Plant mitochondrial phosphate transporters regulate phosphate transport and ATP synthesis. Determining whether they function in abiotic stress response process would shed light on their response to salt stress. We use...Plant mitochondrial phosphate transporters regulate phosphate transport and ATP synthesis. Determining whether they function in abiotic stress response process would shed light on their response to salt stress. We used the CRISPR/Cas9 gene-editing system to mutagenize two mitochondrial phosphate transporters, OsMPT3;1 and OsMPT3;2, to investigate their regulatory roles under salt stress. Two cas9(CRISPR-associated protein9)-free homozygous mutants, mpt33 and mpt30, were confirmed to be stable. Both OsMPT3;1 and OsMPT3;2 were markedly induced by salt stress, and their mutagenesis strongly inhibited growth and development, especially under salt stress. Mutagenesis sharply reduced the accumulation of ATP, phosphate, calcium, soluble sugar, and proline and increased osmotic potential, malondialdehyde, and Na^+ /K^+ ratio under salt stress. Both mutants demonstrate normal growth and development in the presence of ATP, revealing high sensitivity to exogenous ATP under salt stress. The mutants showed lowered rates of Na^+ efflux but also of K^+ and Ca^(2+) influx under salt stress. Mutagenesis of OsMPT3;2 altered the enrichment profiles of differentially expressed genes involved mainly in synthesis of secondary metabolites, metabolism of glycolysis, pyruvate, tricarboxylic acid cycle, in response to salt stress. The mutant displayed significant accumulation differences in 14 metabolites involved in 17 metabolic pathways, and strongly up-regulated the accumulation of glutamine, a precursor in proline synthesis, under salt stress. These findings suggest that the OsMPT3 gene modulates phosphate transport and energy supply for ATP synthesis and triggers changes in accumulation of ions and metabolites participating in osmotic regulation in rice under salt stress, thus increasing rice salt tolerance. This study demonstrates the effective application of CRISPR/Cas9 gene-editing to the investigation of plant functional genes.展开更多
To understand the response characteristics of soybean seedling resistance systems to high-temperature stress,the spring soybean varieties HN44 and HN65 were selected in this study as the experimental materials,as they...To understand the response characteristics of soybean seedling resistance systems to high-temperature stress,the spring soybean varieties HN44 and HN65 were selected in this study as the experimental materials,as they had substantial differences in stress resistance.Soybean physiological status was studied under high-temperature stress.Sand culture was used in the experiment,and soybeans at the seedling stage were treated with high-temperature stress.The results showed that the activity of antioxidases and osmotic regulatory substance contents in soybean increased under high-temperature treatment for a certain period of time,the activity of antioxidases and osmotic regulatory substance contents decreased after continuous treatment,and high temperatures continuously increased malondialdehyde contents in the two varieties.At high temperatures,the antioxidant activity and osmotic regulator contents of HN44 were higher than those of HN65,and the malondialdehyde contents of HN44 were higher than those of HN65.The above results showed that HN44 soybeans exhibited significantly higher resistance to high-temperature stress than HN65 variety.展开更多
基金supported by the National Key Research and Development Program of China(2016YFC0501203)the National Genetically Modified Organism Project(2016ZX08010005-9)。
文摘Plant mitochondrial phosphate transporters regulate phosphate transport and ATP synthesis. Determining whether they function in abiotic stress response process would shed light on their response to salt stress. We used the CRISPR/Cas9 gene-editing system to mutagenize two mitochondrial phosphate transporters, OsMPT3;1 and OsMPT3;2, to investigate their regulatory roles under salt stress. Two cas9(CRISPR-associated protein9)-free homozygous mutants, mpt33 and mpt30, were confirmed to be stable. Both OsMPT3;1 and OsMPT3;2 were markedly induced by salt stress, and their mutagenesis strongly inhibited growth and development, especially under salt stress. Mutagenesis sharply reduced the accumulation of ATP, phosphate, calcium, soluble sugar, and proline and increased osmotic potential, malondialdehyde, and Na^+ /K^+ ratio under salt stress. Both mutants demonstrate normal growth and development in the presence of ATP, revealing high sensitivity to exogenous ATP under salt stress. The mutants showed lowered rates of Na^+ efflux but also of K^+ and Ca^(2+) influx under salt stress. Mutagenesis of OsMPT3;2 altered the enrichment profiles of differentially expressed genes involved mainly in synthesis of secondary metabolites, metabolism of glycolysis, pyruvate, tricarboxylic acid cycle, in response to salt stress. The mutant displayed significant accumulation differences in 14 metabolites involved in 17 metabolic pathways, and strongly up-regulated the accumulation of glutamine, a precursor in proline synthesis, under salt stress. These findings suggest that the OsMPT3 gene modulates phosphate transport and energy supply for ATP synthesis and triggers changes in accumulation of ions and metabolites participating in osmotic regulation in rice under salt stress, thus increasing rice salt tolerance. This study demonstrates the effective application of CRISPR/Cas9 gene-editing to the investigation of plant functional genes.
基金Supported by the National Key Research and Development Program(2018YFD1000903)。
文摘To understand the response characteristics of soybean seedling resistance systems to high-temperature stress,the spring soybean varieties HN44 and HN65 were selected in this study as the experimental materials,as they had substantial differences in stress resistance.Soybean physiological status was studied under high-temperature stress.Sand culture was used in the experiment,and soybeans at the seedling stage were treated with high-temperature stress.The results showed that the activity of antioxidases and osmotic regulatory substance contents in soybean increased under high-temperature treatment for a certain period of time,the activity of antioxidases and osmotic regulatory substance contents decreased after continuous treatment,and high temperatures continuously increased malondialdehyde contents in the two varieties.At high temperatures,the antioxidant activity and osmotic regulator contents of HN44 were higher than those of HN65,and the malondialdehyde contents of HN44 were higher than those of HN65.The above results showed that HN44 soybeans exhibited significantly higher resistance to high-temperature stress than HN65 variety.
