The use of growth retardants to improve microtuber formation by potato (Solanum tuberosum)

The growth retardant chlormequat stimulated microtuber formation by a recalcitrant cultivar of potato (Solanum tuberosum), but reduced microtuber fresh weight in a cultivar that tuberised readily in its absence. Inhibition of microtuber growth by high concentrations of chlormequat was confirmed using a different in vitro system where all cultivars tuberised in the absence of growth retardants.Alternative growth retardants were tested. Daminozide also had a detrimental effect on microtuber fresh weight, but ancymidol and paclobutrazol did not inhibit microtuber growth at the concentrations required for stimulation of tuberisation by recalcitrant cultivars. In addition, 10^-5 M ancymidol and paclobutrazol inhibited premature sprouting of microtubers in vitro.Abbreviations BA benzyladenine - DMSO dimethyl sulphoxide - FW fresh weight - PAR photosynthetically active radiation - SE standard error     

Promotion of asparagus shoot and root growth by growth retardants

Plantlets regenerated from shoot-tip culture of Asparagus officinalis L. possessed weak shoots and roots. Various combinations of auxins and cytokinins did not improve the plantlets. Incorporation of a number of growth retardants, viz. ancymidol, B-995, phosfon, Amo 1618, cycocel and paclobutrazol, promoted growth of stronger shoots and roots. The effectiveness of the growth retardants varied, with ancymidol being most effective and cycocel least effective.The response to ancymidol was prevented by exogenous GA₃ and GA_4/7. GA_1/3 and GA_4/7-like activities were detected in asparagus shoot-tip culture and these activities were reduced by the presence of the growth retardants ancymidol, Amo-1618, and cycocel.     

The influence of relative humidity on vitrification, growth and morphology of Gypsophila paniculata L.

A vitrification scoring system was devised that comprised a visual assessment of vitrification in vitro followed by transplanting of plantlets ex vitro and recording of plantlet survival rates. This proved to be a simple method of predicting survival of plantlets ex vitro and demonstrated that vitrification severity is more important than acclimitization procedures in promoting survival ex vitro. Growth of normal plantlets in liquid medium in an environment where plantlets are able to transpire made it clear that lack of transpiration as opposed to high water availability is the primary cause for induction of vitrified growth. Finally, small decreases in relative humidity in vitro were found to be sufficient to increase plantlet transpiration to the extent that vitrification is greatly reduced and survival ex vitro increased. Small differences in relative humidity also influenced plantlet morphology with plantlets grown at a higher relative humidity being larger and consisting of more shoots than plantlets grown at a lower relative humidity. It is suggested that in vitro relative humidity may be manipulated to produce plantlets of a particular morphology and should be measured in tissue culture experiments and included in the materials and methods sections of papers to ensure correct interpretation of results.     

Number of air exchanges,sucrose concentration,photosynthetic photon flux,and differences in photoperiod and dark period temperatures affect growth of Rehmannia glutinosa plantlets in vitro

Rehmannia glutinosa plantlets were cultured for 4 weeks under different culture conditions to determine the optimum environment for in vitro growth and ex vitro survival. Plantlet growth increased with an increasing number of air exchanges of the culture vessel, exhibiting greatest shoot weight, total fresh weight, leaf area, and chlorophyll content at 4.4 h⁻¹ of air exchanges. High sucrose concentration (30 g l⁻¹) increased root weight but reduced shoot growth. Net photosynthetic rates of the plantlets were greatest when sucrose was not added to the medium. On the other hand, ex vitro survival of the plantlets was not influenced by sucrose concentration. In the experiment on difference in photoperiod and dark period temperatures (DIF) and photosynthetic photon flux (PPF), plantlet growth increased as DIF and PPF levels increased. Particularly, increasing PPF level had a more distinctive effect on plantlet growth than increasing DIF level. The interaction of DIF × PPF was also significant, showing the greatest plantlet growth in positive DIF (+8 DIF) and a high PPF (210 μmol m⁻² s⁻¹). In conclusion, the results of this experiment suggest that increased number of air exchanges of the culture vessel, decreased sucrose concentration, and positive DIF in combination with high PPF level enhanced growth and acclimatization of Rehmannia glutinosa plantlets. This revised version was published online in June 2006 with corrections to the Cover Date.     

