Optimization of Sterilizing Method of Larvae and Cyst of Heterodera Schachtii and the Possibility of their Application in Sugar Beet Seedlings in In vitro Condition

In this research, after optimization of sterilizing cyst and larvae of second stage of Heterodera schachtii, the possibility of using nematode on seedlings of sugar beet (Beta vulgaris L.) in in vitro condition was studied for developing larvae to cyst. For this purpose, non sterile cysts were extracted from infected soil and hatched into zinc chloride solution with concentration of 0.5gl-1. Then, for preparation of sterile second stage larvae, several sterilizing treatments were used . Mean comparisons were performed between sterilized live larvae number by Duncan's method. Results showed that 70% ethanol for 1 minute followed by 5% sodium hypochlorite for 5 minutes and 0.1% sodium hypochlorite for 20 minutes were the best treatments for disinfecting cysts and larvae, respectively. In the next step, two nematode susceptible sugar beet varieties were applied to produce cyst from the larvae in in vitro culture. PGoB medium containing different hormonal compositions was used to produce hairy roots and inoculation of seedling with sterilized larvae. After nematode inoculation tests, were the stained cysts were observed under stereomicroscope and counted 40 days later. Five to twelve cysts were formed on the roots of each seedling from two varieties. As a result, it seems that this technique can be used for sugar beet germplasm evaluation to screen nematode resistant genotypes in in vitro controlled condition.