The regulation of cambial division and secondary xylem differentiation in xanthium by auxins and gibberellin

Cambial division continued in decapitated Xanthium plants without concomitant xylem fiber differentiation. The application of indoleacetic acid to these plants did not affect the production of cambial derivatives or induce xylem fiber differentiation. When naphthaleneacetic acid was applied either to the second internode or to the stump of a lateral shoot, xylem fiber differentiation was induced in the newly formed cambial derivatives on the xylem side of the cambium in the stem. When naphthaleneacetic acid was applied unilaterally, xylem fiber differentiation was restricted to that side of the stem in the first internode and hypocotyl. Naphthaleneacetic acid also enhanced the production of cambial derivatives. Gibberellic acid enhanced cambial derivative production but did not affect the differentiation of xylem fibers. Similar numbers of cambial derivatives were produced in some naphthaleneacetic acid-treated plants in which xylem fiber differentiation was induced and in gibberellic acid-treated plants which did not differentiate xylem. When naphthaleneacetic acid was applied 72 hours after decapitation, the oldest of the cambial derivatives on the xylem side failed to develop into fibers although younger cells did. These results suggest that auxin has its direct effect on the induction of xylem differentiation rather than the induction of divisions prerequisite to differentiation.     

The Control of Vascular Branching in Coleus 2. The Corner Traces

Corner trace connections are less well defined than those ofthe side bundle in Coleus, the locations of branch points, branchpartners, and number of connections made by a corner trace beingmore variable. The auxin balance between corner traces was alteredby leaf removal and by application of exogenous auxin. Branchingof new strands was shifted toward the pre-existing strand withthe lower auxin flux, but only within a narrow range of developmentalstages and with the imposition of a large auxin imbalance. Branchingoccurred only in nodal regions, as in control plants. Thus,auxin balance can be made to control xylem strand branching,but it does not account fully for the control of vascular branchingin intact plants. In the intact pattern, corner trace branchesappear to be directed toward the pre-existing strand with thehigher auxin flux. It is proposed that, in the vicinity of astrand with high flux, auxin is transported laterally withinthe nodal vascular cambium, facilitating vessel differentiationbetween strands in the derivatives of the vascular cambium.These vessels comprise the connections between traces. Coleus, vascular differentiation, vascular anatomy, vascular branching, vascular patterns, auxin, auxin balance, node     

XYLARY UNION BETWEEN THE NEW SHOOT AND OLD STEM DURING TERMINAL BUD BREAK IN POPULUS DELTOIDES

Bursting terminal buds and subtending stems of Populus deltoides Bartr. ex Marsh. plants were examined at several stages to determine the pattern of xylary union between the 1st- and 2nd-yr growth increments. Metaxylem vessels differentiated first in traces serving the basalmost leaf in the bud. As successively younger leaves began growth, metaxylem vessels differentiated in their traces. The site of metaxylem reactivation was in the second or third internode beneath the bud (i.e., in the 1st-yr stem), and subsequent differentiation progressed bidirectionally in each set of traces as the leaves they served expanded. Traces leading either to abscised leaf positions on the 1st-yr stem or to bud-scale leaf positions were reactivated by the tangential “spread” of activity from adjacent traces serving expanding leaves. The new elements were all secondary xylem vessels, as were those of the basipetal trace components, although they were functionally continuous with the metaxylem vessels that differentiated acropetally. Xylem fibers were initiated in the same position and differentiated in the same sequence as vessels. However, fiber differentiation lagged behind that of vessels. Whereas vessel differentiation was associated with leaf expansion, fiber differentiation was associated with leaf maturation. As each leaf matured in sequence, the primary-secondary transition zone advanced acropetally to the bud base and then in the new shoot until it attained a positional relation with leaf maturation comparable to that of 1st-yr plants.     

Histochemical Study of Xylem Cells in In Vitro Culture of Iris sibirica L.

