Developments of SRAP and SCAR Molecular Markers Associated with Sex in Bama Hemp
In this study,Bama hemp was used as experimental material,the specific fragments linked or related to hemp sex traits were screened using sequence-related amplified polymorphism(SRAP)molecular marker technol-ogy.The results showed that the primer ISSR839 had abundant polymorphisms and clear bands in hemp male and female individuals,then the primer ISSR839 was used to detect the genetic stability between male and female indi-vidual plants and to construct extreme male and female DNA pools.In addition,among 560 pairs of SRAP primers,24 pairs of primers could amplify different bands between male and female DNA pools,among which,the primer pair me11/em12 could steadily amplify a specific fragment of 1 431 bp from the male DNA pool,while this fragment could not be amplified in the corresponding female DNA pool.To convert this specific band into a stable SCAR(sequence characterized amplified region)marker,2～3 bases were extended at the 5'and 3'ends based on the me11/em12 primer pair,resulting in the specific SCAR primer HXF/HXR.This primer was validated in male and female single plants of hemp.The results showed that a unique 1 436 bp band could be amplified in male plants,named SCAR-1436,while the corresponding female plants did not amplify this band.Cloning,sequencing,and comparison of this specific fragment revealed no homologous sequences in the NCBI database.The results of this study can provide a technical means for the sex identification of hemp plants at the seedling stage,reasonably control the ratio of male to female plants,and improve hemp seed yield.
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