Physiological changes and growth of micropropagated chile ancho pepper plantlets during acclimatization and post-acclimatization

Little is known about physiological changes that occur with micropropagated chile ancho pepper (Capsicum annuum L. cv. San Luis) plantlets during acclimatization. Plantlets were transferred to ex vitro conditions to study selected physiological changes and growth performance during acclimatization and post-acclimatization. The physiology of the plantlets was characterized by measuring leaf gas exchange and water status. Plant growth was determined by assessing plant height, leaf number, total leaf area, relative growth rate (RGR), and leaf, root, and stem dry matter (DM). Chile pepper plantlets became acclimatized within 6 days after transplantation. During this period, physiological adjustments occurred, which were critical for plantlet survival. After initial ex vitro transplanting, plantlets experienced water deficit [leaf wilting and reduced relative water content (RWC)], which corresponded with reduced stomatal conductance (g _s) and transpiration (E), and an increase in stomatal resistance (r _s). Thus, leaf stomata that developed in vitro were functional ex vitro. Because of this stomatal control, plantlets minimized transplant shock, recovered and survived. Prior to transplanting, plantlets were photomixotrophic, as indicated by low photosynthetic rates (A). During acclimatization, RWC, g _s, E, and A were significantly lower two days after transplanting. However, within 6 days after transplanting, plantlets recovered and became photoautotrophic – attaining high A, g _s, and E. Water use efficiency was initially low during the first days after transplanting, but increased dramatically at the end of the acclimatization period in part due to increased A. The stabilization and improvement of plantlet water status and gas exchange during acclimatization and post-acclimatization closely correlated with increased plantlet growth. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effects of in vitro treatments on leaf area growth of potato transplants during acclimatisation

The importance of leaf area of in vitro propagated potato (Solanum tuberosum L.) plantlets for further growth during acclimatisation and the after-effects of in vitro treatments on growth were examined. The in vitro treatments included different levels of alar, nitrogen or mannitol or different temperatures during the last in vitro phase, the rooting phase. Leaf area or ground cover was recorded one day after planting to soil and at the end of the first phase of ex vitro growth, the acclimatisation phase. Regression analysis showed that leaf area of a transplant at the end of acclimatisation phase was positively influenced by leaf area of the same plantlet at the beginning of the phase. The relative increase in leaf area during acclimatisation (increase/early leaf area) was linearly related to the inverse of the early leaf area, indicating almost comparable relative increases for plantlets having larger early leaf areas, but more variable responses for plantlets having smaller early leaf areas. In vitro treatments mainly affected leaf area of transplants through their effects on early leaf area. Adding alar, reducing nitrogen and reducing temperature increased leaf area. Reducing mannitol increased ground cover. A lower nitrogen concentration and higher temperature in some cultivars had slight negative effects on the relative increase in leaf area after acclimatisation. For nitrogen these negative effects were less significant than the positive effects through early leaf area. Results stress the importance of manipulation of leaf area in vitro to enhance plant performance in later stages of growth. This revised version was published online in June 2006 with corrections to the Cover Date.     

PP333对离体苹果苗的生长及其生理的影响

在生根培养基中加入0.5或2.0PPm PP333, 明显地减少了苹果离体新梢的鲜重和干重、叶片鲜重和叶面积,增加了比叶重、根的鲜重和干重及根梢鲜重和干重比,并促进了根的形成。PP333对单位叶重量的叶绿素含量没有影响,而增加了单位叶面积的叶绿素含量。经PP333处理后,叶片中的淀粉含量、过氧化物酶活性、蛋白质和游离氨基酸的含量均明显高于对照,可溶性糖与对照差异不明显。     

Arbuscular mycorrhizal fungi influence water relations, gas exchange, abscisic acid and growth of micropropagated chile ancho pepper (Capsicum annuum) plantlets during acclimatization and post-acclimatization

Little is known about the role of arbuscular mycorrhiza fungi (AMF) on physiological changes of micropropagated plantlets during acclimatization and post-acclimatization. Using chile ancho pepper (Capsicum annuum L. cv. San Luis), measurements were made of water relations, gas exchange, abscisic acid (ABA), plantlet growth and AMF development. Plantlets had low photosynthetic rates (A) and poor initial growth during acclimatization. Relative water content (RWC) decreased during the first days after transfer from tissue culture containers to ex vitro conditions. Consequently, transpiration rates (E) and stomatal conductance (gs) declined, confirming that in vitro formed stomata were functional and able to respond ex vitro to partial desiccation--thus avoiding excessive leaf dehydration and plant death. Colonization by AMF occurred within 3 days after inoculation. Colonized plantlets had lower leaf ABA and higher RWC than noncolonized (NonAMF) plantlets during peak plant dehydration (6 days after plant transfer)--and a higher A and gs as early as days 5 and 7. During post-acclimatization [after day 8, when RWC increased and stabilized], A increased in all plantlets; however, more dramatic changes occurred with AMF plantlets. Within 48 days, 45% of the roots sampled of inoculated plantlets were colonized and had extensive arbuscule development. At this time, AMF plantlets also had greater E, A, leaf chlorophyll, leaf elemental N, P and K, leaf dry biomass and leaf area, fruit production and differences in carbon partitioning [lower root/shoot ratio and higher leaf area ratio] compared with NonAMF plantlets. Rapid AMF colonization enhanced physiological adjustments, which helped plantlets recover rapidly during acclimatization and obtain greater growth during post-acclimatization.     

