In Vitro Regeneration of Mat Sedge (Cyperus pangorei Rottb)

An in vitro protocol was developed for regeneration of Cyperus pangorei that may supplement enough raw materials for the mat weaving community. Callus was initiated from inflorescence explants on Murashige and Skoog’s (MS) medium supplemented with 5 and 10 μM each of 2, 4-D, 2, 4, 5-T and CPA. Development of numerous de novo spikelets from immature inflorescence explants grown in (10 μM) 2, 4, 5-T was observed. MS with 5 μM Kn and 100 ml l^?1 Coconut milk (CM) promoted shoot regeneration from calli. Calli from 2,4-D and CPA medium sub-cultured on medium containing 5 μM BAP, 5 μM Kn, 1 μM IAA and 100 ml l^?1 CM produced extensive and rapid rhizogenesis with wiry and scaly roots. Micropropagation using rhizome buds on MS medium with BAP, Kn and Zeatin at 10 μM concentrations resulted in shoot release and multiplication by breaking the bud dormancy. An average of 10 shoots per explant was produced in 10 μM BAP, whereas (10 μM) Kn and (10 μM) Zeatin induced only single shoot formation. The shoots were transferred to rooting media comprising 10 μM IAA with 1 μM BAP or Kn and then acclimatized. The results accomplished were found to be useful in developing a complete in vitro regeneration protocol towards the mass production of Cyperus species, which may provide a basis for further genetic improvements that may prove its use as an alternative natural fibre resource in commercial applications.     

Plantlet Regeneration via Multiple Shoot Induction in Indian Cultivars of Lucerne (Medicago sativa L)

An efficient protocol has been developed for in vitro plant regeneration via multiple shoot induction in lucerne (Medicago sativa L). Shoot tips from in vitro grown 5–6 days old seedlings of 3 cultivars, LLC-3, Chetak and RL-88 were used as explants for multiple shoot induction on MS medium supplemented with cytokinins. Maximum of 14 shoots per apical meristem were observed in case of cv Chetak on MS medium supplemented with BAP (12.6 μM) and KN (9.3 μM). Shoot elongation on MS medium supplemented with GA (5.8 μM), while root induction was achieved on MS medium supplemented with IAA (11.4 μM) and activated charcoal (2.0 g l^?1). Tissue raised plants showed 75% survival after transfer to soil under field conditions.     

Callus induction and shoot organogenesis from anther cultures of Curcuma attenuata Wall

Curcuma attenuata is a highly valued ornamental. This study provides the first report on C. attenuata shoot organogenesis and plant regeneration. Immature anthers derived from 5 to 7?cm long inflorescences were isolated and cultured on different variations of Murashige and Skoog (MS) media to induce callus and then shoot organogenesis. When the 2-mm long anthers in which microspores were at the uninucleate developmental stage were cultured in the dark on MS medium containing 13.6???M 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.3???M kinetin (KT) for 15?days and then transferred to 40???mol?m^?2?s^?1 fluorescent light for 30?days, the percentage callus induction reached 33.3?%. After callus was transferred to various differentiation media and cultured in the light, 33.1?% of all callus cultures could differentiate into adventitious shoots on MS medium supplemented with 22.0???M 6-benzyladenine (BA), 0.53???M ??-naphthaleneacetic acid (NAA) and 1.4???M thidiazuron (TDZ) after culturing for 60?days. Over 95?% of plantlets survived after transplanting plantlets into trays with a mixture of sand and perlite (2: 1) for 20?days. Chromosome number, determined from the root tips of young plantlets, indicated that all plantlets were diploid (2n?=?84).     

