Efficient Plant Regeneration from Rice Protoplasts Through Somatic Embryogenesis

Abdullah, Rohana; Cocking, E. C.; Thompson, John A.

doi:10.1038/nbt1286-1087

Cited by 195 publications

(57 citation statements)

References 18 publications

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“…Production of seed material from crosses and plant regeneration has been described (11,12). Seeds were surface-sterilized by washing in 70% (vol/vol) ethanol for 10 min and 10% (vol/vol) domestic bleach for 30 min, followed by six washes in sterile distilled water.…”

Section: Methodsmentioning

confidence: 99%

Detection of DNA "fingerprints" of cultivated rice by hybridization with a human minisatellite DNA probe.

Dallas

1988

Proc. Natl. Acad. Sci. U.S.A.

176

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A human minisatellite DNA probe detects several restriction fragment length polymorphisms in cultivars of Asian and African rice. Certain fragments appear to be inherited in a Mendelian fashion and may represent unlinked loci. The hybridization patterns appear to be cultivar-specific and largely unchanged after the regeneration of plants from tissue culture. The results suggest that these regions of the rice genome may be used to generate cultivar-specific DNA fingerprints. The demonstration of similarity between a human minisatellite sequence and polymorphic regions in the rice genome suggests that such regions also occur in the genomes of many other plant species.The detection of restriction fragment length polymorphisms (RFLPs) by single-copy or cDNA probes has allowed the construction of linkage maps in some crop plants (1, 2) and the diagnostic identification of strains of commercial fungi (3). In these studies, the probes detecting RFLPs were isolated from random collections of single-copy DNA or cDNA sequences of each species. In cases where the level of RFLP detected by single-copy DNA sequences is low, hypervariable regions that are dispersed throughout the genome may provide more informative RFLP markers. Furthermore, such regions will be systematically detectable if they share sufficient sequence similarity between distantly related species. The approach taken in this study was to determine whether human minisatellite probes (4) can detect RFLPs in cultivated rice. The human minisatellite family (4, 5) consists of many dispersed arrays of short (10-to 50-basepair) tandem direct repeats that contain variants ofa common "core" sequence. The arrays exhibit a high degree of length variability, probably due to changes in the copy number of tandem repeats. The "polycore" probes, 33.6 and 33.15 (4), were chosen due to their ability to detect polymorphic regions in vertebrate genomes (4-9). The Asian and African cultivated rices, Oryza sativa and Oryza glaberrima, were chosen for their small genome size (0.6 pg ofDNA per haploid genome; ref. 10) and the availability of crosses and regenerated material. The former was expected to facilitate the detection of low levels of core sequence similarity. MATERIALS AND METHODSRice seeds were obtained from the International Rice Research Institute (Manila, Philippines) and from the Botany Department, University of Nottingham. Production of seed material from crosses and plant regeneration has been described (11, 12). Seeds were surface-sterilized by washing in 70% (vol/vol) ethanol for 10 min and 10% (vol/vol) domestic bleach for 30 min, followed by six washes in sterile distilled water. Seeds were germinated on MSO agar (13) and grown for 3-6 weeks (30'C, 18-hr light cycle). Genomic DNA was isolated by a rapid method devised for fungi (14). Restriction enzyme digests contained 3-5 pug of DNA and 10 units of restriction enzyme with the supplier's recommended buffer (Bethesda Research Laboratories), as well as 2 tig of RNase per ml and 4 mM spermidine trihydrochl...

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Section: Methodsmentioning

confidence: 99%

Detection of DNA "fingerprints" of cultivated rice by hybridization with a human minisatellite DNA probe.

Dallas

1988

Proc. Natl. Acad. Sci. U.S.A.

176

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“…Isolation and culturing of the protoplasts followed the methods proposed by Abdullah et al (1986). Six days after subculturing, protoplasts were isolated from the suspension cells.…”

Section: Isolation and Culturing Of The Protoplastsmentioning

confidence: 99%

Production of bacterial blight resistant lines from somatic hybridization between Oryza sativa L. and Oryza meyeriana L.

Yan

Qian

Gong

et al. 2004

J. Zheijang Univ.-Sci.

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Novel bacterial blight (BB) resistance gene(s) for rice was (were) introduced into a cultivated japonica rice variety Oryza sativa (cv. 8411), via somatic hybridization using the wild rice Oryza meyeriana as the donor of the resistance gene(s). Twenty-nine progenies of somatically hybridized plants were obtained. Seven somatically hybridized plants and their parents were used for AFLP (amplified fragment length polymorphism) analysis using 8 primer pairs. Results confirmed that these plants were somatic hybrids containing the characteristic bands of both parents. The morphology of the regenerated rice showed characters of both O. sativa and O. meyeriana. Two somatic hybrids showed highest BB resistance and the other 8 plants showed moderate resistance. The new germplasms with highest resistance have been used in the rice breeding program for the improvement of bacterial blight resistance.

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“…For transformation, 1 mL of protoplast suspension was incubated with 10 pg of circular plasmid DNA, 50 pg/mL calf thymus carrier DNA, and an equal volume of 30% polyethylene glycol 4000 (Sigma, St. Louis, MO). The mixture was incubated for 30 min, diluted with CPW13 medium (Abdullah et al, 1986), washed 3 times in CPW13 media, with centrifugation between each wash, before being resuspended in simplified KPR liquid media (Zhang and Wu, 1988) to a final density of 1 O6 protoplasts/mL. Aliquots (0.1 mL) of this final suspension were incubated by thin-layer culture in 96-well plates (Krackeler Scientific, Inc., Albany, NY) before subsequent assays for GUS activity (Jefferson, 1987;Zhang and Wu, 1988).…”

Section: Culture Transformation and Visualization Of Gus Activity Imentioning

confidence: 99%

Isolation of an efficient actin promoter for use in rice transformation.

et al. 1990

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We have characterized the 5' region of the rice actin 1 gene (Actl) and show that it is an efficient promoter for regulating the constitutive expression of a foreign gene in transgenic rice. By constructing plasmids with 5' regions from the rice Actl gene fused to the coding sequence of a gene encoding bacterial P-glucuronidase, we demonstrate Pthat a region 1.3 kilobases upstream of the Actl translation initiation codon contains all of the 5'-regulatory elements necessary for high-leve1 P-glucuronidase (GUS) expression in transient assays of transformed rice protoplasts. The rice Actl primary transcript has a noncoding exon separated by a 5' intron from the first coding exon. Fusions that lack this Actl intron showed no detectable GUS activity in transient assays of transformed rice protoplasts. Deletion analysis of the Act 7 5' intron suggests that the intron-mediated stimulation of GUS expression is associated, in part, with an in vivo requirement for efficient intron splicing.

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Efficient Plant Regeneration from Rice Protoplasts Through Somatic Embryogenesis

Cited by 195 publications

References 18 publications

Detection of DNA "fingerprints" of cultivated rice by hybridization with a human minisatellite DNA probe.

Detection of DNA "fingerprints" of cultivated rice by hybridization with a human minisatellite DNA probe.

Production of bacterial blight resistant lines from somatic hybridization between Oryza sativa L. and Oryza meyeriana L.

Isolation of an efficient actin promoter for use in rice transformation.

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