Pea Root Regeneration After Tip Excisions at Different Levels: Polarity of New Growth

The roots of light-grown pea seedlings (Pisum sativum L. cv.Alaska) were excised at 250, 500 or 1500 µm from the body/capjuncture. Tips were sampled for 7 d after excision to monitorthe polarity and structure of the regeneration response. Inroots excised at 250 µm a new single apex regeneratedwithout swelling or any sign of repair. The new root cap appearedidentical to the control. After the 500 µm excision tworesponses occurred. In 45% of the roots examined, a single newapex regenerated, in 55% two or three new apices regeneratedto form a dichotomized or trichotomized root in the same longitudinalaxis as the primary root. In roots excised at 1500 µm,one, two or three lateral roots formed adjacent to the xylemat right angles to the primary root axis. In most instancesthe new roots developed triarch xylem. The discussion concentrateson the relationship of the differentiation state of the vascularcylinder and the pericycle, at the excision site, to the regenerationresponse. Root regeneration, Pisum sativum, Pericycle, Root apex     

Polarity of IAA Effect on Sieve-Tube and Xylem Regeneration in Coleus and Tomato Stems

A technique is described for the processing of regenerated xylem and sieve tubes from the same wound area for microscopic and quantitative comparison.

Regeneration was examined in internodes of 2 developmental stages in Coleus: internode 2, elongating, characteristic of primary growth; and internode 5, non-elongating, characteristic of secondary growth.

Transport of indoleacetic acid (IAA) in excised number 5 internodes of Coleus is strictly polar, in a basipetal direction, judging by a regeneration bioassay involving both sieve tube strands and xylem cells. Similar results were obtained with tomato.

If isolated number 5 Coleus internodes are not treated with hormone, they regenerate no xylem cells and a small number of sieve tube strands. With increasing concentrations of IAA added apically, the number of regenerated sieve tube strands (and, with higher concentrations, of xylem cells) increases progressively up to 1% IAA, the highest concentration applied.

Internode 2 of Coleus regenerates fewer xylem cells or sieve tube strands than internode 5, whether on the otherwise intact plant or with a given concentration of IAA added apically. The amount of regenerated xylem increases with added apical IAA, except that the highest concentration gives no further increase. The number of xylem cells regenerated in intact plants occurs at the same interpolated IAA concentration as in number 5 internodes. No concentration of IAA tried provided replacement of intact number of sieve tube strands in internode 2.

IAA can exert a regenerative stimulus on both xylem and sieve tubes in the area immediately adjacent to the site of its application.

     

辽东栎幼苗生长对种子大小和子叶去除处理的响应

该文在温室内盆播条件下,设置不去除子叶(对照)、去除1/4子叶(轻度子叶去除)、1/2子叶(中度子叶去除)和3/4子叶(重度子叶去除) 4个处理,研究了种子大小和子叶去除处理对辽东栎幼苗生长及其形态学可塑性的影响。结果表明:(1)大种子幼苗株高、基茎、叶片数、单株叶面积、总干质量和根冠比在所有处理均大于小种子幼苗,除株高外,上述其他参数在轻度子叶去除处理的不同大小种子幼苗间均差异显著(P0.05),前者总干质量在中度子叶去除处理与后者差异也显著(P0.05)。大种子幼苗比叶面积和比根长在各处理均大于小种子幼苗,比枝长则相反。(2)随着子叶去除强度增大,大、小种子幼苗株高、基茎、叶片数、单株叶面积和总干质量等均呈减小趋势,株高、基径、叶片数和单株叶面积在重度子叶去除处理均显著小于对照和轻度子叶去除处理;小种子幼苗对子叶损失反应更为敏感,其株高、叶片数、单株叶面积和总干质量在中度子叶去除处理也显著小于对照(P0.05);小种子幼苗根冠比随子叶去除强度增大而增大,在中度和重度子叶去除处理显著大于对照和轻度子叶去除处理(P0.05)。(3)随着子叶去除强度增大,大小种子幼苗的比叶面积和比根长均逐渐增大,但大种子幼苗比枝长持续减小,轻度子叶去除处理的小种子幼苗比枝长最大。     