GROWTH,PHOTOSYNTHESIS AND MAINTENANCE METABOLIC COST IN THE DIATOM PHAEODACTYLUM TRICORNUTUM AT VERY LOW LIGHT LEVELS 1

The compensation point for growth of Phaeodactylum tricornutum Bohlin is less than 1 μmol. m^?2s^?1. Growth at low PFDs (<3.5 μmol. m^?2.s^?1) does not appear to reduce the maximum quantum efficiency of photosynthesis (ø_m) or to greatly inhibit the potential for light-saturated, carbon-specific photosynthesis (P_m^c). The value for ø_m in P. tricornutum is 0.10–0.12 mol O₂-mol photon^?1, independent of acclimation PFD between 0.75 and 200 μmol.m^?2.s^?1 in nutrient-sufficient cultures. P_m^c in cells of P. tricornutum acclimated to PFDs <3.5 μmol m^?2?s^?1 is approximately 50% of the highest value obtained in nutrient-sufficient cultures acclimated to growth-rate-saturating PFDs. In addition, growth at low PFDs does not severely restrict the ability of cells to respond to an increase in light level. Cultures acclimated to growth at lees than 1% of the light-saturated growth rate respond rapidly to a shift-up in PFD after a short initial lag period and achieve exponential growth rates of 1.0 d^?1 (65% of the light- and nutrient-saturated maximum growth rate) at both 40 and 200 μmol.m^?2.s^?1     

Efficient In vitro Plant Regeneration Protocol from Leaf Explant of Jatropha curcas L — A Promising Biofuel Plant

An efficient in vitro plant regeneration from leaf-disc culture of Jatropha curcas L has been established. Adventitious shoot buds along with callus were induced from leaves of 2-year-old J. curcas plants cultured on Murashige and Skoog’s (MS) medium supplemented with TDZ (2 μM) BAP (2 μM) and IBA (1 μM), wherein 63.3% leaf explants responded. The multiplication of shoots was achieved from the adventitious shoot buds after transferring them to shoot induction medium. The highest number of shoots (9.7/explant) was achieved after 6 weeks of culture on MS medium containing 3 μM of BAR The welldeveloped shoots were rooted on MS medium supplemented with IBA (1.5 μM) with the rooting frequency of 53.3%. Addition of phloroglucinol (200 μM) to the medium enhanced the frequency of rooting to 76.7%. Regenerated plantlets were successfully transferred to field after initial acclimatization.     

Zeatin and TDZ-induced Shoot Proliferation and Use of Bioreactor in Clonal Propagation of Medicinal Herb, Roseroot (Rhodiola rosea L)

A procedure for in vitro propagation of roseroots (Rhodiola rosea L), a medicinal plant, was developed using a RITA bioreactor system containing liquid medium, combined with a gelled medium. Wild roseroot clones: ‘RCi’, ‘RC2’ and ‘RC3’ were established on a basal medium (BM) from in vitro-germinated seedlings on half-strength Murashige and Skoog (MS) salts. TDZ at 2–4 μM supported shoot proliferation but inhibited shoot elongation of ‘RCi’ shoots on gelled medium. Clones differed significantly with respect to multiplication rate with ‘RCi’ producing the most shoots per explant on gelled BM with 2 μM zeatin. In a bioreactor system, TDZ supported rapid shoot proliferation at lower concentration (0.5 μM) but induced hyperhydricity at more than 0.5 μM. Bioreactor-multiplied hyperhydric shoots of all clones when transferred to gelled medium containing 1–2 μM zeatin produced normal shoots within 4 wk of culture. Shoots were rooted in vitro on BM void of growth regulators. Almost all (9U to 95%) in vitro plantlets survived when transferred to potting medium.     

Enhanced in vitro multiplication of Nothapodytes nimmoniana Graham using semisolid and liquid cultures and estimation of camptothecin in the regenerated plants

Nothapodytes nimmoniana (Icacinaceae) yields camptothecin (isoquinoline alkaloid) which is a potent anti-cancer drug. The major objectives of the present study were to develop an efficient protocol for mass propagation of N. nimmoniana using liquid medium and to compare regeneration with semisolid cultures; as also to quantify the amount of camptothecin in regenerated plants. Adventitious shoots were induced from the callus derived from nodal explants on semisolid and liquid Murashige and Skoog (MS) medium supplemented with 1.0, 2.0, 5.0 and 10.0???M 6-benzylaminopurine or kinetin or 2-isopentenyl adenine (2-iP). The highest number of adventitious shoots was regenerated on medium supplemented with 2.0???M BAP. Compared to semisolid medium (41.9 shoots per explant), liquid medium (165.9 shoots per explant) was found suitable for shoot induction and shoot multiplication. Shoots were rooted on MS semisolid medium of one-fourth strength containing IBA (2.4???M) and IAA (5.7???M). The plantlets were acclimatized in a growth chamber at 25°C, 60% relative humidity, with 16-h photoperiod (40???mol?m^?2?s^?1). The camptothecin content was determined in ex vitro plants using HPLC. The analysis revealed that the leaves and stems of ex vitro plants had a considerable amount of camptothecin and these plants could be used as a raw material for camptothecin extraction.     

