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Characterization of the Tomato Prosystemin Promoter:Organ-specific Expression,Hormone Specificity and Methyl Jasmonate Responsiveness by Deletion Analysis in Transgenic Tobacco plants

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Journal of Integrative plant Biology 2012年第1期54卷 15-32页

作者： Hamlet Avilés-Arnaut John Paul Délano-Frier Center of Research and Advanced Studies (Cinvestav) at Irapuato:Unit for plant Biotechnology and genetic engineering

Tomato systemin is a bioactive peptide that regulates the systemic activation of wound-responsive genes. It is released from its 200 amino acid precursor called prosystemin. Initial tissue-localization and hormone-induced expression assays indicated that the tomato prosystemin gene （SIPS） accumulates mainly in floral tissues and in response to exogenous abscisic acid and methyl jasmonate （MeJA） treatments, respectively. Later, the promoter regions of the PS gene in tomato （Solanum lycopersicum L. cv. Castlemart）, pepper （Capsicum annuum） and potato （Solanum tuberosum） were isolated and an in silico analysis of the SIPS promoter revealed an over-representation of stress- and MeJA-responsive motifs. A subsequent 5＇ deletion analysis of the SIPS promoter fused to the/^-glucuronidase reporter （GUS） gene showed that the -221 to ＋40 bp proximal SIPS promoter region was sufficient to direct the stigma, vascular bundle-specific and MeJA-responsive expression of GUS in transgenic tobacco plants. Important ***-specific, light- and MeJA-responsive cis-elements were also present in this region. These findings provide relevant information regarding the transcriptional regulation mechanisms of the SIPS promoter operating in transgenic tobacco plants. They also suggest that its Ussue-specificity and inducible nature could have wide applicability in plant biotechnology.

关键词： cis-regulatory elements flower specific jasmonic acid methyl jasmonate promoter analysis prosystemin wound-responsive genes.

Effort and Contribution of T-DNA Insertion Mutant Library for Rice Functional Genomics Research in China:Review and Perspective

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Journal of Integrative plant Biology 2012年第12期54卷 953-966页

作者： Yuxiao Chang Tuan Long Changyin Wu national Key laboratory of Crop genetic Improvement and national Center of plant Gene Research(Wuhan) Huazhong Agricultural University

With the completion of the rice （Oryza sativa L.） genome-sequencing project, the rice research community proposed to characterize the func- tion of every predicted gene in rice by 2020. One of the most effective and high-throughput strategies for studying gene function is to employ genetic mutations induced by insertion elements such as T-DNA or transposons. Since 1999, with support from the Ministry of Science and Technology of China for Rice Functional Genomics Programs, large-scale T-DNA insertion mutant populations have been generated in Huazhong Agricultural University, the Chinese Academy of Sciences and the Chinese Academy of Agricultural Sciences. Currently, a total of 372,346 mutant lines have been generated, and 58,226 T-DNA or Tos17 flanking sequence tags have been isolated. Using these mutant resources, more than 40 genes with potential applications in rice breeding have already been identified. These include genes involved in biotic or abiotic stress responses, nutrient metabolism, pollen development, and plant architecture. The functional analysis of these genes will not only deepen our understanding of the fundamental biological questions in rice, but will also offer valuable gene resources for developing Green Super Rice that is high-yielding with few inputs even under the poor growth conditions of many regions of Africa and Asia.

关键词： Flanking sequence tags functional genomics insertion site rice T-DNA insertion mutant library.

The tRNA 3'-end Processing Enzyme tRNase Z2Contributes to Chloroplast Biogenesis in Rice

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Journal of Integrative plant Biology 2013年第11期55卷 1104-1118页

作者： Tuan Long Dong Guo Dong He Wenjie Shen Xianghua Li national Key laboratory of Crop genetic Improvement and national Center of plant Gene Research (Wuhan) Huazhong Agricultural University

