B. Van Le et al., High frequency shoot regeneration from trifoliate orange (Poncirus trifoliata L. Raf.) using the thin cell layer method, CR AC S III, 322(12), 1999, pp. 1105-1111
Citations number
45
Categorie Soggetti
Multidisciplinary,"Experimental Biology
Journal title
COMPTES RENDUS DE L ACADEMIE DES SCIENCES SERIE III-SCIENCES DE LA VIE-LIFE SCIENCES
A method for a high frequency and direct in vitro bud regeneration of a woo
dy species, the trifoliate orange (Poncirus trifoliata L. Raf), was designe
d. Transverse thin cell layer (tTCL) explants excised from the stem interno
des of 1-year-old young plants of P. trifoliata regenerated bud in vitro on
a medium containing 6-benzylaminopurine (BAP 1-50 mu M) and N-phenyl-N'-1,
2,3-thidiazol-5-ylurea (thidiazuron, TDZ) (0.1-10 mu M). The optimal concen
trations for bud induction were 25 mu M BAP and 1 mu M TDZ leading to 87 an
d 72 % of responsive tTCLs and 21 and 15 buds per tTCL, respectively. A hig
her percentage of responsive fTCLs and a higher frequency of bud regenerati
on were obtained with BAP and TDZ combined. With a combination of 10 mu M B
AP and 1 mu M TDZ, 90 % of responsive tTCLs forming 37 buds per tTCL were o
btained. Shoot elongation occurred after a transfer onto a medium containin
g 1 mu M GA(3). Rooting of individual shoot was induced using 5 mu M NAA. O
ne hundred per cent of rooted shoots developed normally after transfer to t
he greenhouse; no phenotype variation was observed. High numbers of regener
ated viable plants can be produced directly without callus formation from t
TCL after 9 weeks of culture. (C) 1999 Academie des sciences/Editions scien
tifiques et medicales Elsevier SAS.