Thidiazuron-induced high-frequency direct shoot organogenesis of <Emphasis Type="Italic">Cannabis sativa</Emphasis> L.

Induction of high-frequency shoot regeneration using nodal segments containing axillary buds from a 1-yr-old mother plants of Cannabis sativa was achieved on Murashige and Skoog (MS) medium containing 0.05–5.0 μM thidiazuron. The quality and quantity of regenerants were better with thidiazuron (0.5 μM thidiazuron) than with benzyladenine or kinetin. Adding 7.0 μM of gibberellic acid into a medium containing 0.5 μM thidiazuron slightly increased shoot growth. Elongated shoots when transferred to half-strength MS medium supplemented with 500 mg l⁻¹ activated charcoal and 2.5 μM indole-3-butyric acid resulted in 95% rooting. The rooted plants were successfully acclimatized in soil. Following acclimatization, growth performance of 4-mo-old in vitro propagated plants was compared with ex vitro vegetatively grown plants of the same age. The photosynthesis and transpiration characteristics were studied under different light levels (0, 500, 1,000, 1,500, or 2,000 μmol m⁻² s⁻¹). An increase in photosynthesis was observed with increase in the light intensity up to 1,500 μmol m⁻² s⁻¹ and then decreased subsequently at higher light levels in both types of plants. However, the increase was more pronounced at lower light intensities below 500 μmol m⁻² s⁻¹. Stomatal conductance and transpiration increased with light intensity up to highest level (2000 μmol m⁻² s⁻¹) tested. Intercellular CO₂ concentration (C _i) and the ratio of intercellular CO₂ concentration to ambient CO₂ (C _i/C _a) decreased with the increase in light intensity in both in vitro as well as ex vitro raised plants. The results show that in vitro propagated and hardened plants were functionally comparable to ex vitro plants of same age in terms of gas and water vapor exchange characteristics, within the limits of this study.     

Direct shoot regeneration from intact leaves of Arnebia euchroma (Royle) Johnston using thidiazuron

The present study highlights the importance of preculture time and concentration of TDZ (thidiazuron) for direct regeneration from in vitro leaves (attached to shoots) in Arnebia euchroma. Shoot buds proliferated to form multiple shoots on MS medium (Murashige and Skoog medium) with 5.0 μM Kn. Different additives viz. ascorbic acid, PVP (polyvinylpyrrolidone), PVPP (polyvinylpolypyrrolidone) or activated charcoal (50, 100 and 250 mg/l each) were used to check the phenolic exudations. Direct shoot regeneration was obtained when shoots were initially precultured for 40 days on medium with a higher concentration of TDZ (20.0 μM) and then transferred to a lower concentration (5.0 μM TDZ). The identity of shoot buds was confirmed by histological studies. Regenerated shoots were cultured for 30 days on medium containing Kn (5.0 μM) for proliferation and then transferred to IBA (0.25 μM)‐containing medium for rooting. Rooted plantlets were transferred to greenhouse with 45–50% survival.     

