Concentrations Of NAA Research Articles

Somatic embryogenesis and indirect regeneration in Mirabilis jalapa Linn.

Indirect somatic embryogenesis and indirect regeneration protocol was developed for the first time from leaf and nodal explants of Mirabilis jalapa Linn. Initially callus was induced in leaf and nodal explants cultured on MS medium supplemented with various concentrations of 2, 4-D, NAA and Kn, after 3 weeks of culture. Maximum amount of mean fresh and dry weight (mg) of callus(189.6 ± 0.25, 7.6 ± 0.23); (198.4± 0.34, 9.2 ± 0.23)was obtained from leaf and nodal explants, respectively when cultured on MS media supplemented with combinations of 2,4-D(3 mg/L) and NAA(5mg/L).At high concentrations of NAA (5 mg/L), somatic embryos are induced from the nodal explants after 4 weeks of culture. Shoot buds were induced from the leaf and nodal calluses when cultured on combinations of TDZ & BAP after 3 weeks of culture. Maximum number of shoot buds (08 ± 0.17 & 06 ± 0.09) were produced from leaf and nodal calluses cultured on MS + TDZ (0.5mg/L) +BAP (2 mg/L).When micro shoots were transferred to rooting media, the best rooting percentage (100%) was obtained on MS + IAA (1.0 mg/L) + IBA (2.0 mg/L). The raised plantlets when transferred to soil for acclimatization, 75% plantlets survived in field conditions.

Bioactive compounds in hyperhydric and normal micropropagated shoots of Aronia melanocarpa (Michx.) Elliott

An efficient in vitro propagation protocol was developed for the production of bioactive compounds from shoot cultures of Aronia melanocarpa (Michx.) Elliott. Nodal explants obtained from mature field-grown plants of A. melanocarpa were cultured on Murashige and Skoog (MS) medium supplemented with 0.25, 0.5, 1.0 or 2.0mgl−1 thidiazuron (TDZ) alone or in combination with 0.1 or 0.5mgl−1 α-naphthaleneacetic acid (NAA) for shoot multiplication. Among the various concentrations of TDZ studied, the maximum frequency of shoot multiplication (85.2%), mean number of (10.4) shoots per explant and mean shoot length (3.2cm) were obtained on MS medium fortified with 0.5mgl−1 TDZ. The culture medium containing TDZ with low concentration of NAA produced more shoots than with high NAA concentration. The highest frequency of shoot multiplication (98.9%), mean number of (19.8) shoots per explant and mean shoot length (3.8cm) were obtained on MS medium fortified with 0.5mgl−1 TDZ and 0.1mgl−1 NAA. Hyperhydric shoots were observed on MS medium fortified with 2.0mgl−1 TDZ and 0.1mgl−1 NAA. The highest frequency of rooted shoots (100%), mean number of (10.6) roots per shoot and mean root length (5.4cm) were obtained on 1/2 MS medium fortified with 1.0mgl−1 IBA after 30 days of culture. The in vitro-developed plantlets were successfully acclimatized in the greenhouse with 100% survival. The highest contents of carotenoids, α-tocopherol and fatty acids were found in leaves collected from greenhouse-grown in vitro plants followed by normal and hyperhydric leaves obtained from in vitro-regenerated shoots. All-E-lutein was the most abundant carotenoid (65.1–191.4μgg−1 fresh weight) in A. melanocarpa leaf samples. Linolenic acid was the most abundant fatty acid in A. melanocarpa leaf samples tested, comprising 45.91–50.04% of total fatty acid.

Optimizing Tuber Set and Size Distribution for Potato Seed (Solanum tuberosum L.) Expressing Varying Degrees of Apical Dominance

