Objective To investigate the main proteinases responsible for CD16b shedding under different stimulators. Methods HEK293 cell line stably expressing CD16b was constructed by lentivirus system. The cell line was then ...Objective To investigate the main proteinases responsible for CD16b shedding under different stimulators. Methods HEK293 cell line stably expressing CD16b was constructed by lentivirus system. The cell line was then overexpressed with a disintegrin and metalloproteinase 10 （ADAM10） or ADAM17, sup- pressed with short hairpin RNA of ADAM10 or ADAM I 7, and reconstituted with ADAM 10 or ADAM17, respectively. After each treatment, the cell line was stimulated with ionomycin or phorbol 12-myristate- 13-acetate （PMA） for 12 hours. The soluble CD 16b released from cell membrane was detected by immuno- precipition and immunoblot. Quantitation was then implemented to compare the amount of soluble CD 16b in cell supernatant after stimulation. Results HEK293 cell line stably expressing CD16b was successfully established. When CDI6b ex- pressing cell line was overexpressed with ADAM 10, shedding of CD 16b was increased after stimulation with ionomycin but not PMA; when the cell line overexpressed with ADAM I7, shedding of CDI6b was increased after stimulation with PMA but not ionomycin. Similarly, when ADAM10 was suppressed by short hairpin RNA, CD 16b shedding was decreased after stimulation with ionomycin; when ADAM 17 was suppressed by short hairpin RNA, CD16b shedding was decreased after stimulation with PMA. The shedding of CD16b was increased again when CD16b expressing cell line was reconstituted with ADAM10 and stimulated by ionomycin or reconstituted with ADAM 17 and stimulated by PMA. Conclusions Both ADAM10 and ADAM17 could shed CD16b, but they possess differed prefer- ences. ADAM10 is the main sheddase under stimulation of ionomycin, while ADAM17 is the main sheddase under stimulation of PMA.展开更多
Non-hydraulic root signals(nHRS)are affirmed as a unique positive response to soil drying,and play a crucial role in regulating water use efficiency and yield formation in dryland wheat production.Strigolactones(SLs)c...Non-hydraulic root signals(nHRS)are affirmed as a unique positive response to soil drying,and play a crucial role in regulating water use efficiency and yield formation in dryland wheat production.Strigolactones(SLs)can enhance plant drought adaptability.However,the question of whether strigolactones enhance grain yield and water use efficiency by regulating nHRS and antioxidant defense systems in dryland wheat remains unanswered.In this study,pot experiments were conducted to investigate the effects of strigolactones on nHRS,antioxidant defense system,and grain yield and water use efficiency in dryland wheat.The results showed that external application of SLs increased drought-induced abscisic acid(ABA)accumulation and activated an earlier trigger of nHRS at 73.4% field capacity(FC),compared to 68.5%FC in the control group(CK).This phenomenon was mechanically associated with the physiological mediation of SLs.The application of SLs significantly enhanced the activities of leaf antioxidant enzymes,reduced ROS production,and mitigated oxidative damage to lipid membrane.Additionally,root biomass,root length density,and root to shoot ratio were increased under strigolactone treatment.Furthermore,exogenous application of SLs significantly increased grain yield by 34.9%under moderate drought stress.Water use efficiency was also increased by 21.5% and 33.3% under moderate and severe drought conditions respectively,compared to the control group(CK).The results suggested that the application of strigolactones triggered earlier drought-sensing mechanism and improved the antioxidant defense ability,thus enhancing grain yield and water use efficiency in dryland wheat production.展开更多
基金Supported by the National Natural Science Foundation of China (30872287)
文摘Objective To investigate the main proteinases responsible for CD16b shedding under different stimulators. Methods HEK293 cell line stably expressing CD16b was constructed by lentivirus system. The cell line was then overexpressed with a disintegrin and metalloproteinase 10 （ADAM10） or ADAM17, sup- pressed with short hairpin RNA of ADAM10 or ADAM I 7, and reconstituted with ADAM 10 or ADAM17, respectively. After each treatment, the cell line was stimulated with ionomycin or phorbol 12-myristate- 13-acetate （PMA） for 12 hours. The soluble CD 16b released from cell membrane was detected by immuno- precipition and immunoblot. Quantitation was then implemented to compare the amount of soluble CD 16b in cell supernatant after stimulation. Results HEK293 cell line stably expressing CD16b was successfully established. When CDI6b ex- pressing cell line was overexpressed with ADAM 10, shedding of CD 16b was increased after stimulation with ionomycin but not PMA; when the cell line overexpressed with ADAM I7, shedding of CDI6b was increased after stimulation with PMA but not ionomycin. Similarly, when ADAM10 was suppressed by short hairpin RNA, CD 16b shedding was decreased after stimulation with ionomycin; when ADAM 17 was suppressed by short hairpin RNA, CD16b shedding was decreased after stimulation with PMA. The shedding of CD16b was increased again when CD16b expressing cell line was reconstituted with ADAM10 and stimulated by ionomycin or reconstituted with ADAM 17 and stimulated by PMA. Conclusions Both ADAM10 and ADAM17 could shed CD16b, but they possess differed prefer- ences. ADAM10 is the main sheddase under stimulation of ionomycin, while ADAM17 is the main sheddase under stimulation of PMA.
基金financially supported by the National Key Research and Development Program of China(2022YFF1302804)the National Natural Science Foundation of China(31901123).
文摘Non-hydraulic root signals(nHRS)are affirmed as a unique positive response to soil drying,and play a crucial role in regulating water use efficiency and yield formation in dryland wheat production.Strigolactones(SLs)can enhance plant drought adaptability.However,the question of whether strigolactones enhance grain yield and water use efficiency by regulating nHRS and antioxidant defense systems in dryland wheat remains unanswered.In this study,pot experiments were conducted to investigate the effects of strigolactones on nHRS,antioxidant defense system,and grain yield and water use efficiency in dryland wheat.The results showed that external application of SLs increased drought-induced abscisic acid(ABA)accumulation and activated an earlier trigger of nHRS at 73.4% field capacity(FC),compared to 68.5%FC in the control group(CK).This phenomenon was mechanically associated with the physiological mediation of SLs.The application of SLs significantly enhanced the activities of leaf antioxidant enzymes,reduced ROS production,and mitigated oxidative damage to lipid membrane.Additionally,root biomass,root length density,and root to shoot ratio were increased under strigolactone treatment.Furthermore,exogenous application of SLs significantly increased grain yield by 34.9%under moderate drought stress.Water use efficiency was also increased by 21.5% and 33.3% under moderate and severe drought conditions respectively,compared to the control group(CK).The results suggested that the application of strigolactones triggered earlier drought-sensing mechanism and improved the antioxidant defense ability,thus enhancing grain yield and water use efficiency in dryland wheat production.