tRNase Z （TRZ） is a ubiquitous endonuclease that removes the 3＇-trailer from precursor tRNAs during maturation. In yeast and animals, TRZ regulates the cell cycle via its （t）RNA processing activity; however, its physiological function in higher plants has not been well characterized. This study describes the identification of a rice （Oryza sativa） TRZ2 mutant; plants homozygous for the osatrz2 mutation were albinos with deficient chlorophyll content. A microscopic analysis of the mutant plants revealed that the transition of proplastids to chloroplasts was arrested at an early stage, and the number and size of the plastids in callus cells was substantially decreased. A genetic complementation test and an RNA interference analysis confirmed that disruption of OsaTRZ2 was responsible for the mutant phenotype. OsaTRZ2 is expressed in all rice tissues, but is preferentially expressed in leaves, sheathes, and calli. OsaTRZ2 was subcellularly localized in chloroplasts, and displayed tRNA 3＇-end processing activity in both in vitro and in vivo assays. In the osatrz2 mutants, transcription of plastid-encoded and nucleus- encoded RNA polymerases was severely reduced and moderately increased, respectively. These results suggest that the tRNA 3＇ processing activity of OsaTRZ2 contributes to chloroplast biogenesis.

关键词： Chloroplast biogenesis rice tRNA processing tRNase Z.

Transcriptional silencing and developmental reactivation of cry1Ab gene in transgenic rice

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Science China(Life Sciences) 2002年第1期45卷 68-78页

作者：吴刚崔海瑞舒庆尧叶恭银夏英武高明蔚 Illimar ALTOSAAR 李毅 Institute of Nuclear-Agricultural Sciences Zhejiang University Hangzhou 310029 China Department of plant Protection Zhejiang University Hangzhou 310029 China Department of Biochemistry University of Ottawa K1N 6N5 Canada Peking-Yale Joint Center for Plant Molecular Genetics and Agrobiotechnology National Laboratory of Protein Engineering and Plant Genetic Engineering College of Life Sciences Peking University Beijing 100871 China

One transgenic rice line lacking Cry1Ab expression product was screened in the progenies of Agrobacterium-transformed transgenic rice variety Zhong 8215 with a cry1Ab gene under field releasing conditions by using GUS histochemical assay and Western blot. Molecular hybridization results revealed that the cry1Ab gene was silenced in the transgenic rice variety Zhong 8215 and two copies of ubiquitin promoter were integrated into the rice genome. The silencing of cry1Ab gene in transgenic rice was found to be due to the methylation of the ubiquitin promoter as revealed by methylation analysis. Meanwhile, different concentrations of demethylation reagent 5-azacytidine combining with different treatment time were employed to treat the silenced transgenic rice seeds. The results indicated that 5-azacytidine could reactivate 8%-30% of the silenced transgenic rice plants and the expression level of the reactivated cry1Ab transgene could reach as high as 0.147% of the total soluble protein. Treatment with low concentration of 5-azacytidine(45 mg/L for 1 d and 2 d) could lead to the highest reactivation ratio and the highest expression level of the cry1Ab gene.

关键词： transgenic rice, cry1Ab gene, silencing, reactivation.

A novel gene delivery system targeting cells expressing VEGF receptors

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Cell Research 1999年第1期9卷 11-25页

作者： LI JUN MIN JUN SONG HAN YI HUANG PEI KUN TIAN SHU MIN QU MIN YAO HUI QIU JIANG DA FANG WAN JING CHU LUO CHENG XIAO GU JIAN REN GU( national Labomtory for Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai 200032,China)(national laboratory of 1national laboratory for Oncogenes and Related Genes Shanghai Cancer Institute Shanghai 200032 China 1national laboratory for Oncogenes and Related Genes Shanghai Cancer Institute Shanghai 200032 China 1national laboratory for Oncogenes and Related Genes Shanghai Cancer Institute Shanghai 200032 China 1national laboratory for Oncogenes and Related Genes Shanghai Cancer Institute Shanghai 200032 China 1national laboratory for Oncogenes and Related Genes Shanghai Cancer Institute Shanghai 200032 China 1national laboratory for Oncogenes and Related Genes Shanghai Cancer Institute Shanghai 200032 China 1national laboratory for Oncogenes and Related Genes Shanghai Cancer Institute Shanghai 200032 China 1national laboratory for Oncogenes and Related Genes Shanghai Cancer Institute Shanghai 200032 China 2national laboratory of protein engineering and plant genetic engineering Peking University Beijing 100871 China 2national laboratory of protein engineering and plant genetic engineering Peking University Beijing 100871 China 1national laboratory for Oncogenes and Related Genes Shanghai Cancer Institute Shanghai 200032 China

