scholarly journals An improved suppression subtractive hybridization technique to develop species-specific repetitive sequences from Erianthus arundinaceus (Saccharum complex)

2018 ◽  
Vol 18 (1) ◽  
Author(s):  
Fan Yu ◽  
Yongji Huang ◽  
Ling Luo ◽  
Xueting Li ◽  
Jiayun Wu ◽  
...  
Keyword(s):  
Suppression Subtractive Hybridization ◽  
Repetitive Sequences ◽  
Subtractive Hybridization ◽  
Hybridization Technique ◽  
Saccharum Complex ◽  
Erianthus Arundinaceus ◽  
Species Specific

scholarly journals Efficient anchoring of Erianthus arundinaceus chromatin introgressed into sugarcane by specific molecular markers

2020 ◽  
Author(s):  
Yongji Huang ◽  
Jiayun Wu ◽  
Xueting Li ◽  
Fan Yu ◽  
Xuguang Hu ◽  
...  
Keyword(s):  
Molecular Markers ◽  
High Throughput ◽  
Repetitive Sequence ◽  
Repetitive Sequences ◽  
Distribution Patterns ◽  
Subtractive Hybridization ◽  
High Accuracy ◽  
Wild Relatives ◽  
45S Rdna ◽  
Erianthus Arundinaceus

Abstract Erianthus arundinaceus is a valuable gene reservoir for sugarcane improvement. However, insufficient molecular markers for high-accuracy identification and tracking of the introgression status of E. arundinaceus chromatin impede sugarcane breeding. Fortunately, suppression subtractive hybridization (SSH) technology provides an excellent opportunity for development of high-throughput E. arundinaceus-specific molecular markers at a reasonable cost. In this study, we constructed a SSH library of E. arundinaceus. In total, 288 clones E. arundinaceus-specific repetitive sequences were screened out and their distribution patterns on chromosomes were characterized by fluorescence in situ hybridization (FISH). A subtelomeric repetitive sequence Ea086 and a diffusive repetitive sequence Ea009, plus 45S rDNA-bearing E. arundinaceus chromosome repetitive sequence EaITS were developed as E. arundinaceus-specific molecular markers, namely Ea086-128, Ea009-257, and EaITS-278, covering all the E. arundinaceus chromosomes for high-accuracy identification of putative progeny. Both Ea086-128 and Ea009-257 were successfully applied to identify the authenticity of F1, BC1, BC2, BC3, and BC4 progeny between sugarcane and E. arundinaceus. In addition, EaITS-278 was a 45S rDNA-bearing E. arundinaceus chromosome-specific molecular marker for rapid tracking the inherited status of this chromosome in sugarcane background. Three BC3 progeny had apparently lost the 45S rDNA-bearing E. arundinaceus chromosome. We reported herein a highly effective and reliable SSH-based technology for discovery of high-throughput E. arundinaceus-specific sequences bearing high potential as molecular markers. Given its reliability and savings in time and efforts, the method is also suitable for development of species-specific molecular markers for other important wild relatives to accelerate introgression of wild relatives into sugarcane.

scholarly journals Efficient anchoring of Erianthus arundinaceus chromatin introgressed into sugarcane by specific molecular markers

2020 ◽  
Author(s):  
Yongji Huang ◽  
Jiayun Wu ◽  
Xueting Li ◽  
Fan Yu ◽  
Xuguang Hu ◽  
...  
Keyword(s):  
Molecular Markers ◽  
High Throughput ◽  
Repetitive Sequence ◽  
Repetitive Sequences ◽  
Distribution Patterns ◽  
Subtractive Hybridization ◽  
High Accuracy ◽  
Wild Relatives ◽  
45S Rdna ◽  
Species Specific

Abstract Background: Erianthus arundinaceus is a valuable gene reservoir for sugarcane improvement. However, insufficient molecular markers for high-accuracy identification and tracking of the introgression status of E. arundinaceus chromatin impede sugarcane breeding. Fortunately, suppression subtractive hybridization (SSH) technology provides an excellent opportunity for development of high-throughput E. arundinaceus-specific molecular markers at a reasonable cost. Results: In this study, we constructed a SSH library of E. arundinaceus. In total, 288 clones E. arundinaceus-specific repetitive sequences were screened out and their distribution patterns on chromosomes were characterized by fluorescence in situ hybridization (FISH). A subtelomeric repetitive sequence Ea086 and a diffusive repetitive sequence Ea009, plus 45S rDNA-bearing E. arundinaceus chromosome repetitive sequence EaITS were developed as E. arundinaceus-specific molecular markers, namely Ea086-128, Ea009-257, and EaITS-278, covering all the E. arundinaceus chromosomes for high-accuracy identification of putative progeny. Both Ea086-128 and Ea009-257 were successfully applied to identify the authenticity of F1, BC1, BC2, BC3, and BC4 progeny between sugarcane and E. arundinaceus. In addition, EaITS-278 was a 45S rDNA-bearing E. arundinaceus chromosome-specific molecular marker for rapid tracking the inherited status of this chromosome in sugarcane background. Three BC3 progeny had apparently lost the 45S rDNA-bearing E. arundinaceus chromosome. Conclusions: We reported herein a highly effective and reliable SSH-based technology for discovery of high-throughput E. arundinaceus-specific sequences bearing high potential as molecular markers. Given its reliability and savings in time and efforts, the method is also suitable for development of species-specific molecular markers for other important wild relatives to accelerate introgression of wild relatives into sugarcane.

Suppression subtractive hybridization as a basis to assess Mycoplasma agalactiae and Mycoplasma bovis genomic diversity and species-specific sequences

Microbiology ◽  
2005 ◽  
Vol 151 (2) ◽  
pp. 475-489 ◽  
Author(s):  
Marc S. Marenda ◽  
Evelyne Sagné ◽  
François Poumarat ◽  
Christine Citti
Keyword(s):  
Suppression Subtractive Hybridization ◽  
Type Strain ◽  
Subtractive Hybridization ◽  
Genomic Diversity ◽  
Mycoplasma Species ◽  
Mycoplasma Bovis ◽  
Dna Fragments ◽  
Mycoplasma Agalactiae ◽  
Field Isolates ◽  
Species Specific

The phylogenically related Mycoplasma agalactiae and Mycoplasma bovis species are two ruminant pathogens difficult to differentiate and for which a limited amount of sequence data are available. To assess the degree of genomic diversity existing between and within these mycoplasma species, sets of DNA fragments specific for M. bovis type-strain PG45 or for M. agalactiae type-strain PG2 were isolated by suppression subtractive hybridization and used as probes on a panel of M. agalactiae and M. bovis field isolates. Results indicated that approximately 70 % of the DNA fragments specific to one or the other type strain are represented in all field isolates of the corresponding species. Only one M. bovis isolate, which was first classified as M. agalactiae, reacted with 15 % of the PG2-specific probes, while several M. agalactiae isolates reacted with 15 % of the PG45-specific probes. Sequence analyses indicated that most of the genomic diversity observed within one species is related to ORFs with (i) no homologies to proteins recorded in the databases or (ii) homologies to proteins encoded by restriction modification systems. Reminiscent of gene transfer as a means for genomic diversity, a PG45-specific DNA fragment with significant homologies to a central protein of an integrative conjugative element of Mycoplasma fermentans (ICEF) was found in most M. bovis field isolates and in a few M. agalactiae isolates. Finally, sequences encoding part of DNA polymerase III were found in both sets of M. agalactiae- and M. bovis-specific DNA fragments and were used to design a species-specific PCR assay for the identification and differentiation of M. agalactiae and M. bovis.

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