Cultivating <Emphasis Type="Italic">in vitro</Emphasis> coconut palms (<Emphasis Type="Italic">Cocos nucifera</Emphasis>) under glasshouse conditions with natural light,improves <Emphasis Type="Italic">in vitro</Emphasis> photosynthesis nursery survival and growth

Plantlets of coconut were cultured in vitro under three different ambient conditions including a standard culture room, a culture room inside a glasshouse with natural light but controlled temperature, and a standard glasshouse with natural light and natural fluctuations of temperature. Plantlets from the 3 treatments were compared in terms of growth, plant survival as well as net photosynthesis and efficiency of PSII (Fv/Fm ratio) both at the end of the in vitro stage and at 3 stages of ex vitro acclimatization. At the end of the in vitro stage, plantlets cultured in vitro under glasshouse conditions showed the best performance showing the highest photosynthesis rate, dry weight and number of leaves. Plantlets from the standard culture room showed the lowest photosynthesis and growth rate. After 6 months of ex vitro acclimatization, plantlets originally grown in vitro under glasshouse conditions maintained better field survival and growth rates in terms of fresh weight, dry weight and leaf number than plantlets originally grown in vitro in the standard culture room. Although more studies are required to define the reason for this effect, it is clear that the conditions of standard culture rooms are not the best for in vitro cultivation of coconut and perhaps other tropical species.     

Enhanced shoot and cormlet proliferation in liquid cultured gladiolus buds by growth retardants

Gladiolus bud explants propagated in agitated liquid medium, in the presence of the growth retardants daminozide, ancymidol, paclobutrazol or Majic, proliferated into massive bud aggregates without leaves. The buds developed into protocorms and after subculture to agar-solidified medium formed cormlets 8–10 mm in diameter. The corms were not dormant and, after transplanting to pots in the greenhouse, developed into plantlets, 5–6 months after explant isolation.     

PP_(333)对离体苹果苗的生长及其生理的影响

在生根焙养基中加入0.5或2.0ppm PP_333,明显地减少了苹果离体新梢的鲜重和干重、叶片鲜重和叶面积,增加了比叶重、根的鲜重和干重及根梢鲜重和干重比,并促进了根的形成。PP_333对单位叶重量的叶绿素含最没有影响,而增加了单位叶面积的叶绿素含量。经PP_333处理后,叶片中的淀粉含量、过氧化物酶活性、蛋白质和游离氨基酸的含量均明显离于对照,可溶性糖与对照差异不明显。     

Abscisic acid content and growth of spring barley plants treated with triazoles

Effects of triazolium compounds (paclobutrazol, uniconazole, and azovit), applied under laboratory and greenhouse conditions, on growth of barley plants (Hordeum vulgare L., cvs. Moskovskii 2 and Zazerskii 85) and ABA content in their tissues were studied for the first time. At various application techniques, the retardants suppressed growth of axial organs, promoted thickening and strengthening of lower internodes, and caused a considerable increase in ABA content. These changes were observed already on the next day after spraying plants with retardants and maintained for two weeks after the spraying treatment. At the same time, the retardant action sustained for 42 days when the retardants were applied to soil. The triazoles had positive effects on plant productivity characteristics, such as the number of filled grains in the main spike, total weight of grain per plant (grain collected from the main and lateral spikes), and the weight of 1000 grains. It was concluded that the retardant effect of triazoles on spring barley plants is related to elevated ABA content in plant tissues over a prolonged growth period.Translated from Fiziologiya Rastenii, Vol. 52, No. 1, 2005, pp. 108–114.Original Russian Text Copyright © 2005 by Chizhova, Pavlova, Prusakova.     

