木贼类营养器官凯氏带类型的新观察

该研究用石蜡切片法比较观察了5种木贼科植物营养器官的内皮层及凯氏带,首次报道了2层内皮层及其凯氏带的形态特征及分布规律,并讨论各种类型的凯氏带及其与厚壁组织的协作防御机制。结果表明:(1)5种木贼的地下茎和根都只有1条凯氏带,其中4种木贼的地上茎有2条凯氏带。(2)木贼类营养器官具有3种凯氏带类型,即2层公共内皮层上各具有1条凯氏带、1层散生内皮层上的1条凯氏带、1层公共内皮层上的1条凯氏带。(3)木贼类地下茎和根都有发达的厚壁组织或致密的表皮。(4)问荆地上茎外侧内皮层具有复合内皮细胞。研究认为,木贼类植物凯氏带数量不能作为分类的依据;地下茎和根虽然只有1条凯氏带,但地下部分都有发达的厚壁组织或(和)与其紧密相连的表皮,推测厚壁组织或(和)表皮可能具有与凯氏带相同的功能;3种类型凯氏带的防御能力由强到弱依次是:2层公共内皮层上的凯氏带 1层散生内皮层上的凯氏带 1层公共内皮层上的凯氏带。     

植物内皮层的分化及其屏障功能研究进展

植物根系最主要的作用之一是从土壤中获取养分并将其运输至地上部。水和营养物质径向穿过根的表皮、皮层、内皮层等所有外部细胞层,才能到达中柱,以供地上部代谢所需。其中,内皮层细胞在发育过程中会经历两个特殊的分化阶段,分别形成凯氏带和木栓层两种扩散屏障,二者在控制养分获取与流失方面起着重要的作用。该文就近年来国内外有关植物内皮层分化过程及其屏障功能方面的研究进展进行了综述,以期对深入探索内皮层屏障在植物生长发育和逆境适应中的作用提供参考,为植物育种工作开辟新的思路。     

大豆初生根凯氏带对铜离子的通透性

采用在根内生成有色铜沉淀的方法研究大豆（Glycine max）初生根凯氏带对铜离子的通透性。用真空泵抽取浓度为200μmol·L^–1的CuSO4溶液进入根中,然后在重力作用下从根基部灌注400μmol·L^–1的K4[Fe（CN）6]溶液,两种物质在根内相遇即可产生棕色的Cu2[Fe（CN）6]沉淀,根据沉淀的位置来确定铜离子所经过的途径。结果表明：Cu^2＋可以穿过内皮层凯氏带,在木质部导管壁以及凯氏带至木质部之间的细胞壁处产生棕色沉淀,侧根发生的部位也产生了大量的沉淀;当抽取K4[Fe（CN）6]溶液后再灌注CuSO4溶液,发现Cu^2＋仍然可以穿过凯氏带,并在凯氏带外侧以及外皮层细胞的细胞壁处产生棕色沉淀。研究结果证明凯氏带并不是一个可以完全阻止离子进出的完美屏障。     

华山松根与针叶凯氏带的比较研究

应用冰冻切片、酶解分离、荧光显微技术和傅里叶红外光谱分析(FTIR)等手段,对华山松初生根和针叶内皮层凯氏带进行了分离、显微结构特征和化学成分的比较。研究结果表明：针叶凯氏带的“网格”结构比较整齐,大小较一致,排列也较规则,同时在“网格”的纵向壁上具有明显的初生纹孔场。而初生根凯氏带网状结构的大小、排列均不规则,在其“网格”的纵向壁上的初生纹孔场不明显。根据FTIR的检测结果显示：初生根凯氏带中木栓质和木质素的含量均高于针叶,而纤维素的含量则明显低于针叶;两者细胞壁蛋白的含量基本相同。本文的研究结果为深入探讨植物地下部分和地上部分凯氏带的生理功能提供新的佐证。     

