ABSTRACT
The present work aims to establish an efficient protocol for in vitro regeneration of peanut (Arachis hypogaea) cultivar L14. The study showed that de-embryonated cotyledon was a suitable explant for shoot multiplication on MS medium containing 4 mg/L BAP. The highest number of shoots per explant obtained after 4 weeks of culture was up to 6.8 shoots. Shoots in vitro were able to produce a large number of approximately 11 roots on MS medium supplemented with 0.5 mg/L NAA. These results will be very useful in establishing an in vitro regeneration protocol for peanut cultivar L14 during gene transfer in the next studies to improve their disease resistance.
Subject(s)
Arachis , In Vitro TechniquesABSTRACT
Aim: Abelmoschus moschatus have been extensively used in traditional medicine as well as in perfume industries. The primary goal of the present research was to develop an efficient plant regeneration protocol of Abelmoschus moschatus from aseptic seedling explants such as cotyledon, internode, leaf and root. Methodology: The seeds of Abelmoschus moschatus were surface sterilized with 0.1% mercuric chloride and 70% ethanol were cultured on ½ MS basal media for developing aseptic seedlings Aseptic seedling explants were cultured on different concentrations of auxins for callus induction. Later callus was transferred on to different concentrations of cytokinins for shoot regeneration and for in vitro, rooting different concentrations of auxins were used. Finally, such in vitro developed plantlets were acclimatized. Results: Half strength MS medium with 1% sucrose was used for raising aseptic seedlings. Maximum of 92% response of callus induction was obtained from leaf explants on MS medium + 2 mg/L 2, 4-dichlorophenoxyacetic acid. An average of 2.4 shoots per callus were observed on MS + 2 mg/L benzyl-6-aminopurine from leaf explant. The regenerated shoots were best rooted on 1/2 MS + 0.5 mg/L indole-3-butyric acid. The regenerated plantlets were established with 70% survival. Conclusion: An efficient plant regeneration protocol of Abelmoschus moschatus was developed.
ABSTRACT
Effects of N6-benzyladenine (BA) or thidiazuron (TDZ) on adventitious shoot regeneration and axillary shoot multiplication of Sedum sarmentosum was investigated. Leaf and shoot tip explants obtained from in vitro-grown shoots of S. sarmentosum were cultured on Murashige and Skoog (MS) medium supplemented with 0, 2.0, 4.0 or 8.0 µM BA or TDZ. Of the two cytokinins studied, BA was found to be more responsive as compared to TDZ with respect to number of shoots produced per explant. High frequency of shoot regeneration (92.2%) with a mean of 12.3 shoots was produced when the leaf explants were cultured on MS medium supplemented with 8.0 µM BA. The highest number of shoots (25.4) was obtained when shoot tip explants were cultured on MS medium devoid of cytokinins after 35 days of culture. For root induction, regenerated shoots were cultured on MS medium supplemented with 0, 2.0, 4.0 or 8.0 µM indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or α-naphthaleneacetic acid (NAA). The highest number of (17.6) roots per shoot was obtained on MS medium supplemented with 2.0 µM IBA after 30 days of culture. Regenerated plantlets were successfully acclimatized in the greenhouse with 100% survival rate.
ABSTRACT
It was evaluated the antibacterial activity of seven β-lactam antibiotics against A. tumefaciens strain EHA105, living in indica rice calli. For detection of persistent Agrobacterium in callus tissues, homogenates of infected calli were spread over LB agar plates to count colony forming units per gram of calli. Agrobacterium growth was completely suppressed during plant regeneration at 250 mg/l of all of the antibiotics tested. Also it was appraised the effect of β-lactams on callus growth and plant regeneration. A similar tendency of increased calli fresh weight with 100 mg/l and 250 mg/l of all antibiotics was proven. But the use of 500 mg/l caused decreasing of callus growth. About 80% of calli formed shoots in a month when we used 100 mg/l or 250 mg/l of β-lactam during the regeneration stage. Two morphogenic responses were distinguished during regeneration stage: somatic embryogenesis and adventitious shoots organogenesis.
Se evaluó la actividad antibacteriana de siete antibióticos β-lactámicos sobre la cepa EHA105 de Agrobacterium tumefaciens inoculada en callos de arroz. Para detectar el Agrobacterium persistente en los callos, se cultivó en medio LB sólido un extracto de callos infectados, y se contó el número de unidades formadoras de colonias por gramo de callo. La bacteria se eliminó totalmente en la fase de regeneración utilizando 250 mg/l de cualquiera de los antibióticos probados. Se evaluó además el efecto de los β-lactámicos sobre el crecimiento de los callos y la regeneración de plantas. El incremento del peso de los callos fue similar cuando se utilizó 100 y 250 mg/l de antibiótico, pero disminuyó significativamente cuando la concentración se elevó a 500 mg/l. La regeneración de plantas ocurrió a partir del 80% de los callos cultivados con 100 y 250 mg/l de antibiótico. En la regeneración se distinguieron dos respuestas morfogénicas: embriogénesis somática y organogénesis adventicia.
