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The original version is available at http://www.springerlink.com/content/l0788jw70644 ; The aim of this work was to study the effects of colchicine application on chromosome doubling and androgenic response in anther and microspore culture of different bread wheat genotypes. Colchicine was applied during a mannitol stress pretreatment or during the first 48 h of culture at concentrations of 0, 150 and 300 mg l−1. When colchicine was applied during stress pretreatment, the percentage of doubling depended on genotype and concentration. A significant increase in doubling was observed with 300 mg l−1 in the low androgenic responding cv. Caramba. Colchicine incorporation during the first hours of culture improved percentage of doubling in all genotypes, in both anther and microspore culture. Application of 300 mg l−1 colchicine improved the percentage of doubling in the two low responding genotypes, to 118% of control in DH24033, and 75% in Caramba in microspore and anther culture, respectively. Concerning the androgenic response, the effect of colchicine on embryo formation and percentage of green plants depended on the genotype and on the culture method. In cv. Pavon, a 2- and a 3-fold increase in percentage of embryogenesis and green plants, respectively, were obtained with 300 mg l−1 colchicine in microspore culture. However, no significant differences in these two variables were observed in anther culture. The number of green doubled haploid (DH) plants reflects the index of success of the procedure. Regardless of the culture method, when colchicine was incorporated during the first hours of culture, the number of green DH plants increased significantly in three of four genotypes. These results confirm the usefulness of colchicine application during the first hours of culture in wheat breeding programs. ; MS was the recipient of a fellowship from the Ministerio de Educación y Ciencia of the Spanish Government. We thank the Instituto de Agrobiotecnología y Recursos Naturales (UPNA-CSIC) for the use of the PA flow cytometer. The research was supported by Proyects from the Plan Nacional de Recursos y Tecnologías Agroalimentarias AGL2002-04139-C02-02 and AGL2005-07195-C02-01. ; Peer reviewed

12 Pags.- 4 Figs.- 2 Tabls. ; [EN] In recent years, spelt (Triticum aestivum (L.) ssp. Spelta) has become an added-value alternative crop to modern wheat. Spanish spelt constitutes a unique separate gene pool from central European germplasm. The availability of spelt Doubled Haploid (DH) production protocol is a great advantage to speed up breeding programs. This is the first study evaluating the ability of DH plant production, by anther culture, of five Spanish spelt landraces and three F5 lines derived from Spanish spelt x bread wheat crosses. Two central European commercial varieties were also included in the analysis. DH plants were obtained from all material with the exception of one F5 line. The Spanish spelt landraces produced more embryos/100 anthers (73-166.3) than the two European varieties (8.6-22.2). The main bottleneck in the Spanish germplasm was the high number of albino plants regenerated, with percentage of green plants lower than 13% in three of the landraces. Nevertheless, up to 15.6 and 1.8 green plants/100 anthers were obtained from the Spanish and the central European germplasm, respectively. A great variation in the percentage of spontaneous chromosome doubling was obtained, with 4 lines producing around 80% and 2 lines less than 15%. The ovary genotype used for anther co-culture is a critical factor to increase the efficiency of the system. Bread wheat 'Caramba' ovaries increased almost 6 times the number of green plants as compared to spelt landrace 'BG-1987' ovaries. This study shows that DH plants can be produced efficiently from Spanish spelt to be used in breeding programs. ; [AR] (Triticum aestivum (L.) ssp. با اً م علوی )spelt( در سال اّی اخیر، گ ذٌم پ ضَاى برای گ ذٌم هذرى گسی ای با ارزش افس دٍ ضذ است. گ ذٌم پ ضَاى اسپا یًایی دارای خسا Spelta) شیً ه حٌصر ب فرد هجسا از شرم پلاسن اّی ار پٍای هرکسی است. برای سرعت دادى ب بر اًه اّی بسیار هفیذ است. (DH) اصلاح صًاد، دردسترض ب دَى دست رَالعول ت لَیذ اّپل ئَیذ هضاعف گ ذٌم پ ضَاى با ر شٍ کاضت بساک از 5 رقن ب هَی DH ایي خًستیي پص طٍّی است ک ت اَ اًیی ت لَیذ گیا گ ذٌم اًى را ارزیابی هیک ذٌ. ایي x هطتق از تلاقی گ ذٌم پ ضَاى اسپا یًایی F 3 ه رَد لایي 5 )landrace( تحلیل د رقن تجاری ار پٍای هرکسی را یًس ضاهل ب دَ. از وّ ه اَد )گیا یّ( هطالع ضذ ،ُ ب استث اٌء یک ب دست آهذ. ب ازای 100 بساک، در رقن اّی ب هَی گ ذٌم پ ضَاى اسپا یًایی DH گیا اّى ، F لایي 5 8( بیطتر ب دَ. گل گَا /6-22/ 73 ( در هقایس با د رقن ار پٍای هرکسی ) 2 - تعذاد ج یٌي ت لَیذی ) 166.3 باززایی ضذ ب دَ در حالیک ک )albino( اصلی در شرم پلاسن اسپا یًایی زیاد ب دَى تعذاد گیا اّى زال درصذ گیا اّى سبس در س رقن ب هَی کوتر از 13 % ب دَ. با ایي جٍ دَ، از شرم پلاسن اّی اسپا یًایی 1 گیا سبس ب ازای 100 بساک ب دست آهذ. درصذ د /8 15/ ار پٍای هرکسی، ب ترتیب تا حذ 6 تغییرات )spontaneous chromosome doubling ( برابر ضذى خ دَ ب خ دَی کر هٍ زَ مٍ اّ co-( 2 لایي کوتر از 15 % داضت ذٌ. در ر شٍ وّرا -ُکطتی % زیادی طًاى داد، ب ط رَی ک 4 لایي 80 بساک، ش تًَیپ ج یٌي ه رَد استفاد عاهل ه وْی در افسایص ساها است. ج یٌي گ ذٌم اًى )culture افسایص داد. "BG- تعذاد گیا اّى سبس را تا 6 برابر ج یٌي رقن ب هَی گ ذٌم پ ضَاى " 1987 "Caramba" را از گ ذٌم DH ایي پص صٍّ طًاى هی د ذّ ک برای استفاد در بر اًه اّی اصلاح صًاد، هی ت اَى گیا اّى پ ضَاى اسپا یًایی با کارآیی بالا ت لَیذ کرد. ; This work was supported by Projects AGL2013-46698-R and AGL2016-77211-R from the National Plan for Agrofood Resources and Technology of Spain (Plan Nacional de Recursos y Tecnologías Agroalimentarias) of Spain and the Regional Government of Aragon (Diputación General de Aragón) (Grupo A06). ; Peer reviewed