大豆初生根凯氏带对铜离子的通透性

采用在根内生成有色铜沉淀的方法研究大豆(Glycine max)初生根凯氏带对铜离子的通透性。用真空泵抽取浓度为200 μmol·L^–1 的CuSO₄溶液进入根中, 然后在重力作用下从根基部灌注400 μmol·L^–1的K₄[Fe(CN)₆]溶液, 两种物质在根内相遇即可产生棕色的Cu₂[Fe(CN)₆]沉淀, 根据沉淀的位置来确定铜离子所经过的途径。结果表明: Cu²⁺可以穿过内皮层凯氏带, 在木质部导管壁以及凯氏带至木质部之间的细胞壁处产生棕色沉淀, 侧根发生的部位也产生了大量的沉淀; 当抽取K₄[Fe(CN)₆]溶液后再灌注CuSO₄溶液, 发现Cu²⁺仍然可以穿过凯氏带, 并在凯氏带外侧以及外皮层细胞的细胞壁处产生棕色沉淀。研究结果证明凯氏带并不是一个可以完全阻止离子进出的完美屏障。     

植物凯氏带的研究进展

凯氏带是植物内皮层细胞径向壁和横向壁的带状增厚部分 ,近年来对其细微结构、化学成分、生理功能以及在植物体的存在部位等方面的研究工作已成为国内、外的研究热点。本文就凯氏带上述诸方面的研究进展 ,作一详尽的论述 ,同时还着重对凯氏带研究的新方法和新技术作了评述。最后对凯氏带的进一步研究工作提供新的启示     

植物凯民带的研究进展

凯氏带是植物内皮层细胞径向壁和横向壁的带状增厚部分,近年来对其细微结构、化学成分、生理功能以及在植物体的存在部位等方面的研究工作已成为国内、外的研究热点。本文就凯氏带上述诸方面的研究进展,作一详尽的论述,同时还着重对凯氏带研究的新方法和新技术作了评述。最后对凯氏带的进一步研究工作提供新的启示。     

白皮松针叶内皮层在盐胁迫中的屏障作用研究

应用荧光显微技术、傅里叶变换显微红外光谱分析(FTIR)、扫描电镜及X-射线能谱微区分析等手段,对白皮松(Pinus bungeana)子叶、初生叶及2a生针叶内皮层细胞径向壁的显微结构特征、化学成分,以及在叶子横切面上Na和Cl的微区分布进行分析。通过荧光显微观察发现,白皮松子叶内皮层不具凯氏带,而初生叶及2a生针叶均存在凯氏带加厚现象。根据FTIR的检测结果显示:子叶内皮层细胞径向壁不含木栓质或极少,2a生针叶内皮层细胞径向壁木栓质含量高于初生叶。对相应区域的X射线微区分析表明,子叶内皮层对Na和Cl在质外体运输中不起障碍作用,而初生叶与2a生针叶内皮层阻碍Na和Cl以质外体途径进入维管组织。研究结果表明:具凯氏带加厚的内皮层细胞壁中木栓质含量决定其在质外体运输过程中的生理功能。     

单子叶植物根的内皮层及其观察方法

裸子植物和具有少量次生长的双子叶植物根的内皮层细胞通常都具有径向壁和横壁加厚而成的凯氏带结构。但多数单子叶植物和少数双子叶植物(没有次生生长的种类)根的内皮层细胞只有在发育的早期(幼根)具有凯氏带,发育的后期(较老的根)内皮层细胞壁内敷贴了一整层栓质层,随后细胞壁又经过了五面加厚(只有邻接皮层一面的壁没有加厚)或六面加厚(即全部细胞壁都加厚),加厚部分都经过了木质化,因此这时就看不到凯氏带了。由于内皮层细胞内敷贴了一整层栓质层,这类植物根的维管柱似乎被一层不透水的套子所隔开了。实际不然,因为并不是全部内皮层细胞都具有五面或六面加厚的壁,通常有一部分内皮层细胞,就是在根的横切面上靠近木质部角端的那一部分内皮层细胞,仍有未栓质化、未加厚、只有凯氏带的纤维素薄壁,这一部分内皮层细胞     

白皮松针叶内皮层中具凯氏带的证据

利用荧光显微镜对白皮松（Pinus bungeana Zuce.)针叶内皮层结构进行深入观察,确认内皮层径向壁和横向壁上具有能够激发荧光的带状结构;用纤维素酶和果胶酶对内皮层充分酶解分离后,首次从叶中得到类似于根部内皮层特有的网状结构。应用傅立叶变换红外光谱分光术（FTIR）对网状结构进行化学成分鉴定,其吸收光谱的分析结果表明,针叶内皮层细胞壁带状结构部分含有木质素,木栓质,纤维素和细胞壁蛋白等,而这些成分与针叶维管组织的成分不尽相同,上述证据表明,白皮松针叶内皮层细胞壁具有与根部内皮层相似的凯氏带结构。     

