We report here the construction of a new set of chromosome segment substitution lines (CSSLs) in rice, in which the genome of the elite japonica cultivar C418 has been introgressed into the background of the elite indica cultivar 9311. This set was developed by marker aided selection, based on 136 SSR and in/del markers. The introgressed chromosomal segments presented in the 108 CSSLs covered 98.3% of the cultivar C418 genome. The CSSL set was used to genetically analyze variation for 1000-grain weight (TGW) and related traits in two contrasting environments, and led to the identification of 41 quantitative trait loci (QTLs), five of which were expressed in both environments. More detailed mapping of qTGW7 showed that it is cosegregated with RM22034 on the short arm of chromosome 7. The CSSLs varied phenotypically with respect to a number of agronomic traits besides TGW. CSSL populations would be effective in identifying QTLs for these various traits, and could provide germplasm relevant for crop improvement.
Diagnostics based on circulating circular RNA (circRNA) is an emerging field for noninvasive molecular diagnosis, owing to the stable circular structure of circRNA. However, the uniquely circular structure still poses a challenge for circRNA quantification in the research community. Here, we verified the discrepancy in the circRNA quantification by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and reverse transcription-droplet digital polymerase chain reaction (RT-ddPCR), which is caused by the rolling reverse transcription (RT) product originating from the circular structure. In addition, we detected the stability of cell-free circRNA in serum/ plasma and determined the pre-analysis sampling procedure that will compromise the quantification of circRNA. Our results showed that prolonged RT incubation time resulted in multiple PCR products from circular RNA, which will reduce the accuracy of circRNA quantification by RT-qPCR technology, whereas ddPCR can address this limitation and could be a good alternative to qPCR for circRNA quantification. CircRNA showed a high stability in promptly separated serum/ plasma but not in delay-separated samples.
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