title: Characterization of Genomic Inheritance of Intergeneric Hybrids between Ascocenda and Phalaenopsis Cultivars by GISH, PCR-RFLP and RFLP.
abstract: BACKGROUND
  The intergeneric hybrids between Ascocenda John De Biase 'Blue' and Phalaenopsis Chih Shang's Stripes have been generated to introduce the blue color into the Phalaenopsis germplasm in prior study. In order to confirm the inheritance in hybrid progenies, genomic in situ hybridization (GISH) and restriction fragment length polymorphism (RFLP) analysis were conducted to confirm the intergeneric hybridization status.
  
  
  RESULTS
  GISH analysis showed the presence of both maternal and paternal chromosomes in the cells of the putative hybrids indicating that the putative hybrid seedlings were intergeneric hybrids of the two parents. Furthermore, twenty-seven putative hybrids were randomly selected for DNA analysis, and the external transcribed spacer (ETS) regions of nrDNA were analyzed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and RFLP analyses to identify the putative hybrids. RFLP analysis showed that the examined seedlings were intergeneric hybrids of the two parents. However, PCR-RFLP analysis showed bias to maternal genotype.
  
  
  CONCLUSIONS
  Both GISH and RFLP analyses are effective detection technology to identify the intergeneric hybridization status of putative hybrids. Furthermore, the use of PCR-RFLP analysis to identify the inheritance of putative hybrids should be carefully evaluated.
authors.affiliation: Kaohsiung District Agricultural Research and Extension Station, Pingtung 900, Taiwan.
authors.forename: Wen-Lin
authors.initials: WL
authors.lastname: Liu
authors.affiliation: Department of Nursing, Meiho University, Pingtung 912, Taiwan.
authors.forename: Huei-Chuan
authors.initials: HC
authors.lastname: Shih
authors.affiliation: Kaohsiung District Agricultural Research and Extension Station, Pingtung 900, Taiwan.
authors.forename: I-Szu
authors.initials: IS
authors.lastname: Weng
authors.affiliation: Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung 804, Taiwan.
authors.forename: Ya-Zhu
authors.initials: YZ
authors.lastname: Ko
authors.affiliation: Kaohsiung District Agricultural Research and Extension Station, Pingtung 900, Taiwan.|National Pingtung University of Science and Technology, Pingtung 912, Taiwan.
authors.forename: Chi-Chu
authors.initials: CC
authors.lastname: Tsai
authors.affiliation: Research Center for Biodiversity, China Medical University, Taichung 404, Taiwan.
authors.forename: Chang-Hung
authors.initials: CH
authors.lastname: Chou
authors.affiliation: Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung 804, Taiwan.|Department of Biomedical Science and Environment Biology, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
authors.forename: Yu-Chung
authors.initials: YC
authors.lastname: Chiang
mesh_terms: D001483:Base Sequence; D021901:DNA, Intergenic / Q000235:genetics*; D018745:Genome, Plant; D006824:Hybridization, Genetic; D017403:In Situ Hybridization / Q000379:methods*; D040582:Inheritance Patterns; D008969:Molecular Sequence Data; D029595:Orchidaceae / Q000145:classification / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
chemical_list: D021901:DNA, Intergenic