Production of ?super-males? of asparagus by anther culture and its detection with SSR-ESTs

E. CARMONA MARTÍN; J. GIL; J. J. REGALADO ; R. MORENO; E. MADRID; C. L. ENCINA

PLANT CELL TISSUE AND ORGAN CULTURE

SPRINGER

Lugar: Berlin; Año: 2016 vol. 124 p. 119 - 135

0167-6857

Anther culture is used to develop asparagus??super-male?? (di-haploids) in asparagus, which can beused to develop ??all-male?? varieties, by crossing them withsuitable females; their progenies will be formed only bymales which is advantageous for producers. This reportdescribe a new anther culture protocol adapted to ??Moradode Hue´tor??, a Spanish tetraploid landrace, and studied thedifferent factors involved in callus proliferation successfrom anther explants such as the microspore developmentstage, or the type of stress used to induce the symmetricdivision of the microspores, to obtain a high success rate(90 %). For plantlets regenerates from anther culture(PRACs) regeneration we develop a proliferation mediasupplemented with a combination of pCPA and BA able toinduce callus proliferation and plantlet regeneration in thesame step in a 50 % of calli, simplifying the procedure.The high percentage of heterozygous male recovery,originated from somatic cells, is an important problem inthe anther culture, and to elucidate the origin of PRACs wehave combined different tools: ploidy analysis, characterizationwith the linked sex-marker Asp1-T7 and with ESTSRRs.We can establish that 50 % of PRACs obtained inthis work were regenerated from diploid microspores of??Morado de Hue´tor??, regenerating diploid, di-diploid andtetra-diploid plantlets. The di-diploids males (MMmm)would generate a ratio male:female of 5:1 (83.3 %) and thetetra-diploid males (MMMMmmmm) a ratio male:female of69:1 (98.6 %), so the tetra-diploid males could be considered??super-males?? and be used to develop ??all-male??varieties of ??Morado de Hue´tor??.