28 Pags.- 6 Tabls.- 1 Fig. This Document is Protected by copyright and was first published by Frontiers. All rights reserved. it is reproduced with permission (Creative Commons Attribution License). ; The use of doubled haploids in onion breeding is limited due to the low gynogenesis efficiency of this species. Gynogenesis capacity from Spanish germplasm, including the sweet cultivar Fuentes de Ebro, the highly pungent landrace BGHZ1354 and the two Valenciana type commercial varieties Recas and Rita, was evaluated and optimized in this study. The OH-1 population, characterized by a high gynogenesis induction, was used as control. Growing conditions of the donor plants were tested with a one-step protocol and field plants produced a slightly higher percentage of embryogenesis induction than growth chamber plants. A one-step protocol was compared with a twostep protocol for embryogenesis induction. Spanish germplasm produced a 2 to 3 times higher percentage of embryogenesis with the two-step protocol, Recas showing the highest percentage (2.09%) and Fuentes de Ebro the lowest (0.53%). These percentages were significantly lower than those from the OH-1 population, with an average of 15% independently of the protocol used. The effect of different containers on plant regeneration was tested using both protocols. The highest percentage of acclimated plants was obtained with the two-step protocol in combination with Eco2box (70%), whereas the lowest percentage was observed with glass tubes in the two protocols (20- 23%). Different amiprofos-methyl (APM) treatments were applied to embryos for chromosome doubling. A similar number of doubled haploid plants were recovered with 25 or 50 μM APM in liquid medium. However, the application of 25 μM in solid medium for 24 h produced the highest number of doubled haploid plants. Somatic regeneration from flower buds of haploid and mixoploid plants proved to be a successful approach for chromosome doubling, since diploid plants were obtained from the 4 regenerated lines. In this study, doubled haploid plants were produced from the four Spanish cultivars, however further improvements are needed to increase their gynogenesis efficiency. ; O. Fayos was a recipient of a predoctoral fellowship from the Spanish Ministry of Economy and Competitiveness FPI (INIA-FEDER). This work was supported by the Spanish Ministry of Economy and Competitiveness INIA-FEDER (RTA2011-00118- C02-01), by the Agreement EEAD-CSIC with CITA-Aragón for Onion Doubled Haploid Production and the Aragon Government (Group A16 and Group A06). ; Peer reviewed

The original publication is available at http://www.springerlink.com/content/lg4126j81q8x15m2/fulltext.pdf ; In order to analyse the genetic control of anther culture response in barley, a doubled haploid (DH) population from the cross between a medium responsive cultivar 'Dobla' and the model cultivar 'Igri' was produced. A linkage map was constructed with 91 markers. A sub-population of 41 lines was characterised for different components of the anther culture response, and was used for quantitative trait loci (QTL) analysis. The vrs1 locus region on chromosome 2H, which determines inflorescence row type, was coincident with the largest putative QTL for number of embryos (nEMB) and albino plants. A region of chromosome 6H was associated with QTLs for nEMB and green plants. QTLs for number and percentage of green plants were located on the long arm of chromosome 5H. Therefore, new QTLs for main components of barley anther culture response were identified on chromosomes 2H, 5H and 6H, indicating that anther culture response in barley could be controlled by relative few genes of large effect. This work is a useful step towards the identification of new regions on the barley genome that could be associated with fundamental biological process implicated in the anther culture response. ; X-W Chen was recipient of a fellowship from the Ministerio de Educación y Cultura from the Spanish government. The research was supported by Projects PB97-1159, and AGL2001-1631 from Plan Comisión Interministerial de Ciencia y Tecnología of Spain. ; Peer reviewed