EFFECTS OF LEAF EXCISION ON FLOWERING OF XANTHIUM APICAL BUDS IN CULTURE UNDER INDUCTIVE AND NONINDUCTIVE PHOTOPERIODS

Apical buds of Xanthium were grown in aseptic culture under short-day cycles known to induce flowering in the intact plants or under “light-break” conditions known to prevent flowering. The total light provided in each 24-hr cycle was the same under the two photoperiods. Various numbers of leaves were excised from the apical buds. Excision of leaves did not change the response to photoperiod: even with all leaves excised the apical buds cultured under short-day conditions reached the same average floral stage as the control buds, and those under light-break conditions all remained vegetative. Fresh weight was not significantly changed by the excisions, either. However, excision of the young leaves resulted in an increase in the number of new leaves developed by the apical bud during the two-week culture period.     

BOSTONIA PERPLEXA GEN. ET SP. NOV., A CALAMOPITYAN AXIS FROM THE NEW ALBANY SHALE OF KENTUCKY

Bostonia perplexa, gen. et sp. nov., was collected from the Lower Mississippian Falling Run member of the Sanderson Formation. The single short segment of an axis, preserved as a petrifaction, contains at least three vascular columns, each with both primary and secondary tissues. Primary xylem is two or three ribbed, and contains several mesarch protoxylem strands. Gymnospermous secondary xylem is characterized by both uniseriate and multiseriate rays. The ground tissue is parenchymatous except for a few clusters of sclerotic cells. In its apparent polystelic nature, the specimen superficially resembles members of the Pennsylvanian to Permian Medullosaceae. All evidence currently available, however, leads to the conclusion that this species should be placed in the Upper Devonian to Lower Mississippian Calamopityaceae. It has not been determined with certainty whether the species is polystelic (in the sense of the Medullosaceae), or whether the apparent polystely is the result of stelar branching proximal to the level of branch divergence.     

Perfusion of onion root xylem vessels: a method and some evidence of control of the pH of the xylem sap

We describe a method for perfusing the xylem in the stele of excised onion roots with solutions of known composition under a pressure gradient. Tracer studies using [¹⁴C] polyethylene glycol 4000 and the fluorescent dye, Tinopal CBSX, indicated that perfusing solutions passed exclusively through the xylem vessels. The conductance of the xylem was small over the apical 100 mm of the root axis but increased markedly between 100 and 200 mm. Unbuffered perfusion solutions supplied in the range pH 3.7–7.8 emerged after passage through the xylem adjusted to pH 5.2–6.0, indicating the presence of mechanisms for absorbing or releasing protons. This adjustment continued over many hours with net proton fluxes apparently determined by the disparity between the pH of the perfusion solution and the usual xylem sap pH of about 5.5. Mild acidification of the xylem sap by buffered perfusion solutions increased the release of ⁸⁶Rb (K⁺) and ³⁵SO₄ ^2- from the stelar tissue into the xylem stream. The ion-transporting properties of onion roots seemed little changed by excision from the bulbs, or by removal of the apical zones of the root axis. The pH of sap produced by root pressure resembles that found in the outflow solutions of perfused root segments.     

Unusual stomatal behaviour on partial root excision in wheat seedlings

The excision of four out of five primary roots of wheat (Triticum durum Desf.) seedlings often leads to an enhanced rate of transpiration. Surprisingly this enhancement could be maintained for several hours after root excision and was particularly likely to occur at low irradiances or high atmospheric humidity. This long‐term enhancement could not be explained in terms of conventional hydropassive stomatal effects. Elevated rates of transpiration were associated with and possibly caused by increased cytokinin concentrations in shoots of plants with partially excised roots. The single root remaining after excision was able to maintain an adequate water uptake for the continued enhanced transpiration, after only a short transient reduction in leaf water content. The enhanced capacity for water uptake by the remaining root was confirmed by measuring the water flow from detached roots at negative hydrostatic pressure. Even without additional suction, flow from the reduced root system increased about 1.5 h after the start of treatment, suggesting an increase in membrane permeability for water. Although abscisic acid (ABA) concentrations in the roots increased after the root excision treatment, there was no evidence for any enhanced concentration in the xylem sap. The possible role that this accumulation of ABA in roots may have in the apparent increase in hydraulic conductivity after root excision is discussed.     