In this study, lignin content data are presented for annual regenerant Iris sibirica plants, comparable to those in six-year-old intact plants. The structure of the shoots of Iris sibirica grown on artificial nutrient media was studied by the histochemical method. Features of the formation of the xylem in Iris sibirica on artificial nutrient media were revealed. Regenerants very quickly developed a complex system consisting of vascular bundles containing sieve tubes, vessels and tracheids, and hydrocyte systems. Hydrocytes of Iris sibirica were tracheids with lignified thickening, but, in contrast to tracheids and vessels of xylem (they are formed based on procambium or cambium—special lateral primary or secondary meristem), hydrocytes differentiated from the cells of permanent tissues (like phellogen), which probably possessed meristematic activity at the time of differentiation. In Iris sibirica hydrocytes covered the vascular bundle by the thick layer and strung along it up to a certain height. High lignin content in young regenerant Iris sibirica plants was due to the formation of the dense tissue from lignified tracheal elements. The study of the differentiation of xylem elements under controlled conditions can serve as a model for our understanding of wood formation processes.

     

POLARITY OF INDUCTION AND PATTERN OF PRIMARY PHLOEM FIBER DIFFERENTIATION IN COLEUS

The differentiation of primary phloem fibers was studied in Coleus blumei on a quantitative basis. The pattern of fiber differentiation in intact, untreated plants was found to be in the acropetal direction (from a mature internode to a young one). The youngest internodes to differentiate primary phloem fibers were those with cambial activity. In plants grown in the winter, fibers started to differentiate in internodes closer to leaf #2 than in spring-grown plants. A wound changes the pattern of fiber differentiation surrounding it. A wound in which the tissues above and below it were separated with parafilm, prevented fiber differentiation in the tissues directly below the wound, and caused more fiber differentiation in the tissues above and lateral to it. Under wounds with no parafilm separation, few or many fibers differentiated depending on the angle of the wound. The number of fibers under diagonal wounds was five to nine times more than under a horizontal wound. By excision experiments it was found that mature leaves were the source of induction of fiber differentiation. Leaves that produced induction caused fiber differentiation in the internode below them but did not cause fiber differentiation in the internode above. The induction, which can flow through a wound and cause fiber differentiation in at least two internodes below the source, is a polar induction in the basipetal direction (i.e., in the direction from the leaves to the root). Phloem fibers differentiated only in the vascular strands and not from the parenchyma cells between the strands. Therefore, they follow the new regenerative sieve and vessel elements in the pre-existing vascular strands, but do not follow them in their regeneration between the longitudinal strands.     

A Rapid Digestive Technique to Expose Networks of Vascular Elastic Fibers for Sem Observation

The NaOH sonication digestion technique permits rapid isolation and exposure of intact networks of elastic fibers in vascular tissue for 3-dimensional observation with the SEM. The configuration of the network of elastic fibers within the vascular wall of large elastic arteries (aorta) is generally agreed to be a flexible framework through which smooth muscle cells and collagenous fibers are interwoven. However, the configuration of elastic fiber networks in muscular arteries, medium sized veins and smaller vessels remains unknown. When the lengthy standard biochemical elastin purification techniques were applied to vessels containing lesser amounts of elastic tissue and finer elastic fibers, the vessels were completely digested. In contrast, the digestion and sonication technique isolated and exposed intact networks of delicate elastic fibers in blood vessels which do not contain large amounts of elastic tissue. Unfixed vessels were cut into short segments, placed in 0.5 N NaOH and sonicated for 20-40 min. The specimens were rinsed in deionized distilled H₂O, then autoclaved for 30 min. The tissue was rinsed a second time, fixed and processed routinely for SEM. Elastic stains and enzymatic digestion with chromatographically purified elastase and collagenase confirmed that the digestion and sonication technique produced clean, isolated networks of elastic fibers. Knowledge of the configuration of the networks of elastic fibers in different vessels enhances understanding of distensibility characteristics of individual vessels and serves as a baseline for studying alterations in the elastic framework which occur during aging and disease processes such as atherosclerosis, arterial hypertension and aneurysms.     

