Identification Of The Sex Of Buffalograss By The RAPD Molecular Marker And Isozvme Zymogram

Buffalograss (Buchloe dactyloides(Nutt.) Engelm), a perennial grass, belong to the tribe Eragrostoideae of Gremineae family, originated in the arid steppe of North America, covering from Montana to northern Mexico. It is the best forage in the dry season because of the drought tolerant. It is one of good turfgrass because of tolerance of low maintenance, drought.Buffalograss is a dicling plant, that mean there is no difference in morphological features at the early vegetative growth stages (female, male, monoecious) which bring a big trouble to find some useful materials. In this study, we tried to identify the sex of buffalograss using RAPD marker and Isozyme zymogram at early vegetative growth stages and found a special band of male buffalograss which could be used to identify the male buffalograss plant. PPO isozyme zymagram from younger buffalograss leaves’could be used to identify the sex of plant stands. The result could help the breeder identify the sex of buffalograss stands in early stage and find useful materials. The details were listed as following:(1)23identified sex buffalograss (nine were male, ten were female and four were hermaphrodite) for continuous3years were research materials in this experiment. Of these,. Screening of300RAPD primers identified a primer, named S211(5’-ttccccgcga-3’). which is capable of identifying male plants. The specific fragment was cloned, sequenced and submitted to the Gene bank database (Accession Number JN982469).Wrhen the S211primer used to identify the sex of188young buffalograss plants during their first growing season, the correctly identified rate of male is85.8%.(2) Some sex identified bufolograss leaves and spikes also used as the experimental materials to analysis of superoxide dismutase (SOD), peroxidase (POD) isozymes and polyphenol oxidase (PPO) expression through polyacrylamide gel electrophoresis method. The results showed that there was a special band in male-spike’s POD and PPO zymogram analysis, which named.Rf0.12and Rf0.15respectively; A special band named Rf0.12was found in both leaves and spikes’PPO zymogram of monoecious plant analysis. Leaf POD and PPO zymogram analysis could be used for sex determination at early growth stages of Buchloe dactyloides.