Salt and drought limit the range of applications of perennial ryegrass(Lolium perenne L.), which is one of the important turf and forage grasses. Previous studies have suggested that pyrroline-5-carboxylate reductase(...Salt and drought limit the range of applications of perennial ryegrass(Lolium perenne L.), which is one of the important turf and forage grasses. Previous studies have suggested that pyrroline-5-carboxylate reductase(P5CR) might play a central role in proline accumulation in plants that are responsive to stresses. In the present study, the Lolium perenne L. pyrroline-5-carboxylate reductase(LpP 5CR) gene was cloned from leaves of the cultivar ‘Derby' using the RACE technique. The full-length LpP 5CR gene was 1 047 bp in length, which comprised an open reading frame(ORF) of 840 bp in size. Sequence alignment revealed that the putative Lp P5 CR had a 94.3% similarity to Ta P5 CR. q RT-PCR displayed that the mR NA levels of the LpP 5CR gene were almost the same as that in the roots, stems, and leaves of perennial ryegrass seedlings subjected to normal condition or NaC l treatment for 1 h. Moreover, the transcription level of LpP 5CR was up- or down- regulated with Na Cl, polyethylene glycol(PEG), cold, or abscisic acid(ABA) treatment for 3 to 48 h. In addition, confocal microscopy localized the GFP-Lp P5 CR fusion protein to the cytoplasm of onion epidermal cells. These findings suggest that LpP 5CR encodes a cytoplasmic P5 CR protein that plays an important role in the response of perennial ryegrass to various stresses.展开更多
African swine fever virus(ASFV),as a member of the large DNA viruses,may regulate autophagy and apoptosis by inhibiting programmed cell death.However,the function of ASFV proteins has not been fully elucidated,especia...African swine fever virus(ASFV),as a member of the large DNA viruses,may regulate autophagy and apoptosis by inhibiting programmed cell death.However,the function of ASFV proteins has not been fully elucidated,especially the role of autophagy in ASFV infection.One of three Pyrroline-5-carboxylate reductases(PYCR),is primarily involved in conversion of glutamate to proline.Previous studies have shown that depletion of PYCR2 was related to the induction of autophagy.In the present study,we found for the first time that ASFV E199 L protein induced a complete autophagy process in Vero and HEK-293 T cells.Through co-immunoprecipitation coupled with mass spectrometry(Co IP-MS)analysis,we firstly identified that E199 L interact with PYCR2 in vitro.Importantly,our work provides evidence that E199 L down-regulated the expression of PYCR2,resulting in autophagy activation.Overall,our results demonstrate that ASFV E199 L protein induces complete autophagy through interaction with PYCR2 and down-regulate the expression level of PYCR2,which provide a valuable reference for the role of autophagy during ASFV infection and contribute to the functional clues of PYCR2.展开更多
基金The National High Technology Research and Development Program of China(Grant No.2011AA100209)Jilin Provincial Department of Education(Grant No.2013-23)supported this study
文摘Salt and drought limit the range of applications of perennial ryegrass(Lolium perenne L.), which is one of the important turf and forage grasses. Previous studies have suggested that pyrroline-5-carboxylate reductase(P5CR) might play a central role in proline accumulation in plants that are responsive to stresses. In the present study, the Lolium perenne L. pyrroline-5-carboxylate reductase(LpP 5CR) gene was cloned from leaves of the cultivar ‘Derby' using the RACE technique. The full-length LpP 5CR gene was 1 047 bp in length, which comprised an open reading frame(ORF) of 840 bp in size. Sequence alignment revealed that the putative Lp P5 CR had a 94.3% similarity to Ta P5 CR. q RT-PCR displayed that the mR NA levels of the LpP 5CR gene were almost the same as that in the roots, stems, and leaves of perennial ryegrass seedlings subjected to normal condition or NaC l treatment for 1 h. Moreover, the transcription level of LpP 5CR was up- or down- regulated with Na Cl, polyethylene glycol(PEG), cold, or abscisic acid(ABA) treatment for 3 to 48 h. In addition, confocal microscopy localized the GFP-Lp P5 CR fusion protein to the cytoplasm of onion epidermal cells. These findings suggest that LpP 5CR encodes a cytoplasmic P5 CR protein that plays an important role in the response of perennial ryegrass to various stresses.
基金supported by the National Key Research and Development Program of China(Grant No. 2017YFD0502303)the Key Research and Development Program of Guangdong Province (2020B020222001)+2 种基金China Postdoctoral Science Foundation (grant no. 2019M652922)the Natural Science Foundation of Guangdong Province (grant no.2019A1515012006)the Chief expert Project of Agricultural Industry Technology system in Guangdong Province (2019KJ128)。
文摘African swine fever virus(ASFV),as a member of the large DNA viruses,may regulate autophagy and apoptosis by inhibiting programmed cell death.However,the function of ASFV proteins has not been fully elucidated,especially the role of autophagy in ASFV infection.One of three Pyrroline-5-carboxylate reductases(PYCR),is primarily involved in conversion of glutamate to proline.Previous studies have shown that depletion of PYCR2 was related to the induction of autophagy.In the present study,we found for the first time that ASFV E199 L protein induced a complete autophagy process in Vero and HEK-293 T cells.Through co-immunoprecipitation coupled with mass spectrometry(Co IP-MS)analysis,we firstly identified that E199 L interact with PYCR2 in vitro.Importantly,our work provides evidence that E199 L down-regulated the expression of PYCR2,resulting in autophagy activation.Overall,our results demonstrate that ASFV E199 L protein induces complete autophagy through interaction with PYCR2 and down-regulate the expression level of PYCR2,which provide a valuable reference for the role of autophagy during ASFV infection and contribute to the functional clues of PYCR2.