Influence of the excision shock on the protein metabolism ofVicia faba L. meristematic root cells

UsingVicia faba root meristems we have shown that protein synthesis was dramatically changed after excision. The amino-acid incorporation dropped to 13% of the level in the unexcised control. This downshift was a direct consequence of the breakdown of polysomes which are converted into monosomes. In order to perform an analysis of the protein pattern by two-dimensional gel electrophoresis, endogenous proteolytic activity, which is high in broad bean root, had to be inhibited. Therefore, several protease inhibitors were tested and a very efficient inhibitor pool was obtained which could be used during the preparation of meristematic cell extracts. Protein-pattern analysis showed important differences between the unexcised control and excised apices. The number of proteins synthesized after excision droped from 250 in the control to 80, as a consequence of polysome breakdown. Futhermore, we present evidence that new and apparently specific proteins are synthesized in response to this excision shock.Abbreviations NEM N-ethylmaleimide - PMSF phenylmethylsulfonyl fluoride - TLCK N-tosyl-L-lysin chloromethyl ketone - TPCK N-tosyl-L-phenylalanine chloromethyl ketone     

Micropropagation through excised root culture of Clitoria ternatea and comparison between in vitro–regenerated plants and seedlings

An efficient protocol has been developed for high‐frequency shoot regeneration and plant establishment of Clitoria ternatea – a potential medicinal legume. Adventitious shoots were regenerated from young excised root segments of aseptic seedlings on Murashige and Skoog (MS) medium supplemented with various concentrations of 6‐benzyladenine (BA), kinetin, α‐naphthalene acetic acid (NAA) or 2,4‐dichlorophenoxy acetic acid (2,4‐D) either singly or in various combinations. The highest frequency (100%) of shoot regeneration and maximum number (16.4 ± 0.24) of shoots per explant was obtained on MS medium supplemented with 20 μm BA and 2.0 μm NAA. Organogenic calli were produced on a medium containing 2,4‐D (10 or 20 μm ) and BA (5.0 μm ). The calli were differentiated into multiple shoots on MS medium supplemented with 2.5–10 μm BA and 2.0 μm NAA. The microshoots were rooted on half‐strength MS medium supplemented with 5.0 μm indole‐3‐butyric acid and transplanted successfully in field conditions. After 12 months of transfer to ex vitro conditions, the performance of micropropagated plants were evaluated on the basis of some physiological and biochemical parameters and compared with the in vivo–grown plants of the same age. The sodium dodecyl sulphate polyacrylamide gel electrophoresis protein profile was same between regenerated and naturally growing shoots. Total soluble protein in aerial part as well as in seeds of in vitro–regenerated and in vivo–grown plants was almost the same. The mitotic study showed normal chromosomal movement and numbers (2x = 16).     

A Correlation between Structure and Function in the Root of Zea mays

The exudation rates of fluid and potassium ions from isolatedmaize roots were determined before and after excision of certainlengths of root tip. The results of this study suggest thatexcised maize roots possess the ability to absorb potassium(and presumably chloride) ions and concomitant amounts of waterover a considerable distance (10 cm) from the tip. Moreover,the observed power of absorption of ions and water into thetranslocatory pathway decreases in passing from the tip towardsthe base of the root. Both light and electron microscope techniques were used to examinethe anatomy of primary roots similar to those used in the physiologicalexperiments. The principal observation was that the xylem vesselsnear the root tip contain membrane-bounded cytoplasm with organelles.The number of mature xylem vessels, i.e. without cytoplasm,progressively increased in transverse sections cut from 1 to10 cm from the root tip; above 10 cm from the root tip all ofthe xylem vessels were found to be completely mature. It isevident that prima facie a connexion exists between this singleaspect of root anatomy and fluid exudation from excised roots. The uptake of tritiated water by roots and its transport intoexudates was examined. These data were analysed on the assumptionthat the exchange of external labelled water with the exudatewas achieved by the fluid exudation itself; this analysis indicatedthat an operational volume, similar to that of the total xylemvolume within the root, must become labelled during the formationof the exudate.     

