Direct shoot organogenesis from in vitro-derived mature leaf explants of pistachio

Abstract

An efficient system was developed for direct plant regeneration from in vitro-derived leaf explants of Pistacia vera L. cv. Siirt. The in vitro procedure involved four steps that included (1) induction of shoot initials from the regenerated mature leaf tissue, (2) regeneration and elongation of shoots from the shoot initials, (3) rooting of the shoots, and (4) acclimatization of the plantlets. The induction of shoot initials was achieved on an agarified Murashige and Skoog (MS) medium with Gamborg vitamins supplemented in different concentrations of benzylaminopurine (BA) and indole-3-acetic acid (IAA). The best medium for shoot induction was a MS medium with 1 mgl −1 IAA and 2 mgl −1 BA. Numerous shoot primordia developed within 2-3 wk on the leaf margin and the midrib region, without any callus phase. In the second step, the shoot clumps were separated from the leaf explants and transferred to a MS medium supplemented with 1 mgl −1 BA, resulting in a differentiation of the shoot initials into well-developed shoots. The elongated shoots (>3 cm long) were rooted on a full-strength MS basal medium supplemented with 2 mgl −1 of indole-3-butyric acid in the third stage. Finally, the rooted plants were transferred to soil with an 80% success rate. This protocol was utilized for the in vitro clonal propagation of this important recalcitrant plant species.

Novel Methods in Micropropagation of Pistachio

“…Once shoot buds were obtained from axenic leaves, the use of only BA on the proliferation medium proved to be more bene fi cial than the combination of IAA and BA. The results reported by Tilkat and Onay ( 2009 ) and Tilkat and co-workers (2009b) showed consistency with other studies where BA and IAA promoted the proliferation and elongation of shoots in buds from Ladebouria hyacinthiana (Turakhia and Kulkarni 1988 ) leaves. The protocol for forcing the regenerated leaves for organogenesis summarized above on the female cultivar "Siirt" showed consistency with the pistachio male cultivar "Atli" (Tilkat and Onay 2009 ;Tilkat et al 2009b ) .…”

“…The details of the method are as follows: Healthy mature tips of 30-year-old male P. vera L. "Siirt" were collected from the pistachio research institute in Gaziantep province, a province of southeastern Turkey, and were used as the source of material (Tilkat and Onay 2009 ) . To establish aseptic shoot cultures, apical shoot tips were disinfested by the protocol described above, and cultured on a modi fi ed Murashige and Skoog (MS) (Murashige and Skoog 1962 ) medium (initiation medium) with Gamborg vitamins (Gamborg et al 1968 ) supplemented with 1.0 mg L −1 BA, 30 g L −1 sucrose and 5.5 g L −1 agar.…”

“…Gharyal and Maheshwari ( 1990 ) reported 36% direct shoot regeneration from petioles on a B5 medium supplemented with 0.5 mg L −1 IAA and 1 mg L −1 BA. However, 31.9% of direct shoot regeneration was obtained in female cultivar ("Siirt") from culturing lea fl et explants on a MS medium supplemented with 1 mg L −1 IAA and 2 mg L −1 BA (Tilkat and Onay 2009 ) . Shoots subcultured on the 1:2 IAA:BA media also proliferated rapidly, but the medium regenerated hyperhydric shoots.…”

Shoot organogenesis in leaf explants of Hydrangea macrophylla ‘Hyd1’ and assessing genetic stability of regenerants using ISSR markers

“…This indicated that specific BA/IBA ratios were more optimal for shoot formation than lower or higher ones. The strong influence of increasing BA concentrations on shoot formation in treatments relative to fixed IBA or NAA concentrations was described in a regeneration study of pistachio too (Tilkat and Onay 2009), and similarly for somatic embryogenesis and organogenesis in callus cultures of Albizzia richardiana King (Tomar and Gupta 1988). IBA was more effective than NAA on Euphorbia esula shoot induction (Xu et al 2008).…”

Organogenesis from in vitro-derived leaf and internode explants of Hoya wightii ssp. palniensis -a vulnerable species of Western Ghats

“…Shoot regeneration from in vitro derived explants have been reported in other plants such as Decalepis hamiltonii by Giridhar et al (2004); Vaccinium corymbosum (Billings et al 1988); Pyrus communis (Leblay et al 1991); Elaegnus angustifolia (Economou and Maloupa, 1995); Pistachio vera L. cv. Siirt (Tilkat and Onay, 2009); Morus alba (Chitra and Padmaja, 2005); Phellodendron amurense (Azad et al 2005). This is the first report on indirect organogenesis and the establishment of complete plantlets from the regenerated leaf and internode explants of H. wightii ssp.…”