Fluorescence in situ hybridization of single copy transgenes in rice chromosomes

Kharb, Pushpa; Dong, Jiaqi; Islam-Faridi, M. Nurul; Stelly, David M.; Hall, Timothy C.

doi:10.1007/s11627-001-0001-6

Cited by 10 publications

(4 citation statements)

References 42 publications

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“…In plant species with bigger genomes, the application of cDNA sequences as FISH probes allowed chromosome identification, comparative analysis of chromosome structure between species, and identification of gene order along the chromosomes. The method called single-gene FISH was successfully used in rice (Kharb et al 2001 ), maize (Wang et al 2006 ), barley (Karafiátová et al 2013 ), wheat and its wild relatives; i.e., Aegilops caudata , Ae. markgrafii, and Ae.…”

Section: Introductionmentioning

confidence: 99%

The Agropyron cristatum karyotype, chromosome structure and cross-genome homoeology as revealed by fluorescence in situ hybridization with tandem repeats and wheat single-gene probes

et al. 2018

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Key messageFluorescence in situ hybridization with probes for 45 cDNAs and five tandem repeats revealed homoeologous relationships of Agropyron cristatum with wheat. The results will contribute to alien gene introgression in wheat improvement.AbstractCrested wheatgrass (Agropyron cristatum L. Gaertn.) is a wild relative of wheat and a promising source of novel genes for wheat improvement. To date, identification of A. cristatum chromosomes has not been possible, and its molecular karyotype has not been available. Furthermore, homoeologous relationship between the genomes of A. cristatum and wheat has not been determined. To develop chromosome-specific landmarks, A. cristatum genomic DNA was sequenced, and new tandem repeats were discovered. Their distribution on mitotic chromosomes was studied by fluorescence in situ hybridization (FISH), which revealed specific patterns for five repeats in addition to 5S and 45S ribosomal DNA and rye subtelomeric repeats pSc119.2 and pSc200. FISH with one tandem repeat together with 45S rDNA enabled identification of all A. cristatum chromosomes. To analyze the structure and cross-species homoeology of A. cristatum chromosomes with wheat, probes for 45 mapped wheat cDNAs covering all seven chromosome groups were localized by FISH. Thirty-four cDNAs hybridized to homoeologous chromosomes of A. cristatum, nine hybridized to homoeologous and non-homoeologous chromosomes, and two hybridized to unique positions on non-homoeologous chromosomes. FISH using single-gene probes revealed that the wheat-A. cristatum collinearity was distorted, and important structural rearrangements were observed for chromosomes 2P, 4P, 5P, 6P and 7P. Chromosomal inversions were found for pericentric region of 4P and whole chromosome arm 6PL. Furthermore, reciprocal translocations between 2PS and 4PL were detected. These results provide new insights into the genome evolution within Triticeae and will facilitate the use of crested wheatgrass in alien gene introgression into wheat.Electronic supplementary materialThe online version of this article (10.1007/s00122-018-3148-9) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

The Agropyron cristatum karyotype, chromosome structure and cross-genome homoeology as revealed by fluorescence in situ hybridization with tandem repeats and wheat single-gene probes

et al. 2018

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“…FISH detection of transgenes in cereal crops has been reported in wheat, barley, triticale (Pedersen et al, 1997), oat (Leggett et al, 2000;Svitashev et al, 2000) and maize (Castro and Williams, 1997). FISH applications for rice genome research have been conducted in many studies (Jiang et al, 1995(Jiang et al, , 1996a(Jiang et al, , 1996bDong et al, 2001;Kharb et al 2001;Jin et al, 2002). Indeed, a great deal of information has been obtained from the visualization of transgenes by FISH on metaphase or interphase spreads, and this has led to supplement molecular interpretation of the position of the inserted genes based on PCR, southern blot hybridization, sequencing, etc.…”

