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A New Farnesyl Diphosphate Synthase Gene from Taxus media Rehder:Cloning,Characterization and Functional complementation

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Journal of Integrative Plant Biology 2006年第6期48卷 692-699页

作者： Zhi-Hua Liao Min Chen Yi-Fu Gong Zhu-Gang Li Kai-Jing Zuo Peng Wang Feng Tan Xiao-Fen Sun Ke-Xuan Tang State Key Laboratory of Genetic Engineering School of Life Sciences Fudan-SJTU-Nottingham Plant Biotechnology R＆D Center Morgan-Tan International Center for Life Sciences Fudan University Shanghai 200433 China Laboratory of Natural Products and Metabofic Engineering Institute of Biotechnology Chongqing Sweetpotato Research Center School of Life Sciences Southwest University Chongqing 400715 China Plant Biotechnology Research Center Fudan-SJTU-Nottingham Plant Biotechnology R＆D Center School of Agriculture and Biology Shanghai Jiaotong University Shanghai 200030 China Faculty of Life Sciences and Biotechnology Ningbo University Ningbo 315211 China

Farnesyl dlphosphate synthase （FPS; EC 2.***.1.10） catalyzes the production of 15-carbon farnesyl dlphosphate which Is a branch-point Intermediate for many terpenoids. This reaction Is considered to be a ratelimiting step In terpenold biosynthesis. Here we report for the first time the cloning of a new full-length cDNA encoding farnesyl dlphosphate synthase from a gymnosperm plant species, Taxus media Rehder, designated as TmFPS1. The full-length cDNA of TmFPS1 （GenBank accession number： AY461811） was 1 464 bp with a 1 056-bp open reading frame encoding a 351-amino acid polypeptlde with a calculated molecular weight of 40.3 kDa and a theoretical pl of 5.07. Biolnformatlc analysis revealed that TmFPS1 contained all five conserved domains of prenyltransferases, and showed homology to other FPSs of plant origin. Phylogenetlc analysis showed that farnesyl dlphosphate synthases can be divided Into two groups： one of prokaryotic origin and the other of eukaryotic origin. TmFPS1 was grouped with FPSs of plant origin. Homologybased structural modeling showed that TmFPS1 had the typical spatial structure of FPS, whose most prominent structural feature Is the arrangement of 13 core helices around a large central cavity In which the catalytic reaction takes place. Our blolnformatic analysis strongly suggests that TmFPS1 is a functional gene. Southern blot analysis revealed that TmFPS1 belongs to a small FPSgene family in T. media. Northern blot analysis indicated that TmFPS1 is expressed in all tested tissues, Including the needles, stems and roots of T. media. Subsequently, functional complementatlon with TmFPS1 in a FPS-deflclent mutant yeast demonstrated that TmFPS1 did encode farnesyl dlphosphate synthase, which rescued the yeast mutant. This study will be helpful In future Investigations aiming at understanding the detailed role of FPS In terpenold biosynthesis flux control at the molecular genetic level.

关键词： farnesyl diphosphate synthase rapid amplification of cDNA ends Taxus media yeast complementation

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PtHAK5, a candidate for mediating high-affinity K^+ uptake in the halophytic grass, Puccinellia tenuiflora

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Frontiers of Agricultural Science and Engineering 2018年第1期5卷 108-117页

作者： Haili YANG Weidan ZHANG Weiwei CHAI Wenying WANG Li GAO Jing ZHANG Yongping WANG Suo-Min WANG State Key Laboratory of Grassland Agro-ecosystems/College of Pastoral Agriculture Science and Technology Lanzhou University

Puccinellia tenuiflora is a typical salt-excluding halophytic grass with strong salt-tolerance, which enhances tolerance by restricting Na^+influx as well as having a strong selectivity for K^+ over Na^+. The HAK5 K^+ transporters generally modulate effective K^+acquisition in plants, especially under low K^+ condition. In this study,Pt HAK5 from P. tenuiflora was isolated by RT-PCR and characterized using yeast complementation. The results showed Pt HAK5 consisted of 784 amino acids and shared over 80% homology with the identified high-affinity K^+ transporter HAK5 from other higher plants. The expression of PtHAK5 rescued the K^+ -uptake-defective phenotype of yeast strain CY162. In conclusion, PtHAK5 is a candidate for mediating high-affinity K^+ uptake under low K^+ conditions.