Xylem pressure response in maize roots subjected to osmotic stress: determination of radial reflection coefficients by use of the xylem pressure probe

The absolute pressure in conducting xylem vessels of roots of 2-week-old, slowly transpiring intact maize plants (bathed in nutrition medium) was determined to be +0·024 ± 0·044 MPa using the xylem pressure probe. When the roots were subjected to osmotic stress (NaCI, KCI or sucrose), the xylem pressure decreased immediately and became more negative. However, the response of xylem pressure to osmotic stress was considerably attenuated, indicating that the radial reflection coefficients, σ₁₃ of the maize root for these solutes were rather low (between 0·2 and 0·4 depending on the concentration of the osmoticum). The low values of a, may be caused (partly) by unstirred layer effects. In repeated osmoticum/nutrition regimes a complex pattern of changes in xylem pressure was observed which was apparently linked to the interplay between transpiration and (passive and/or active) solute loading of the xylem. These processes were not observed when the roots were subjected to osmotic stress after excision. In this case, a biphasic response was observed comparable to that found for excised roots using the root pressure probe.     

<Emphasis Type="Italic">In vitro</Emphasis> organogenesis and antioxidant enzymes activity in <Emphasis Type="Italic">Acanthophyllum sordidum</Emphasis>

The effect of various hormonal combinations on callus formation and regeneration of shoot and root from leaf derived callus of Acanthophyllum sordidum Bunge ex Boiss. has been studied. Proteins and activity of antioxidant enzymes were also evaluated during shoot and root organogenesis from callus. Calli were induced from leaf explants excised from 30-d-old seedlings grown on Murashige and Skoog medium containing 4.52 μM 2,4-dichlorophenoxyacetic acid + 4.65 μM kinetin. Maximum growth of calli and the most efficient regeneration of shoots and roots occurred with 2.69 μM 1-naphthalene acetic acid (NAA), 2.69 μM NAA + 4.54 μM thidiazuron and 2.46 μM indole-3-butyric acid. Protein content decreased in calli and increased significantly during regeneration of shoots from callus. Superoxide dismutase activity decreased in calli comparing to that of seedlings, then increased in regenerated shoots and roots. High catalase activity was detected in seedlings and regenerated shoots, whereas high peroxidase activity was observed in calli and regenerated roots.     

Cavitation studies on whole Ricinus plants by acoustic detection

Summary Acoustic detection has been used to investigate the incidence of cavitation in whole potted Ricinus plants subjected to water stress by withholding water. Cavitation proceeded rather slowly and was detectable before and during wilting. Techniques which restricted water uptake more drastically such as root cooling or overlapping cuts induced more rapid click production and wilting; a response already described for excised leaves. When water stress was removed by rewatering, or rewarming a cooled root system, cavitation soon ceased. This response was more sluggish of over-delayed.Cavitation in aging leaves on well watered plants has also been examined. Despite the onset of senescence over many days there was no evidence that dry patches, which often develop extensively, are a consequence of water shortage induced by xylem blockage. Leaves, falling naturally by abscission in still air, were often remarkably turgid with water potentials similar to those of healthy attached leaves. Only after losing water was cavitation apparent, as usual for excised mature leaves. Sometimes more persistent leaves did cavitate in situ, just before abscission, showing that in normal leaves xylem blockage can occasionally precede leaf fall by several hours.     

Patterns of Regeneration in Mutilated Seedlings of Garden Balsam, Impatiens balsamina L.

Seedlings of garden balsam, Impatiens balsamina, form an encirclingswollen ring, the collet, basal to the hypocotyl. From this,four lateral roots emerge promptly after germination. When thisstructure is excised, it regenerates as a basal encircling swellingfrom which four lateral roots with root hairs emerge. Repetitionof the removal of this structure results again in the regenerationof a similar complex. Eventually, after 3–4 excisions,the pattern is broken and lateral roots occur sporadically alongthe hypocotyl. Mutilations of seedlings of the garden balsamindicate that the regeneration will not occur in the absenceof cotyledonary tissue. This suggests a control site for thisregenerative phenomenon but no mechanism for this control isadvanced. Impatiens balsamina, balsam, lateral roots, regeneration, collet     

