Aquaporins play important regulatory roles in improving plant abiotic stress tolerance.To better understand whether the Os PIP1 genes collectively dominate the osmotic regulation in rice under salt stress,a cluster ed...Aquaporins play important regulatory roles in improving plant abiotic stress tolerance.To better understand whether the Os PIP1 genes collectively dominate the osmotic regulation in rice under salt stress,a cluster editing of the Os PIP1;1,Os PIP1;2 and Os PIP1;3 genes in rice was performed by CRISPR/Cas9 system.Sequencing showed that two mutants with Cas9-free,line 14 and line 18 were successfully edited.Briefly,line 14 deleted a single C base in both the Os PIP1;1 and Os PIP1;3 genes,and inserted a single T base in the Os PIP1;2 gene,respectively.While line 18 demonstrated an insertion of a single A base in the Os PIP1;1gene and a single T base in both the Os PIP1;2 and Os PIP1;3 genes,respectively.Multiplex editing of the Os PIP1 genes significantly inhibited photosynthetic rate and accumulation of compatible metabolites,but increased MDA contents and osmotic potentials in the mutants,thus delaying rice growth under salt stress.Functional loss of the Os PIP1 genes obviously suppressed the expressions of the Os PIP1,Os SOS1,Os CIPK24 and Os CBL4 genes,and increased the influxes of Na+and effluxes of K^(+)/H^(+)in the roots,thus accumulating more Na+in rice mutants under salt stress.This study suggests that the Os PIP1 genes are essential modulators collectively contributing to the enhancement of rice salt stress tolerance,and multiplex editing of the Os PIP1 genes provides insight into the osmotic regulation of the PIP genes.展开更多
Alfalfa(Medicago sativa L.),when exposed to abiotic stress such as salinity,suffers significant losses in yield and productivity.The present study evaluated the salinity tolerance of 12 alfalfa cultivars in vitro usin...Alfalfa(Medicago sativa L.),when exposed to abiotic stress such as salinity,suffers significant losses in yield and productivity.The present study evaluated the salinity tolerance of 12 alfalfa cultivars in vitro using five concentrations of sodium chloride(NaCl),ranging from 0 to 250 mmol L^(−1).The results obtained in the current study revealed that the Saudi cultivars,Kasimi and Hassawi,and the German cultivar(Berlin)had the highest salinity tolerance in terms of germination percentage(GP),corrected germination rate index(CGRI),days to reach 50%germination(GT_(50)),and ability to form cotyledonary and true leaves.Under mmol L^(−1) NaCl,the Saudi cultivar Kasimi cultivar showed GP,CGRI,and GT_(50) of 55.20%,123.15,and 3.77 days,respectively.Similarly,the German cultivar(Berlin)showed GP,CGRI,and GT_(50) of 50.06%,86.61,and 5.17 days,respectively.These findings might reveal a pivotal aspect in salt tolerance in alfalfa.Our results will help to select salt-tolerant alfalfa cultivars that could thrive in arid and semi-arid areas with salinity problems.展开更多
基金supported by the National Key Research and Development Program of China (2021YFF1000402-2)the Key Project of Transgenic Crops Cultivation (2016ZX08010005-9)。
文摘Aquaporins play important regulatory roles in improving plant abiotic stress tolerance.To better understand whether the Os PIP1 genes collectively dominate the osmotic regulation in rice under salt stress,a cluster editing of the Os PIP1;1,Os PIP1;2 and Os PIP1;3 genes in rice was performed by CRISPR/Cas9 system.Sequencing showed that two mutants with Cas9-free,line 14 and line 18 were successfully edited.Briefly,line 14 deleted a single C base in both the Os PIP1;1 and Os PIP1;3 genes,and inserted a single T base in the Os PIP1;2 gene,respectively.While line 18 demonstrated an insertion of a single A base in the Os PIP1;1gene and a single T base in both the Os PIP1;2 and Os PIP1;3 genes,respectively.Multiplex editing of the Os PIP1 genes significantly inhibited photosynthetic rate and accumulation of compatible metabolites,but increased MDA contents and osmotic potentials in the mutants,thus delaying rice growth under salt stress.Functional loss of the Os PIP1 genes obviously suppressed the expressions of the Os PIP1,Os SOS1,Os CIPK24 and Os CBL4 genes,and increased the influxes of Na+and effluxes of K^(+)/H^(+)in the roots,thus accumulating more Na+in rice mutants under salt stress.This study suggests that the Os PIP1 genes are essential modulators collectively contributing to the enhancement of rice salt stress tolerance,and multiplex editing of the Os PIP1 genes provides insight into the osmotic regulation of the PIP genes.
基金funded by the Researchers Supporting Project No.(RSP-2021/390),King Saud University,Riyadh,Saudi Arabia.
文摘Alfalfa(Medicago sativa L.),when exposed to abiotic stress such as salinity,suffers significant losses in yield and productivity.The present study evaluated the salinity tolerance of 12 alfalfa cultivars in vitro using five concentrations of sodium chloride(NaCl),ranging from 0 to 250 mmol L^(−1).The results obtained in the current study revealed that the Saudi cultivars,Kasimi and Hassawi,and the German cultivar(Berlin)had the highest salinity tolerance in terms of germination percentage(GP),corrected germination rate index(CGRI),days to reach 50%germination(GT_(50)),and ability to form cotyledonary and true leaves.Under mmol L^(−1) NaCl,the Saudi cultivar Kasimi cultivar showed GP,CGRI,and GT_(50) of 55.20%,123.15,and 3.77 days,respectively.Similarly,the German cultivar(Berlin)showed GP,CGRI,and GT_(50) of 50.06%,86.61,and 5.17 days,respectively.These findings might reveal a pivotal aspect in salt tolerance in alfalfa.Our results will help to select salt-tolerant alfalfa cultivars that could thrive in arid and semi-arid areas with salinity problems.