Reduced irradiance and applied auxin influence carbohydrate relations in Pinus banksiana cuttings during propagation

Untreated and indole-3-butyrie acid-treated (IBA) cuttings from 90-day-old Pinus banksiana Lamb, stock plants were propagated under normal greenhouse irradiance (max. 900 $$mol m^-2 s^-1) and shade (max. 120 $$mol m^-2 s^-1) to determine effects on adventitious rooting and on reducing sugar and starch concentrations in needles and basal stems. In one experiment, cuttings were assessed at days 15 and 25 of propagation for basal 1-cm stem fresh weight, proportion rooted, number of roots and longest root length. In a second experiment with cuttings, basal 1-cm stem fresh weight and concentrations of reducing sugar and starch in needles and basal stems were measured each day for the first 10 days of propagation. Carbohydrate measurements were also made for seedling stock plants as controls for the second experiment. Carbohydrate data for cuttings were primarily evaluated based on net (cutting minus seedling) concentrations, to correct for changes in cuttings not related to adventitious rooting. Increase of basal stem fresh weight and rooting of cuttings, based on all measured variables, occurred in the order: light + IBA > light > shade + IBA > shade. The best rooting required the greater irradiance. Compared to results from cuttings in the light, shading resulted in lesser accumulations of reducing sugars and starch in needles and basal stems. Reducing sugar: starch concentration ratios were significantly greater in shade- vs light-propagated cuttings, IBA treatment did not offset the effects of shade on rooting or on reducing sugar and starch concentrations or ratios. Overall, the results suggested that decreased reducing sugar and starch concentrations and/or their increased ratios are associated with shade-induced poor rooting of P. banksiana cuttings.     

Rejuvenation of Sequoia sempervirens by Repeated Grafting of Shoot Tips onto Juvenile Rootstocks in Vitro: Model for Phase Reversal of Trees

Repeated grafting of 1.5-centimeter long shoot tips from an adult Sequoia sempervirens tree onto fresh, rooted juvenile stem cuttings in vitro resulted in progressive restoration of juvenile traits. After four successive grafts, stem cuttings of previously adult shoots rooted as well, branched as profusely, and grew with as much or more vigor as those of seedling shoots. Reassays disclosed retention for 3 years of rooting competence at similar levels as originally restored. Adventitious shoot formation was remanifested and callus development was depressed in stem segments from the repeatedly grafted adult. The reversion was associated with appearance and disappearance of distinctive leaf proteins. Neither gibberellic acid nor N⁶-beneyladenine as nutrient supplements duplicated the graft effects.     

UV-B response of cucumber seedlings grown under metal halide and high pressure sodium/deluxe lamps

UV-B-sensitive (Poinsett) and -insensitive (Ashley) cultivars of cucumber ( Cucumis sativus L.) were grown in growth chambers at 600 μmol m⁻²s⁻¹ of photosynthetically active radiation provided by metal halide (MH) or high pressure sodium/deluxe (HPS/DX) lamps. Plants were irradiated 15 days from seeding for 6 h per day under 18. 2 kJ m⁻² day⁻¹ of biologically effective UV-B (UV-B_BE) radiation. One of the most pronounced effects of UV-B was a 27 to 78% increase in phenylalanine ammonialyase (PAL) activity. UV-B also increased total polyamines. Catalase and superoxide dismutase varied greatly in their response to UV-B. There were no interactive effects on PAL or catalase activity, or total polyamines. There was a UV × PAR source interaction for superoxide dismutase activity. UV-B increased chlorosis and decreased height, dry weight and leaf area. Stem elongation, biomass production, leaf enlargement and chlorosis were greater under HPS/DX lamps than under MH lamps. Chlorosis was greater in Poinsett than in Ashley and in lower leaves than in upper ones. Aside from chlorosis, there were no interactive effects of UV-B, PAR source or cultivar on any of the growth parameters measured, suggesting that the growth response of cucumber seedlings to UV-B is unaffected by PAR source or cultivar. Similarly, except for SOD activity, the biochemical response to UV-B was also not influenced by PAR source or cultivar.     

INITIATION AND DEVELOPMENT OF ROOTS IN JUVENILE AND MATURE LEAF BUD CUTTINGS OF FICUS PUMILA L.

