Application of shed-microspore culture technique on different pepper (Capsicum annuum L.) genotypes and identification of microspore development stages

Pepper plant (Capsicum annuum L.) which belongs to Solanaceae family has high economic importance because of the proper climatic comditions and high amount of production on wide fields (Including greenhouses) in our region. Especially producing in the greenhouses and extensive usage of F1 vegetable seeds make pepper breeding very populer. Further development of high yielding pepper breds with high quality fruits and resistance against pests and diseases; also, protection of those breds are highly important in our region which has high potential on pepper producing. Along with classical plant breeding methods; biotechnological methods are implemented in our country to produce high-grade plant breds because of the difficulties and slowness on hybrid producing, seed importation from other countries and time-consuming breeding process. Plant tissue culture is the leading application amongst all biotechnological applications about plant breeding. With plant tissue culture applications, new plant breds can be produced and protected much easier and faster. Within plant tissue culture techniques, methods serving plant breeding researches by producing haploid plants have furthermore importance. In our country pepper must be bred as high-grade, high yielding, resitant to pests and diseases, at the same time suitable to future consumption habits as it is produced widely in the high amounts. Plant breeding researches overwhelmingly aim to produce high-grade, resistant and early grown plants as F1 breds. Biotechnological applications can fasten and ease the production of new plant breds. Especially in Vitro androgenesis has been in use in order to produce homogeneous genetic line from heterozygote materials and to produce F1 hybrids (Collonier 2001). This makes breeding process work faster and reveals characteristic controlled by recessive genes which takes long time or impossible to detect with classical plant breeding methods. Researches show that, bacterial infections caused by donor plants can affect haploid plant production or embriyos in classical anther and microspore culture methods negatively; also, some pepper genotypes don't respond to classical anther and microspore culture threatments. Shed-microspore culture technique is a brand new technique developed in order to eliminate these problems by using a new combined culture medium and a different method to isolate anthers and microspores. Anthers preserving immature microspores still in the phase of pollen mithosis I are desirable starting materials for the haploid plant production by androgenesis. Shed-microspore culture technique can be implemented by in vitro cultivation of these anthers in the new double-layer culture medium as they provide shelter to the immature microspores inside theirselves in the beginning. In this work, all the steps of the shed-microspore technique are explained. Commercial varieties of three different pepper genotypes were used as vegetable material. The relationship between the anther length and anthocyanin levels of these cultivars and the developmental stages of microspores in anthers were determined. Embryo development potentials in the liquid nutrient environment were investigated by determining the numbers of embryos and haploid plants that these three different genotypes were forming and their susceptibility to shed-microspore culture technique.