The OsSPL10 gene has previously been reported to positively regulate trichome development and negatively regulate salt and drought stress tolerance in rice. However, it is not clear whether this gene can be used for gene editing to create new germplasm of glabrous leaf and salt-tolerant rice. In this study, we created six rice mutants by CRISPR/Cas9-mediated editing of OsSPL10 from 'Xinfeng 2', 'Xinkedao 31', and 'Xindao 25', the main rice cultivars along the Yellow River. Visual observation and scanning electron microscopy verified that the mutants lacked trichomes on the leaves and glumes, and the expression of glabrous marker genes OsHL6, OsGL6, and OsWOX3B in mutants was down-regulated compared with that in the wild type. The net photosynthetic rate, stomatal conductance, and transpiration rate of flag leaves in the mutants were significantly higher than those in the wild type. In addition, the survival rates of the mutants were much higher than that of the wild type after 7 days of treatment with 200 mmol/L NaCl. The results of quantitative real-time polymerase chain reaction (qRT-PCR) further verified that compared with the wild type, the mutants demonstrated down-regulated expression of the salt stress-related gene OsGASR1 and up-regulated expression of OsNHX2 and OsIDS1. Statistical analysis of agronomic traits showed that the mutants had increased plant height and no significant changes in yield-related traits compared with the wild type. The six spl10 mutants created in this study not only had glabrous leaves and glumes but also demonstrated enhanced tolerance to salt stress, serving as new germplasm resources for directional breeding of rice along the Yellow River.
Oryza/physiology* ; CRISPR-Cas Systems/genetics* ; Salt Tolerance/genetics* ; Gene Editing/methods* ; Plant Proteins/genetics* ; Rivers ; Plant Leaves/genetics* ; Mutation ; Plants, Genetically Modified/genetics* ; China

AIM:To study the effect of hsa-miR-218 on cervical cancer HeLa cell growth and the underlying molecular mechanism .METHODS:The lentivirus expression vector pmiR-218 targeting to hsa-miR-218 was constructed . pmiR-218 was transfected into HeLa cells .The number of viable HeLa cells was counted by the method of Trypan blue ex-clusion.The inhibitory rate of cell activity was detected by WST-8 assay.The expression of LIM and SH3 protein 1 (LASP1) at mRNA and protein levels was determined by real-time PCR and Western blot.The interaction between miR-218 and LASP1 was examined using a luciferase reporter assay .RESULTS:The lentivirus expression vector pmiR-218 tar-geting to hsa-miR-218 was constructed successfully and confirmed by DNA sequencing .Over-expression of miR-218 inhibi-ted the activity of HeLa cells with the inhibitory rates of 15%, 26%and 65%at 24 h, 48 h and 72 h, respectively .The difference between transfection group and blank control /negative control group was statistically significant .The luciferase activity was reduced when co-transfection with miR-218 mimics and LASP1-3’ UTR plasmid.The relative expression of miR-218 was increased after transfection with pmiR-218.Over-expression of miR-218 down-regulated the LASP1 expression at mRNA and protein levels by 25%and 75%respectively.Compared with blank control group and negative control group , the difference was statistically significant (P<0.05).CONCLUSION:pmiR-218 effectively inhibits the growth of HeLa cells in a time-dependent manner.miR-218 targets to the 3’UTR of LASP1, thus down-regulating the expression of LASP1 in HeLa cells .