Adventitious root formation (ARF) was studied in woody leaf bud cuttings of Ficus pumila L., creeping fig. Juvenile cuttings rooted easily, whereas only mature cuttings treated with indole-3-butyric acid (IBA) attained any rooting success. In the rooting process, both juvenile and mature material exhibited dedifferentiation of phloem ray parenchyma, root initial formation, primordia differentiation, and root elongation. The early stages of adventitious rooting were most critical since few primordia were observed in mature controls. The stages leading up to root primordia differentiation and elongation occurred more rapidly in IBA-treated juvenile vs. mature cuttings; however, time differences in both types between first observable roots and maximum rooting were comparable. Root primordia differentiated from basal callus of some cuttings, but neither these nor the few primordia in mature controls elongated into well-developed roots. Anatomical differences between the juvenile and mature material did not account for rooting disparity, nor did presence of perivascular fibers, sclereids, and laticifers retard rooting.     

Movement and distribution of 14C-indole-3-acetic acid in branches and rooted cuttings of Douglas-fir

The transport of ¹⁴C-indole-3-acetic acid in branch terminals and stems of rooted cuttings of Pseudotsuga menziesii (Mirb.) Franco was studied to determine if the plagiotropic growth of cuttings might result from an accumulation of basipetally transported auxin in the morphologically upper side of cuttings stems. Twenty-four h after application of 10 μl of ¹⁴C-IAA solution to the cut surface of decapitated, rooted cuttings, nearly twice as much activity was detected in extracts of tissue from the morphologically upper than from the lower halves of the stems. A similar distribution of activity was observed in horizontal branch terminals and in branch terminals which had been tied vertically for 2 weeks. The magnitude of the difference in activity between the 2 sides of the stem was greater in the horizontal than in the vertical branches.
There was no significant difference in transport through the upper and lower sides of excised stem segments from cuttings or branch terminals. In segments from rooted cutting stems, however, significantly more radioactivity from ¹⁴C-IAA donor blocks was detected in the lower than in the upper halves of segments.     

Auxins or Sugars: What Makes the Difference in the Adventitious Rooting of Stored Carnation Cuttings?

Cold storage of cuttings is frequently applied in the vegetative propagation of ornamental plants. Dianthus caryophyllus was used to study the limiting influences of auxin and sugars on adventitious root formation (ARF) in cuttings stored at 5°C. Carbohydrate levels during storage were modulated by exposing cuttings to low light or darkness. The resulting cuttings were treated (or not) with auxin and planted, and then ARF was evaluated. Carbohydrate levels in the cuttings were monitored and the influence of light treatment on indole-3-acetic acid (IAA) and zeatin (Z) in the basal stem was investigated. Dark storage for up to 4 weeks increased the percentage of early rooted cuttings and the final number and length of adventitious roots, despite decreased sugar levels in the stem base. Light during cold storage greatly enhanced sugar levels, particularly in the stem base where the Z/IAA ratio was higher and ARF was lower than observed in the corresponding dark-stored cuttings. Sugar levels in nonstored and dark-stored cuttings increased during the rooting period, and auxin application enhanced the accumulation of sugars in the stem base of nonstored cuttings. Auxin stimulated ARF most strongly in nonstored, less so in light-stored, and only marginally in dark-stored cuttings. A model of auxin-sugar interactions in ARF in carnation is proposed: cold storage brings forward root induction and sink establishment, both of which are promoted by the accumulation of auxin but not of sugars, whereas high levels of sugars and probably also of cytokinins act as inhibitors. Subsequent root differentiation and growth depend on current photosynthesis.     

The consequences of freezing temperatures followed by high irradiance on in vivo chlorophyll fluorescence and growth in Picea abies

Picea abies (L.) Karst. plants, propagated by cuttings, were subjected to one night of freezing temperatures (-5°C), high irradiance (1 200 or 1 800 μmol m⁻² s⁻¹), or freezing temperatures followed by high irradiance. The treatments were applied at bud burst, at time of shoot elongation, and when the shoots had ceased to elongate. The maximum quantum yield of photosynthesis, F_v/F_m, dry weight of branches and needles, and length and survival of shoots were measured. F_v/F_m and growth decreased after a night of freezing temperatures followed by high irradiance, at the time of bud burst and shoot elongation. High irradiance alone influenced F_v/F_m, but not growth. Freezing temperatures affected F_v/F_m, and growth at the time of shoot elongation. F₀ increased after a night of freezing temperatures and decreased with age of the current-year needles. It was concluded that the use of short-term measurements of chlorophyll fluorescence induction to predict changes in growth after a night of frost and subsequent high light was not a reliable method.     