切根贴膜对黑土区杨树农田防护林光合特性的影响

切根贴膜简单易行,是减轻林带胁地效应的有效方法,但可能会对林带造成不良影响.本文以黑龙江省黑土区农田防护林为研究对象,以不切根贴膜为对照,研究距树干0.5、1、2、3 m切根贴膜处理对杨树光合特性的影响.结果表明: 切根贴膜对杨树叶片含水率的影响较小.处理后杨树净光合速率峰、谷值出现的时间在7月较对照提前,日均值明显低于对照,各处理间差异显著.8—9月,处理后杨树净光合速率日均值仍低于对照,但降低幅度有所减小,距树干3 m处切根贴膜的杨树净光合速率与对照相比无显著差异.处理后杨树蒸腾曲线出现多种形式,距树干2、3 m处切根贴膜杨树蒸腾速率日均值较对照无显著变化,而距树干0.5、1 m处切根贴膜的杨树蒸腾速率在7月下降幅度比光合速率下降幅度更大,导致水分利用效率高于对照.切根贴膜处理没有改变杨树叶片水分利用效率的日变化趋势,但对杨树叶片水分利用效率的日均值产生了明显影响,且这种影响可能是负向的.综合分析,在距树干3 m处切根贴膜对杨树的光合特性影响较小,明显优于其他处理.     

Xylem and cell turgor pressure probe measurements in intact roots of glycophytes: transpiration induces a change in the radial and cellular reflection coefficients

Xylem probe measurements in the roots of intact plants of wheat and barley revealed that the xylem pressure decreased rapidly when the roots were subjected to osmotic stress (NaCl or sucrose). The magnitude of the xylem pressure response and, in turn, that of the radial reflection coefficients (σ_r) depended on the transpiration rate. Under very low transpiration conditions (darkness and high relative humidity), σ_r assumed values of the order of about 0·2–0·4. The σ_r values of excised roots were also found to be rather low, in agreement with data obtained using the root pressure probe of Steudle. For transpiring plants (light intensities at least 10 μmol m^?2 s^?1; relative humidity 20–40%) the response was nearly 1:1, corresponding to radial reflection coefficients of σ_r= 1. Further increase of the light intensity to about 400 μmol m^?2 s^?1 resulted in a slight but significant decrease of the σ_r values to about 0·8. Similar measurements on maize roots confirmed our previous results (Zhu et al. 1995, Plant, Cell and Environment 18, 906–912) that, in intact transpiring plants at low light intensities of about 10 μmol m^?2 s^?1 and at relative humidities of 20–40% as well as in excised roots, the xylem pressure response was much less than expected from the external osmotic pressure (σ_r values 0·3–0·5). In contrast to wheat and barley, very high light intensities (about 700 μmol m^?2 s^?1) were needed to shift the radial reflection coefficients of maize roots to values of about 0·9. Osmotically induced xylem pressure changes were apparently linked to changes in turgor pressure in the root cortical parenchyma cells, as shown by simultaneous measurements of xylem and cell turgor pressure. In analogy to the σ_r values of the respective glycophytes, the σ_c values of the root cortical cells of wheat and barley were close to unity, whereas σ_c for maize was significantly smaller (about 0·7) under laboratory conditions. When the light intensity was increased up to about 700 μmol m^?2 s^?1 the cellular reflection coefficient of maize roots increased to about 0·95. In contrast to the σ_r values, the σ_c values of the three species investigated remained almost unchanged when the leaves were exposed to darkness and humidified air or when the roots were cut. The transpiration-dependent (species-specific) pattern of the cellular and radial reflection coefficients of the root compartment of the three glycophytes apparently resulted from (flow-dependent) concentration-polarization and sweep-away effects in the roots of intact plants. The data could be explained straightforwardly terms of theoretical considerations outlined previously by Dainty (1985, Acta Horticulturae 171, 21–31). The far-reaching consequences of this finding for root pressure probe measurements on excised roots, for the occurrence of pressure gradients under transpiring conditions, and for the non-linear flow-force relationships in roots found by other investigators are discussed.     