Regulation of a plant aquaporin by a Casparian strip membrane domain protein‐like

The absorption of soil water by roots allows plants to maintain their water status. At the endodermis, water transport can be affected by initial formation of a Casparian strip and further deposition of suberin lamellas and regulated by the function of aquaporins. Four Casparian strip membrane domain protein‐like (CASPL; CASPL1B1, CASPL1B2, CASPL1D1, and CASPL1D2) were previously shown to interact with PIP2;1. The present work shows that CASPL1B1, CASPL1B2, and CASPL1D2 are exclusively expressed in suberized endodermal cells, suggesting a cell‐specific role in suberization and/or water transport regulation. When compared with wild‐type plants, and by contrast to caspl1b1*caspl1b2 double loss of function, caspl1d1*caspl1d2 double mutants showed, in some control or NaCl stress experiments and not upon abscisic acid (ABA) treatment, a weak enlargement of the continuous suberization zone. None of the mutants showed root hydraulic conductivity (Lp_r) phenotype, whether in control, NaCl, or ABA treatment conditions. The data suggest a slight negative role for CASPL1D1 and CASPL1D2 in suberization under control or salt stress conditions, with no major impact on whole root transport functions. At the molecular level, CASPL1B1 was able to physically interact with PIP2;1 and potentially could influence the regulation of aquaporins by acting on their phosphorylated form.     

Development of the Casparian strip in primary roots of maize under salt stress

The Casparian strip in the endodermis of vascular plant roots appears to play an important role in preventing the influx of salts into the stele through the apoplast under salt stress. The effects of salinity on the development and morphology of the Casparian strip in primary roots of maize (Zea mays L.) were studied. Compared to the controls, the strip matured closer to the root tip with increase in the ambient concentration of NaCl. During growth in 200 mM NaCl, the number and the length of the endodermal cells in the region between the root tip and the lowest position of the endodermal strip decreased, as did the apparent rate of production of cells in single files of endodermal cells (the rate of cell formation being equal to the rate at which cells are lost from the meristem). The estimated time required for an individual cell to complete the formation of the strip after generation of the cell in the presence of 200 mM NaCl was not very different from that required in controls. Thus, salinity did not substantially affect the actual process of formation of the strip in individual cells. The radial width of the Casparian strip, a morphological parameter that should be related to the effectiveness of the strip as a barrier, increased in the presence of 200 mM NaCl. The mean width of the lignified region was 0.92 m in distilled water and 1.33 m in 200 mM NaCl at the lowest position of the strip. The mean width of the strip relative to that of the radial wall at this position was significantly greater after growth in the presence of 200 mM NaCl than in the controls, namely, 20.5% in distilled water and 33.9% in 200 mM NaCl. These observations suggest that the function of the strip is enhanced under salt stress.     

The Casparian strip in pea epicotyls: Effects of light on its development

The Casparian strip, which is specific to roots, was studied in the epicotyls of dark-grown seedlings of pea (Pisum sativum L.) where it was found to have the same morphology and properties as the strip in roots. In dark-grown seedlings, the distance between the upper-most position of the Casparian strip and the bending point of the hook (about 37 mm) did not change during growth of the seedlings. In the uppermost 0.5-mm region of the region in which the Casparian strip could be detected by fluorescence microscopy, the plasma membrane was not firmly attached to the cell wall. The development of the Casparian strip continued for about 42 h after dark-grown seedlings were transferred to the light, indicating that (i) the cells that have been determined to form the Casparian strip in darkness form the strip in the light, and that (ii) it takes about 42 h for the cells to complete formation of the strip. Cells in the hook of dark-grown seedlings did not form a Casparian strip when such seedlings were transferred to the light. The Casparian strip was formed in rapidly elongating internodes of light-grown seedlings when the seedlings were transferred to darkness. Light did not control the formation of the Casparian strip in roots.Abbreviation PBS phosphate-buffered saline     

Effects of brefeldin A on the development of the Casparian strip in pea epicotyls