Plant emergence, apical dominance, tuber set, and size are affected by the physiological age of seed tubers, which can substantially impact overall crop value. This study investigated the efficacy of seed spacing (15, 25, and 35 cm) and 1-naphthaleneacetic acid (NAA) seed treatments in altering these variables in cv. Ranger Russet to improve yield and tuber size distribution of seed expressing low (2.8 stems/seed piece) and high (4.8–5.4 stems/seed piece) apical dominance. Age primed, high-stem seed produced more tubers per plant and per ha than non-aged seed; however, tuber number per ha from both seed lots fell to the same extent with decreasing plant density. Importantly, tuber set per plant increased substantially more for the physiologically older, high-stem seed as plant spacing increased. Average tuber weight also increased with decreasing plant density but the response was greatest for the physiologically younger, low-stem seed. Regardless of seed age, marketable yields were comparable at 25- and 35-cm spacing. Tuber size distributions from the 2.8-stem seed shifted from oversize (>340-g) tubers to higher percentage 113–284-g tubers as spacing decreased from 35 to 15 cm. The 5.4-stem seed produced less undersize ( 284-g tubers when planted at 35-cm spacing. Adjusting in-row spacing relative to seed age and expected stem numbers improved tuber size distribution and value. However, because plants from older seed set more tubers in response to decreasing plant density than younger seed, average tuber weight and size distribution never matched the younger seed at any spacing. Restoration of apical dominance by treatment of seed with NAA was more effective in this regard. Depending on seed age, NAA delayed plant emergence (22–74 %) and decreased stem (24–38 %) and tuber numbers per plant (8–13 %). Stem numbers from age-primed seed fell from 4.8 to 3.0 as NAA concentration increased. Marketable yields were not affected by seed age but decreased slightly (7.3 %, P 284-g) tubers, resulting in a yield profile approaching that of the non-treated younger seed. Although seed spacing and NAA treatments are effective techniques for altering tuber size distribution to maximize crop value in relation to seed age and expected stem numbers, tuber age had a small but residual effect on productivity beyond that attributable to apical dominance.

Detection of Normal Aging Effects on Human Brain Metabolite Concentrations and Microstructure with Whole-Brain MR Spectroscopic Imaging and Quantitative MR Imaging.

Knowledge of age-related physiological changes in the human brain is a prerequisite to identify neurodegenerative diseases. Therefore, in this study whole-brain (1)H-MRS was used in combination with quantitative MR imaging to study the effects of normal aging on healthy human brain metabolites and microstructure. Sixty healthy volunteers, 21-70 years of age, were studied. Brain maps of the metabolites NAA, creatine and phosphocreatine, and Cho and the tissue irreversible and reversible transverse relaxation times T2 and T2' were derived from the datasets. The relative metabolite concentrations and the values of relaxation times were measured with ROIs placed within the frontal and parietal WM, centrum semiovale, splenium of the corpus callosum, hand motor area, occipital GM, putamen, thalamus, pons ventral/dorsal, and cerebellar white matter and posterior lobe. Linear regression analysis and Pearson correlation tests were used to analyze the data. Aging resulted in decreased NAA concentrations in the occipital GM, putamen, splenium of the corpus callosum, and pons ventral and decreased creatine and phosphocreatine concentrations in the pons dorsal and putamen. Cho concentrations did not change significantly in selected brain regions. T2 increased in the cerebellar white matter and decreased in the splenium of the corpus callosum with aging, while the T2' decreased in the occipital GM, hand motor area, and putamen, and increased in the splenium of the corpus callosum. Correlations were found between NAA concentrations and T2' in the occipital GM and putamen and between creatine and phosphocreatine concentrations and T2' in the putamen. The effects of normal aging on brain metabolites and microstructure are region-dependent. Correlations between both processes are evident in the gray matter. The obtained data could be used as references for future studies on patients.

A Longitudinal Assessment of Structural and Chemical Alterations in Mixed Martial Arts Fighters

Growing evidence suggests that temporally proximal acute concussions and repetitive subconcussive head injuries may lead to long-term neurological deficits. However, the underlying mechanisms of injury and their relative time-scales are not well documented in human injury models. The current study therefore investigated whether biomarkers of brain chemistry (magnetic resonance [MR] spectroscopy: N-acetylaspartate [NAA], combined glutamate and glutamine [Glx], total creatine [Cre], choline compounds [Cho], and myo-inositol [mI]) and structure (cortical thickness, white matter [WM]/subcortical volume) differed between mixed martial artists (MMA; n = 13) and matched healthy controls (HC) without a history of contact sport participation (HC; n = 14). A subset of participants (MMA = 9; HC = 10) returned for follow-up visits, with MMA (n = 3) with clinician-documented acute concussions also scanned serially. As expected, MMA self-reported a higher incidence of previous concussions and significantly more cognitive symptoms during prior concussion recovery. Fighters also exhibited reduced memory and processing speed relative to controls on neuropsychological testing coupled with cortical thinning in the left posterior cingulate gyrus and right occipital cortex at baseline assessment. Over a 1-year follow-up period, MMA experienced a significant decrease in both WM volume and NAA concentration, as well as relative thinning in the left middle and superior frontal gyri. These longitudinal changes did not correlate with self-reported metrics of injury (i.e., fight diary). In contrast, HC did not exhibit significant longitudinal changes over a 4-month follow-up period (p > 0.05). Collectively, current results provide preliminary evidence of progressive changes in brain chemistry and structure over a relatively short time period in individuals with high exposure to repetitive head hits. These findings require replication in independent samples.

