Protocol for augmented shoot organogenesis in selected variety of soybean [Glycine max L. (Merr.)].

Development of a reproducible, versatile and efficient in vitro plant regeneration system is highly warranted for Indian soybean varieties for their mass multiplication in view of their commercial significance. Accordingly a protocol for direct shoot organogenesis in soybean variety JS 335 has been developed. Using cotyledonary node explants significant organogenic responses, mean shoot number and shoot length were observed when these were incubated on MS medium supplemented with 0.89 µM Benzyladenine (BA) and 5 μg/L triacontanol (TRIA) where in 9.3 ± 0.5 shoots were obtained. TRIA at 5 μg /L able to produce 6.8 ± 0.5 shoot buds in presence of 0.98 µM IBA and 0.89 µM BA. Highest mean shoot buds (14.0 ± 0.5 and 9.0 ± 0.5) and mean shoot length (4.6 ± 0.3 and 10.0 ± 0.7) were obtained when cotyledonary node and shoot tip explants were cultured on MS medium containing 0.14 µM gibberellic acid (GA3), 0.89 µM BA and 5 μg/L TRIA. Moreover, TRIA supported highest mean root number (6.3±0.5) and root length (21.5 ± 0.57 cm). Field survival of in vitro derived plants of TRIA treatment was 70% and the overall growth and seed yield was also significantly better than control plants. This protocol may be used for improving the in vitro regeneration of soybean variety JS 335 for transformation studies.

Photoperiod influences endogenous indoleamines in cultured green alga Dunaliella bardawil.

Effect of light intensity and photoperiod on growth, indoleamines and carotenoid production was studied in unicellular green algae D. bardawil. Maximum biomass and carotenoid contents were found when cultures were grown in light (intensity of 2.0 Klux) at a photoperiod of 16/8h light and dark cycle. There was a profound influence of tested photoperiod conditions of light:dark viz. 8:16, 10:14, and 12:12 hr, continuous light on indoleamines (SER and MEL) production as estimated by HPLC and confirmed by mass spectral data obtained from LC-MS-ESI studies. Serotonin level increased from 908 to 1765 pg/g fresh wt with increase in light duration and melatonin level increased from 267 to 584 pg/g fresh wt during increase in dark phase. Carotenoids production was high in continuous light than other tested conditions.

Antioxidant property of Decalepis hamiltonii Wight & Arn.

Aromatic edible root of D. hamiltonii was subjected to the extraction of the antioxidant rich fraction. Different parts of root namely whole tuber, peel, tuber without peel and medullary portion were extracted with dichloromethane (European Patent No. W02005063272). The extract was found to contain flavor compound 2-hydroxy-4-methoxybenzaldehyde (2H4MB), which was identified by TLC and GC. Medullary portion was found to be rich in 2H4MB, (73.73 mg g(-1) dry tissue) followed by peel, containing 68.34 mg g(-1) 2H4MB. Different concentration of dichloromethane extracts were subjected for antioxidant assay by DPPH (1,1 dihydroxy 2-picryl hydrazyl) method, this has shown 44, 46.7% radical scavenging activity in case of medullary, peel extracts and 67.3% in case of pure 2-hydroxy-4-methoxybenzaldehyde at 100 ppm concentration, whereas ascorbic acid used as standard showed 94.3% activity. In beta-carotene linoleate model system (b-CLAMS) 43.46 and 45.7% antioxidant activity was observed in medullary and peel extracts at 100 ppm concentrations respectively, whereas standard 2-hydroxy-4-methoxybenzaldehyde exhibited 69.64% at 100 ppm and BHA (butylated hydroxyl anisole) 90.1% activity also at 100-ppm level. Similarly hydroxyl radical scavenging activity was found to be 48.36, 46.86, 48.26 and 73.60% in whole tuber, medullary, peel and standard 2-hydroxy-4-methoxy benzaldehyde respectively at 100 ppm levels. This is the first report on the antioxidant activity of D. hamiltonii. Results have shown that 2H4MB is one of the major constituents responsible for antioxidant activity. Hence the extract of D. hamiltonii can be utilized for the production of antioxidant rich fractions required for various health benefits.

In vitro clonal propagation of bird eye chilli (Capsicum frutescens Mill.).

An efficient and highly reproducible protocol for micropropagation of bird eye chilli Capsicum frutescens was attempted. Murashige and Skoog (MS) medium containing 0.5-3.0 mgl(-1) of 6-benzyladenine (BA), 2-isopentenyl adenine (2iP), kinetin and 0.5-2.0 mg l(-1) of indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) along with 1 gl(-1) activated charcoal (AC) were used for shoot regeneration from both shoot tip and nodal explants. Shoot tip explants (100%) grew well on medium containing 1 mgl(-1) of kinetin and 1 mgl(-1) of IBA. Shoot proliferation (1-3) from nodal explants was effective on this medium. The regenerated shoots with 4-7 nodes had further growth upon sub-culturing onto kinetin (1 mgl(-1)) and IBA (1 mgl(-1)) and rooted simultaneously. The rooted plants were transferred to pots after hardening under controlled conditions. The survival percentage in pots was 80-90%.

Efficient clonal propagation method for Decalepis hamiltonii, an endangered shrub, under the influence of phloroglucinol.

A highly efficient two stage protocol was developed for induction of multiple shoots from single node in vitro shoot tip explants of Decalepis hamiltonii. It was found that phloroglucinol (PG) had synergistic effect on shoot multiplication when added with N6-benzyladenine and gibberellic acid. This protocol uses PG for both multiple shoot induction from nodal explants, elongation of primary shoots and initiation of adventitious shoot formation from primary shoots, which was more in presence of triacontanol (TRIA). Maximum number of shoots per culture was observed on the medium containing N6-benzyladenine (1.1 microM; BA), GA3 (5.8 microM) and PG (800 microM). Sub-culturing of the shoots onto MS medium containing optimum concentration of BA (5.6 microM), PG (200 microM) and TRIA (0.011 microM) produced elongated shoots along with secondary shoot formation. The long shoots were rooted on alpha-naphthalene acetic acid (5.38 microM; NAA) and PG (400 microM) containing medium. The rooted plantlets were hardened and their field survival rate was 80-90%.

Production of 2-hydroxy-4-methoxybenzaldehyde in roots of tissue culture raised and acclimatized plants of Decalepis hamiltonii Wight & Arn., an endangered shrub endemic to Southern India and evaluation of its performance vis-a-vis plants from natural habitat.

Axillary buds obtained from field grown plants of D. hamiltonii were used to initiate multiple shoots on Murashige and Skoog's medium (MS) supplemented with 2 mg L(-1) 6-benzyl aminopurine (BA) and 0.5 mg L(-1) indole-3-acetic acid (IAA). Profuse rooting was achieved when the actively growing shoots were cultured on MS medium supplemented with 1.0 mg l(-1) indole-3-butyric acid (IBA). Regenerated plants were grown successfully in the plains, in contrast to wild growth in high altitudes and rocky crevices of hilly regions. Roots of different sizes from one-year-old tissue culture raised field grown plants had the same profile of 2-hydroxy-4-methoxybenzaldehyde as that of wild plants. A maximum of 0.14% and 0.12% 2-hydroxy-4-methoxybenzaldehyde was produced in roots of one year old tissue culture derived plants and greenhouse grown plants respectively.