Leaf vascular dimensions associated with freeze tolerance in bahiagrass (Paspalum notatum)

Foliage damage as a result of individual freeze events is a major limitation to the expansion of bahiagrass ( Paspalum notatum ) pastures and hay production in Southeastern USA. Greater tolerance to such freeze events would allow production deeper into the fall and winter and allow expansion of this species into colder regions. While it has been reported that small cells are more tolerant to freeze damage, this possibility has not been explored in bahiagrass. Specifically, the hypothesis was examined that xylem vessels with smaller diameter in the midrib of leaves are associated with freeze tolerance among bahiagrass genotypes. Vascular bundle diameter was also measured as a possible index of xylem cell size. A total of eight bahiagrass genotypes were eventually studied representing four freeze-sensitive and four freeze-tolerant lines. There was a clear distinction in xylem cell size between the freeze-sensitive and the freeze-tolerant lines. The freeze-tolerant genotypes had xylem element cells that were significantly smaller than the freeze-sensitive genotypes. Averaged across three leaf positions and all genotypes, the xylem element diameter for the freeze-sensitive lines was 222 μm and for the freeze-tolerant lines was only 164 μm. A similar difference was observed in overall vascular bundle diameter with freeze-sensitive lines having a mean of 1168 μm and the freeze-tolerant lines a mean of 917 μm. These results indicated that the diameter of the xylem cells in the vascular midrib of bahiagrass may be an important variable influencing the sensitivity among genotypes to freeze damage.     

Abnormal `wrinkled' cell walls and retarded development of transgenic Arabidopsis thaliana plants expressing endo-1,4-β-glucanase (cell) antisense

Transgenic Arabidopsis thaliana plants expressing cell antisense exhibit reduced levels of cell mRNA and protein compared with wild-type plants. The former display significant alterations in their phenotype. cell antisense plants have shorter stems and roots and are mechanically weaker than their wild-type counterparts. In cell antisense plants, the cell wall structure is markedly disrupted: both fluorescent confocal microscopy and scanning electron microscopy revealed `wrinkled' cell walls, thus indicating that CEL1 plays an important role in cell wall relaxation during cell growth and expansion. In cell antisense plants, the number of xylem elements per bundle is smaller than in the wild-type. In addition, both xylem elements and interfascicular fibers are significantly less lignified in the former. It is suggested that in A. thaliana, abnormal cell wall deposition affected by CEL1 depletion is associated not only with cell growth, but also with the differentiation process in the vascular and supporting tissues.Equal contributors     

A rapid digestive technique to expose networks of vascular elastic fibers for SEM observation

The NaOH sonication digestion technique permits rapid isolation and exposure of intact networks of elastic fibers in vascular tissue for 3-dimensional observation with the SEM. The configuration of the network of elastic fibers within the vascular wall of large elastic arteries (aorta) is generally agreed to be a flexible framework through which smooth muscle cells and collagenous fibers are interwoven. However, the configuration of elastic fiber networks in muscular arteries, medium sized veins and smaller vessels remains unknown. When the lengthy standard biochemical elastin purification techniques were applied to vessels containing lesser amounts of elastic tissue and finer elastic fibers, the vessels were completely digested. In contrast, the digestion and sonication technique isolated and exposed intact networks of delicate elastic fibers in blood vessels which do not contain large amounts of elastic tissue. Unfixed vessels were cut into short segments, placed in 0.5 N NaOH and sonicated for 20-40 min. The specimens were rinsed in deionized distilled H2O, then autoclaved for 30 min. The tissue was rinsed a second time, fixed and processed routinely for SEM. Elastic stains and enzymatic digestion with chromatographically purified elastase and collagenase confirmed that the digestion and sonication technique produced clean, isolated networks of elastic fibers. Knowledge of the configuration of the networks of elastic fibers in different vessels enhances understanding of distensibility characteristics of individual vessels and serves as a baseline for studying alterations in the elastic framework which occur during aging and disease processes such as atherosclerosis, arterial hypertension and aneurysms.     

