Populus euphratica Olive is the only tree species that can grow in the saline land and also survive cold winters in northwest China, and it plays a very important role in stabilizing the vulnerable ecosystem there. A ...Populus euphratica Olive is the only tree species that can grow in the saline land and also survive cold winters in northwest China, and it plays a very important role in stabilizing the vulnerable ecosystem there. A cell suspension culture was initiated from callus derived from plantlets of Populus euphratica. Cold acclimation was induced (LT50 of 17.5 ℃) in cell suspension at 45 ℃ in the dark for 30 days and the freezing tolerance increased from LT50 of 12.5 ℃ in nonacclimated cells to LT50 of 17.5 ℃ in cold-acclimated cells. Microvacuolation, cytoplasmic augmentation and accumulation of starch granules were observed in cells that were cold-acclimated by exposure to low temperatures. Several qualitative and quantitative changes in proteins were noted during cold acclimation. Antibodies to carrot extracellular (apoplastic) 36 kD antifreeze protein did not cross react on immunoelectroblots with extracellular proteins in cell suspension culture medium of Populus euphratica, indicating no common epitopes in the carrot 36 kD antifreeze protein and P. euphratica extracellular proteins. The relationship of these changes to cold acclimation in Populus euphratica cell cultures was discussed.展开更多
旨在筛选高山植物喜马拉雅紫茉莉调控类黄酮等次生代谢产物合成的MYB转录因子(MhMYB)基因家族。基于喜马拉雅紫茉莉愈伤组织低温转录组测序数据库,利用PlantTFDB 3.0和BLASTP等软件对MhMYB序列进行筛选与鉴定,使用Web Logo 3.0、MEGA 7....旨在筛选高山植物喜马拉雅紫茉莉调控类黄酮等次生代谢产物合成的MYB转录因子(MhMYB)基因家族。基于喜马拉雅紫茉莉愈伤组织低温转录组测序数据库,利用PlantTFDB 3.0和BLASTP等软件对MhMYB序列进行筛选与鉴定,使用Web Logo 3.0、MEGA 7.0和Mev等生物信息学软件对MhMYB转录因子进行结构域、系统进化及低温诱导表达水平分析。研究筛选出34个喜马拉雅紫茉莉MhMYB家族成员,分别为1R-MYB(2个)、R2R3-MYB(30个)和3R-MYB(2个)3类转录因子;亚细胞定位预测大多数定位于细胞核中;进化分析显示,MhMYB家族成员可分为15个亚类,基于拟南芥AtMYB家族成员的生物学功能,预测MhMYB的S2、S7、S20亚家族成员可能参与调控类黄酮等次生代谢产物合成;转录组数据分析显示,低温可诱导MhMYB6、MhMYB15、MhMYB62基因表达水平增加;共表达分析发现,MhMYB6、MhMYB15、MhMYB62对类黄酮合成途径基因起到一定调控作用。初步研究表明,MhMYB6、MhMYB15、MhMYB62可参与调控低温诱导喜马拉雅紫茉莉类黄酮的累积。展开更多
The recombinant expression vector pET43.1b-AFP, which contains full encoding region of a carrot 36 kD antifreeze protein (AFP) gene was constructed. The recombinant was transformed into expression host carrying T7 RNA...The recombinant expression vector pET43.1b-AFP, which contains full encoding region of a carrot 36 kD antifreeze protein (AFP) gene was constructed. The recombinant was transformed into expression host carrying T7 RNA polymerase gene (DE3 lysogen) and induced by 1 mmol稬-1 IPTG (isopropyl--D-thiogalactoside) to express 110 kD polypeptide of AFP fusion protein. The analysis of product solubility revealed that pET43.1b-AFP was predominately soluble, and the expressed amount reached the maximum after the IPTG treatment for 3 h.展开更多
microRNAs(miR NAs) are important post-transcriptional regulators of gene expression in plant abiotic stress responses. Ammopiptanthus mongolicus is a typical evergreen woody xerophyte, which makes it an ideal model sy...microRNAs(miR NAs) are important post-transcriptional regulators of gene expression in plant abiotic stress responses. Ammopiptanthus mongolicus is a typical evergreen woody xerophyte, which makes it an ideal model system for studying drought tolerance in woody plants. The response of mi RNAs to drought stress is still unknown in this plant. In this research, we obtained 34 695 665 raw reads from two small RNA libraries constructed from control and drought-treated A. mongolicus seedlings by the Illumina deep sequencing technology. Length analysis revealed that reads of 20, 21, and 24 nucleotides accounted for the majority of the small RNAs in the two libraries. Sequence analyses identified 65 conserved mi RNA sequences from 190 members of 31 mi RNA families and 20 potential novel mi RNAs from 17 families that were differentially expressed between the two libraries. The expression patterns of 25 of these mi RNAs were significantly different in the two libraries, but only am-mi R4 a-c was downregulated during drought stress, the other 24 were upregulated. The expression trends determined by the Hi Seq sequencing and by q RT-PCR were similar. Furthermore, 35 target genes for the conserved mi RNA were markedly changed and 52 target genes for the potential novel mi RNAs were predicted and their functions were annotated by computational analysis. Our results provide new molecular evidence for understanding the molecular mechanisms of drought response and tolerance in A. mongolicus.展开更多
基金the National Natural Science Foundation of China (Grant No. 30271067)Fok Ying Tung Education Foundation (71030)+1 种基金 Key Teachers Foundation of the Educational Ministry of China and the State Key Basic Research and Development Plan of China (G199901600
文摘Populus euphratica Olive is the only tree species that can grow in the saline land and also survive cold winters in northwest China, and it plays a very important role in stabilizing the vulnerable ecosystem there. A cell suspension culture was initiated from callus derived from plantlets of Populus euphratica. Cold acclimation was induced (LT50 of 17.5 ℃) in cell suspension at 45 ℃ in the dark for 30 days and the freezing tolerance increased from LT50 of 12.5 ℃ in nonacclimated cells to LT50 of 17.5 ℃ in cold-acclimated cells. Microvacuolation, cytoplasmic augmentation and accumulation of starch granules were observed in cells that were cold-acclimated by exposure to low temperatures. Several qualitative and quantitative changes in proteins were noted during cold acclimation. Antibodies to carrot extracellular (apoplastic) 36 kD antifreeze protein did not cross react on immunoelectroblots with extracellular proteins in cell suspension culture medium of Populus euphratica, indicating no common epitopes in the carrot 36 kD antifreeze protein and P. euphratica extracellular proteins. The relationship of these changes to cold acclimation in Populus euphratica cell cultures was discussed.