Subject(s)
Agrobacterium tumefaciens , Anti-Bacterial Agents , Oryza , Agrobacterium , Embryonic Development , Plant Somatic Embryogenesis Techniques , RegenerationABSTRACT
Objective:To determine the effects of different cytokinins at various concentrations on in vitro shoot multiplication of an important medicinal plant. Methods: Nodal explants (1.5-2.0 cm) of Sophora tonkinensis were used. Multiple shoots were induced from nodal explants cultured on the Murashige and Skoog (MS) medium supplemented with 0.0, 0.5, 1.0, 2.0, 4.0, 8.0, or 16.0 μmol 2-isopentyladenine (2iP), N6 benzyladenine, kinetin or thiadiazuron. Results: Among the four investigated cytokinins, 2iP showed the best response for shoot multiplication. Maximum shoot induction (75%) was achieved on the MS medium supplemented with 2.0 μmol 2iP, with a mean number of 5.0 shoots per explant. In comparison to other cytokinins tried, 2iP showed the highest shoot elongation with a mean shoot length of 4.8 cm. Root initiation was observed within 15 d within the transfer of shoots onto the MS basal medium, and the rooting percentage was 100%with a mean number of 5.4 roots per shoot and root length of 6.2 cm over a period of 4 weeks. The healthy plants, hardened and transferred to a greenhouse for proper acclimatization, exhibited 100%survival. Conclusions:It can be summarized that 2iP is the optimal plant growth regulator for Sophora multiplication.
ABSTRACT
An attempt was made to standardize the protocol for the shoot regeneration via caulogenesis in Pteris vittata L. employing leaf primordium explants. Calli were induced on Murashige and Skoog’s (MS) and Parkers and Thompson’s (P and T) media supplemented with different combinations of 2, 4-dichlorophenoxyaceticacid (2, 4-D), 6-benzylaminopurine (BAP), a-naphthalene acetic acid (NAA) and Indole - 3-acetic acid (IAA). A combination of full strength MS medium with 2, 4-D (2.26 µM) and BAP (2.22 µM) was found to be ideal for profuse callusing (80%) against other combinations. More shoot differentiation (2.8±0.06) was achieved from the calli on one-fourth strength of P and T media fortified with BAP (4.44 µM) and NAA (2.68 µM) when compared to other combinations but statistically not significant.
ABSTRACT
Exacum wightianum Arn. (Gentianaceae) is an endemic medicinal plant from the Nilgiri hills, Western Ghats, Tamil Nadu. Indirect regeneration of E. wightianum was obtained through organogenesis in callus culture. Axial bud explants were found to be best suited for callus induction on MS medium supplemented with BA + NAA (2.0+0.03 mg/L). Multiple shoots originated from callus obtained from the axial buds. were multiplied by subculture on the same medium. Maximum shoot regeneration was obtained on MS medium supplemented with BA with NAA (2.0+0.5 mg/L), and up to 25shoots was observed within 2 weeks.
ABSTRACT
Cistus creticus L. ssp. creticus is a medicinal aromatic shrub native in Crete (Greece). The protocol described in this paper provides optimal levels of growth regulators required to obtain high regeneration rates of Cistus in vitro. Micropropagation has been achieved through rapid proliferation of shoot-tips on Murashige and Skoog (MS) basal medium supplemented with 0.2 mg l-1 6-benzylaminopurine (BAP). After four weeks shoots were transferred to MS medium without growth regulators for further development and rooting. The highest percentage of regenerated shoots was obtained with 0.1 mg l-1 TDZ and 0.1 mg l-1 NAA after 4 weeks. Elongation and rooting was readily achieved when multiple shoots more than 1 cm in length were singled out and cultured on the MS medium without growth regulators. The plantlets were successfully adapted and grew vigorously in greenhouse conditions. This is the first report of shoot regeneration in the genus Cistus. The regeneration protocol developed in this study provides a basis for further investigation of the medicinally active constituents of this elite medicinal plant.
ABSTRACT
In the present study we examined the possibility of propagating different Cyclamen species (C. africanum Boiss. and Reut., C. cilicium Boiss. and Heldr., C. coum Mill., C. hederifolium Ait., C. persicum Mill., C. purpurascens Mill.) including some of their subspecies and cultivars in vitro using explants of adult plants. For this purpose two protocols have been applied to eleven genotypes combined with mostly four explant types (placentas with ovules, leaves, petioles and peduncles). The use of these protocols has given rise to either somatic embryo-like structures and/or adventitious shoots in all genotypes. This way it was possible to propagate each of the examined genotypes in vitro using explants of adult plants in a time less than one year. These results may be used in breeding and propagation of Cyclamen as an ornamental plant and as a medicinal plant.