Influence of spectral quality on the growth response of intact bean plants to brassinosteroid, a growth-promoting steroidal lactone. I. Stem elongation and morphogenesis

The growth response of bean ( Phaseolus vulgaris L. 'Pinto') plants treated with 5 μg of brassinosteroid (BR) in the bean second-internode assay was measured in a controlled environment under 3 radiation sources: cool white fluorescent (CWF), far-red (FR) fluorescent and incandescent (INC) lamps. Growth comparisons were made under equal levels (90 μmol s^-1 m^-2) of photosynthetic photon flux density provided by CWF or INC lamps and equal levels of far-red (28 W m^-2, 700–800 nm) radiation provided by the same INC or FR lamps. Treatment of the second internode with BR produced a sequential increase in elongation, curvature, and swelling under normal bioassay conditions (CWF lamps), as observed previously with brassins. In addition, BR induced marked splitting of the treated internode provided that ample photosynthate was available. Spectral quality had a differential effect on internodal elongation. Under CWF lamps, internodes, 6 days after BR-treatment, were 2–3 times longer than those of controls; under INC or FR lamps they were 30–60% shorter than those of controls. In all cases, BR-treatment greatly stimulated accumulation of photosynthate in the treated internode, as indicated by fresh and dry weights and stem diameter measurements. This suggests a possible mobilization role for BR in the intact plant. Brassinosteroid also partially overcame the natural inhibitory effects of CWF radiation on stem elongation.     

Effect of carbon dioxide, fertilization and irrigation on loblolly pine branch morphology

 Foliage and wood parameters of branches of 12-year-old loblolly pine (Pinus taeda L.) trees were characterized after 21 months of exposure to fertilizer, irrigation and elevated CO₂ treatments. Branches of loblolly pine trees were enclosed in plastic chambers and exposed to ambient, ambient +175 and ambient +350 umol mol^–1 CO₂ concentrations. Measurements of foliage and wood at the fascicle, flush and branch levels were made at the end of the 21 month study period. The +350 CO₂ treatment did not significantly increase fascicle radius or length but did increase the number of fascicles on the first flush. Fertilization significantly increased fascicle radius and length, while irrigation significantly increased number of fascicles and flush length of first flush. The +350 CO₂ treatment also significantly increased flush length of the first flush. Significant interaction of fertilization and irrigation with CO₂ was observed for fascicle length. Significant interactions of fertilization and irrigation were also observed for flush length, number of fascicles and fascicle length. Observed increases in fascicle radius, fascicle length, number of fascicles and flush length may have been responsible for the significantly higher flush leaf area observed for the all three treatments. Also, a combination of fertilization and irrigation increased leaf area by 82% compared to that in the control when averaged across CO₂ treatments. At the branch level +350 CO₂ treatment significantly increased shoot length but not the number of flushes on the branch. In general with the exception of bark density and total number of needle scales, neither fertilization nor irrigation had any significant effect on other branch level parameters. Results from this study indicate that with ‘global change’ an increase in CO₂ alone may increase leaf area via an increase in flush length and number of fascicles. Combining increases in CO₂ with fertilization and irrigation could greatly enhance leaf area which when coupled to observed increases in net photosynthesis as a result of elevated CO₂ could greatly increase productivity of loblolly pine trees. Received: 22 August 1996 / Accepted: 5 March 1997     

Growth analysis of UV-B-irradiated cucumber seedlings as influenced by photosynthetic photon flux source and cultivar