STEM ANATOMY AND ASPECTS OF DEVELOPMENT IN TOMATO

Aspects of anatomical development were correlated with internodal growth in tomato plants, variety ‘Yellow Plum,’ grown for more than 3 months. Internodal length was measured weekly in control plants and those harvested for anatomical study. Gross structure indicated progressive development with increasing age. Primary xylem and phloem first mature in distinct strands and the strands are joined laterally by procambium to form a continuous vascular cylinder. Primary phloem occurs on the outer periphery of the procambium between the early-formed vascular strands. Successive periclinal divisions in the procambium during internode elongation give rise to pronounced radial seriations of the cells. Procambial derivatives are included in the cylinder of thick-walled, lignified vascular cells that become prominent after elongation ceases. Secondary xylem is of greater radial width in the stem sectors which include protoxylem. During early secondary growth, vessels develop in the secondary xylem only in these sectors. Nucleate fibers and rays constitute the remainder of the secondary xylem. The rays exhibit an organization noted in other plants of reduced growth habit. Some of these interpretations do not agree with those described for tomato in earlier studies, and they are discussed in relation to pertinent aspects of development.     

两种苦丁茶代用品叶结构和后含物与正品的异同

大叶冬青、枸骨叶的结构与苦丁茶基本相似:背腹叶;栅栏组织数层;外韧维管束;小维管束具维管束鞘;气孔仅分布于下表皮,有无规则型和轮列型两种;叶内有纤维、石细胞。叶内都含有单宁、淀粉、脂类及少量草酸钙结晶。叶结构主要区别在于苦丁茶叶主脉维管束成心形的环,另两种呈弧形;枸骨叶缘有横跨叶断面的大纤维束,而另两种叶缘仅少量纤维。     

铁筷子(毛茛科)营养器官的结构及其系统学意义

应用解剖学方法,对铁筷子(Helleborus thibetanusFranch.)(毛茛科)营养器官的结构进行了研究。结果表明,铁筷子根的初生结构观察到三原型、四原型和六原型。营养器官中的维管束在横切面上木质部中的导管分子不呈“V”字形排列;根状茎的次生结构由外向内为表皮、皮层和维管柱,髓射线发达。茎的初生结构中多个维管束排列成环状,维管束鞘分化不明显,节部为单隙三迹,叶迹分别来自于3条维管束或同一条维管束。叶为两面叶,表皮细胞不规则;气孔器只分布于下表皮,为毛茛科典型的无规则型气孔。从铁筷子营养器官的解剖学特点来看,与毛茛科其它植物基本相同,但在营养器官中维管束木质部不呈“V”字形、维管束鞘分化不明显、节部具单叶隙等特征上与其它毛茛科植物不同。     

PETIOLE DEVELOPMENT AND XYLEM DIFFERENTIATION IN XANTHIUM REPRESENTED BY THE PLASTOCHRON INDEX

Petiole development and formation of xylem vessels have been investigated in Xanthium leaves from early ontogeny to maturity. Kinetics of growth was presented in terms of absolute and relative elemental rates of elongation. The process of vascularization was assessed by the number of differentiated xylem vessels. The leaf plastochron index (LPI) developed by Erickson and Michelini (1957) was used for designating the various stages of development. An exponential increase in petiole length was observed between the LPIs –3 and +4 indicating a constant relative rate of 0.20 or 20% increase per day. After cessation of lamina elongation at LPI 8, petiole elongation continued for an additional 5 day period, to LPI 9.5. Relative elemental rate analysis revealed that the basipetal pattern of elongation was maintained throughout the leaf development. At a specific plastochron age, the only growth was due to the petiole elongation. Leaves which ceased elongating had not completed their internal development, since the process of xylem formation continued for several plastochrons, or about 8 days. The highest rate of xylem formation was ten vessels per day at LPI 5. On the average, about five xylem vessels differentiated per day in the middle portion of a Xanthium petiole. Mature petioles contained an average of 218 xylem vessels. About 12 canals of schizogenous origin preceeded the development of the vascular tissue.     