THE INTERACTION BETWEEN INORGANIC CARBON ACQUISITION AND LIGHT SUPPLY IN PALMARIA PALMATA (RHODOPHYTA)1

The dependence of the carbon concentrating mechanism of Palmaria palmata (L.) Kuntze on the growth light level was examined 1) to determine whether or not there is a threshold photon flux density (PFD) at which the inorganic carbon uptake mechanism can operate and 2) to attempt to quantify the relative energetic costs of acclimation to the two different limiting factors, PFD and dissolved inorganic carbon (DIC) concentration. Plants were grown at six PFDs: 5, 25, 50, 75, 95, and 125 μmol photons. m^?2.s^?1. Growth rates increased with increasing PFD from 5 to 50 μmol photons. m^?2. s^?1 and were light-saturated at 75, 95, and 125 μmol photons. m^?2. s^?1 Values of δ¹³C increased continuously with increasing growth PFD and did not saturate over the range of light levels tested. Time-resolved fluorescence characteristics indicated a progressive photoacclimation below 50 μmol photons. m^?2. s^?1. Analysis of chlorophyll fluorescence induction showed three levels of light use efficirncy associated with growth at 5 or 25, 50, and >75 μmol photons. m^?2. s^?1. The light-haruesting efficiency was inversely proportional to the effectiveness of DIC acquisition in plants grown at the six PFDs. These data were interpreted to indicate that there is a physiological tradeoff between photosynthetic efficiency and bicarbonate use in this species.     

Rapid In Vitro Propagation of Eclipta alba (L) Hassk by Shoot Tip Culture

An efficient and rapid tissue culture system employing shoot tip explants has been developed for Eclipta alba (L) Hassk, an important medicinal plant of the family Asteraceae. The highest shoot regeneration frequency (95%) as well as the maximum number (32.2 ± 0.4) of shoots was recorded on MS medium amended with BA (5 μM) and NAA (0.5 μM). The regenerated shoots rooted best on MS medium supplemented with 0.2 μM IBA. The in vitro developed plantlets were acclimatized successfully with 100 % survival.     

Enhanced multiplication and improved <Emphasis Type="Italic">ex vitro</Emphasis> acclimatization of <Emphasis Type="Italic">Decalepis arayalpathra</Emphasis>

The proposed work describes a protocol for high-frequency in vitro regeneration through nodal segments and shoot tips in Decalepsis arayalpathra, a critically endangered medicinal liana of the Western Ghats. Nodal segments were more responsive than shoot tips in terms of shoot proliferation. Murashige and Skoog’s (MS) basal medium supplemented with 5.0 μM 6-benzyladenine (BA) was optimum for shoot initiation through both the explants. Among different combinations of plant growth regulators and growth additive screened, MS medium added with 5.0 μM BA + 0.5 μM indole-3-acetic acid + 20.0 μM adenine sulphate effectuated the highest response: 11.8 shoots per nodal segment and 5.5 shoots per shoot tip with mean shoot length of 9.2 and 4.8 cm, respectively. Half-strength MS medium with 2.5 μM α-naphthalene acetic acid was optimum for in vitro root induction. The plantlets with the well developed shoot and root were acclimatized in Soilrite? with 92 % survival rate in the field conditions. During acclimatization, chlorophyll content, net photosynthetic rate, stomatal conductance, and transpiration rate were gradually changed in dependence of formation of new leaves. Further, the changes in activities of antioxidant enzymes, i.e., superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) as well as activity of carbonic anhydrase were also observed: a continuous rise in SOD activity, but a rise and fall in the activities of CAT, APX, and GR were also noticed. Maximum fresh mass (3.1 g plant^-1), dry mass (0.35 g plant^-1) of roots and 2-hydroxy-4-methoxybenzaldehyde content of 9.22 μg cm^-3(root extract) were recorded after 8 weeks of acclimatization.     

Improved micropropagation of Gypsophila paniculata with bioreactor and factors affecting ex vitro rooting in microponic system

