Adventitious shoot regeneration from the leaves of some pear varieties (Pyrus spp.) grown in vitro

The pear (Pyrus spp.) is one of the most important temperate fruit crops. A complete protocol for adventitious shoot regeneration was developed from the leaves of four pear varieties grown in vitro: Abbe Fetel, Yali, Packham’s Triumph and Aikansui, and the Chinese rootstock variety Duli. Shoot explants were collected from the field and cultured in vitro in Murashige and Skoog (MS) media supplemented with 1.0 mg·L⁻¹ 6-benzylaminopurine (BA) and 0.1 mg·L⁻¹ indole-3-butyric acid (IBA). After four weeks, leaf explants of all 5 varieties grown in vitro were excised and cultured in MS media supplemented with 0.0 mg·L⁻¹, 0.2mg·L⁻¹, 0.5 mg·L⁻¹, 1.0 mg·L⁻¹ and 2.0 mg·L⁻¹ naphthaleneacetic acid (NAA) and 5.0 mg·L⁻¹ BA or with 1.0 mg·L⁻¹, 2.0 mg·L⁻¹ and 4.0 mg·L⁻¹ thidiazuron (TDZ). The cultures were maintained in darkness for 21 days for shoot induction in the shoot induction medium (IM), then transferred to the shoot expression medium (EM) in 1.0 mg·L⁻¹ TDZ without any auxins and kept in a growth room at (25±2)°C under a 16/8 h light/dark photoperiod regime for 8 weeks. Finally, the shoots were transferred to the MS shoot elongation medium (SEM) supplemented with 0.2 mg·L⁻¹ BA, 0.1 mg·L⁻¹ IBA and 0.2 mg·L⁻¹ gibberellic acid (GA₃). A combination of TDZ and NAA had a significant effect on the number of shoot regenerations in all 5 tested varieties. The maximum mean number of shoots and maximum number of shoots per leaf obtained from Yali variety were 11.8 (P⩽0.001) and 22, followed by Aikansui with 6.6 (P⩽0.001) and 4.6, and Duli with 8 (P⩽0.001) and 12, all arising from the combination of 0.2 mg·L⁻¹ NAA with 1.0 mg·L⁻¹ TDZ. For Packham’s Triumph and Abbe Fetel, the maximum mean number of shoots and maximum number of shoots per leaf were 5.6 (P⩽0.001), 4.8 and 8 (P⩽0.001), and 11, respectively, from the combination of NAA (1.0 mg·L⁻¹) and TDZ (2.0 mg·L⁻¹). Abbe Fetel was the only variety which produced significantly higher adventitious shoots from the two different combinations of 1.0 mg·L⁻¹ NAA and 5.0 mg·L⁻¹ BA (P⩽0.05), and 2.0 mg·L⁻¹ NAA and 5.0 mg·L⁻¹ BA (P⩽0.01). Some of the most prominent problems associated with shoot proliferation and regeneration were also observed and discussed in this paper.