The Control of Vascular Branching in Coleus 1. The Side Bundle

Xylary branching at the proximal end of a differentiating sidebundle was modified by surgical alteration of the surroundingleaf traces and manipulation of their auxin fluxes. Incisionsthrough one corner trace, with the other pre-existing traceintact, resulting in xylem differentiation in the branch ofthe newly formed side bundle toward only the severed trace.Application of IAA to the cut trace allowed xylem branchingof the new strand in both directions. With sufficient auxinimbalance created by increasing the concentration of the appliedIAA, the new xylem strand branched away from the higher auxinsource. Auxin relations were thus able to regulate the courseof differentiation of vascular strands, but their role in regulatingbranching patterns in intact plants may be questioned. Xylembranched exclusively toward an incised trace only when the auxinflux of the incised trace was virtually eliminated. Phloem andprocambium of the differentiating strand were unaffected bythis treatment. Coleus, vascular differentiation, vascular anatomy, vascular branching, vascular patterns, auxin, auxin balance, node     

Regulatory effect of cytokinin on secondary xylem fiber formation in an in vivo system

The regulatory effect of cytokinin on the formation of secondary xylem fibers was studied in the hypocotyl of young Helianthus annuus L. plants. Positive correlation was found between the kinetin supplied (0.25-0.5 micrograms/gram) to the growth medium and the rate of fiber formation within and between the vascular bundles. Reducing the root originated cytokinin supply, either by root removal or by lowering the transpiration rate, diminished the number of newly formed secondary xylem fibers. This decrease was considerably reversed in the presence of 0.5 microgram/gram kinetin. Early pulse exposure of kinetin had a temporary promoting effect on fiber differentiation at low concentrations and a permanent inhibitory effect at high concentration.     

A cell wall protein down-regulated by auxin suppresses cell expansion in Daucus carota (L.)

We investigated the function of the auxin-regulated cell wall gene DC 2.15, a member of a small gene family, present in Daucus carota (L.) and other plants. Cultured cells derived from carrot hypocotyls transformed by the DC 2.15 cDNA in antisense direction were ten-fold longer than wild-type cells, indicating a function of the corresponding protein in suppression of cell expansion. The analysis of carrot plants expressing the DC 2.15 gene in antisense direction showed that the corresponding protein and/or related proteins probably are involved in leaf and vascular bundle development. The antisense plants generally displayed a retarded growth phenotype and delayed greening in comparison to wild-type plants. The asymmetric architecture of the wild-type leaves was degenerated in the DC 2.15 antisense plants and the leaves showed a torsion within and along their major vein. The vascular bundles showed a lowered ratio of the phloem/xylem area in cross sections of the leaf middle vein whereas the bundle sheath and the cambium showed no obvious phenotype. Expression of a promoter-GUS construct was found primarily in vascular bundles of stems, leaves and in the nectar-producing flower discs. The observed pleiotropic antisense phenotype indicates, by loss of function, that one or several related cell wall proteins of this gene family are necessary to realize several complex developmental processes.     

ACAULIS5 controls Arabidopsis xylem specification through the prevention of premature cell death

