Micropropagation and root culture of Turkish endemic Astragalus chrysochlorus (Leguminosae)

Efficient micropropagation and root culture protocols were developed for the endemic Astragalus chrysochlorus Boiss. & Kotschy. A high frequency of shoot formation (100%) and maximum multiplication (13 shoots per hypocotyl explant) were achieved on Murashige and Skoog (MS) media supplemented with 0.5 mg/L trans-Zeatin riboside (ZR). By using single regenerated hypocotyl explants, rooting was achieved at a rate of 93% on MS medium containing 2% sucrose without growth regulators. High frequency callus initiation and growth were achieved when single hypocotyl explants were inoculated on MS medium supplemented with 0.5 mg/L 2,4-Dichlorophenoxyacetic acid. Plant regeneration through indirect organogenesis was achieved on MS medium supplemented with 0.5 mg/L ZR. Root cultures were successfully established in liquid MS medium containing 0.5 mg/L alpha-Naphthaleneacetic acid. The optimised in vitro propagation, callus culture, and root culture protocols offer the possibility to use cell/root culture techniques for vegetative propagation and secondary metabolism studies on Astragalus L. species

IDENTIFICATION AND PRODUCTION OF PHENOLIC NICOTIFLORIN IN ASTRAGALUS CHRYSOCHLORUS CALLUS

Astragalus chrysochlorus Boiss. & Kotschy (2n = 16) is one of the rare Turkish endemic species and it is listed in the Red Data Book of Turkish Plants as endangered. This species has been used traditionally for its wound healing properties and a crude ethanol extract prepared from the roots exhibits antioxidant and cytotoxic activities. In this study, a detailed phytochemical analysis was performed on A. chrysochlorus calli that resulted in the isolation of a major constituent. The purified molecule's structure elucidation was completed by spectral methods [nuclear magnetic resonance (NMR) and mass spectrometry (MS)], which revealed a rarely encountered the flavonoid in the Astragalus genus, nicotiflorin. In order to increasing nicotiflorin content in the callus cultures, the effects of culturing time and elicitor treatment were investigated. The HPLC analyses showed that the maximal production of nicotiflorin occurred with long-term cultured (13 years old) callus as 4775 mu g/g dry weight (DW), whereas it was 132 mu g/g DW for short-term cultured (2 months old) ones. Then, the 24 h treatment of the yeast extract that was used as biotic elicitor had negative effect on the production of nicotiflorin. The data obtained from this study could be significant for the mass production of nicotiflorin from long term in vitro cultured A. chrysochlorus callus

Proteomic analysis of early responsive resistance proteins of wheat (Triticum aestivum) to yellow rust (Puccinia striiformis f. sp tritici) using ProteomeLab PF2D

WOS: 000317715600004Wheat (Triticum aestivum L.) yellow rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most destructive diseases of wheat worldwide. To clarify the molecular details and components of the resistance response in wheat offers further possibilities to combat yellow rust. In this study, differentially regulated early response proteins in wheat leaves infected by Pst isolates were investigated by proteomic approaches. Total proteins extracts from leaves harvested at 24 hour post inoculation (hpi) were separated by two dimensional liquid chromatography system, ProteomeLab PF2D. Following PF2D analysis, six hundred and thirty-seven protein peaks were compared one by one between protein patterns obtained from pathogen-and mock-inoculated leaf tissue. Among those differentially expressed 33 proteins were identified in Pst-infected plants as compared with mock-inoculated controls by nanoLC-ESI-MS/MS. Six proteins were exhibited homology to fungal proteins. Two fungal proteins, including E3 ubiquitin protein ligase and Ubiquitin-like protein, are important members of ubiquitin-proteasome system which the importance of the its proteolytic function in regulating the virulence of pathogenic fungi has just been realized recently. Other identified 27 proteins were host proteins in response to Pst and classified in five groups based on their roles in diverse biological processes. The results indicated that identified defence related proteins such as pathogene related protein 1 and 4 (PR1, PR4), Glutathione S transferase (GST) are major component for systemic acquired resistance (SAR) which is one of the strong disease resistance form in plants and appears within several days following the initial pathogen attack.TUBITAK, COST programmeTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [109T293]This study was supported by TUBITAK, COST programme 109T293 project. We thank Kadir Akan, Ayse Yildiz and Lutfi Cetin for their help during plant inoculation and sampling. We also thank Abdulmecit Gokce and Yavuz Ozturk for their technical support for PF2D as well as Konca Bulut and Rahmi Buyukkeskin for their experimental assistance

Identification of an AFLP marker linked with yellow rust resistance in wheat (Triticum aestivum L.)

An important application of molecular markers in plant systems involves improvement in the efficiency of conventional plant breeding by carrying out indirect selection through molecular markers linked to the traits of interest. AFLP analysis was used to identify molecular markers associated with yellow rust resistance in wheat (Triticum aestivum L.) in this study. DNA isolated from the selected yellow rust tolerant and susceptible F 2 individuals derived from a cross between Izgi2001 (resistant) and ES14 (susceptible) at seedling and adult stage was used for bulked segregant analysis combined with AFLP. From the screening of 34 PstI/MseI AFLP primer combinations, the AFLP marker P-GAC/M-ACG (133 bp) was identified and presented in the resistant parent and resistant F 2 individuals but not in the susceptible ones. Future research will obtain more adjacent sequences associated with the polymorphic M-ACG/P-GAC (133bp) marker by the PCR walking method to design SCAR markers for wheat breeding programs