PENGARUH NAA DAN BAP TERHADAP EKSPLAN PEGAGAN (Centella asiatica (L.) Urb.)

Pegagan ( Centella asiatica (L.) Urb.) representing one of theplant which good of as drug. Plantof pegagan ( Centella asiatica (L.) Urb.) good of to launch the urine, degrading blood pressure and quicken to heal the hurt. Ready of the seed for the crop of drug which require to be paid attention by quality from itself seed. One of the alternative to get the uniform seed and a flash in the pan is with the technique of tissue culture. Tissue culturelaboratory research was conducted Researchand Development Center for Medicinal Plantsand Traditional MedicineTawangmangu. Research Method use the Random Device of Complete Group at MS (Murashige Skoog) media with the treatment ofplant growth regulator the NAA concentration 0, 1, 3 and 5 mg / l and BAP concentration 0, 1, 2, 3 and 4 mg / l). Result of research show the combination of giving of NAA 1 until 3 mg / l and BAP 1 until 5 mg / l of is condition of explan experience of the change become the callus. Treatment of combination NAA 3 mg / l and BAP 4 mg / l give the best result to callus forming with the quicker callus forming time that is 25 day. Keywords : Pegagan, Centella asiatica (L.) Urb., tissue culture, NAA, BAP

Mass propagation of microtubers from suspension cultures of Pinellia ternata cells and quantitative analysis of succinic acid in Pinellia tubers

The conditions for efficient tuber production from suspension cultures of Pinellia ternata cells which is one of medicinal herbs were established and succinic acid in tubers propagated in vitro was determined. Leaf explants formed white nodular structures and off-white calluses at a frequency of 90.6 % when cultured on Murashige and Skoog medium supplemented with 0.54 μM α-naphthaleneacetic acid (NAA); however, this frequency declined substantially with increasing NAA concentration, up to 16.2 μM, at which the frequency reached zero percent. In combination treatments with 4.44 μM 6-benzyladenine (BA) and NAA, however, the frequency of white nodular structure and off-white callus formation from leaf explants did not decrease, even at 16.2 μM NAA. Suspension cultures of P. ternata cells were established from leaf-derived off-white calluses in MS liquid medium containing 5.4 μM NAA and 4.44 μM BA. Upon plating onto MS basal medium, over 90 % of cell aggregates gave rise to microtubers and developed into plantlets. Regenerated plantlets were transplanted in potting soil and grown to maturity in a growth chamber, with a survival rate of >90 %. The highest succinic acid content in suspension culture-derived microtubers was 45 g/kg of extract. Compared with wild P. ternata medicinal tubers, the succinic acid content was very similar. The in vitro P. ternata microtuber proliferation system established in this study is thus an efficient alternative for the mass production of medicinal tubers.

Accumulation of biologically active phenolic acids in agitated shoot cultures of three Hypericum perforatum cultivars: ‘Elixir’, ‘Helos’ and ‘Topas’

