Plant regeneration from petal protoplast culture ofPetunia hybrida

Abstract

Morphologically normal plants have been regenerated from petal protoplasts of petunia (Petunia hybrida) flower. Maximum protoplast yields from petal tissues were obtained within 2 days after anthesis. Protoplasts were cultured on modified Murashige and Skoog's medium in which NH4NO3 and Fe.EDTA concentrations were reduced to 1/3 (7mM) and 1/10 (10 ~tM), respectively. After plating, protoplasts gradually reduced pigment density, and plastids developed near the nucleus. In premitotic petunia petal cells, the nucleus moved from the periphery to the central region of the cell. The first cell divisions were detected after 6-10 days of culture initiation, and the average division frequency was 15% in the best culture condition. The results indicated that the time of the first cell division and cell division frequency were closely related to flower age after anthesis. More than a hundred plants with morphologically normal shoots and roots have been obtained. Those plants grew vigorously in soil.

Protoplast isolation and plant regeneration of different genotypes of Petunia and Calibrachoa

“…An incubation period of 4 h, as proposed by Oh and Kim (1994), gave very poor yields. Frearson et al (1973) and Renaudin et al (1990) reported that overnight enzyme incubation was successful for isolation of Petunia protoplasts.…”

“…Protoplasts from Petunia have been isolated from different sources of tissues, such as petals (Oh and Kim 1994), ovules (Potrykus 1971), mesophylls (Taguchi et al 1993), and pollen (Bajaj and Davey 1974). For Calibrachoa, protoplast isolation from callus (Power et al 1980), suspension cultures (Ford-Logan and Sink 1988), and mesophyll tissues have been reported.…”

Protoplast Isolation and Shoot Regeneration from Protoplast-Derived Callus of Petunia hybrida Cv. Mirage Rose

“…Various methods of isolating and culturing protoplasts in Petunia hybrida have been reported since a few decades ago [ 11 , 12 , 13 , 14 , 15 , 16 ]. However, shoot regeneration from a protoplast-derived callus has thus far remained challenging, although a few studies have reported genotype-dependent shoot regeneration from protoplast-derived calli [ 16 ], with most genotypes being minor or of no commercial importance.…”

“…Indeed, successful protoplast isolation with high yield and reproducibility often requires an optimal digestion enzyme dose and digestion time [ 16 , 17 , 18 ]. In addition, the concentration of sucrose as a carbon source also plays a vital role in the success of protoplast culture [ 13 , 14 , 15 , 16 ]. Unfortunately, a protocol suitable for one cultivar might not be suitable for other cultivars.…”

Medicinal and Aromatic Plants IX

“…They are used for several purposes, e.g., the isolation ofmutants (Colijn et al 1979) and the study ofprimary metabolism (De Gucht 1993), secondary metabolism (see below), protoplast isolation (e.g., Phillips and Darrell1992), or regeneration (e.g., Oh and Kim 1994). They are used for several purposes, e.g., the isolation ofmutants (Colijn et al 1979) and the study ofprimary metabolism (De Gucht 1993), secondary metabolism (see below), protoplast isolation (e.g., Phillips and Darrell1992), or regeneration (e.g., Oh and Kim 1994).…”