A growth analysis was made of ultraviolet-B (UV-B)-sensitive (Poinsett) and insensitive (Ashley) cultivars of Cucuumis satives L. grown in growth chambers at 600 μmol m⁻² s⁻¹ of photosynthetic photon flux (PPF) provided by red- and far-red-deficient metal halide (MH) or blue- and UV-A-deficient high pressure sodium/deluxe f HPS/DX) lamps. Plants were irradiated 6 h daiiy with 0.2 f-UV-B) or 18.2 C+UV-B) kJ m⁻² day⁻¹ of biologically effective UV-B for 8 or 15 days from time of seeding. In general, plants given supplemental UV-B for 15 days showed lower leaf area ratio (LARs, and higher specific leaf mass (SLM) mean relative growth rate (MRGR) and net assimilation rate (NAR) than that of control plants, but they showed no difference in leaf mass ratio (LMR), Plants grown under HPS/DX lamps vs MH lamps showed higher SLM and NAR. lower LAR and LMR. hut no difference in MRGR. LMR was the only growth parameter affected by cultivar: at 15 days, it was slightly greater in Poinsett than in Ashley. There were no interactive effects of UV-B. PPF source or cultivar on any of the growth parameters determined, indicating that the choice of either HPS/DX or MH lamps should not affect growth response to UV-B radiation. This was true even though leaves of UV-B-irradiated plants grown under HPS/DX lamps have been shown to have greater chlorosis than those grown under MH lamps.     

Dependence of Root Growth on Photosynthesis in Populus tremula

Hoot elongation of rooted cuttings of aspen, Populus tremula L., was followed on moist filter paper. The time course of root growth during treatments which decrease or interrupt the supply of carbohydrates to the roots was studied. At light intensities below 4000 lux the rate of root elongation was found to berelated to the light intensity over the leaves. In the dark, root growth decreased within 24 hours and the roots stopped growing completely during the second or third day. Excision of the leaves or steam-killing of the tissue in a stem section below the leaves was followed by root growth stoppage within 24 hours. These results imply that the rate of root growth under certain conditions is determined by the supply of carbohydrates from the leaves.     

Treatment of bean cuttings by chestnut extracts during rooting modifies their IAA-oxidase activity

The IAA-oxidase activity in bean cuttings ( Phaseolus vulgaris L. cv. Contender) treated with extracts from juvenile and adult chestnut ( Castanea sativa Mill.) was studied in the light of the effect of the extracts on the rooting activity of some auxins. Extracts from adult chestnut increased IAA-oxidase activity in bean cuttings. Extracts from juvenile chestnut inhibited IAA-oxidase activity only slightly, but substantially reduced the IAA-oxidase activity of bean cuttings pre-treated with adult chestnut extracts. These findings provide evidence that there is a relationship between the IAA-oxidase system and the effect of chestnut extracts on IAA-induced rhizogenesis in bean cuttings.     

Phytochrome involvement in stimulation and alleviation of inhibition of plant growth by malformin

Stimulation of stem elongation on green cuttings of Phaseolus aureus by malformin occurred only in red light and was specifically reversible by subsequent treatment with far red radiation. Inhibition of stem elongation of etiolated cuttings by malformin in the dark was alleviated by red light and was repeatedly reversible with far red irradiation. A direct or indirect effect of malformin on phytochrome action was suggested.     

A Mung Bean Assay for Malformin-induced Growth Stimulation

A bioassay employing green or etiolated cuttings of Phaseolus aureus Roxb. was developed for determining malformin-induced growth stimulation in light. Growth enhancement of green cuttings was more rapid and relatively greater than that of etiolated cuttings. Cuttings from green seedlings responded less as seedlings aged; those from etiolated seedlings responded more. Malformin also stimulated the growth of green or etiolated seedlings in light. Most growth enhancement induced by malformin occurred in the upper 1 cm of the stem. Using green cuttings, malformin stimulated stem elongation relatively more when cotyledons, leaves, or especially apical buds were removed. Although malformin failed to stimulate elongation of 2-cm stem sections “floated” on solutions in Petri dishes, it stimulated elongation of sections when they were upright. High concentrations of indoleacetic acid inhibited growth enhancement by malformin. When gibberellin and malformin were combined, growth enhancement was nearly additive.     

