caa a22 4500 445324031 CHVBK 20180317142714.0 cr unu---uuuuu 170323e20110301xx s 000 0 eng 10.1007/s10681-010-0303-z doi (NATIONALLICENCE)springer-10.1007/s10681-010-0303-z Genetic, quantitative and microscopic evidence for fusion of haploid nuclei and growth of somatic calli in cultured ms10 35 tomato anthers [Elektronische Daten] [Patricia Corral-Martínez, Fernando Nuez, José Seguí-Simarro] In plant breeding, androgenic doubled haploids represent powerful tools to save time and resources for pure line generation. While in many species efficient protocols are known, in tomato (Solanum lycopersicum), the knowledge on the induction of androgenesis is still very scarce, and little is known about the particularities of this highly recalcitrant species. The only known method capable of yielding haploid/doubled haploid tomato plants is anther culture. However, this method has important limitations, including low efficiency of haploid induction and a low proportion of spontaneously doubled haploids. To understand these limitations better, we have analyzed the process of callus formation in anthers of tomato lines carrying the ms10 35 gene for male-sterility, using light and electron microscopy, flow cytometry and genetic analysis with morphological and molecular markers. Our results demonstrate that haploid, doubled haploid and diploid calli occur in tomato anthers, although at different frequencies. Diploid calli derived either from somatic cells or from the fusion of two genetically different haploid nuclei account for more than 90% of the total of calli produced. Somatic calli are derived from the stubs of connective tissue present in the interlocular septa of anthers. This growth is markedly increased in the ms10 35 mutants, which explains their higher callogenic rates than standard tomato lines. Together, our results reveal serious drawbacks that explain the low efficiency of anther-derived, doubled haploid production in tomato, and stress the need for alternatives towards doubled haploidy. The Author(s), 2010 Androgenesis nationallicence Anther culture nationallicence Doubled haploid nationallicence Male sterility nationallicence Meiocyte nationallicence Nuclear fusion nationallicence Solanum lycopersicum nationallicence Corral-Martínez Patricia COMAV—Universidad Politécnica de Valencia, CPI, Edificio 8E—Escalera I, Camino de Vera, s/n, 46022, Valencia, Spain aut Nuez Fernando COMAV—Universidad Politécnica de Valencia, CPI, Edificio 8E—Escalera I, Camino de Vera, s/n, 46022, Valencia, Spain aut Seguí-Simarro José COMAV—Universidad Politécnica de Valencia, CPI, Edificio 8E—Escalera I, Camino de Vera, s/n, 46022, Valencia, Spain aut Euphytica Springer Netherlands 178/2(2011-03-01), 215-228 0014-2336 178:2<215 2011 178 10681 https://doi.org/10.1007/s10681-010-0303-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10681-010-0303-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Corral-Martínez Patricia COMAV—Universidad Politécnica de Valencia, CPI, Edificio 8E—Escalera I, Camino de Vera, s/n, 46022, Valencia, Spain aut NATIONALLICENCE 700 1- Nuez Fernando COMAV—Universidad Politécnica de Valencia, CPI, Edificio 8E—Escalera I, Camino de Vera, s/n, 46022, Valencia, Spain aut NATIONALLICENCE 700 1- Seguí-Simarro José COMAV—Universidad Politécnica de Valencia, CPI, Edificio 8E—Escalera I, Camino de Vera, s/n, 46022, Valencia, Spain aut NATIONALLICENCE 773 0- Euphytica Springer Netherlands 178/2(2011-03-01), 215-228 0014-2336 178:2<215 2011 178 10681 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer