The nuclear-encoded factors and the photosynthetic apparatus have been studied extensively during chloroplast biogenesis.However,many questions regarding these processes remain unanswered,particularly in perennial woo...The nuclear-encoded factors and the photosynthetic apparatus have been studied extensively during chloroplast biogenesis.However,many questions regarding these processes remain unanswered,particularly in perennial woody plants.As a model material of woody plants,poplar not only has very significant value of research,but also possesses economic and ecological properties.This study reports the Populus trichocarpa DJ-1C(PtrDJ1C)factor,encoded by a nuclear gene,and a member of the DJ-1 superfamily.PtrDJ1C knock-out with the CRISPR/Cas9 system resulted in different albino phenotypes.Chlorophyll fluorescence and immunoblot analyses showed that the levels of photosynthetic complex proteins decreased significantly.Moreover,the transcript level of plastid-encoded RNA polymerase-dependent genes and the splicing efficiency of several introns were affected in the mutant line.Furthermore,rRNA accumulation was abnormal,leading to developmental defects in chloroplasts and affecting lignin accumulation.We concluded that the PtrDJ1C protein is essential for early chloroplast development and lignin deposition in poplar.展开更多
A 5＇ flanking region of the well-conserved Toll/interleukin-1 receptor domain （TIR）-encoding sequence was isolated from the genomic DNA ofMelampsora magnusiana Wagner resistant clones of hybrid triploid poplars [（...A 5＇ flanking region of the well-conserved Toll/interleukin-1 receptor domain （TIR）-encoding sequence was isolated from the genomic DNA ofMelampsora magnusiana Wagner resistant clones of hybrid triploid poplars [（Populus tomentosa × P. bolleana） × P. tomentosa]. Sequencing results and alignment analysis show that the obtained TIR-specific promoter （named as PtTIRp01） was 1,732 bp in length; moreover 3＇ region of the PtTIRp01 contains a responds to the 5＇ composition of TIR-NBS type gene PtDRG02, of 747 bp long 5＇ region of TIR-NBS type gene PtDRG02 and its 398 bp complete TIR-encoding sequence, which significantly corindicating that the obtained TIR-specific promoter region consists upstream region of promoter （985 bp）. It was found that the 5＇ region of TIR-NBS type gene PtDRG02 was characterized in the downstream region of the transcriptional start, named as 5＇-untranslated region （5＇ UTR）, consisting of one 93 bp 5＇-untranslation exon, one 213 bp intron and one 441 bp TIR-encoding open reading frame （ORF）. In addition, several putative cis-acting motifs were present in the obtained TIR-specific promoter of PtDRG02, including one TATA box, one GC-rich, one AT-rich, one P-box, one 3-AF1 binding site, two CAAT boxes, two GT-1 motifs, three typical W-boxes, four I-boxes, and one multi-cis-acting fragment （MCF）. The latter contains five types of regulatory elements （E4, G-box, ABRE motif, box 1 and HVA 1 s）, most of which were homologous to the cis-acting regulatory elements involved in the activation of defense genes in plants. Thus, it can be suggested that TIR-specific promoter might be a pathogen-inducible promoter and be necessary for the inducible expression of defense-related genes. Key words Toll/interleukin- 1 receptor domain, promoter, Cis-acting element, poplar展开更多
The VirE2-interaction protein 1(VIP1)serves as a regulator of mitogen-activated protein kinase 3(MPK3)-mediated stress gene modulation under biotic stress,which in turn activates the MPK3 pathway in Arabidopsis.The mo...The VirE2-interaction protein 1(VIP1)serves as a regulator of mitogen-activated protein kinase 3(MPK3)-mediated stress gene modulation under biotic stress,which in turn activates the MPK3 pathway in Arabidopsis.The mode of action of the VIP1 protein in Populus in response to biotic stress remains unknown.In this study,we cloned the full-length cDNA of the PtVIP1 gene from Populus trichocarpa(accession number of GenBank:KY793105).The VIP1 protein harboured a conserved bZIP(basic leucine zipper)domain located in the C-terminus.The VIP1 subcellular localization assay indicated that the VIP1 protein was present in the cytoplasm and nucleus under normal conditions,and that an increase in the amount of the protein in the nucleus occurred after treatment with flg22,the elicitor-active epitope of flagellin which triggers the innate immune response in plants.Transgenic Populus plants overexpressing VIP1 genes(PtVIP1 of Populus;or AtVIP1 of Arabidopsis,as positive control)were generated to investigate the role of VIP1 in vivo.The expression of poplar pathogenesis-related protein 1(PR1)genes was upregulated in transgenic-PtVIP1 or AtVIP1 poplar plants.The transgenic poplar plants overexpressing PtVIP1 or AtVIP1 also showed enhanced resistance to Brenneria salicis infection.These results suggest that the VIP1 protein accumulates in the nucleus in response to biotic stress,and that the pathogen resistance of transgenic VIP1 poplar may be associated with the induced expression of PR1 genes in response to pathogen challenge.展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.32201516,91954202)the Youth Top-notch Talent Program of Hebei Education Department(BJK2022028)+1 种基金National Training Program of Innovation and Entrepreneurship for Undergraduates(Grant Nos.S202110022037,G202010022075)the funding of Hebei North University(XJ2021013)。
文摘The nuclear-encoded factors and the photosynthetic apparatus have been studied extensively during chloroplast biogenesis.However,many questions regarding these processes remain unanswered,particularly in perennial woody plants.As a model material of woody plants,poplar not only has very significant value of research,but also possesses economic and ecological properties.This study reports the Populus trichocarpa DJ-1C(PtrDJ1C)factor,encoded by a nuclear gene,and a member of the DJ-1 superfamily.PtrDJ1C knock-out with the CRISPR/Cas9 system resulted in different albino phenotypes.Chlorophyll fluorescence and immunoblot analyses showed that the levels of photosynthetic complex proteins decreased significantly.Moreover,the transcript level of plastid-encoded RNA polymerase-dependent genes and the splicing efficiency of several introns were affected in the mutant line.Furthermore,rRNA accumulation was abnormal,leading to developmental defects in chloroplasts and affecting lignin accumulation.We concluded that the PtrDJ1C protein is essential for early chloroplast development and lignin deposition in poplar.