The rooting ability of shoots raised 'in vitro' from the apple rootstock A2 in juvenile and in adult growth phase

The apple rootstock A2 can be readily propagated in vitro both in the juvenile and in the adult growth phase. Shoots were produced by meristem tip culture from the apple rootstock A2 in different growth phases. The influence of growth phases and different concentrations of PG and IBA was investigated as to rooting percentage, survival percentage, number of roots per rooted shoot, root length, shoot length and formation of callus. IBA at 15 μ M without PG gave a significantly lower rooting percentage than 5 and 10 μ M IBA. PG together with IBA stimulated rooting, the optimum concentrations of PG being, however, not the same for the different growth phases. For the adult growth phase, 10⁻⁴ M PG promoted rooting, whereas 10⁻³ M PG markedly inhibited rooting. In the juvenile growth phases, both 10⁻⁴ and 10⁻³ M PG stimulated rooting. PG at 10⁻⁴ M also increased the number of roots. The longest roots were obtained at 10⁻³ M PG and 5 μ M IBA. PG at 10⁻³ M reduced callus formation at all IBA concentrations used. Neither shoot length nor root length influenced the survival percentage.     

A relationship between irradiation, carbohydrates and rooting in cuttings of Pisum sativum

Rooting ability was studied for cuttings derived from pea plants ( Pisum sativum , L. cv. Alaska) grown in controlled environment rooms. When the cuttings were rooted at 70 μmol m⁻² s⁻, ¹ (photosynthetic photon flux density) or more, a stock plant irradiance at 100 μmol m⁻² s⁻¹ decreased rooting ability in cuttings compared to 5 μmol m⁻², s⁻¹, However, cuttings rooted at 160 μmol m⁻² s⁻¹ formed more roots compared to 5 (μmol m⁻² s⁻¹. Although a high irradiance increased the number of roots formed, it could not overcome a decreased potential for root formation in stock plants grown at high irradiance. Light compensation point and dark respiration of cuttings decreased by 70% during the rooting period, and the final levels were strongly influenced by the irradiance to the cuttings. Respiratory O₂ uptake decreased in the apex and the base of the cutting from day 2 onwards, whereas a constant level was found in the leaves. Only the content of extractable fructose, glucose, sucrose and starch varied during the early part of the rooting period. We conclude that the observed changes in the cuttings are initiated by excision of the root system, and are not involved in the initiation of adventitious roots.     

Vegetative propagation of adult Eucalyptus grandis X urophylla and comparison of growth between micropropagated plantlets and rooted cuttings

Methods for the production of micropropagated plantlets and rooted cuttings were developed and used to vegetatively multiply adult Eucalyptus grandis X urophylla. Rooting success was less than 5% when cuttings excised from twigs of 3-year-old trees were used. The rooted cuttings were grown in the greenhouse as explant- or cutting-donors and maintained at a height of 30 to 100 cm by trimming back periodically. Good rooting success (95%) of cuttings was obtained for epicormic shoots produced from donor plants after trimming 5 times. Explants of both apical and axillary buds taken from the donor plants produced multiple shoots when cultured in vitro. In vitro multiple shoot production was optimal on MS medium containing 0.1 mg/l BA and 0.01 mg/l NAA averaging 13.7 shoots per explant in a 40-day culture period. Shoot elongation was accelerated on a modified MS medium containing half strength potassium nitrate and sucrose. Elongated shoots excised at approximately 1.5 cm in length were successfully rooted on media with NAA or IBA concentrations ranging from 0.1 to 10 mg/l. Root formation was optimal on medium consisting of full strength MS basal macro elements and vitamins, half strength micro elements, 1% sucrose and supplemented with 0.3 mg/l IBA. In the field test, no significant differences were found in tree height and DBH between micropropagated plantlets and rooted cuttings at 1 and 3 years old, with the exception at 2 years old. A considerable difference arose between the 2 types of vegetative propagules in physiological response to flowering, caused by dissimilar degrees of rejuvenation.Abbreviations BA Benzyl-Aminopurine - NAA Naphthalene Acetic Acid - IBA Indole-3-Butyric Acid - MS medium Murashige and Skoog's medium - DBH Diameter at Breast Height     

Effects of Nutrients and Light on Growth and Root Formation in Pisum sativum Cuttings

Cuttings obtained from seedlings of Pisum sativum L. were rooted in water solution. Shoot growth continued after excision and shoot length increased considerably before roots emerged. Increase in dry weight was strongly dependent on light supply. Continued growth was dependent on supply of mineral nutrients to the rooting solution. Mineral nutrients had no or slight influence on the number of roots formed on cuttings from stock plants grown in fertilized soil, but the growth in length of the roots was dependent on the presence of calcium in the solution. Root formation was dependent on photosynthetic products formed after excision. No roots were formed on cuttings kept in the dark. The number of roots increased with increasing irradiance given to the leafy part of the cutting. At a low level of irradiance sucrose supply through the rooting medium increased the number of roots. Light given to the basal part of the cuttings had a strongly inhibitory effect on the number of roots formed. It is concluded that the carbohydrate level easily becomes a limiting factor for root formation in growing pea cuttings. Availability of mineral nutrients influences in the first place the growth of the shoots.     