Agitated shoot cultures of three Hypericum perforatum cultivars: ‘Elixir’, ‘Helos’ and ‘Topas’ were established and maintained on Linsmaier and Skoog (LS) and Murashige and Skoog (MS) media containing varying concentrations (0.1–3.0 mg l−1) of α-naphthaleneacetic acid (NAA) and 6-benzyladenine (BA). In methanolic extracts of the biomass, the amount s of free phenolic acids and cinnamic acid were determined by HPLC. Six of the seventeen compounds analyzed were detected in all the extracts: 3,4-dihydroxyphenylacetic acid, p-coumaric acid, protocatechuic acid, syringic acid, and two depsides—chlorogenic and neochlorogenic acids. The amounts of individual compounds and the total amount of phenolic acids depended on the media variants used and the increases ranged from 1.2- to 15.0-fold and from 1.2- to 1.9-fold, respectively. The maximum total amounts in the biomass of ‘Elixir’, ‘Helos’ and ‘Topas’ were 212, 222, and 191 mg 100 g−1 DW, respectively. The main metabolites in all the cultivars were neochlorogenic acid (max. 118.81 mg 100 g−1 DW) and 3,4-dihydroxyphenylacetic acid (max. 129.29 mg 100 g−1 DW). Three metabolites that also accumulated in high amounts were protocatechuic acid (max. 21.43 mg 100 g−1 DW), syringic acid (max. 23.63 mg 100 g−1 DW) and chlorogenic acid (max. 18.79 mg 100 g−1 DW). The richest s ource of the estimated compounds and hence a potential biotechnological platform were the shoots of ‘Helos’ cultivated on LS and MS media containing low concentrations of NAA and BA (0.1–1.0 mg l−1).

Interactive Effects of Growth Regulators, Carbon Sources, pH on Plant Regeneration and Assessment of Genetic Fidelity Using Single Primer Amplification Reaction (SPARS) Techniques in Withania somnifera L.

An improved and methodical in vitro shoot morphogenic approach through axillary bud multiplication was established in a drug yielding plant, Withania somnifera L. Effects of plant growth regulators [6-benzyladenine (BA), kinetin (Kin), 2-isopentenyladenine (2iP), and thidiazuron (TDZ)] either singly or in combination with α-napthalene acetic acid (NAA), indole-3-butyric acid (IBA), and indole-3-acetic acid (IAA) in Murashige and Skoog (MS) medium were tested. The highest regeneration frequency (90 %) with optimum number of shoots (32 ± 0.00)/explant were obtained on MS medium fortified with 2.5 μM 6-benzyladenine (BA) and 0.5 μM NAA and 30 g/l sucrose at pH 5.8. Among the tried TDZ concentrations, 0.5 μM resulted in maximum number of shoots (20.4 ± 0.40)/explant after 4 weeks of exposure. The proliferating shoot cultures established by repeated subculturing of the mother explants on the hormone-free medium produced the highest shoot number (29.4 ± 0.40) with shoot length (6.80 ± 0.12 cm)/explant at fourth subculture passage, which a decline in shoot proliferation was recorded. Different concentrations of NAA were tested for ex vitro rooting of microshoots. The maximum percentage of rooting 100 % with maximum roots (18.3 ± 0.1) was achieved in soilrite when basal portion of the microshoots were treated with 200 μM (NAA) for 15 min per shoot. The plantlets went through hardening phase in a growth chamber, prior to ex vitro transfer. The PCR-based single primer amplification reaction (SPAR) methods which include random amplified polymorphic DNA (RAPD) and direct amplification of minisatellite DNA (DAMD) markers has been used for assessment of genetic stability of micropropagated plantlets. No variation was observed in DNA fingerprinting patterns among the micropropagated and the donor plants illustrating their genetic uniformity.

Efficient in vitro culture protocols for propagating Phalaenopsis ‘Cool Breeze’

Phalaenopsis orchids have high economic value in the floriculture industry as cut flowers and potted plants throughout the world. Plant tissue culture technology is being widely used for large scale plant multiplication of Phalaenopsis to feed into this industry. In order to increase the efficiency of this technology, four experiments were undertaken: Plantlet regeneration from seeds, nodes or leaves, and hardening of the regenerated plants. In the first experiment, seed germination was examined in three media (half MS, Chen and Vacin?Went) of which the Chen medium had the best result (83.4%) in comparison to the other two media. In the second experiment, nodes on the flower stalk were studied for their shoot formation potential to different concentrations of BA and NAA, The highest frequency of shoot regeneration was achieved on MS containing 4 mg/l BA and 1 mg/l NAA while in vitro derived leaves formed clusters of somatic embryos directly when cultured on MS containing TDZ at different concentrations (0.5, 1, 2 and 3 mg/l). The embryos turned green and developed into protocorm?like bodies after 7 weeks of culture followed by plantlet regeneration. The highest plantlet regeneration from the leaf?derived embryos was obtained from MS supplemented with 3 mg/l TDZ. Finally, regenerated plants from (seeds, nodal explants and leaves) were compared in two medium for hardening, regenerated plant from nodal explants showed the highest survival rate (100%) on the medium containing cocopeat, coal, industrial cartridge and the bites of yonolit (1 : 1 : 2 : 4).Plant Tissue Cult. & Biotech. 24(2): 191-203, 2014 (December)

