A plant DNA minipreparation: Version II

Dellaporta, Stephen L.; Wood, Jonathan; Hicks, James

doi:10.1007/bf02712670

Cited by 6,542 publications

(3,484 citation statements)

References 2 publications

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“…The progeny seedlings were inoculated with a suspension of V. inaequalis field conidia (4×10 5 conidia/ml) as described by Gianfranceschi et al (1996). Assessment of susceptibility and resistance was performed macroscopically after 10-12 days and rated in six classes (Chevalier et al 1991) DNA extraction, bulked segregant analysis and RAPD amplification DNA was extracted as described by Dellaporta's protocol (Dellaporta et al 1983), with minor modifications following Koller et al (1994).…”

Section: Plant Materialsmentioning

confidence: 99%

Molecular markers linked to the apple scab resistance gene Vbj derived from Malus baccata jackii

et al. 2004

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Breeding for scab-resistant apple cultivars by pyramiding several resistance genes in the same genetic background is a promising way to control apple scab caused by the fungus Venturia inaequalis. To achieve this goal, DNA markers linked to the genes of interest are required in order to select seedlings with the desired resistance allele combinations. For several apple scab resistance genes, molecular markers are already available; but until now, none existed for the apple scab resistance gene Vbj originating from the crab apple Malus baccata jackii. Using bulk segregant analysis, three RAPD markers linked to Vbj were first identified. These markers were transformed into more reliable sequence-characterised amplified region (SCAR) markers that proved to be codominant. In addition, three SSR markers and one SCAR were identified by comparing homologous linkage groups of existing genetic maps. Discarding plants showing genotype-phenotype incongruence (GPI plants) plants, a linkage map was calculated. Vbj mapped between the markers CH05e03 (SSR) and T6-SCAR, at 0.6 cM from CH05e03 and at 3.9 cM from T6-SCAR. Without the removal of the GPI plants, Vbj was placed 15 cM away from the closest markers. Problems and pitfalls due to GPI plants and the consequences for mapping the resistance gene accurately are discussed. Finally, the usefulness of co-dominant markers for pedigree analysis is also demonstrated.

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Section: Plant Materialsmentioning

confidence: 99%

Molecular markers linked to the apple scab resistance gene Vbj derived from Malus baccata jackii

et al. 2004

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“…DNA was extracted from the frozen leaf tissue according to Dellapporta et al [9] with minor modification. Following extraction, the DNA was dissolved in TE buffer (10 mM Tris base, 0.1 mM EDTA).…”

Section: Dna Extractionmentioning

confidence: 99%

Fine mapping and marker-assisted selection (MAS) of a low glutelin content gene in rice

Wang

Liu

et al. 2005

Cell Res

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Rice with low glutelin content is suitable as functional food for patients affected with diabetes and kidney failure. The fine mapping of the gene(s) responsible for low glutelin content will provide information regarding the distribution of glutelin related genes in rice genome and will generate markers for the selection of low glutelin rice varieties. Following an SDS-PAGE screen of rice germplasm from Taihu Valley of China, Japonica selection W3660 is identified to be a novel mutant characterized with low glutelin content. For fine mapping the mutant gene for low glutelin content, F 2 and F 3 populations were derived from a cross between W3660 and Jingrennuo. SDS-PAGE analysis of the total endosperm protein showed that the low glutelin content trait was controlled by a single dominant nuclear gene. Genetic mapping, using SSRs, located this gene to chromosome 2, in the region between SSR2-001/SSR2-004 and RM1358. The distances of the two markers to the target gene were 1.1 cM and 3.8 cM respectively. By semi-quantitative RT-PCR analysis, the transcripts of GluB4/GluB5 genes located within the region do not change. However, GluB5 gene located proximal to SSR2-001/SSR2-004 was specifically reduced. SSR profiles of seven Japonica varieties were compared with that of W3660 for loci in the relevant genetic region. The markers SSR2-004 and RM1358 were used for markerassisted selection. The selection efficiencies of SSR2-004 and RM1358 were 96.8% and 92.7% respectively. This provides a standard starting point for the breeding of low glutelin content rice varieties in China.

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“…Genomic DNA was extracted from the leaf tissue collected from putative transgenic (T 0 generation) rice plants grown in greenhouse following the procedure of Dellaporta et al (1983). The 575 bp region of nptII fragment was amplified with nptII-specific primer sequences: 5′-T C C G C T T G C T G A A A AT G T C C -3 ′ , a n d 5 ′ -CTGTTGTGCCCAGTCATAGC-3′; and 478 bp region of g u s w i t h g u s -s p e c i f i c p r i m e r s e q u e n c e s : 5 ′ -G A A G T T C AT C T G C A C C A C C G -3 ′ ; a n d 5 ′ -GGTGCTCAGGTAGTGGTTGT-3′.…”

Section: Pcr Analysesmentioning

confidence: 99%

Agrobacterium-mediated genetic transformation of commercially elite rice restorer line using nptII gene as a plant selection marker

Chakraborty

Reddy

Narasu

et al. 2016

Physiol Mol Biol Plants

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Transformation of commercially important indica cultivars remains challenging for the scientific community even though Agrobacterium-mediated transformation protocols for a few indica rice lines have been well established. We report successful transformation of a commercially important restorer line JK1044R of indica rice hybrid JKRH 401. While following existing protocol, we optimized several parameters for callusing, regeneration and genetic transformation of JK1044R. Calli generated from the rice scutellum tissue were used for transformation by Agrobacterium harboring pCAMBIA2201. A novel two tire selection scheme comprising of Geneticin (G418) and Paramomycin were deployed for selection of transgenic calli as well as regenerated plantlets that expressed neomycin phosphotransferase-II gene encoded by the vector. One specific combination of G418 (30 mg l −1 ) and Paramomycin (70 mg l −1 ) was very effective for calli selection. Transformed and selected calli were detected by monitoring the expression of the reporter gene uidA (GUS). Regenerated plantlets were confirmed through PCR analysis of nptII and gus genes specific primers as well as dot blot using gus gene specific as probe.

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A plant DNA minipreparation: Version II

Cited by 6,542 publications

References 2 publications

Molecular markers linked to the apple scab resistance gene Vbj derived from Malus baccata jackii

Molecular markers linked to the apple scab resistance gene Vbj derived from Malus baccata jackii

Fine mapping and marker-assisted selection (MAS) of a low glutelin content gene in rice

Agrobacterium-mediated genetic transformation of commercially elite rice restorer line using nptII gene as a plant selection marker

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