Anatomical features of pepper plants (Capsicum annuum L.) grown under red light-emitting diodes supplemented with blue or far-red light

Pepper plants (Capsicum annuum L. cv., Hungarian Wax) were grown under metal halide (MH) lamps or light-emitting diode (LED) arrays with different spectra to determine the effects of light quality on plant anatomy of leaves and stems. One LED (660) array supplied 90% red light at 660 nm (25nm band-width at half-peak height) and 1% far-red light between 700-800nm. A second LED (660/735) array supplied 83% red light at 660nm and 17% far-red light at 735nm (25nm band-width at half-peak height). A third LED (660/blue) array supplied 98% red light at 660nm, 1% blue light between 350-550nm, and 1% far-red light between 700-800nm. Control plants were grown under broad spectrum metal halide lamps. Plants were gron at a mean photon flux (300-800nm) of 330 micromol m-2 s-1 under a 12 h day-night photoperiod. Significant anatomical changes in stem and leaf morphologies were observed in plants grown under the LED arrays compared to plants grown under the broad-spectrum MH lamp. Cross-sectional areas of pepper stems, thickness of secondary xylem, numbers of intraxylary phloem bundles in the periphery of stem pith tissues, leaf thickness, numbers of choloplasts per palisade mesophyll cell, and thickness of palisade and spongy mesophyll tissues were greatest in peppers grown under MH lamps, intermediate in plants grown under the 660/blue LED array, and lowest in peppers grown under the 660 or 660/735 LED arrays. Most anatomical features of pepper stems and leaves were similar among plants grown under 660 or 660/735 LED arrays. The effects of spectral quality on anatomical changes in stem and leaf tissues of peppers generally correlate to the amount of blue light present in the primary light source.     

Evaluation of aeroponics for clonal propagation of Caralluma edulis,Leptadenia reticulata and Tylophora indica – three threatened medicinal Asclepiads

The present study explores the potential of aeroponic system for clonal propagation of Caralluma edulis (Paimpa) a rare, threatened and endemic edible species, Leptadenia reticulata (Jeewanti), a threatened liana used as promoter of health and Tylophora indica (Burm.f.) Merill, a valuable medicinal climber. Experiments were conducted to asses the effect of exogenous auxin (naphthalene acetic acid, indole-3-butyric acid, indole-3-acetic acid) and auxin concentrations (0.0, 0.5, 1, 2, 3, 4 or 5gl⁻¹) on various root morphological traits of cuttings in the aeroponic chamber. Amongst all the auxins tested, significant effects on the length, number and percentage of rooting was observed in IBA treated nodal cuttings. Cent per cent of the stem cuttings of C. edulis rooted if pre-treated with 2.0 gl⁻¹ of IBA for 5 min while 97.7 % of the stem cuttings of L. reticulata and 93.33 % of stem cuttings of Tylophora indica rooted with pre-treatment of 3.0 gl⁻¹ of IBA for 5 min. Presence of at least two leaves on the nodal cuttings of L. reticulata and T. indica was found to be a prerequisite for root induction. In all the species, the number of adventitious roots per cutting and the percentage of cuttings rooted aeroponically were significantly higher than the soil grown stem cuttings. Shoot growth measured in terms of shoot length was significantly higher in cuttings rooted aeroponically as compared to the cuttings rooted under soil conditions. All the plants sprouted and rooted aeroponically survived on transfer to soil. This is the first report of clonal propagation in an aeroponic system for these plants. This study suggests aeroponics as an economic method for rapid root induction and clonal propagation of these three endangered and medicinally important plants which require focused efforts on conservation and sustainable utilization.