ONTOGENETIC ANALYSIS OF TREE ROOTS IN ACER RUBRUM AND BETULA PAPYRIFERA

The development of long (6-20 m) tree roots was reconstructed from serial transverse sections taken 10-20 cm apart. Measurements of primary xylem diameter showed that the tips of long roots of red maple (Acer rubrum L.) and paper birch (Betula papyrifera Marsh.) are initially small, but they increase in diameter with increasing distance from the stem. A repeating pattern of the width of the first annual ring marks length increments of long roots. The pattern of ring width is paralleled by a pattern of primary xylem diameter. Our interpretation is that primary xylem diameter increases during spring and early summer, stays constant during late summer and decreases in autumn. After injuries to large root tips, new large lateral tips are formed that replace the parent tip. Thus, a single long root is actually formed by a succession of these replacement roots. In a series of sections there may be sudden changes in primary xylem diameter as the successive sections pass from parent root to replacement root at points of injury. These changes in birch are associated with changes in a number of protoxylem poles.     

Lateral Transport of Ions into the Xylem of Corn Roots: II. Evaluation of a Stelar Pump

When an excised corn (Zea mays) root pretreated with chloride was exposed for 10 minutes to pulse labeling with ³⁰Cl and then transferred to unlabeled chloride, the activity in the xylem exudate reached a maximum about 4 minutes after pulse labeling was discontinued and then declined sharply. The rate at which labeled chloride was transported across the root into the xylem and basipetally therein was on the order of 75 to 250 centimeters per hour. Consequently, symplasmic movement of chloride in corn roots is fast and may not be rate-limiting in transfer from the root surface to the xylem. Experiments on pulse labeling with ²²Na gave similar results. A large fraction of the absorbed ²²Na was not translocated into the exudate but was tightly sequestered in a cell compartment, probably the vacuole.     

Effect of root formation on 14C-glycine uptake and incorporation into protein by attached and excised cotyledons of Helianthus annuus

Root formation was found to extend the life-span of excised cotyledons of Helianthus annuus L. markedly. Excised cotyledons of 12-day-old plants attained longer life-span, higher root number and total root length than cotyledons excised before or after 12 days. Protein content of attached cotyledons reached a maximum level 12 days after the commencement of germination followed by a decrease. Cotyledons excised 8 days after sowing showed maximum level of protein content 44 days after excision followed by a decrease. The increase was correlated with the full development of roots. The incorporation of ¹⁴C-glycine into protein followed a pattern similar to the protein content, both in attached and detached cotyledons.     

Compensatory abilities depending on seasonal timing of thallus excision of the kelp Undaria pinnatifida cultivated in Matsushima Bay, northern Japan

Improved cultivation technology for the kelp Undaria pinnatifida is greatly needed to increase production to meet increasing commercial demand. A previous cultivation trial indicated that the crop yield of U. pinnatifida sporophytes could be increased greatly by thallus excision in late February due to compensatory growth of the remaining tissues. To develop this potential new cultivation technology, it is essential to identify the time period during which this kelp can compensate and its physiological responses to thallus excision. In this study, U. pinnatifida sporophytes were excised at about 30 cm above the meristem at the beginning of January, February, March, and April, respectively. Morphological features, photosynthesis, nutrient uptake, and carbon and nitrogen contents of excised kelps were measured and compared with these parameters in control kelps grown without excision. Both experimental and control kelps were farmed together in Matsushima Bay, northern Japan. The kelps excised in January and February showed significant increases in the lengths and dry weights of the blade, photosynthetic rates, nutrient uptake rates, and carbon and nitrogen contents compared with the control kelps, and the growth phase was prolonged for at least 1 month. No significant increases were found in dry weights and carbon and nitrogen contents of sporophylls until early April, which indicated that the maturation period was delayed. At the end of this experiment, the nitrogen contents of sporophyll tissues formed after excisions were significantly lower than those of tissues formed before excisions. In contrast, the kelps excised in March and April showed no significant increases in morphological and physiological parameters compared with control kelps. These results suggest that U. pinnatifida sporophytes exhibited great compensation when excisions were conducted during the growing phase in January and February but not in March and April when the maturation phase had started. The regulation of resource allocation to growth and maturation after thallus excisions in January and February likely results in prolongation of the growth phase and maturation phase in excised kelps.