Study of Oil Palm (Elaeis guineensis Jacq) In Vitro Embryogenesis using Young Leaves Explants

This study reported in vitro embryogenesis of oil palm using young leaves as explants. Explants were grown in solid modified MS or Eeuwens medium containing different concentrations of NAA and 2,4-D, i.e. media C1, C2, C3, C4 and C5, M1, M2, M3 and M4, to induce embryogenic calli. Compact and pearly-white, globular calli were obtained from the youngest leaf explants 28 weeks after culture.C1 media (MS medium + 107.41 µM of NAA + 100 mg.L-1 of asparagine + 100 mg.L of glutamine-1) produced the highest percentage of calli formation (30.56%), whereas C4 media (C1 supplemented with 67.86 µM of 2,-D ) was the optimal media for embryogenic callus induction. Direct embryoids were obtained from slightly older leaf explants on the C3 media containing NAA after 36 weeks of culture. However, four subcultures using the same medium with gradual reduction of auxin concentration were not successful to develop embryogenic callus and embryoid cells during the course of this study.

Biomass Yield and Steviol Glycoside Production in Callus and Suspension Culture of Stevia rebaudiana Treated with Proline and Polyethylene Glycol.

Enhanced production of steviol glycosides (SGs) was observed in callus and suspension culture of Stevia rebaudiana treated with proline and polyethylene glycol (PEG). To study their effect, yellow-green and compact calli obtained from in vitro raised Stevia leaves were sub-cultured on MS medium supplemented with 2.0mg l(-1) NAA and different concentrations of proline (2.5-10mM) and PEG (2.5-10%) for 2weeks, and incubated at 24 ± 1°C and 22.4μmol m(-2) s(-1) light intensity provided by white fluorescent tubes for 16h. Callus and suspension culture biomass (i.e. both fresh and dry weight content) was increased with 5mM proline and 5% PEG, while at further higher concentrations, they got reduced. Further, quantification of SGs content in callus (collected at 15th day) and suspension culture (collected at 10th and 15th day) treated with and without elicitors was analysed by HPLC. It was observed that chemical stress enhanced the production of SGs significantly. In callus, the content of SGs increased from 0.27 (control) to 1.09 and 1.83% with 7.5mM proline and 5% PEG, respectively, which was about 4.0 and 7.0 times higher than control. However, in the case of suspension culture, the same concentrations of proline and polyethylene glycol enhanced the SG content from 1.36 (control) to 5.03 and 6.38%, respectively, on 10th day which were 3.7 times and 4.7 times higher than control.

In vitro plant regeneration of Aster scaber via somatic embryogenesis

We established an in vitro plant regeneration system via somatic embryogenesis of Aster scaber, an important source of various biologically active phytochemicals. We examined the callus induction and embryogenic capacities of three explants, including leaves, petioles, and roots, on 25 different media containing different combinations of α-naphthalene acetic acid (NAA) and 6-benzyladenine (BA). The optimum concentrations of NAA and BA for the production of embryogenic calli were 5.0 μM and 0.05 μM, respectively. Media containing higher concentrations of auxin and cytokinin (such as 25 μM NAA and 25 μM BA) were suitable for shoot regeneration, especially for leaf-derived calli, which are the most readily available calli and are highly competent. For root induction from regenerated shoots, supplemental auxin and/or cytokinin did not improve rooting, but instead caused unwanted callus induction or retarded growth of regenerated plants. Therefore, plant growth regulator-free medium was preferable for root induction. Normal plants were successfully obtained from calli under the optimized conditions described above. This is the first report of the complete process of in vitro plant regeneration of A. scaber via somatic embryogenesis.