A comparative study on growth and morphology of wasabi plantlets under the influence of the micro-environment in shoot and root zones during photoautotrophic and photomixotrophic micropropagation

The growth of wasabi (Wasabia japonica Matsumura) plantlets under different micro-environments inside culture vessels in photoautotrophic micropropagation (PA) and photomixotrophic micropropagation (PM) conditions were compared. After 28 days of culture, dry weight, relative growth rate, leaf area, and leaf chlorophyll contents of plantlets in PA were greater than those in PM. The number of leaves did not differ significantly between PA and PM conditions. PA promoted root growth and development with a greater number of roots, root length, root diameter, root fresh weight, root dry weight, and root xylem vessel system. Dissolved oxygen concentration in PA culture medium sharply decreased after 7 days of culture and then recovered. In PM culture medium, no significant fluctuation of dissolved oxygen concentration was apparent. The net photosynthetic rates of plantlets in PA were much higher than those in PM and increased with culture time. In contrast, the net photosynthetic rates of wasabi plantlets in PM kept a low and constant value during the culture period. With the presence of gas exchange membranes attached to the vessel lids, the detected vapor pressure deficit was higher in PA than in PM conditions. Higher stomatal density and larger stomatal aperture on the abaxial and adaxial surfaces of the leaves in PM medium promoted leaf water loss following ex vitro conditions. Thus, PA is applicable for producing healthy wasabi transplants.     

Water relations and growth of rose plants cultured in vitro under various relative humidities

Stomatal malfunctioning is one of the main reasons why plants desiccate when transferred from in vitro to greenhouse conditions. In order to overcome this problem in Rosa hybrida cv. Madame G. Delbard (R) Deladel, two techniques, bottom cooling and water vapour permeable lid, were used. Both methods aimed to increase the vapour pressure gradient between leaf and atmosphere and consequently to improve plant transpiration.The results showed that these techniques increased leaf resistance to dehydration and improved stomatal regulation. Water relations of treated plantlets were similar to those generally observed in hardened plants: lower leaf water and osmotic potentials, and lower leaf water content than in the control ones. Osmotic adjustment occurred in treated plantlets maintaining turgor pressure. Each technique also induced some effects on growth during the rooting phase: with bottom cooling, roots were shorter, with permeable lids, apices were necrosed.These results are discussed in terms of physiological causes and in terms of effect during the following acclimatization.Abbreviations AWC absolute water content - DW dry weight - FW fresh weight     

Effect of ethrel,chlormequat chloride and paclobutrazol on growth and pyrethrins accumulation in <Emphasis Type="Italic">Chrysanthemum cinerariaefolium</Emphasis> Vis.

Pyrethrins and flower yield of pyrethrum (Chrysanthemum cinerariaefolium Viz.) plants were determined after application of ethrel, chlormequat chloride and paclobutrazol. Ethrel at 50, 100, 250 and 500 mg l^−l produced a significant positive effect on pyrethrins level, decreased plant height, while 50 and 100 mg l^−l significantly increased fresh and dry flower yield. Chlormequat chloride at 1000 and 2000 mg l^−l and paclobutrazol (80 and 160 mg l^−l) increased pyrethrins level, single flower weight and decreased plant height and flower yield. ¹⁴C-acetate incorporation studies further substantiated positive effect of growth retardants on pyrethrins biosynthesis. The effect of growth retardants on pyrethrins seems to be mediated through its effect on biosynthesis.     

Effects of growth retardants on bulbil production by Narcissus twin-scales

Narcissus twin-scale propagules, cut from various positions in the parent bulb, were incubated with gibberellic acid (GA₃) or growth retardants. Bulbil production was inhibited in all cases by GA₃ at 10–100 mg/litre. Chlormequat chloride, chlorphonium chloride and paclobutrazol also inhibited bulbil production; in the case of paclobutrazol, outer twin-scales were more sensitive to lower concentrations of the retardant than inner ones. Ancymidol enhanced bulbil production (in numbers by up to 15%) in the outermost twin-scales, and had no stimulatory effect on the inner twin-scales. Treatments affected bulbil numbers and bulbil lengths about equally.     

Distinguishing the effects of light and temperature variations on the growth, development, multiplication potential and ex vitro survival rates of in vitro cassava

