Abstract: Wild almond is a woody species, which is difficult to propagate either generatively by seed or by vegetative methods (grafting or cuttings) and also considered as Endangered (EN) in Albania based on IUCN criteria. As a wild relative of cultivated fruit trees, this species represents a source of genetic variability and can be very important in breeding programs and cultivation. For this reason, it would be of interest to use an effective method of in vitro mid-term conservation, which involves strategies to slow plant growth through physicochemical alterations of in vitro growth conditions. Multiplication of wild almond was carried out using zygotic embryos, as primary explants, with the purpose to develop a successful propagation protocol. Results showed that zygotic embryos can proliferate through direct or indirect organogenesis. During subculture, stage was obtained a great number of new plantlets identical to mother plants derived from the zygotic embryos. All in vitro plantlets obtained from subcultures underwent in vitro conservation by minimal growth in low temperature (4ºC) and darkness. The efficiency of this technique was evaluated for 3, 6, and 10 months of conservation period. Maintenance in these conditions reduced micro cuttings growth. Survival and regeneration rates for each period were evaluated and resulted that the maximal time of conservation without subculture on 4ºC was 10 months, but survival and regeneration rates were significantly reduced, specifically 15.6% and 7.6%. An optimal period of conservation in these conditions can be considered the 5-6 months storage, which can lead to 60-50% of survival and regeneration rates. This protocol may be beneficial for mass propagation, mid-term conservation, and for genetic manipulation of wild almond.
Abstract: Vernonia divergens Benth., commonly known as
“Insulin Plant” (Fam: Asteraceae) is a potent sugar killer. Locally the
leaves of the plant, boiled in water are successfully administered to a
large number of diabetic patients. The present study evaluates the
putative anti-diabetic ingredients, isolated from the in vivo and in
vitro grown plantlets of V. divergens for their antimicrobial and
anticancer activities. Sterilized explants of nodal segments were
cultured on MS (Musashige and Skoog, 1962) medium in presence of
different combinations of hormones. Multiple shoots along with
bunch of roots were regenerated at 1mg l-1 BAP and 0.5 mg l-1 NAA.
Micro-plantlets were separated and sub-cultured on the double
strength (2X) of the above combination of hormones leading to
increased length of roots and shoots. These plantlets were
successfully transferred to soil and survived well in nature. The
ethanol extract of plantlets from both in vivo & in vitro sources were
prepared in soxhlet extractor and then concentrated to dryness under
reduced pressure in rotary evaporator. Thus obtainedconcentrated
extracts showed significant inhibitory activity against gram
negative bacteria like Escherichia coli and Pseudomonas
aeruginosa but no inhibition was found against gram positive
bacteria. Further, these ethanol extracts were screened for in vitro
percentage cytotoxicity at different time periods (24 h, 48 h and 72 h)
of different dilutions. The in vivo plant extract inhibited the growth of
EAC mouse cell lines in the range of 65, 66, 78, and 88% at 100, 50,
25 & 12.5μg mL-1 but at 72 h of treatment. In case of the extract of in
vitro origin, the inhibition was found against EAC cell lines even at
48h. During spectrophotometric scanning, the extracts exhibited
different maxima (ʎ) - four peaks in in vitro extracts as against single
in in vivo preparation suggesting the possible change in the nature of
ingredients during micropropagation through tissue culture
techniques.
Abstract: Shoots, with three leaves, of Paphiopedilum 'Delrosi'
were used as explants for multiple shoot induction. Modified
Hyponex medium was supplemented with thidiazuron (TDZ), N6-
benzyladenine (BA) or kinetin (Kn) alone and in combinations with
2,4-dichlorophenoxyacetic acid (2,4-D). All explants were cultured
for 15 weeks. It was found that TDZ alone at the concentration of
0.45μM or in combination with 4.52μM 2,4-D and 8.88μM BA in
combination with 13.56μM 2,4-D promoted multiple shoots. The
highest shoot sprouting efficiencies (80.0, 90.0 and 80.0%) and new
shoot numbers (1.5, 1.3 and 1.1) were obtained, respectively. Fresh
weight, height, numbers of leaf and root of new shoots and initial
explants were discussed.