A single dose of a neuron-binding human monoclonal antibody improves brainstem NAA concentrations, a biomarker for density of spinal cord axons, in a model of progressive multiple sclerosis.

BackgroundIntracerebral infection of susceptible mouse strains with Theiler’s murine encephalomyelitis virus (TMEV) results in chronic demyelinating disease with progressive axonal loss and neurologic dysfunction similar to progressive forms of multiple sclerosis (MS). We previously showed that as the disease progresses, a marked decrease in brainstem N-acetyl aspartate (NAA; metabolite associated with neuronal integrity) concentrations, reflecting axon health, is measured. We also demonstrated stimulation of neurite outgrowth by a neuron-binding natural human antibody, IgM12. Treatment with either the serum-derived or recombinant human immunoglobulin M 12 (HIgM12) preserved functional motor activity in the TMEV model. In this study, we examined IgM-mediated changes in brainstem NAA concentrations and central nervous system (CNS) pathology.Findings1H-magnetic resonance spectroscopy (MRS) showed that treatment with HIgM12 significantly increased brainstem NAA concentrations compared to controls in TMEV-infected mice. Pathologic analysis demonstrated a significant preservation of axons in the spinal cord of animals treated with HIgM12.ConclusionsThis study links drug efficacy of slowing deficits with axon preservation and NAA concentrations in the brainstem in a model of progressive MS. HIgM12-mediated changes of NAA concentrations in the brainstem are a surrogate marker of axon injury/preservation throughout the spinal cord. This study provides proof-of-concept that a neuron-reactive human IgM can be therapeutic and provides a biomarker for clinical trials.Electronic supplementary materialThe online version of this article (doi:10.1186/s12974-015-0303-y) contains supplementary material, which is available to authorized users.

Monitoring glutamate levels in the posterior cingulate cortex of thyroid dysfunction patients with TE-averaged PRESS at 3 T

Backgrounds and ObjectivesPatients with thyroid dysfunction frequently have neuropsychiatric complaints such as lack of concentration, poor memory, depression, anxiety and mania, which suggest brain dysfunction. However, the underlying process of this dysfunction remains unclear. Recent studies of the glutamatergic system have offered important insight into the neuropsychiatric process. Thus, this study investigates changes in glutamate concentration in patients with thyroid dysfunction. It also clarifies whether Glu levels are related to thyroid hormones via proton magnetic resonance spectroscopy. Methods36 untreated patients with thyroid dysfunction (18 hyperthyroidism patients and 18 hypothyroidism patients) and 18 age- and gender-matched controls were included in this study. The posterior cingulate cortex was examined by MRS with TE-averaged PRESS at 3T. The intensity of glutamate, choline, N-acetylaspartate, and creatine was assessed using jMRUI v4.0 software. ResultsWe found a significant difference among hyper-/hypo- and control groups in Glu (P=0.003) and Cho (P=0.015). The concentrations of glutamate increased (P=0.006) in the posterior cingulate cortex in patients with hypothyroidism and significantly decreased (P=0.002) in hyperthyroidism patients relative to controls. There were no difference in the concentrations of choline between hyperthyroidism patients and controls (P=0.679). Versus the hyperthyroidism group, the hypothyroidism group showed increased glutamate (P=0.018) and choline (P=0.001) in the posterior cingulate cortex. There was no significant difference in the concentrations of NAA or creatine across the three groups (P>0.05). The Glu level correlates with TT3 (P=0.000) and FT3 (P=0.022). ConclusionThe signal intensity of glutamate shows significant differences in the region of the posterior cingulate cortex in patients with thyroid dysfunction. This change indicates a potential role of glutamate in the brain dysfunction experience by patients with thyroid hormone disorders.

