Screening and analysis of sex-related ISSR molecular marker in Idesia polycarpa Maxim. var. vestita Diels

Abstract:

In order to screen the sex-related ISSR molecular marker in Idesia polycarpa Maxim. var. vestita Diels，the inter-simple sequence repeat-polymerase chain reaction (ISSR-PCR) system was established and optimized．And we found the best ISSR-PCR reaction system with total volume of 25μL was 55 ng DNA template， 1U Taq polymerase，2μL dNTPs tendency（2.5 mmol/L)，2μL primers (10μmol/L)，2.5μL 10 x Taq Buffer (including magnesium ion)and then adding ultrapure water to make up total 25μL．With this system we discovered only one ISSR primer——UBC841，which could amplify one stable female-special fragment，with size of 250-300bp．By sequencing the female-special fragment，we found the fragment was made up of many fragments with similar sizes，which indicated the female-special fragment showed by PAGE  electrophoresis was a mixed bands．Even so，the primer UBC841 still can be used for sex-related markers to identify the gender of Idesia polycarpa Maxim. var. vestita Diels．