Studies on the mass production of high-quality plantlets in Gypsophila paniculata L. using a bioreactor and microponic system (a hydroponic system in which micropropagation shoots are planted) indicated that both aeration treatments, in which bioreactors were aerated from the top of explants by sparger (AS) and by tub (AT), were more effective than unaerated treatment for shoot proliferation and growth, and the maximum shoots (15.7 shoots per explant) with low hyperhydricity rate (2.9%) were found in the AS group. The ex vitro culture was more efficient for rooting when compared to the in vitro culture; the better shoot and root growth was obtained in the ex vitro culture, with rooting rate reaching 100% after 20 d of culture, but only 65% of in vitro shoots rooted; all stomata of ex vitro shoots closed, and their length was more than their width, but the stomata in in vitro shoots were all opened, the length close to the width. Furthermore, the stomata numbers were less in ex vitro (67.8) than in vitro (267.2). The survival rate of ex vitro plants reached 83.3% when plantlets derived in vitro and ex vitro were transferred to pots, while only 23.3% of in vitro plantlets survived. During ex vitro rooting with the microponic system, foam as the supporter material, 90 μmol?m^?2?s^?1 of light, and 80 shoots of planting density were favorable for shoot and root growth. The combination of bioreactor and microponic systems is an efficient way to produce high-quality plantlets of G. paniculata. Their application can reduce costs during large-scale industrial production.     

Anatomical and Biochemical Changes Associated with In Vitro Rhizogenesis in Dendrocalamus giganteus

Successful micropropagation protocol of a difficult-to-root bamboo species, Dendrocalamus giganteus (10–15 years old) along with the analysis of anatomical and biochemical changes during in vitro rhizogenesis was accomplished. Proliferated axillary shoots from nodal segments of 10–15 years old field culms exhibited shoot necrosis during multiple shoot formation phase and was controlled by subculturing in modified MS liquid medium having 825 mg ^l?1 NH₄NO₃, 3800 mg ^l?1 KNO₃, 740 mg ^l?1 MgSO₄ and 9% coconut water, 26.64 μM 6-benzylaminopurine (BA) and 0.46 μM kinetin. These multiple shoots proliferated from field grown culms, failed to root and hence callus was induced on MS solid medium containing 4.44 μM BA, 4.52 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 5.37 μM naphthalene acetic acid (NAA). Organogenesis from the callus was achieved upon transfer to MS medium with 11.10 μM BA and 2.32 pM kinetin. The callus-derived shoots multiplied on modified MS medium were rooted the best (91%) by culturing 3 days on MS medium having glucose (0.5%), sucrose (2.5%) and 98.41 μM indolebutyric acid (IBA) and subsequently to IBA-free MS medium containing 3% sucrose. Studies on peroxidase and IAA oxidase activity and endogenous free- and bound-IAA content showed that IAA oxidase and peroxidase oxidize endogenous IAA resulting in root initials formation. Anatomical studies confirmed the root primordia formation from 3^rd day of IBA treatment and primordia were visible over the surface on 8^th to 10^th day. However, the shoot necrosis symptoms which started on 6^th day of treatment intensified by 10^th day leading to the death of the whole shoot system by 12^th–15^th day. Nevertheless, on the root formation medium with 9.84 μM IBA, new shoot buds were emerged and showed shoot growth in 60% of the rooted cultures, which were successfully acclimatized in shade-house with 100% survival. The present study establishes rooting of callus-derived shoots as the best way for the successful propagation of the difficult-to-root bamboo, D. giganteus when compared to axillary bud proliferated shoots.     

Improved production of <Emphasis Type="Italic">Oroxylum indicum</Emphasis> (L.) Kurz by altering the salts of the medium

The present study was conducted to test the effects of KNO₃, KH₂PO₄, and CaCl₂ on shoot multiplication, root proliferation, and accumulation of phytochemicals in in vitro cultures of Oroxylum indicum. The results indicate that modifying the MS salt formulation in relation to particular inorganic nutrients highly affected shoot multiplication, root proliferation, and accumulation of flavonoids in in vitro cultures. A concentration of 0.60 g L^?1 CaCl₂ resulted in the highest frequency of shoot regeneration (5.6 shoots per explant). A concentration of 0.40 g L^?1 CaCl₂ resulted in the highest frequency of root regeneration (7.8 roots per shoot). Modifications of the concentrations of inorganic salts were also found to be advantageous for production media for both multiple shoots and shoot-derived root in vitro cultures. Multiple shoots generated on shoot induction medium with a concentration of 0.60 g L^?1 CaCl₂ and roots generated on root induction medium with a concentration of 1.5 g L^?1 KNO₃ yielded about a five times higher flavonoid level than cultures generated on control medium respectively.     