MULTIPLIKASI TANAMAN NILAM (Pogostemon cablin Benth) var. TAPAK TUAN SECARA IN VITRO

ABSTRACTTissue culture is a technique widely used for propagation and improvement of crop quality. One way of plant propagation can be done with in vitro techniques. The purpose of the study was to determine the combinations and concentrations of NAA and BAP are appropriate for plant propagation patchouli. Dependent variable is the time of initiation of callus and shoots, callus texture, color callus, shoot number, and the number of roots patchouli. Independent variable is the concentration of growth regulators NAA and BAP. The method in this study is the experimental method. Data were analyzed by two-way Anova test followed by Duncan’s Multiple Range Test (DMRT). The results showed the stem explants concentrations of NAA and BAP best to induce callus is without NAA and BAP (control), 0.1 ppm NAA without BAP, NAA + BAP 0.5 ppm 0, while the leaf explants is there a 0.1 ppm NAA without BAP, NAA 0.1 ppm BAP + 0.5 ppm, 0.5 ppm BAP without NAA and NAA + BAP 0:01 ppm 1 ppm. Highest number of shoots obtained at a concentration of 0 ppm NAA concentration and NAA and 0.01 for leaf explants at a concentration of 0:01 ppm NAA + BAP 1 ppm and 0 ppm NAA + BAP 0.5 ppm for stem explants. Highest number of roots was found in 0:01 ppm for NAA concentration and leaf explants at a concentration of 0.1 ppm NAA + BAP and NAA 00:01 0 ppm ppm + 0.5 ppm BAP stem explants. Results of statistical analysis showed that there were significant differences of NAA and BAP concentrations on plant propagation patchouli (Pogostemon cablin Benth).
 Keywords: tissue culture, NAA, BAP, plant nilam (Pogostemon cablin Benth)

High frequency organogenesis in hypocotyl, cotyledon, leaf and petiole explants of broccoli (Brassica oleracea L. var. italica), an important vegetable crop.

Broccoli (Brassica oleracea L. var. italica) is an important, nutritionally rich vegetable crop, but severely affected by environmental stresses, pests and diseases which cause massive yield and quality losses. Genetic manipulation is becoming an important method for broccoli improvement. In the present study, a reproducible and highly efficient protocol for obtaining organogenesis from hypocotyl, cotyledon, leaf and petiole explants of broccoli (Brassica oleracea L. var. italica cv. Solan green head) has been developed. Hypocotyl and cotyledon explants were used from 10 to 12days old aseptically grown seedlings whereas leaf and petiole explants were excised from 18 to 20days old green house grown seedlings and surface sterilized. These explants were cultured on shoot induction medium containing different concentration and combination of BAP and NAA. High efficiency shoot regeneration has been achieved in hypocotyl (83.33%), cotyledon (90.11%), leaf (62.96%) and petiole (91.10%) explants on MS medium supplemented with 3.5mg/l BAP + 0.019mg/l NAA 2.5mg/l BAP + 0.5mg/l NAA, 4.0mg/l BAP + 0.5mg/l NAA and 4.5mg/l BAP + 0.019mg/l NAA respectively. Petiole explants showed maximum shoot regeneration response as compared to other explants. MS medium supplemented with 0.10mg/l NAA was found best for root regeneration (100%) from in vitro developed shoots. The regenerated complete plantlets were transferred to the pots containing cocopeat and successfully acclimatized. This optimized regeneration protocol can be efficiently used for genetic transformation in broccoli. This is the first comparative report on multiple shoot induction using four different types of explants viz. hypocotyl, cotyledon, leaf and petiole.

IAA and zeatin controls cell division and endoreduplication process in quiescent center cells of Allium cepa root

Role of endogenous IAA and zeatin level in regulation of cell division and endoreduplication processes was studied in quiescent center cells of Allium cepa using colchicine by considering morphological and cytological parameters. This resulted in formation of c-tumor at root tips, which contained endoreduplicated cells. Different concentrations of NAA and BAP (1–250 μM) were exogenously applied to these endoreduplicated root tips to depolyploidize endoreduplicated quiescent center cells. Exogenous NAA treatment showed negative response on depolyploidization of quiescent center cells of root tips. However, elongation of cells, and decrease in nucleus size was observed after exogenous NAA treatment. There was no change in length of endoreduplicated roots, and growth of root at the tip of c-tumor. Exogenous BAP showed positive response in depolyploidization of quiescent center cells of root tips. There was remarkable increase in length of roots, growth of root at tip of c-tumor and decrease in cell size and nucleus size after BAP treatment. It induced cytokinesis process in endoreduplicated meristematic cells and started cell division and differentiation process in quiescent center cells. Endogenous IAA and zeatin measured from normal, endoreduplicated and depolyploidized quiescent center root tip cells suggested that when endogenous IAA/zeatin ratio is high in meristematic cells then cell cycle shifts towards endoreduplication process but when it decreases cell cycle shifts towards cell division process.