Summary The Casparian strip, a structure that is present in roots, is also present in epicotyls of dark-grown pea seedlings. In a dark-grown epicotyl, the cells in each stage of the development of the Casparian strip have been suggested to be lined up basipetally in the region 3 to 37 mm below the bending point of the hook, in order of the developmental stage. Brefeldin A (BFA), a specific inhibitor of secretory transport, was administrated at 200 M. to dark-grown pea epicotyls for 2 h via a thread passed through the epicotyl 40 mm below the bending point. The basipetal sequence of development of the modification of the cell wall at the Casparian strip, as judged by fluorescence microscopy, stopped 5 h after the start of 2 h treatment with BFA and resumed after 30 h. This basipetal sequence of development did not stop in control seedlings. Electron micrographs of endodermal cells in epicotyls treated with BFA showed striking morphological changes in the Golgi stacks and the ER. Histological examination made 20 h after the start of the experiment revealed that the basipetal sequence of development of the cell wall modification stopped at a point which was present at 25.2 ± 1.6 mm (mean with SD, n=5) from the bending point of the hook at the start while the basipetal sequence of development of the tight adhesion of the plasma membrane to the cell wall at the Casparian strip stopped 0.9 ± 0.5 mm (mean with SD, n=5) below this point. These results indicate the involvement of secretory transport not only in the introduction of the modification of the cell wall but also in the completion of the tight adhesion of the plasma membrane.Abbreviations BFA brefeldin A - PBS phosphate-buffered saline - ER endoplasmic reticulum     

Comparative investigation of primary and tertiary endodermal cell walls isolated from the roots of five monocotyledoneous species: chemical composition in relation to fine structure

The chemical composition of isolated endodermal cell walls from the roots of the five monocotyledoneous species Monstera deliciosa Liebm., Iris germanica L., Allium cepa L., Aspidistra elatior Bl. and Agapanthus africanus (L.) Hoffmgg. was determined. Endodermal cell walls isolated from aerial roots of M. deliciosa were in their primary developmental state (Casparian bands). They contained large amounts of lignin (6.5% w/w) and only traces of suberin (0.5% w/w). Endodermal cell walls isolated from the other four species were in their tertiary developmental state. Lignin was still the more abundant cell wall polymer with amounts ranging from 3.8% (w/w, A. cepa) to 4.5% (w/w, I. germanica). However, compared to endodermal cell walls in their primary state of development (Casparian bands), tertiary endodermal cell walls contained significantly higher amounts of suberin, ranging from 1.8% (w/w, I. germanica) to 3.0% (w/w, A. africanus). Thus, chemical characterization of endodermal cell walls from five different species revealed that lignin was the dominant cell wall polymer in the Casparian band of M. deliciosa, whereas tertiary endodermal cell walls contained, in addition to lignin, increasing amounts of suberin (I. germanica, A. cepa, A. elatior and A. africanus). Besides the two biopolymers lignin and suberin, cell wall carbohydrates in the range of between 40 and 60% were also quantified. The sum of all cell wall compounds investigated by gas chromatography resulted in a recovery of 50–80% of the dry weight of the isolated cell wall material. Quantitative chromatographic results in combination with microscopic studies are consistent with the existence of a distinct suberin lamella and lignified tertiary wall deposits. From these data it can be concluded that the barrier properties of the endodermis towards the apoplastic transport of ions and water will increase from primary to tertiary endodermal cell walls due to their increasing amounts of suberin. Received: 23 August 1997 / Accepted: 28 January 1998     

The Casparian Strip of Clivia miniata Reg. Roots: Isolation,Fine Structure and Chemical Nature*

