In vitro bulblet regeneration from immature embryos of Muscari azureum

A high frequency bulblet regeneration was achieved for endemic and endangered ornamental plant Muscari azureum using immature embryos. Immature embryos of M. azureum were cultured on callus induction medium consisting of N6 mineral salts and vitamins, 400 mg/L casein + 40 g/L sucrose + 2 g/l L-proline, 2 mg/L 2,4-D and 2 g/L gelrite. Then, embryogenic callus clusters were transferred to bulblet induction medium consisting of MS mineral salts and vitamins containing different concentrations and combinations of N6-benzylamino-purine (BAP), kinetin (KIN), thidiazuron (TDZ), zeatin, indole-3-acetic acid (IAA), -naphthaleneacetic acid (NAA), 30 g/L sucrose and 7 g/L agar. Prolific bulblets multiplication (over 13 bulblets/embryo) was achieved from immature embryos after 5 - 6 months of culture initiation. Well-developed bulblets were excised and individually rooted on ½ strength MS medium supplemented with 1 mg/l IBA, 0.5 g/l activated charcoal, 20 g/l sucrose and 6 g/l agar and acclimatized.Key words: Muscari azureum, bulblet, micropropagation, immature embryo

In vitro micro-propagation of endangered ornamental plant-Neotchihatchewia isatidea (Boiss.) Rauschert

The ornamental plant, Neotchihatchewia isatidea, is an endangered species of Turkey and threatened by complete extinction in the future. Therefore, in vitro multiplication of this species can be valuable forcommercial production and germplasm conservation. Immature embryos of N. isatidea were cultured for initiation on Murashige and Skoog medium (MS) supplemented with N6-benzylamino-purine (BAP)and -naphthaleneacetic acid (NAA). Shoot primordia were visible within 5 - 6 weeks and the shoot primordia later developed into normal shoots 10 - 12 weeks after the culture initiation on calli developedfrom immature embryos. Shoot tips were also excised from developed plantlets for direct shoot organogenesis and cultured on MS shoot induction medium supplemented with BAP (0.5, 1.0 and 2.0mg/l), kinetin (KIN) (0.5, 1.0 and 2.0 mg/l) and thidiazuron (TDZ) (0.05, 0.10 and 0.50 mg/l). Direct multiple shoots from shoot tips developed in most media tested. High shoot multiplication (3.73), high rooting(53 %) number of root per shoot (3.66) and survival ratio (46.6 %) were achieved

TDZ x IBA induced shoot regeneration from cotyledonary leaves and in vitro multiplication in safflower (Carthamus tinctorius L.)

A high frequency adventitious shoot regeneration protocol for safflower (Carthamus tinctorius L.) cv. Dinçer with high yield (Turkish cultivar) using cotyledonary leaves of 10 day-old seedlings were optimized by studying the influence of different combinations of thidiazuron (TDZ) and indole-3-butyric acid (IBA). Cotyledonary leaves were cultured on Murashige and Skoog medium (MS) supplemented with different concentrations and combinations of TDZ and IBA. The highest percentage of regenerated shoots (33.33%) and the highest number of shoots per explant (6.5) occurred on a MS medium containing 0.5 mg/l TDZ and 0.25 mg/l IBA. Furthermore, cotyledonary nodes and meristem tips of 14 day-old seedlings were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of N6-benzylamino-purine (BAP) alone or combination of -naphthalene acetic acid (NAA). Direct multiple shoots from cotyledonary nodes and meristem tips developed within 18 - 21 days in most media tested. 100% shoot multiplication was achieved from cotyledonary node and meristem tip on a range of MS media supplemented with different concentrations of BAP and NAA

TDZ-induced plant regeneration in Astragalus cicer L.