High-efficiency propagation of Chinese water chestnut [Eleocharis dulcis (Burm.f.) Trin. ex Hensch] using a temporary immersion bioreactor system

Several studies have reported different propagation methods for the Chinese water chestnut [Eleocharis dulcis (Burm.f.) Trin. ex Hensch]. To date, none of these methods have efficiently achieved large-scale plantlet production using minimal labor input. The present study investigated the propagation of Chinese water chestnut shoot tips derived from corm and creeping rhizomes in a temporary immersion bioreactor system (TIBS) compared with propagation using conventional semi-solid medium methods. We evaluated the regeneration capacity of the corm and creeping rhizome shoot tips of E. dulcis Guiti No. 2 sp. nov. and found that culturing under optimized TIBS conditions (15 min of immersion into nutrient medium every 8 h) produced the highest multiplication rate (36.5 times) and shoot-forming capacity (SFC, 68.9), and reduced the vitrification rate. Creeping rhizome shoot tips had slightly higher SFC than corm shoot tips; the highest shoot multiplication rate of 43.7 times in TIBS was obtained using Murashige–Skoog (MS) medium containing 4 mg L−1 6-benzylaminopurine and 0.5 mg L−1 1-naphthaleneacetic acid (NAA). These optimized conditions were applied to corm shoot tips of E. dulcis Guiti No. 1 var. nov., E. dulcis Guiti No. 2 sp. nov., and E. dulcis Guifengti No. 1 sp. nov. Root formation was observed during the shoot multiplication stage on semi-solid MS medium; however, root formation was not noted during the shoot multiplication stage in TIBS. While TIBS produced a higher multiplication rate and SFC indices, there were no significant differences in average number of shoots per explant between TIBS and semi-solid medium. We also evaluated the effects of different immersion frequencies and NAA concentrations on TIBS propagation of E. dulcis Guiti No. 2 sp. nov. shoot tips by measuring the rooting percentage, average number of roots (length ≥ 3 cm) per explant, maximum root length, and root-forming capacity (RFC). The results showed that the medium containing 2.0 mg L−1 NAA at the rooting stage produced the best-developed root systems with the highest rooting percentage, maximum root length, and highest RFC (75.8 %, 8.5 cm, and 3.8, respectively); it also reduced the immersion frequency. The optimized conditions produced plantlets that had a greater capacity to acclimatize after transfer to the greenhouse. E. dulcis Guiti No. 1 var. nov. plantlets had the highest survival rate (95 %), largest diameters, and highest number of creeping rhizomes. The results of this study demonstrate that TIBS is a reliable and efficient method for large-scale propagation of Chinese water chestnut plantlets.

In vitro micropropagation and flowering in Ipomoea sepiaria Roxb. An important ethanomedicinal plant

Article history: Objective: To standardize a protocol for the micropropagation and in vitro flowering of Ipomoea sepiaria(I. sepiaria), an important ethanomedicinal plant. Methods: The nodal cuttings were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of 6-benzyladenine (BA) or Kinetin (Kn; 1.0-4.0 mg/L) alone or in combination with 毩-naphthaleneacetic acid (NAA; 0.2-1.0 mg/L) for shoot proliferation. For rooting ½ MS medium supplemented with indole-3-butyric acid (IBA) or NAA (0.5-3.0 mg/L) was used. When the 45-day-old in vitro derived nodal cuttings were subcultured on MS medium supplemented with 3.0 mg/L BA and 0.5 mg/L NAA and various concentrations of abscisic acid (ABA; 0.5-3.0 mg/L), in vitro flowering was observed. Results: The highest shoot induction response in terms of percent cultures responding and number of shoots per explant was observed on 3.0 mg/L BA and 0.5 mg/L NAA. On this medium 100% cultures responded with an average number of 3.2 shoots per explant. The optimum rooting was observed on 2.0 mg/L IBA. Here 100% shoots rooted with an average number of 5.1 roots per shoot. The optimum in vitro flowering response (38%) was observed on 2.0 mg/L ABA. Conclusion: The present protocol is an efficient method for the rapid multiplication, flowering and conservation of this medicinal plant.