关键词： K+ uptake PtHAK5 Puccinellia tenuiflora yeast complementation

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Cloning and characterization of a gene encoding HMG-CoA reductase from Ganoderma lucidum and its functional identification in yeast

Cloning and characterization of a gene encoding HMG-CoA redu...

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第二届全国食用菌中青年专家学术交流会

作者： Chang-Hua Shang,Fen Zhu,Na Li,Xiang Ou-Yang,Liang Shi,Ming-Wen Zhao~+ and Yu-Xiang Li College of Life Sciences,Nanjing Agricultural University Key Laboratory of Microbiological Engineering of Agricultural Environment,Ministry of Agriculture,Nanjing 210095,Jiangsu,P.R.China

＜正＞A gene encoding 3-hydroxy-3-methylglutaryl-coenzyme A reductase（HMGR） was isolated from a triterpene-producing fungus,Ganoderma *** report provides the complete nucleotide sequence of the full-length cDNA encoding HMGR and its genomic DNA *** cDNA of the HMGR（GenBank Accession Number: EU263989） was found to contain an open reading frame（ORF） of 3681bp encoding a 1226-amino-acid polypeptide,whereas the HMGR genomic DNA sequence（GenBank Accession Number:EU263990 ） consisted of 4262 bp and contained seven exons and six *** deduced amino acid sequence of G lucidum HMGR showed significant homology to the known HMGRs from Ustilago maydis and Crypt5ococcus neoformans and contained four conserved domains.A gene expression analysis showed that the expression level was relatively low in mycelia incubated for 10,12,14 days,and reached the highest level in the *** complementation of Gl-HMGR in a HMGR-deficient mutant yeast strain demonstrated that the cloned cDNA encoded a HMG-CoA reductase.

关键词： cloning Ganoderma lucidum HMG-CoA reductase yeast complementation

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MsYSL1 of Miscanthus sacchariflorus enhances resistance of Arabidopsis to cadmium by mediating metal ion reallocation

MsYSL1 of Miscanthus sacchariflorus enhances resistance of A...

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第七届长三角植物科学研讨会暨青年学术报告会

作者： Cheng Zhang Houming Chen Haipeng Guo Dean Jiang College of Life Sciences Zhejiang University

The Yellow Stripe-Like(YSL) family of transporters mediates the uptake,translocation,and distribution of various mineral elements in vivo by transferring metal ions chelated with phytosiderophore or nicotianamine(NA).However,little is known about the roles of the YSL genes against cadmium in *** this study,we first cloned and characterized a vital member of the YSL gene family,Ms YSL1,from the bioenergy plant Miscanthus *** YSL1 localized in the plasma membrane and was widely expressed throughout the whole seedling with the highest expression level in the *** addition,its expression in the root was stimulated by excess manganese(Mn),cadmium(Cd),and lead,and a shortage of iron(Fe),zinc(Zn),and *** complementation in yeast indicated that Ms YSL1 showed transport activity for Fe(II)-NA and Zn-NA,but not for *** they exhibited no significant differences versus the wild type under normal cultivation conditions,Ms YSL1-overexpressing Arabidopsis lines displayed a higher resistance to Cd accompanied by longer root lengths,lower Cd,Zn,and Mn levels in roots,and higher Cd,Fe,and Mn translocation ratios under Cd ***,genes related to NA synthesis,metal translocation,long-distance transport,and Cd exclusion were highly induced in transgenic lines under Cd ***,MsYSL1 may be an essential transporter for diverse metal-NAs to participate in the Cd detoxification by mediating the reallocation of other metal ions.

关键词： Miscanthus sacchariflorus MsYSL1 yeast complementation Cd tolerance Arabidopsis

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