Two ligand oligopeptides GV1 and GV2 were designed according to the putative binding region of VEGF to its receptors. GV1, GV2 and endosome releasing oligopeptide HA20 were conjugated with poly-L-lysine or protamine and the resulting conjugates could interact with DNA in a noncovalent bond to form a complex. Using pSV2-β-galactosidase as a reporter gene, it has been demonstrated that exogenous gene was transferred into bovine aortic arch-derived endothelial cells (ABAE) andhuman malignant melanoma cell lines (A375) in vitro. In vivo experiments, exogenous gene was transferred into tumor vascular endothelial cells and tumor cells of subcutaneously transplanted human colon cancer LOVO, human malignant melanoma A375 and human hepatoma graft in nude mice. This system could also target gene to intrahepatically transplanted human hepatoma injected via portal vein in nude mice. These results are correlated with theGene delivery system targeting VEGF receptors relevant receptors (flt-1, flk-1/KDR) expression on the targeted cells and tissues.

关键词： VEGF receptors gene delivery system tumor vascular endothelial cells targeting

Reverse genetic Identification of CRN1 and its Distinctive Role in Chlorophyll Degradation in Arabidopsis

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Journal of Integrative plant Biology 2010年第5期52卷 496-504页

作者： Guodong Ren Qian Zhou Shouxin Wu Yufan Zhang Lingang Zhang Jirong Huang Zhenfei Sun Benke Kuai State Key laboratory of genetic engineering Institute of Plant Biology School of Life Sciences Fudan University Shanghai 200433China national laboratory of plant Molecular genetics Institute of Plant Physiology and Ecology Shanghai Institute for Biological SciencesChinese Academy of Sciences Shanghai 200032 China

Recent identification of NYE1/SGR1 brought up a new era for the exploration of the regulatory mechanism of Chlorophyll （Chl） degradation. Cluster analysis of senescence associated genes with putative chloroplast targeting sequences revealed several genes sharing a similar expression pattern with NYE1. Further characterization of available T-DNA insertion lines led to the discovery of a novel stay-green gene CRN1 （Co-regulated with NYE1）. Chl breakdown was significantly restrained in crnl-1 under diversified senescence scenarios, which is comparable with that in acdl-20, but much more severe than that in nyel- 1. Notably, various Chl binding proteins, especially trimeric LHCP II, were markedly retained in crnl-1 four days after dark-treatment, possibly due to a lesion in disassociation of protein-pigment complex. Nevertheless, the photochemical efficiency of PSII in crnl-1 declined, even more rapidly, two days after dark-treatment, compared to those in Col-0 and nyel-1. Our results suggest that CRN1 plays a crucial role in Chl degradation, and that loss of its function produces various side-effects, including those on the breakdown of Ch-protein complex and the maintenance of the residual photosynthetic capability during leaf senescence.

关键词： CHLOROPHYLL ARABIDOPSIS GENES PHOTOSYNTHESIS CLUSTER analysis (Statistics)

Electrocatalytic cleavage of a lignin β-O-4 model compound and coupling with nitrogen-containing aromatics using Prussian blue analogue-derived nickel–cobalt spinel

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Journal of Energy Chemistry 2025年第01期 628-636页

作者： Yi Qi Xueying Chen Bowen Liu Xuliang Lin Xueqing Qiu Yanlin Qin Guangdong Provincial Key laboratory of plant Resources Biorefinery School of Chemical Engineering and Light Industry Guangdong University of Technology Guangdong Provincial laboratory of Chemistry and Fine Chemical engineering Jieyang Center Guangdong Basic Research Center of Excellence for Ecological Security and Green Development Guangdong University of Technology

Electrochemical conversion of lignin for the production of high-value heterocyclic aromatic compounds has great *** demonstrate the targeted synthesis and cation modulation of NiCo2O4spinel nanoboxes,synthesized via cation exchange and calcination *** catalysts exhibit excellent efficacy in the electrocatalytic conversion of lignin model compounds,specifically 2-phenoxy-1-phenylethanol,into nitrogen-containing aromatics,achieving high conversion rates and *** catalysts were synthesized via a cation exchange and calcination oxidation process,using Prussian blue nanocubes as *** porous architecture and polymetallic composition of the NiCo2O4spinel demonstrated superior performance in electrocatalytic oxidative coupling,achieving a 99.2 wt%conversion rate of the 2-phenoxy-1-phenylethanol with selectivities of 37.5 wt%for quinoline derivatives and 31.5 wt%for *** innovations include the development of a sustainable one-pot synthesis method for quinoline derivatives,the elucidation of a multistage reaction pathway involving CAO bond cleavage,hydroxyaldol condensation,and CAN bond formation,and a deeper mechanistic understanding derived from DFT *** work establishes a new strategy for lignin valorization,offering a sustainable route to produce high-value nitrogen-containing aromatics from renewable biomass under mild conditions,without the need for additional reagents.

