E. Prakash et al., Regeneration of plants from seed derived callus of Hybanthus enneaspermus L-Muell., a rare ethnobotanical herb, PL CELL REP, 18(10), 1999, pp. 873-878
A procedure was developed for plant regeneration of Hybanthus enneaspermus,
a rare ethnobotanical herb from the Deccan peninsula in India, through see
d-derived callus. Seeds demonstrated a high induction frequency (69.4+2.8%)
and a high yield (364.4+/-2.5 mg) of light-yellow friable callus on Murash
ige and Skoog's (MS) medium containing 2.6 mu M NAA and 2.2 mu M BA within
4 weeks of incubation. After I year of subculture, yellow friable and light
-green compact calli types were established from initial light-yellow friab
le callus. Shoot differentiation was achieved from light-green compact call
us, but not from yellow friable callus. Shoot differentiation resulted when
light-green compact callus was transferred to MS medium supplemented with
8.8 mu M BA and 2.6 mu M NAA; the highest percentage of calli forming shoot
s (66.6+/-4.8%) and the highest number of shoots (8.9+/-0.3) were achieved
in this medium. Differentiated shoot buds elongated to 4-5 cm within 4 week
s. The addition of casein hydrolysate (500 mg/l) and more potassium phospha
te (1.86 mM) to the culture medium enhanced shoot differentiation. Rooting
was achieved on the shoots using half-strength MS medium containing 4.8 mu
M IBA. About 70% of the plants were established in pots containing pure gar
den soil after 2 weeks of hardening. The regenerated plants were morphologi
cally uniform and exhibited normal seed set.