Articles
INTERGENERIC HYBRIDIZATION, EMBRYO RESCUE AND MOLECULAR DETECTION FOR INTERGENERIC HYBRIDS BETWEEN ASCOCENDA AND PHALAENOPSIS
Article number
829_67
Pages
413 – 416
Language
English
Abstract
Intergeneric hybridization between Ascocenda John De Biase Blue (female) and Phalaenopsis Chih Shang’s Stripes (male) and embryo rescue were used in this study.
Capsules of intergeneric hybrids were harvested after four months of pollination.
The immature embryos with placenta from those capsules were transplanted into artificial tissue culture medium.
About 200-300 intergeneric embryos for each capsule were rescued.
After a one-year subculture and a two-week hardening, intergeneric hybrids were cultivated in a greenhouse.
Different morphologies were found in those intergeneric hybrids including stem high and leaf shape.
To confirm the genetic background of intergeneric hybrids, polymerase chain reaction (PCR) amplified restriction fragment length polymorphism (RFLP) [PCR-RFLP] of an internal transcribed spacer (ITS) of ribosomal DNA was used to analyze the inheritance of hybrids (F1). For the ITS of PCR-RFLP, 60 hybrids were analyzed and the banding pattern of 59 hybrids had biparental inheritance and one hybrid had maternal inheritance.
Unequal crossover or gene conversion occurred to homogenize the repeat sequence of ITS during the intergeneric hybridization on the hybrid with maternal inheritance.
In this study, based on the molecular data, intergeneric hybridization and embryo rescue was successful.
Capsules of intergeneric hybrids were harvested after four months of pollination.
The immature embryos with placenta from those capsules were transplanted into artificial tissue culture medium.
About 200-300 intergeneric embryos for each capsule were rescued.
After a one-year subculture and a two-week hardening, intergeneric hybrids were cultivated in a greenhouse.
Different morphologies were found in those intergeneric hybrids including stem high and leaf shape.
To confirm the genetic background of intergeneric hybrids, polymerase chain reaction (PCR) amplified restriction fragment length polymorphism (RFLP) [PCR-RFLP] of an internal transcribed spacer (ITS) of ribosomal DNA was used to analyze the inheritance of hybrids (F1). For the ITS of PCR-RFLP, 60 hybrids were analyzed and the banding pattern of 59 hybrids had biparental inheritance and one hybrid had maternal inheritance.
Unequal crossover or gene conversion occurred to homogenize the repeat sequence of ITS during the intergeneric hybridization on the hybrid with maternal inheritance.
In this study, based on the molecular data, intergeneric hybridization and embryo rescue was successful.
Authors
C.C. Tsai, Y.C. Chiang, S.C. Huang, W.L. Liu, C.H. Chou
Keywords
moth orchid, internal transcribed spacer, chloroplast DNA, PCR-RFLP
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