Dissemination of cassava tissue culture plantlets is difficult in the arid tropics due to low eux vitro survival rates. Increased in vitro light intensity has been reported to induce high ex vitro survival rates. The results from earlier experiments suggested that it would be worthwhile to analyse the separate effects of in vitro light and of temperature on the in vitro growth pattern as well as differentiate its relation to ex vitro survival. Accordingly, analysis of a range of in vitro light intensities from 0 to 369 μmol^?1 m^?2 photosynthetic photon flux density (PPFD) was conducted both with and without fans to control the heat. Temperature proved stable at low PPFD levels but increased above 204 μmol s^?1 m^?2 when no fans were used. Increased PPFD levels induced larger fresh and dry masses as well as stem thickness. PPFD levels affected the developmental index (senescent leaf numbers) in vitro when it rose above 204 μmol s^?1 m^?2 PPFD. Raised temperature ranges increased the multiplication index (node numbers) in vitro and ex vitro. It increased root number and leaf development (lobe anatomy). As in vitro temperatures of up to 40°C improve multiplication rates and PPFD levels above 101 μmol s^?1 m^?2 were detrimental for ex vitro survival (as low as 60%), it is suggested that simpler and less costly laboratories with low light levels and a wide range of temperature tolerance could be successfully established in the tropics for in vitro cleaning and rapidly multiplying crops like cassava.     

In vitro sucrose concentration affects growth and acclimatization of <Emphasis Type="Italic">Alocasia amazonica</Emphasis> plantlets

Plantlets of Alocasia amazonica were regenerated on the MS medium supplemented with different concentrations (0–9%) of sucrose. An absence of sucrose in the growth medium induced generation of leaves, however, it decreased multiplication. On contrary, sucrose supply of 6% or 9% enhanced multiplication but hampered photoautotrophic growth (generation of leaves). Increasing sucrose supply also increased sugars and starch content and number of stomata and decreased water potential and size of stomata during in vitro growth period. During ex vitro acclimatization, shoot length, root length, leaf number and root number of Alocasia plantlets grown with 3% sucrose, were found to be better among the other studied sucrose concentrations. Under ex vitro acclimatization, number of stomata, contents of various carbohydrates in the leaves were increased but size of stomata decreased with increasing sucrose supply during in vitro growth period. Moreover, water potential of leaves of plantlets, which have been grown with a sucrose concentration other than 3%, was decreased. During in vitro growth, net CO₂ assimilation rate (P_N), transpiration (E), stomatal conductance (g_s) and variable fluorescence to maximum fluorescence ratio (Fv/Fm) were unaffected, however, during acclimatization these were changed and maximum P_N, E, and g_s were observed in the plantlets micropropagated with 3% sucrose. Fv/Fm was decreased severely in the plantlets micropropagated with 6% sucrose during acclimatization. Thus a sucrose concentration of 3% in the medium is appeared to be better among studied concentrations for both in vitro growth and ex vitro acclimatization of A. amazonica plantlets.     

Influence of boron on the growth and biochemical changes in plant growth promoting rhizobacteria (PGPR) inoculated banana plantlets

The effects of rhizobacteria inoculation in modified MS medium containing boron (1 and 10 μM) on the biochemical components, physiological characteristics and mineral content of the in vitro banana plantlets were carried out. The presence of rhizobacteria in the medium supplemented with boron at two concentrations: 1 and 10 μM resulted in an improvement in growth and root biomass compared to the control (uninoculated). Rhizobacteria inoculation also produced an increase in protein, nitrate, soluble nitrogen and chlorophyll contents of the plantlets cultured in MS modified medium containing boron. An increase in percentage of growth (>295%) was shown when boron was applied into medium inoculated with Bacillus sphaericus UPMB10. The effectiveness of inoculation is increased when associated with boron, nitrogen or carbon into the medium. Thus, these bacterial strains could be used as a bioenhancer for growth of in vitro banana plantlets.     

Wood formation during ex vitro acclimatisation in micropropagated true service tree (<Emphasis Type="Italic">Sorbus domestica</Emphasis> L.)

Micropropagated plantlets derived from a superior 90+-year-old slow-growing true service tree (Sorbus domestica L.) have been successfully acclimatised to the ex vitro environment. The temporal pattern of developmental changes was examined in relation to secondary xylem growth during ex vitro acclimatisation. In vitro rooted plantlets already initiated lignification of secondary xylem cells. During early days after transfer to ex vitro conditions, the growth of woody tissue was slow. The most prominent increase in woody tissue development occurred between days 7 and 35. From days 35 to 63, sizes of vessel lumen areas significantly increased. In developing woody tissue, a characteristic diffuse-porous pattern of roughly even vessel area distribution throughout the growing season, typical for mature wood, was not followed. The proportion of woody area occupied by vessels was significantly higher in stems of fully acclimatised plantlets than in stems sampled 35 days after transfer. On day 63 after transfer, a proportion of woody tissue area in fully acclimatised plantlets represented up to 14.8% of the stem area. Early formation of wood during ex vitro acclimatisation provides brittle plantlets with a mechanical support to cope better with deformations and mechanical injuries during handling at the subsequent transplantations.