关键词：

PatU3 plays a central role in coordinating cell division and differentiation in pattern formation of filamentous cyanobacterium Nostoc *** 7120

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Science China(Life Sciences) 2023年第12期66卷 2896-2909页

作者： Lei Yin Zhenggao Zheng Yilin Li Xiying Li Dan Cheng Chunxia Dong Yixuan Liu Jindong Zhao State Key laboratory of protein and plant genetic engineering School of Life SciencesPeking UniversityBeijing 100871China national Teaching Center for Experimental Biology School of Life SciencesPeking UniversityBeijing 100871China Institute of Hydrobiology Chinese Academy of SciencesWuhan 430072China

Spatial periodic signal for cell differentiation in some multicellular organisms is generated according to Turing's principle for pattern *** a dividing cell responds to the signal of differentiation is addressed with the filamentous cyanobacterium Nostoc *** 7120,which forms the patterned distribution of *** show that differentiation of a dividing cell was delayed until its division was completed and only one daughter cell became heterocyst.A mutant of patU3,which encodes an inhibitor of heterocyst formation,showed no such delay and formed heterocyst pairs from the daughter cells of cell division or dumbbell-shaped heterocysts from the cells undergoing *** patA mutant,which forms heterocysts only at the filament ends,restored intercalary heterocysts by a single nucleotide mutation of patU3,and double mutants of patU3/patA and patU3/hetF had the phenotypes of the patU3 *** provide evidence that HetF,which can degrade PatU3,is recruited to cell divisome through its C-terminal domain.A HetF mutant with its N-terminal peptidase domain but lacking the C-terminal domain could not prevent the formation of heterocyst pairs,suggesting that the divisome recruitment of HetF is needed to sequester HetF for the delay of differentiation in dividing *** study demonstrates that PatU3 plays a key role in celldivision coupled control of differentiation.

关键词： cyanobacteria heterocyst differentiation cell division pattern formation PatU3

Complementation analyses of Sinorhizobium meliloti nifA mutant with different originated nifA genes

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Chinese Science Bulletin 2006年第22期51卷 2748-2754页

作者： YAO Zhenhua TIAN Zhexian DAI Xiaomi BECKER Anke LI Jian YAN Haiqin XIAO Yan ZHU Jiabi YU Guanqiao RUVERG Silvia WANG Yiping ZOU Huasong national laboratory of plant Molecular genetics Shanghai Institute of Plant Physiology and Ecology Shanghai Institute for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China national laboratory of protein engineering and plant genetic engineering College of Life Sciences Peking University B eijing 100871 China Lehrstuhl für Genetik Universitat Bielefeld Postfach 100131D-33615 33501 Bielefeld Germany Graduate School of the Chinese Academy of Sciences Beijing 100039China

推断的一个以前的工作 Enterobacter 下水道的 nifA 基因没恢复 Sinorhizobium meliloti nifA 的共生显型变异。在现在的学习， Bra-dyrhizobium 的 twonifA 基因日本嗯并且 Mesorhizobium huakuii 也没恢复 S 的共生显型。meliloti ni... 详细信息

推断的一个以前的工作 Enterobacter 下水道的 nifA 基因没恢复 Sinorhizobium meliloti nifA 的共生显型变异。在现在的学习， Bra-dyrhizobium 的 twonifA 基因日本嗯并且 Mesorhizobium huakuii 也没恢复 S 的共生显型。meliloti nifA 异种。在微数组试验的整个 genomic， 238 基因被发现是在 S 以后表示的差别。meliloti nifA 组成地在它的 nifAmutant 被表示了。相反，当表示 *** 时，仅仅 20， 7 和 9 基因改变了他们的 transcriptional 层次， M。在 Sm nifA 异种的 huakuii 和 Enterobacter 下水道 nifA 基因独立。这些基因包括房子保留，精力和中央中间代谢被分类进几官能基，运输系统和共生。有趣地， nifHoperons，显示出的高表示面对任何一个 B 铺平的基因。japonium 或 M。huakuii NifA，它被随后的紫胶 Z 熔化实验证实。

关键词： NifA 基因表达微阵列生物化学