Changes in photon flux can induce stomatal patchiness

Images of chlorophyll fluorescence were used to detect the occurrence of stomatal patchiness in leaves from eight species under variable photon flux conditions. Pronounced stomatal patchiness was induced within 5–10 min after PFD was changed from intermediate (～450 μmol quanta m^?2 s^?1) to low (～150 μmol quanta m^?2 s^?1) levels. This effect was completely reversible by returning PFD to intermediate levels. The pattern of heterogeneous fluorescence for each leaf was usually similar during repeated applications of medium and low PFD. In three species, stomatal patchiness could only be induced in slightly water-stressed plants. Leaves of more severely water-stressed Xanthium strumarium plants in low air humidity exhibited oscillations in fluorescence that corresponded with oscillatory changes in leaf diffusion conductance for water vapour. Stomatal patchiness was also induced by illuminating dark-adapted leaves with low PFD (below 200–300 μmol quanta m^?2 s^?1). Infiltration of leaves with distilled water showed that heterogeneous chlorophyll fluorescence was caused by changes in stomatal apertures.     

LIGHT DEPENDENCE OF GROWTH AND PHOTOSYNTHESIS IN PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE)1

Phaeodactylum tricornutum Bohlin was maintained in exponential growth over a range of photon flux densities (PFD) from 7 to 230 μmol·m^?2s^?1. The chlorophyll a-specific light absorption coefficient, maximum quantum yield of photosynthesis, and C:N atom ratio were all independent of the PFD to which cells were acclimated. Carbon- and cell-specific, light-satuated, gross photosynthesis rates and dark respiration rates were largely independent of acclimation PFD. Decreases in the chlorophyll a-specific, gross photosynthesis rate and the carbon: chlorophyll ratio and increases of cell- or carbon-specific absorption coefficients were associated with an increase in cell chlorophyll a in cultures acclimated to low PFDs. The compensation PFD for growth was calculated to be 0.5 μmol·m^?2s^?1. The maintenance metabolic rate (2 × 10^?7s^?1), calculated on the basis of the compensation PFD, is an order of magnitude lower than the measured dark respiration rate(2.7 × 10^?6mol O₂·mol C^?1s^?1). Maintenance of high carbon-specific, light-saturated photosynthesis rates in cells acclimated to low PFDs may allow effective use of short exposures to high PFDs in a temporally variable light environment.     

Chlorophyll fluorescence quenching and violaxanthin deepoxidation of FBPase antisense plants at low light intensities and low temperatures

Genetically modified potato (Solanum tuberosum L. cv. Desiree) and tobacco (Nicotiana tabacum cv. Samsun N.N.) plants were used to analyze the effects exerted by the chloroplastic (cp) fructose- 1,6-bisphosphatase (FBPase) on the regulation of light energy discrimination at the level of photosystem II. The cp-FBPase activity was progressively inhibited by an mRNA antisense to this FBPase. The chlorophyll fluorescence quenching parameters of these transgenic plants were compared to those of wild-type and transgenic plants that were acclimated to low temperatures. In particular various lines of the transgenic potato and tobacco plants were exposed to a temperature treatment of 10 and 20°C for 10 days. Light intensities were kept low to reduce photoinhibition so that we could analyze exclusively the effects of a modification in the carbon fixation cycle on the chlorophyll fluorescence quenching parameters. The photon flux densities (PFDs) employed at the level of the middle leaves of all plants were set to two different values of 10 μmol m^?2 s^?1 and 50 μmol m^?2 s^?1. Subsequent to this 10-day acclimation the chlorophyll-fluorescence parameters of all plants were measured. Photoinhibition as expressed by the F_y/F_m ratio was minor in plants subjected to a PFD of 10 μmol m^?2 s^?1. Higher photon fluence rates of 50 μmol m^?2 s^?1 at temperatures of 10°C gave rise to a significant reduction in the F_y/F_m ratios obtained from the transgenic plants which were characterized by a restriction in cp-FBPase capacity to 20% of normal activity. Furthermore, a progressive inhibition of the cp-FBPase activity induced an amplified nonphotochemical quenching of chlorophyll fluorescence with in the genetically manipulated species (except at 10°C and 50 μmol m^?2 s^?1). The increase in nonphotochemical quenching depended upon light and temperature. Photochemical quenching of light quanta within the antisense plants declined relative to that in the wild type. To further characterize the mechanisms producing higher levels of nonphotochemical fluorescence quenching. we analyzed several of the xanthophyll cycle pigments. The deepoxidation state of the xanthophyll cycle pigments in potato plants increased with attenuating FBPase activities under all conditions. For tobacco plants, this elevation of the deepoxidation state was only observed at a PFD of 50 μmol m^?2 s^?1.     

