old Triticum aestivum L. cv. Maris Dove leaves grown under a diurnal light regime. Chloroplasts at all stages of development in the two leaf systems could undergo a State 1 – State 2 transition, except those found in the basal 0.5 cm of the 4-day-old leaf. The ability to physiologically modify the excitation energy distribution between the chlorophyll matrices of the two photosystems developed after the development of mature, fully photochemically competent photosystem 2 units and the appearance of excitation energy transfer between photosystem 2 and photosystem 1. Also, changes in the degree of energetic interaction between the two photosystems, in vivo rates of electron transport and the chlorophyll a/b ratio could not be correlated with the appearance of a State 1 – State 2 transition. Ultrastructural studies demonstrated a 32% increase in the degree of thylakoid appression in chloroplasts at the base of the 8-day-old leaf compared to the situation in the basal 0.5 cm of the 4-day-old leaf. This difference in thylakoid stacking can account for the differing abilities of these two tissues to perform a State 1 – State 2 transition when considered in the context of the distribution of the two photosystems within appressed and non-appressed regions of thylakoid membranes.