期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 23, 期 16, 页码 -出版社
MDPI
DOI: 10.3390/ijms23169435
关键词
sugarcane; Erianthus arundinaceus; species-specific molecular marker; chromosome; suppression subtractive hybridization (SSH); fluorescence in situ hybridization (FISH)
资金
- National Natural Science Foundation of China [32001605]
- Special Fund for Science and Technology Innovation of Fujian Agriculture and Forestry University [KFA17626A, KFA20082A, KHF210001A]
- Guangdong Provincial Team of Technical System Innovation for Sugarcane Sisal Hemp Industry [2019KJ104-04]
In this study, a SSH library of Erianthus arundinaceus was constructed and high-throughput E. arundinaceus-specific molecular markers were developed. These markers have the potential for high-accuracy identification and tracking of the introgression status of E. arundinaceus chromatin in sugarcane breeding.
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 the 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 of 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 F-1, 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 of the inherited status of this chromosome in a 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.
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