Cell size and secondary cell wall patterning are crucial for the proper functioning of xylem vessel elements in the vascular tissues of plants. Through detailed anatomical characterization of Arabidopsis thaliana hypocotyls, we observed that mutations in the putative spermine biosynthetic gene ACL5 severely affected xylem specification: the xylem vessel elements of the acl5 mutant were small and mainly of the spiral type, and the normally predominant pitted vessels as well as the xylem fibers were completely missing. The cell-specific expression of ACL5 in the early developing vessel elements, as detected by in situ hybridization and reporter gene analyses, suggested that the observed xylem vessel defects were caused directly by the acl5 mutation. Exogenous spermine prolonged xylem element differentiation and stimulated cell expansion and cell wall elaboration in xylogenic cell cultures of Zinnia elegans, suggesting that ACL5 prevents premature death of the developing vessel elements to allow complete expansion and secondary cell wall patterning. This was further supported by our observations that the vessel elements of acl5 seemed to initiate the cell death program too early and that the xylem defects associated with acl5 could be largely phenocopied by induction of premature, diphtheria toxin-mediated cell death in the ACL5-expressing vessel elements. We therefore provide, for the first time, mechanistic evidence for the function of ACL5 in xylem specification through its action on the duration of xylem element differentiation.     

Chronology of the differentiation of cotton (Gossypium hirsutum L.) fiber cells

Cotton fibers are single elongated cells that develop from epidermal cells of the ovule. The chronology of fiber differentiation was investigated using cultured ovules. Epidermal cells differentiate into fiber cells approx. 3 d before anthesis. When ovules were cultured on a defined medium, fiber growth could be initiated on ovules any time between 2 d preanthesis and the time of anthesis by adding indole-3-acetic acid and gibberellic acid to the medium. In the absence of phytohormones, fibers did not grow, and when ovules between 2 d preanthesis and anthesis were cultured without hormones past the day of anthesis and hormones then added, most ovules failed to produce fibers. The results define the timing of fiber differentiation from epidermal cells, and also define a window of time when differentiated cells are capable of further development. During this window, fiber cells are latent awaiting appropriate stimulation which in the intact plant is apparently associated with anthesis.Abbreviations GA₃ gibberellic acid - IAA indole-3-acetic acid     

Another evidence for inhibitory effect of auxin in adventitious bud formation of decapitated flax (Linum usitatissimum L.) seedlings

Adventitious buds were formed on the hypocotyls of decapitated flax seedlings. Scanning electron and light microscopic examinations of hypocotyls showed that epidermal cells divided to produce meristematic spots from which several leaf primordia were formed. Between leaf primordia and the original vascular tissues of hypocotyls, new xylem cells were formed which connected them. About 10, 30 and 60% of adventitious buds were formed on upper, middle and basal parts of hypocotyls of decapitated seedlings, respectively. Removal of apical meristem together with longer hypocotyl zero to four cm long below the apical meristem) induced higher percentage of adventitious bud formation in the remaining hypocotyl. When the entire hypocotyl was cut into 16 segments (0.25 cm each) and these segments were cultured on MS medium containing 3% sucrose and 0.8% agar, adventitious buds were mainly formed in the lowest five segments. These results suggested that there was a gradient of inhibitory factor(s) from apical to basal part of hypocotyl with respect to adventitious bud formation. Auxin transport inhibitors, morphactin and TIBA induced adventitious bud formation on intact seedlings by suppressing the basipetal movement of auxin.     

The Differentiation of the Leaf Cells of Wheat and the Development of Its Chloroplasts in the Mesophyll Cells

1. By means of cell separation method, we studied the differentiation of the leaf cells of wheat, Nongda 183 and the development of the chloroplasts in the mesophyll. cells. 2. The differentiation of the cells of the first leaf can be divided into 3 stages. Beginning from the leaf primordium to the fully expanded leaf, the cells are in the stage of division and expansion. When the fully expanded leaf becomes deep green in color, the leaf cells are in the prime of life. When the leaf begins to show yellowish colored spots to its complete withering, the cells are in the stage of senescence. Accompanying these stages, the external form and the internal structure of the cells change also. 3. In the early stage of cell division and expansion, one can observe many 0.5μ × 3.4μ mitochondria-like protoplastids which go through various morphological changes to become chloroplasts. 4. The mesophyll cells of the leaf begin to show the signs of senescence sooner than the epidermal cells and the cells of the vascular bundle. The latter last the longest in the life span of the leaf.