文摘旨在筛选高山植物喜马拉雅紫茉莉调控类黄酮等次生代谢产物合成的MYB转录因子(MhMYB)基因家族。基于喜马拉雅紫茉莉愈伤组织低温转录组测序数据库,利用PlantTFDB 3.0和BLASTP等软件对MhMYB序列进行筛选与鉴定,使用Web Logo 3.0、MEGA 7.0和Mev等生物信息学软件对MhMYB转录因子进行结构域、系统进化及低温诱导表达水平分析。研究筛选出34个喜马拉雅紫茉莉MhMYB家族成员,分别为1R-MYB(2个)、R2R3-MYB(30个)和3R-MYB(2个)3类转录因子;亚细胞定位预测大多数定位于细胞核中;进化分析显示,MhMYB家族成员可分为15个亚类,基于拟南芥AtMYB家族成员的生物学功能,预测MhMYB的S2、S7、S20亚家族成员可能参与调控类黄酮等次生代谢产物合成;转录组数据分析显示,低温可诱导MhMYB6、MhMYB15、MhMYB62基因表达水平增加;共表达分析发现,MhMYB6、MhMYB15、MhMYB62对类黄酮合成途径基因起到一定调控作用。初步研究表明,MhMYB6、MhMYB15、MhMYB62可参与调控低温诱导喜马拉雅紫茉莉类黄酮的累积。
基金the National Natural Science Foundation of China (Grant No.3980011830271067)Fok Ying Tung Education Foundation (71030) and the Key Teachers Foundation of the Education Ministry of China
文摘The recombinant expression vector pET43.1b-AFP, which contains full encoding region of a carrot 36 kD antifreeze protein (AFP) gene was constructed. The recombinant was transformed into expression host carrying T7 RNA polymerase gene (DE3 lysogen) and induced by 1 mmol稬-1 IPTG (isopropyl--D-thiogalactoside) to express 110 kD polypeptide of AFP fusion protein. The analysis of product solubility revealed that pET43.1b-AFP was predominately soluble, and the expressed amount reached the maximum after the IPTG treatment for 3 h.
基金financially supported by the National Natural Science Foundation of China (Grant No. 31270737)111 Project (Grant No. B13007)+1 种基金Program for Changjiang Scholars and Innovative Research Team in University (Grant No. IRT13047)Natural Science Foundation of Beijing (No. 6112016, No.KZ20150020021)
文摘microRNAs(miR NAs) are important post-transcriptional regulators of gene expression in plant abiotic stress responses. Ammopiptanthus mongolicus is a typical evergreen woody xerophyte, which makes it an ideal model system for studying drought tolerance in woody plants. The response of mi RNAs to drought stress is still unknown in this plant. In this research, we obtained 34 695 665 raw reads from two small RNA libraries constructed from control and drought-treated A. mongolicus seedlings by the Illumina deep sequencing technology. Length analysis revealed that reads of 20, 21, and 24 nucleotides accounted for the majority of the small RNAs in the two libraries. Sequence analyses identified 65 conserved mi RNA sequences from 190 members of 31 mi RNA families and 20 potential novel mi RNAs from 17 families that were differentially expressed between the two libraries. The expression patterns of 25 of these mi RNAs were significantly different in the two libraries, but only am-mi R4 a-c was downregulated during drought stress, the other 24 were upregulated. The expression trends determined by the Hi Seq sequencing and by q RT-PCR were similar. Furthermore, 35 target genes for the conserved mi RNA were markedly changed and 52 target genes for the potential novel mi RNAs were predicted and their functions were annotated by computational analysis. Our results provide new molecular evidence for understanding the molecular mechanisms of drought response and tolerance in A. mongolicus.