Section: Introductionmentioning

confidence: 99%

Detection of transgenes in three genetically modified rice lines by fluorescence in situ hybridization

et al. 2010

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Fluorescence in situ hybridization (FISH) using T-DNA probes was applied to localize transgenes onto specific chromosomes and confirm the steady integration of transferred genes in three genetically modified (GM) rice lines, LS28 (event LS30-32-20-1), Cry1Ac1 (event C7-1-9-1) and LS28×Cry1Ac1 (event L/C1-1-3-1), which are a rice leaf blast-resistant single trait GM line, a leaf folder-resistant single trait GM line, and a rice leaf blast-resistant and leaf folder-resistant stacked GM hybrid line, respectively. The FISH signals were clearly detected on the arms of one homologous chromosome pair for LS28, and on the arms of another chromosome pair for Cry1Ac1 when using the transformation vector pSBM AtCK containing the rice leaf blast-resistant gene (LS28) and pMJ-RTB containing the leaf folder-resistant gene (mCry1Ac1) as a probe, respectively. As expected, we detected two pairs of FISH signals, each on the arms of different chromosome pairs in the stacked GM rice line LS28×Cry1Ac1 when using both pSBM AtCK and pMJ-RTB as probes. These results indicate that the transgenes are located at specific homologous loci and show position stability among generations in both single trait and stacked GM rice lines. The usefulness and the necessity of FISH to detect inserted genes in transformed plants will be discussed for the purpose of future studies to develop breeding programs and conduct risk assessment of GM plants.

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“…The procedure has been used in rice, barley and wheat (Anand et al, 2003) to determine copy number and chromosomal location of alien genes (Perdersen et al, 1997). Several transgenes have been reported by using FISH including Xa21 or GUS (Kharb et al, 2001), GFP (Chen et al, 2003), chitinase and β-1,3-glucanase target genes (Anand et al, 2003) and GUS and bar genes (Liu et al, 2020). Gene expression in transformed plants is strongly dependent on the integrity and organization of the transgenic locus.…”

Section: Introductionmentioning

confidence: 99%

Impact of Lycopene Supplemented Canola Meal-Based Diet on Growth Performance, Nutrient Digestibility and Antioxidant Status of Catla Catla Fingerlings

2021

THE JAPS

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The presence of Bt gene, cry1Ac in transgenic lines of Gossypium hirsutum L. was determined by molecular and cytogenetic analysis. Fluorescence in situ hybridization (FISH) was carried out to study the location of transgene on chromosomes. Metaphase chromosomes prepared from root tips and pollen grains showed more condensation, and hence more karyotypically differentiated. The five transgenic lines of cotton showed stable integration of transgene, cry1Ac. Either the lines were heterozygous (CEMB-3010-14, CEMB-3013-10, CEMB-3016-13 and CEMB-3037-2) or homozygous (CEMB-3034-10) showing single and double genes respectively at interphase and metaphase FISH while the control lines showed no fluorescent signal for transgene at all. Metaphase chromosomes prepared from pollens were also analyzed and single copy of cry1Ac was mapped on chromosome 10 which showed the stable inheritance of transgene in the progeny. The information obtained during study may be helpful to understand the possible effects of copy number variation and position effect of transgene in Bt cotton. The study may also help to comprehend the process of transgene inheritance in Bt cotton lines.

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Fluorescence in situ hybridization of single copy transgenes in rice chromosomes

Cited by 10 publications

References 42 publications

The Agropyron cristatum karyotype, chromosome structure and cross-genome homoeology as revealed by fluorescence in situ hybridization with tandem repeats and wheat single-gene probes

The Agropyron cristatum karyotype, chromosome structure and cross-genome homoeology as revealed by fluorescence in situ hybridization with tandem repeats and wheat single-gene probes

Detection of transgenes in three genetically modified rice lines by fluorescence in situ hybridization

Impact of Lycopene Supplemented Canola Meal-Based Diet on Growth Performance, Nutrient Digestibility and Antioxidant Status of Catla Catla Fingerlings

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