The root endodermis of Clivia miniata Reg. was successfully isolated using the cell wall degrading enzymes cellulase and pectinase. The enzymes did not depolymerize those regions of the primary cell walls of anticlinal endodermal root cells where the Casparian strips were located. Since the endodermis of C. miniata roots remained in its primary developmental state over the whole root length, endodermal isolates essentially represented Casparian strips. Thus, sufficient amounts of isolated Casparian strips could be obtained to allow further detailed investigations of the isolates by microscopic, histochemical and analytical methods. Scanning electron microscopy revealed the reticular structure of the Casparian strips completely surrounding the central cylinder of the roots. Whereas in younger parts of the root only the anticlinal cell walls of the endodermis remained intact in the isolates, in older parts of the root the periclinal walls also restricted enzymatic degradation due to the deposition of lignin. Extracts of the isolates with organic solvents did not reveal any wax-like substances which might have been deposited within the cell wall forming a transport barrier, as is the case with cutin and suberin. However, several histochemical and analytical methods (elemental analysis and FTIR spectroscopy) showed that the chemical nature of the Casparian strips of C. miniata roots can definitely be a lignified cell wall. These findings are in complete agreement with studies carried out at the beginning of this century on the chemical nature of the Casparian strips of several other plant species. The implications of these results concerning apoplasmatic transport of solutes and water across Casparian strips are discussed.     

Structure and chemical composition of endodermal and rhizodermal/hypodermal walls of several species

CWM, isolated cell wall material
ECW, isolated endodermal cell walls
G, guaiacyl monomer
H, p-hydroxyphenyl monomer
HCW, isolated hypodermal cell walls
RHCW, isolated rhizodermal and hypodermal cell walls
S, syringyl monomer
XV, isolated xylem vessels

Endodermal cell walls of the three dicotyledoneous species Pisum sativum L., Cicer arietinum L. and Ricinus communis L. were isolated enzymatically and analysed for the occurrence of the biopolymers lignin and suberin. From P. sativum, endodermal cell walls in their primary state of development (Casparian strips) were isolated. Related to the dry weight, these isolates contained equal amounts of suberin (2·5%) and lignin (2·7%). In contrast, the endodermal cell walls of C. arietinum and R. communis, which were nearly exclusively in their secondary state of development, contained significantly higher proportions of suberin (10–20%) and only traces of lignin (1–2%). The results of the chemical analyses were supported by a microscopic investigation of Sudan III-stained root cross-sections, showing a Casparian strip restricted to the radial walls of the endodermis of P. sativum and well-pronounced red suberin lamellae in C. arietinum and R. communis roots. Compared with recently investigated monocotyledoneous species, higher amounts of suberin by one order of magnitude were detected with the secondary state of development of dicotyledoneous species. Furthermore, the carbohydrate and protein contents of primary (Clivia miniata Reg. and Monstera deliciosa Liebm.), secondary (C. arietinum and R. communis) and tertiary endodermal cell walls (Allium cepa L. and Iris germanica L.) were determined. The relative carbohydrate content of secondary endodermal cell walls was low (14–20%) compared with the content of primary (42–50%) and tertiary endodermal cell walls (60%), whereas the protein content of isolated endodermal cell walls was high in primary (13%) and secondary (8%) and low in tertiary endodermal cell walls (0·9–2%). The results presented here indicate that the quantitative chemical composition of primary, secondary, and tertiary endodermal cell walls varies significantly. Finally, cell wall proteins are described as an additional important constituent of endodermal cell walls, with the highest concentrations occurring in primary (Casparian strips) and secondary endodermal cell walls.     

Suberin lamella development in maize seedling roots grown in aerated and stagnant conditions

Hypoxia can stimulate the development of a suberized exodermis in aquatic plants; however, its influence on this aspect of terrestrial root development is sparsely documented. To determine the effects of hypoxia on maize (Zea mays cv. Seneca Horizon) roots, seedlings were grown in vermiculite (VERM), aerated hydroponics (AER), stagnant hydroponics with agar (STAG), or aerated hydroponics with agar (AERAG). The endo- and exodermis were examined for wall modifications. Lateral root emergence and aerenchyma formation were documented qualitatively. The endodermal Casparian band formation was unaffected by treatment. Endodermal and exodermal suberin lamella formation was earliest and most extensive in VERM. Suberization, especially in the exodermis of aerated treatments, was depressed in all hydroponic media. In comparison with AER, STAG exodermal lamellae were increased, but endodermal lamellae were decreased. Since the suberized exodermis forms a barrier to radial oxygen loss from roots to the medium, its stimulation in STAG roots (which also developed extensive aerenchyma) would help retain oxygen in the root. The reduction of endodermal lamellae should facilitate oxygen diffusion into the stele. Clearly, the response to environmental conditions is variable within individual cortical cell layers. Additionally, the observed patterns of lamellae, aerenchyma and lateral root development indicate a tight radial co-ordination of root development.