Hakan Yildirim

Micropropagation of mature male pistachio Pistacia vera L.

Summary Factors affecting the successful rapid proliferation and rooting of male pistachio (Pistacia vera L.) cv. Atlı, were studied. The most suitable type of cytokinin [6 benzyladenine (BA), kinetin (Kin), or thidiazuron (TDZ)], and the effect of eight different concentrations of BA (0.0675, 0.125, 0.25, 0.5, 1.0, 2.0, 4.0, or 8.0 mg l–1) were evaluated to optimise shoot proliferation. The auxins -naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA), and indole-3-butyric acid (IBA), each at 2.0 mg l–1, or different concentrations (0.5, 1.0, 2.0, 4.0, or 8.0 mg l–1) of IBA, and the effect of explant size (1.0, 2.0, 3.0, or 4.0 cm) were assessed for root induction. The highest number of new microshoots per explant (5.64 ± 0.07) was obtained 4 weeks after culturing on Murashige and Skoog (MS) medium supplemented with 1.0 mg l–1 BA. Again, shoot length was highest in 1.0 mg l–1 BA, and decreased as the BA concentration increased. IBA was most effective in promoting root formation. The highest rooting frequency (73%) of microshoots was recorded for explants 4.0 cm in length, 4 weeks after culturing. In vitro-rooted plantlets were transferred to polyethylene pots filled with a 1:1:1 (v/v/v) mixture of soil, sand and peat.This treatment resulted in 90% survival of plantlets, which were acclimatised in a greenhouse.

Somatic embryogenesis of pistachio from female flowers

Summary Induction of somatic embryogenesis from the flower buds of pistachio (Pistacia vera L.) was studied with respect to growth regulators. Sixty-five percent of cultures formed embryogenic calli when placed on modified Murashige and Skoog medium containing 1 mg l–1 benzylaminopurine. In contrast, thidiazuron and α-naphthalene acetic acid reduced embryogenesis. Some of the somatic embryos had fused hypocotyls or multiple cotyledons. Abscisic acid was not necessary for maturation. Fewer somatic embryos germinated with 0.5 mg l–1 abscisic acid, but more converted into plantlets. Separate, morphologically normal, somatic embryos germinated on semi-solid germination medium, and developed into plantlets. Cytological analysis revealed a chromosome number of 2n = 2x = 30, similar to seedlings. The protocol described in this paper, for the induction of somatic embryos indirectly from female flowers of pistachio, represents a first step towards the production of clonal stocks.

IN VITRO MICROGRAFTING OF THE ALMOND CULTIVARS "TEXAS", "FERRASTAR" AND "NONPAREIL"

ABSTRACT A successful micrografting technique for the almond cultivars (cvs) “Texas”, “Ferrastar” and “Nonpareil” was developed using in vitro germinated seedlings as rootstocks and axenic shoot cultures established from mature tree sources as microscions. In vitro germinated seedlings, which developed 14 days after culturing in the modified Murashige and Skoog (MS) medium, were decapitated and used as rootstock. Shoot culture initiation from three almond cvs (“Texas”, “Ferrastar” and “Nonpareil”) was successfully achieved by culturing mature shoot tips from forced nodal buds, about 4–6 mm, on a modified MS medium containing 1 mgL−1 benzyl adenin (BA). Slit micrografting on epicotyl and on hypocotyls were equally successful (83.3% to 100%). Grafting success was dependent on the rootstock type and lenght of the scion. Grafting success varied between 83.33% and 100% depending on the cultivar, when the scion contained 1, 2, and 3 nodes. When almond scions, about 1.5 cm long, were micrografted on germinated seedling and cultured on proliferation medium (PM), the mean shoot length was 19.84 mm, 16.50 mm, 26.93 mm for the cvs “Texas”, “Ferrastar” and “Nonpareil” respectively. Micrografts could be easily cultured on a hormone-free semi-solid MS medium and were potted out after 4 to 6 weeks of culture growth. Rooted micrografted plantlets were successfully acclimatized and transferred to potting mix with 100 % survival. Although low percentages of variation were obtained in tested cvs (3.70%, 6.25% and 10.2% in “Texas”, “Ferrastar” and “Nonpareil”), molecular analysis showed that the developed micrografting technique produces genetically stable plantlets, at least up to 6 months of sub-culturing in cvs “Ferrastar” and “Nonpareil”. The described micrografting technique could be usedfor rejuvenation ofshoot explants of mature elite almond cultivars and it also has potential use in the commercial production of other almond cultivars.

AN EFFECTIVE PROTOCOL FOR IN VITRO GERMINATION AND SEEDLING DEVELOPMENT OF LENTISK (Pistacia lentiscus L.)

Different nutrient media (MS [Murashige and Skoog 1962]; QL [Quoirin and Lepoivre 1977] and WPM [Lloyd and McCown 1980]); plant growth regulators BA (benzil adenin), GA3 (gibberellic acid), IBA (indole-3-butyric-acid), NAA (naftalen acedic acid); and sucrose concentrations were studied to determine the in vitro culture effects on healthier and faster seedling development from mature lentisk (Pistacia lentiscus L.) seeds. After 28 days of culture, the percentage of germinated seeds was the highest (70%) in the full-strength MS medium. The cytokinin BA was superior to other tested treatments in terms of its ability to promote germination of lentisk seeds. When tested at different concentrations, sucrose gave the best results obtained at concentrations of 1–4%, whereas high concentrations (6 and 8%) mainly decreased germination rate and there was no a regular pattern for elongation of the aerial parts of plants. With this described protocol, on average 76.67% seeds germinated 4 weeks after culture. Developed seedlings were satisfactorily acclimatized in sterilized peat, soil and perlite containing compost, with high percentage survival viability was obtained 9 months after transfer to in vivo conditions (93.33%). The results obtained showed that the enriched full-strength MS medium supplemented with 1 mg L BA and 3% sucrose induced homogeneous and healthy seedling development in a period of 4 to 8 weeks of culture.

Odunsu Bitki Türlerinde İn-vitro Aşılama-II (Amaçları)

In-vitro asilama, aksenik kultur kosullarinda minyatur asi kalemlerinin asilanmasini kapsayan ve diger tekniklere nazaran uygulanan en son vejetatif cogaltim tekniklerinden biridir. Bu yontem, surgun ucu kulturu ve asilamanin bazi sinirlayici ozelliklerinin ustesinden gelmekle birlikte, her iki metodun avantajlarini da bir arada bulundurmaktadir. Ilk uygulandigi zamanlarda bazi meyve tur ve cesitlerindeki virus ve benzeri endojen patojenlerin eradikasyonu icin gelistirilen in-vitro asilama, bitki gelisim ve fizyolojisinin farkli alanlarinda cesitli odunsu bitki turlerinde hizla gelismistir. Bunlar bircok odunsu turlerin olgun genotiplerinin klonal cogaltiminda bir on kosul olarak fizyolojik rejuvenasyonu ve asida uyusmazligi da kapsamaktadir. Sonuc olarak in-vitro asilama, yogun olarak kullanilan, diger vejatatif cogaltim metodlarinda bulunan olumsuzluklarin ustesinden gelebilmek icin daha cok dusunulmeyi ve kullanilmayi hak eden bir tekniktir. Doku ve hucreler arasindaki genetik benzerlik ve farkliliklarin ayrintili incelenmesine imkân saglamaktadir. Bu calismanin amaci, 1970’li yillarda turuncgillerde virusten ari bitki uretimi amaciyla baslayan in vitro asilama calismalarinin gunumuze kadar nasil bir surecten gectiginin ve ozellikle meyve turlerinin de dâhil oldugu odunsu bitki turlerinde bitki islahi ve cogaltimi amaciyla ne tur calismalarin yapildiginin ortaya konulmasi ve belli bir duzende sunulmasidir.

In vitro Micrografting of Woody Plant Species-I (Rootstock, Scion, Grafting Technique)

In vitro micrografting is a propagation technique, involving the grafting of relatively miniature cuttings under axenic culture conditions and it is one of the recent developed propagation techniques compared to other conventional vegetative propagation techniques. This method overcomes some of the limitations of shoot tip culture and grafting, while it also keeps together the advantages of both methods. Micrografting was applied for the eradication viruses and pathogens from some fruit species and cultivars during the first application period, but later, the technique was further developed on various woody plant species in different research areas of plant physiology and development. These includes physiological rejuvenation and incompatibility grafting as a prerequisite for the clonally propagation of mature genotypes of many woody species. Consequently, in vitro micrografting is used in large scale propagation and an original technique which needs experience by overcoming the disadvantages of other propagation technique. It also enables to examine in detail the genetic similarities and differences between the tissues and cells. The aims of this study were (1) to review how micrografting studies passed a process from 1970s until today, first started to obtain virus-free plants from citrus; (2) to reveal what kind of work has been presented particularly on the plant breeding and propagation of the woody plant species, including the type of fruit breeding and reproduction and (3) and to present those studies in a specific order.