Modern plant breeding and genetic technologies are used nowadays for breeding vegetables/rapeseed/wheat varieties, including F1-hybrids. Creation of pure lines is one of the basic requirements in F1-hybrids technology. Doubled haploid (DH) lines are completely homozygous lines and anther/isolated microspore culture is an efficient method for DH line production. We have established efficient protocols for doubled haploid production in cucumber (via parthenogenesis induction, Fig1); pepper (via microspore embryogenesis, Fig 2); wheat (via microspore embryogenesis); rapeseed (via microspore embryogenesis) and ornamentals (via microspore embryogenesis).  

Cucumber (C. sativus L.), belonging to the family of cucurbitceae, is one of the most widely cultivated and economically important vegetable. Cucumber productivity can be greatly increased by the use of hybrids as they exhibit high levels of heterosis –the superiority in performance of hybrids compared with their inbred/pure line. In the case of C. sativus, traditional breeding methods often require6-8 years of self-pollination in order to release parental inbred lines. Alternative biotechnological approaches through the androgenesis (in vitro anther/microspore culture), gynogenesis (in vitro ovule/ovary culture), and parthenogenesis(induction of egg cell by pollination with irradiated pollen followed by in vitro haploid embryo rescue)are by far more efficient and sustainable than traditional methods. 

figure 1. In vitro rescued parthenogenic embryo and plant regeneration

Pepper (Capsicum annuum L.) because of its nutritional effects and pharmaceutical application is one of the most important vegetables around the world. Nowadays, F1 hybrids demonstrating unique characteristics are mainly applied seeds, especially in greenhouses. Conventional breeding methods to produce inbred lines through selfing are time and labor consuming. DH technology sheered disadvantages of traditional breeding and made possible achievement to parental lines in one generation. This technology consists of stimulation and germination of haploid plants via anther or microspores culture.

Figure 2. Anther culture of pepper. (A) Embryogenesis (embryoid indicated with arrow) (B) callogenesis (C) Cotyldonary embryoid (D) Regenerated embryoid (E) Regenerated plantlet

Wheat: we have developed a novel and an efficient microspore culture system for haploid/doubled haploid production in wheat in which no particular stress treatment such as heat, cold, and starvation is used for reprogramming microspores. In this simple procedure, the number of green regenerants formed was improved in a wide spectrum of genotypes.

Repeseed: The aim of this research was to study microspore embryogenesis and regeneration in canola (Brassica napus) in order to introduce an applied and efficient protocol for doubled haploid production.