We developed a regeneration protocol using thidiazuron (TDZ) with a high frequency in vitro root induction in Astragalus cicer. High in vitro germination ratio (75%) for hard-seeds of A. cicer was also achieved. For this, hypocotyl and cotyledon explants were cultured on Murashige and Skoog medium supplemented with different concentrations of TDZ. The highest frequency of shoot regeneration (53.3%) was achieved from hypocotyl segments through an initial callus growth stage on MS mediumcontaining 0.25 mg/l TDZ. The shoots were cultured on the different strength (1/1, 3/4, 1/2 and 1/4) of basal Murashige and Skoog medium containing different concentrations of NAA. High rooting (100%)and survival (100%) were achieved using half strength MS medium supplemented with 0.25 and 0.50 mg/l NAA

Efficient in vitro bulblet production of endangered hyacinthella micrantha (Boiss.) chouard

Hyacinthella micrantha (BOISS.) Chouard with their attractive flowers is one of the important endemic and endangered species of Turkey. We aimed to improve a standard in vitro multiplication protocol using basal scales for bulblet production and conservation of the species. Bulb scales of the species collected from natural habitat were cultured in the different basal Murashige and Skoog (MS), Orchimax and Lindeman Orchid Medium embryonic callus induction medium. Then, they were transferred to MS, Orchimax and Lindeman Orchid Medium basal bulblet induction medium supplemented with cytokinins (1.0, 2.0, 4.0 and 8.0 mg/l N6-benzylamino-purine (BAP), Zeatin and 2-isopentyladenine (2-iP) at 16 °C. After ten months of culture initiation, high frequency bulblet regeneration was developed using bulb scales on different basal media containing different concentrations of cytokinintypes. The highest ratio of bulb scales forming shoot/bulblets per explant (87%) and bulblets per explants (5.60) were induced on a Orchimax media supplemented with 8 mg/L Zeatin. MS and Orchimax medium gave the best result in terms of embryonic calli formation and in vitro bulblet production compared to Lindeman Orchid medium. 2,4-D, BAP and Zeatin were more effective than2-iPfor in vitro bulblet production. Also, regenerated bulblets subjected to pre-cold treatments for 3 months at 4-6 ºC resulted in increasing of new mini bulblets (approximately 15%). Regenerated bulblets were acclimatized with high survival ratio (82 %). © 2019, Pakistan Agricultural Scientists Forum. All rights reserved

In vitro micropropagation from immature embryos of the endemic and endangered Muscari muscarimi Medik

An efficient in vitro bulblet production procedure from immature zygotic embryos of endemic and endangered Muscari muscarimi Medik. was described in the current study. Zygotic embryos were first isolated from immature seeds and cultured on different nutrient media compositions supplemented with various combinations of ?-naphthalene acetic acid (NAA), picloram, dicamba, 6-benzylaminopurine (BAP), and thidiazuron (TDZ). The best bulblet regeneration (59 bulblets per explant) was achieved in Murashige and Skoog (MS) medium containing 4 mg/L BAP and 0.5 mg/L NAA after 1 year of culture initiation. Regenerated bulblets were then transferred into MS medium without plant growth regulators for rooting. Bulblets produced well-developed root systems and increased their size on this medium after 2 months. All rooted bulblets were successfully transplanted into a potting mixture and acclimatized to ambient conditions. © TÜBİTAK

Indirect selection of Cre1 gene in winter wheat populations

The nematodes are important biotic constraint in rain-fed wheat production systems. In Turkey, they is found in 75.0% of soil samples in Central Anatolia with the dominant species being Heterodera filipjevi. Yield losses for winter wheat in rain-fed environments are documented between 27.0-46.0 %. A single dominant gene for resistance to H. avenae, designated as Cre1, was assessed in Turkey. It was also found to be effective to Heterodera filipjevi. In this research, a STS based Cre1 marker was applied in a number of segregating wheat populations from F1 to F4 to discriminate Cre1-positive lines among the wheat populations. Results clearly indicated that Marker Assisted Selection (MAS) is functioning effectively, with recovery of Cre1 positive lines up to 88.0 % depending on the cross in early stage of breeding