Abstract
In plant biotechnology, in vitro culture of gametic or sexual cells, microspores or pollen grains, has been described as a successful tool to accelerate genetic improvement, obtaining haploid, homozygotic plants or pure lines in a short time. In chile apple, Capsicum pubescens R and P. Anthers were sown in vitro, and their cytological analysis, locating the meiotic division stage of microspores or pollen grains. Flower buds with diameters from 2.5 to 4.4 mm were pre-incubated at 4°C, in ascorbic and citric acid at 100 and 150 mg-L-1 for 24 h. Five semisolid culture media (A1, A2, A3, A4 and A5) were used, with Murashige and Skoog (1962) salts (MS), modifying iron and vitamin chelates, sucrose, and L-cysteine, 2,4-dichlorophenoxyacetic acid (2,4-D) and Kinetin (Kin). Anthers, in vitro, were plated, in light and dark, for 70 days. Two differentiation media (R1 and R2) were evaluated with 100% MS salts, glycine, kinetin and myo-inositol. The anthers seeded, coincided with the first mitosis of the microspore, the anthers, formed callus in the media (A1) 100 % EDTA-Fe, 0.40 mg-L-1 thiamine, 3 % sucrose) and (A3) 100 % EDTA-Fe, 0.40 mg-L-1 thiamine, 3 % sucrose, 0. 3 mg-L-1 of 2,4-D, and differentiated pro-embryonic structures in (A3) and (A5) 200 % EDTA-Fe, 0.4 mg-L-1 thiamine, 50 mg-L-1 pyridoxine, folic acid, riboflavin and niacin, 0.3 mg-L-1 2,4-D plus 0.3 mg-L-1 Kinetin, as well as roots in (A1). Light influenced the formation of pro-embryos and roots, in the dark callus. The media (R1) and (R2) favored the formation of pro-embryos.
Highlights
Chili bell pepper (Capsicum spp.) is one of the most widely cultivated vegetables in Mexico (SIAP, 2014)
In the results observed in the reductive meiotic division stages of microspores, it was determined that the division stages of microspores in primary stages were strongly associated with the relationship between the size of flower buds, the amount of anthocyanin in anthers and the stage of development of microspores or pollen grains Table 3. Such indicators have been recognized as morphological characteristics of great utility to determine the stages of cell division in the selection and collection of flower bud size and anthers with microspores or pollen grains Figure 1, ensuring the possibility of obtaining potentially embryogenic responses directly or indirectly in different chile bell pepper genotypes, which has been reported by Sibi et al, 1979; Comlekcioglu et al, 2001, Rodeva, 2001; Rodeva et al, 2007; Irikova,2008
In order to obtain haploid, isogenic or pure plants, the particularity of specifying in a timely manner the stages of reductional division of microspores is a process linked to the meiosis of microspores or pollen grains, and in the case of chile, there are reports that locate the most favorable stages, generally during the first mitosis of the microspore (Comlekcioglu et al, 2001); Özkum and Tipirdamaz, 2002; Nowaczyk and Kisiala, 2006; Irikova et al, 2011, Olsewska et al, 2014)
Summary
Chili bell pepper (Capsicum spp.) is one of the most widely cultivated vegetables in Mexico (SIAP, 2014). It is estimated that about 30 species are native to Mexico: five domesticated and 25 wild (Rylski, 1985). Apple chili C. pubescens R. and P. is a species whose fruit is appreciated in America and Mexico for its flavor and high pungency; this species has been increasingly in demand in Mexico and the United States as a result of the diversification of its consumption (Pérez et al, 2004). Apple chile is a species that presents a certain degree of gametophytic self-incompatibility (Saborio and Da Costa, 1992), a phenomenon that somewhat limits the production of inbred lines as hybrid progenitors. The generation of new materials is very low and takes up to five cycles of self-fertilization to achieve a high homozygotic (Pérez, 2002), making it necessary to generate new varieties in a short time
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