Abstract

The genomic relationship between Oryza sativa (2n = 24 AA) and Oryza australinesis (2n = 24 EE) has not been established. Genomic relationship between these two species was assessed by using three strategies: genomic in situhybridization (GISH), meiotic chromosome pairing, pollen and spikelet sterility. The hybrid was produced between these two species at the International Rice Research Institute using embryo rescue technique. The chromosome pairing was examined in pollen mother cells of O. australinesis, O. sativa and the hybrid between O. sativa and O. australinesis. The hybrid was highly sterile with pollen stain ability being 0.05%. Both parents showed regular meiosis with normal chromosome pairing. The F1 hybrid exhibited limited chromosome pairing. On an average, 0 - 4 bivalents and 16 - 24 univalents were recorded at metaphase-1. The most frequent configuration was two bivalent and twenty univalent. The chromosomes of O. australiensis appeared larger and darkly stained. For genomic in situ hybridization, genomic DNA from O. australiensis was used as probe for the mitotic and meiotic chromosomes of the hybrid between O. sativa and O. australiensis. GISH revealed unequivocal discrimination of O. australiensis chromosomes that appeared yellow due to hybridization signal from O. sativa chromosomes that fluoresced red due to counterstaining with propidium iodide (PI). No cross hybridization was examined between the labeled genomic DNA of O. australiensis and the chromosomes of O.sativa. The paired chromosomes were discriminated as autosyndetic andallosyndetic pairing. Meiotic and mitotic chromosomes of the O. australiensis andO. sativa, in the hybrid were discriminated by GISH for the first time. Results showed that both genomes were highly divergent.   Key words: Genomic in situ hybridization, wide hybrid, chromosome pairing.

Highlights

  • Wide hybridization is one of the plant breeding approaches for incorporating alien genetic variation to cultivated species

  • For genomic in situ hybridization, genomic DNA from O. australiensis was used as probe for the mitotic and meiotic chromosomes of the hybrid between O. sativa and O. australiensis

  • Many important characteristics have been incorporated into rice by alien gene transfer for example, grassy stunt resistance from Oryza nivara (Khush, 1977), bacterial blight (BB) resistance (Xa21) from Oryza longistaminata (Khush et al 1990), brown plant hopper (BPH) and white backed plant hopper (WBPH) resistance from Oryza officinalis (Jena and Khush, 1990), blast and bacterial blight resistance from Oryza minuta (Amante et al 1992), cytoplasmic male sterility from Oryza perennis (Dalmacio et al, 1995) and Abbasi et al 1313 bacterial blight resistance from Oryza brachyantha (Brar et al, 1996)

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Summary

Assessment of genomic relationship between Oryza sativa and Oryza australinesis

The genomic relationship between Oryza sativa (2n = 24 AA) and Oryza australinesis (2n = 24 EE) has not been established Genomic relationship between these two species was assessed by using three strategies: genomic in situ hybridization (GISH), meiotic chromosome pairing, pollen and spikelet sterility. The hybrid was highly sterile with pollen stain ability being 0.05% Both parents showed regular meiosis with normal chromosome pairing. No cross hybridization was examined between the labeled genomic DNA of O. australiensis and the chromosomes of O. sativa. Meiotic and mitotic chromosomes of the O. australiensis and O. sativa, in the hybrid were discriminated by GISH for the first time. Results showed that both genomes were highly divergent

INTRODUCTION
Preparation of genomic DNA
Genomic in situ hybridization
Meiotic chromosome preparation
Mitotic chromosome preparation
Pollen sterility
RESULTS AND DISCUSSION
Chromosome pairing
Pollen grain
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