Micropropagation of wild species of the genus Asparagus L. and their interspecific hybrids with cultivated A. officinalis L., and verification of genetic stability using EST-SSRs

An efficient micropropagation method forasparagus species was developed in this study. The methodallows the fast cloning of the elite genotypes from differentasparagus species and the interspecific hybrids obtainedfrom these species. Rhizome bud explants were disinfectedusing 3 g l-1 of benomyl and 20 g l-1 of sodium hypochlorite.Then, they were cultured on Asparagus RhizomeBud Medium 3 (ARBM-3) consisting in modified Murashigeand Skoog medium with salts with EDDHA-Fe(85.7 mg l-1) instead of EDTA-Fe and vitamins, supplementedwith 0.3 mg l-1 NAA, 0.1 mg l-1 KIN, 2 mg l-1ancymidol and 6 % sucrose. Results showed that themethod developed produced high disinfection rates(70?95 %). More than 70 % of the explants developedshoots and the rooting rate on ARBM-3 medium was30?45 %. The rooting rate increased to 60?85 % when theunrooted shoots were subjected to an additional cycle ofrooting, reaching 100 % after two cycles of rooting. Themultiplication was achieved through mechanical divisionof rooted shoot clusters growing in ARBM-3. The acclimatizationrate of the micropropagated plantlets was higherthan 90 %. The micropopagated plantlets were screened forsomaclonal variation using 12 expressed sequence tagderived simple sequence repeat markers. The results confirmedthe character ??true to type?? of the plantlets, indicatingthat the method developed in this study can be usedto successfully micropropagate asparagus species.Fil: Regalado González, Jose Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Consejo Superior de Investigaciones Científicas; EspañaFil: Carmona Martín, E.. Consejo Superior de Investigaciones Científicas; EspañaFil: Castro, P.. Genetic Improvement of Fruits and Vegetables Laboratory; Estados UnidosFil: Moreno, R.. Universidad de Córdoba; EspañaFil: Gil, J.. Universidad de Córdoba; EspañaFil: Encina, C. L.. Consejo Superior de Investigaciones Científicas; Españ

Reference genome sequence of the model plant Setaria

International audienc

Study of the somaclonal variation produced by different methods of polyploidization in Asparagus officinalis L.

Polyploid plants have been induced in different Asparagus officinalis L. breeding programs in order to obtain plants with improved agronomical traits, such as large spear diameter or segregation ratios with a higher number of males. The polyploidization methods can produce somaclonal variation in the polyploid plants obtained and, therefore, unwanted changes in the agronomical traits of the initial elite plants. We used two different polyploidization methods to induce polyploid plants from diploid genotypes of commercial varieties and tetraploid genotypes of the Spanish landrace “Morado de Huétor”. The first method was the culture of rhizome buds in the medium ARBM-3 (Asparagus Rhizome Bud Medium), supplemented with different concentrations of colchicine (0.1–0.75 g l−1) for 10 and 20 days. The best polyploidization rate obtained was 25 % (0.5 g l−1 colchicine for 10 days). The second method was the regeneration of polyploid plants from callus culture, resulting in a polyploidization rate of 40 and 12.5 % for the diploid genotype CM077 and the tetraploid genotype HT156, respectively. Additionally, we have developed a new protocol to separate the mixoploids generated into their different genetic components, obtaining plants with a unique ploidy level. EST-SSRs markers were employed to analyze the genetic stability of polyploidy plants. Somaclonal variation was not detected for polyploidy plants obtained through the culture of rhizome bud explants. Therefore, these polyploid plants should maintain the agronomical traits of the initial elite plants. However, somaclonal variation was detected in the polyploid plants regenerated from callus culture.Fil: Regalado González, Jose Javier. Instituto de Horticultura Subtropical y Mediterránea La Mayora; España. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Carmona Martín, E.. Instituto de Horticultura Subtropical y Mediterránea La Mayora; EspañaFil: Castro, P.. Universidad de Córdoba; EspañaFil: Moreno, R.. Universidad de Córdoba; EspañaFil: Gil, J.. Universidad de Córdoba; EspañaFil: Encina, C. L.. Instituto de Horticultura Subtropical y Mediterránea La Mayora; Españ