文摘A 5＇ flanking region of the well-conserved Toll/interleukin-1 receptor domain （TIR）-encoding sequence was isolated from the genomic DNA ofMelampsora magnusiana Wagner resistant clones of hybrid triploid poplars [（Populus tomentosa × P. bolleana） × P. tomentosa]. Sequencing results and alignment analysis show that the obtained TIR-specific promoter （named as PtTIRp01） was 1,732 bp in length; moreover 3＇ region of the PtTIRp01 contains a responds to the 5＇ composition of TIR-NBS type gene PtDRG02, of 747 bp long 5＇ region of TIR-NBS type gene PtDRG02 and its 398 bp complete TIR-encoding sequence, which significantly corindicating that the obtained TIR-specific promoter region consists upstream region of promoter （985 bp）. It was found that the 5＇ region of TIR-NBS type gene PtDRG02 was characterized in the downstream region of the transcriptional start, named as 5＇-untranslated region （5＇ UTR）, consisting of one 93 bp 5＇-untranslation exon, one 213 bp intron and one 441 bp TIR-encoding open reading frame （ORF）. In addition, several putative cis-acting motifs were present in the obtained TIR-specific promoter of PtDRG02, including one TATA box, one GC-rich, one AT-rich, one P-box, one 3-AF1 binding site, two CAAT boxes, two GT-1 motifs, three typical W-boxes, four I-boxes, and one multi-cis-acting fragment （MCF）. The latter contains five types of regulatory elements （E4, G-box, ABRE motif, box 1 and HVA 1 s）, most of which were homologous to the cis-acting regulatory elements involved in the activation of defense genes in plants. Thus, it can be suggested that TIR-specific promoter might be a pathogen-inducible promoter and be necessary for the inducible expression of defense-related genes. Key words Toll/interleukin- 1 receptor domain, promoter, Cis-acting element, poplar
基金supported by the National Natural Science Foundation of China(Grant No.31570639)the Distinguished Young Scholars Fund of Nanjing Forestry University,the Priority Academic Program Development of Nanjing Forestry University the Poplar Germplasm Nursery Project(Jiangsu Provincial Platform for Conservation and Utilizations for Agricultural Germplasm)
文摘The VirE2-interaction protein 1(VIP1)serves as a regulator of mitogen-activated protein kinase 3(MPK3)-mediated stress gene modulation under biotic stress,which in turn activates the MPK3 pathway in Arabidopsis.The mode of action of the VIP1 protein in Populus in response to biotic stress remains unknown.In this study,we cloned the full-length cDNA of the PtVIP1 gene from Populus trichocarpa(accession number of GenBank:KY793105).The VIP1 protein harboured a conserved bZIP(basic leucine zipper)domain located in the C-terminus.The VIP1 subcellular localization assay indicated that the VIP1 protein was present in the cytoplasm and nucleus under normal conditions,and that an increase in the amount of the protein in the nucleus occurred after treatment with flg22,the elicitor-active epitope of flagellin which triggers the innate immune response in plants.Transgenic Populus plants overexpressing VIP1 genes(PtVIP1 of Populus;or AtVIP1 of Arabidopsis,as positive control)were generated to investigate the role of VIP1 in vivo.The expression of poplar pathogenesis-related protein 1(PR1)genes was upregulated in transgenic-PtVIP1 or AtVIP1 poplar plants.The transgenic poplar plants overexpressing PtVIP1 or AtVIP1 also showed enhanced resistance to Brenneria salicis infection.These results suggest that the VIP1 protein accumulates in the nucleus in response to biotic stress,and that the pathogen resistance of transgenic VIP1 poplar may be associated with the induced expression of PR1 genes in response to pathogen challenge.