Effect of CO2 concentration on the PIC/POC ratio in the coccolithophore Emiliania huxleyi grown under light-limiting conditions and different daylengths

We compared the effect of CO₂ concentration ([CO₂], ranging from ∼5 to ∼34 μmol l⁻¹) at four different photon flux densities (PFD=15, 30, 80 and 150 μmol m⁻² s⁻¹) and two light/dark (L/D) cycles (16/8 and 24/0 h) on the coccolithophore Emiliania huxleyi. With increasing [CO₂], a decrease in the particulate inorganic carbon to particulate organic carbon (PIC/POC) ratio was observed at all light intensities and L/D cycles tested. The individual response in cellular PIC and POC to [CO₂] depended strongly on the PFD. POC production increased with rising [CO₂], irrespective of the light intensity, and PIC production decreased with increasing [CO₂] at a PFD of 150 μmol m⁻² s⁻¹, whereas below this light level it was unaffected by [CO₂]. Cell growth rate decreased with decreasing PFD, but was largely independent of ambient [CO₂]. The diurnal variation in PIC and POC content, monitored over a 38-h period (16/8 h L/D, PFD=150 μmol m⁻² s⁻¹), exceeded the difference in carbon content between cells grown at high (∼29 μmol l⁻¹) and low (∼4 μmol l⁻¹) [CO₂]. However, consistent with the results described above, cellular POC content was higher and PIC content lower at high [CO₂], compared to the values at low [CO₂], and the offset was observed throughout the day. It is suggested that the observed sensitivity of POC production for ambient [CO₂] may be of importance in regulating species-specific primary production and species composition.     

Improved in vitro shoot multiplication and rooting of <Emphasis Type="Italic">Dendrocalamus hamiltonii</Emphasis> Nees et Arn. Ex Munro: production of genetically uniform plants and field evaluation

An efficient in vitro regeneration protocol and field performance of a multipurpose bamboo species Dendrocalamus hamiltonii Nees et Arn. Ex Munro has been demonstrated using single node cuttings taken from the lateral branches of a 20-year-old bush. Axillary buds on the nodal explant sprouted within 10 days of culture on Murashige and Skoog (MS) medium without any plant growth substance. High-frequency proliferation was induced on the propagules (small clusters with 3–5 multiple shoots and rhizomatous portions). Subsequent removal of the shoots (about 1.5 cm) from the rhizomatous portion of propagules (shoot cut) influenced the plantlet formation capacity. A multiplication of about 20-folds was achieved on MS medium supplemented with 8 μM BAP and 1 μM NAA. Rooting efficiency was also markedly enhanced (>90%) when the propagules, following shoot cut, were placed on to MS medium supplemented with 100 μM IBA for 10 days and then transferred to IBA-free medium. This is the first report from this species where 20-fold increment in multiplication was observed at the end of second subculture followed by >90% rooting. The hardened plants, established in the field, exhibited normal growth; their physiological performance has been monitored at 6-month intervals. The rate of photosynthesis increased from 3.55 μmol CO₂ m⁻² s⁻¹ (hardened, ready for field transfer) to 5.44 μmol m⁻² s⁻¹ (6 months of field transfer); following a year of plantation net photosynthesis recorded was 14.0 μmol CO₂ m⁻² s⁻¹ while after 1.5 years it was 12.76 μmol CO₂ m⁻² s⁻¹. These values were compared with those observed for the mother bush. Genetic fidelity of these regenerants was established by RAPD analysis advocating clonal propagation of this species through nodal segment culture and its commercial cultivation.