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Expression and functional analysis of the rice plasma-membrane intrinsic protein gene family

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Cell research 2006年第3期16卷 277-286页

作者： Lei Guo~1 Zi Yi Wang~1 Hong Lin~1 Wei Er Cui~1 Jun Chen~1 Meihua Liu~1 Zhang Liang Chen~(1,2) Li Jia Qu~(1,2) Hongya Gu~(1,2)1 Peking-Yale Joint research center for plant Molecular genetics and AgroBiotechnology,national Laboratoryfor Protein Engineering and plant genetic Engineering,College of Life Sciences,Peking University,beijing 100871,china 2 The national plant gene research center,beijing 100101,china Peking-Yale Joint research center for plant Molecular genetics and AgroBiotechnology National Laboratory for Protein Engineering and Plant Genetic Engineering College of Life Sciences Peking University Beijing China The national plant gene research center Beijing China

Plasma membrane intrinsic proteins (PIPs) are a subfamily of aquaporins that enable fast and controlled translocationof water across the *** this study,we systematically identified and cloned ten PIP genes from *** on thesimilarity of the amino acid sequences they encoded,these rice PIP genes were classified into two groups and designatedas OsPIPI-1 to OsPIP1-3 and OsPIP2-1 to OsPIP2-7 following the nomenclature of PIP genes in ***-PCR analysis identified three root-specific and one leaf-specific OsPIP ***,the expression profile ofeach OsPIP gene in response to salt,drought and ABA treatment was examined in *** on transgenic plantsover-expressing of either OsPIP1 (OsPIPI-1) or OsPIP2 (OsPIP2-2) in wild-type Arabidopsis,showed enhanced toler-ance to salt (100 mM of NaCl) and drought (200 mM of mannitol),but not to salt treatment of higher concentration (150mM of NaCl).Taken together,these data suggest a distinct role of each OsPIP gene in response to different stresses,andshould add a new layer to the understanding of the physiological function of rice PIP genes.

关键词： aquaporins PIP quantitative RT-PCR Oryza sativa organ-specific

Cloning and characterization of two putative seven-transmembrane receptor genes from cotton

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Progress in Natural Science:Materials International 2008年第10期18卷 1225-1231页

作者： Peng Gao Piming Zhao Juan Wang Haiyun Wang Guiling Wang Guixian Xia national Key Laboratory of plant Genomics,Institute of Microbiology,Chinese Academy of Sciences,Datun Road,Chaoyang District,beijing 100101,china national center for plant gene research,beijing 100101,china national Key Laboratory of plant Genomics Institute of Microbiology Chinese Academy of Sciences Datun Road Chaoyang District Beijing 100101 China national center for plant gene research Beijing 100101 China

Using rapid amplification of cDNA ends (RACE)-PCR,two full-length cDNAs encoding putative seven-transmembrane receptors (designated Gh7TMpRI and Gh7TMpR2) were cloned from cotton *** blot and an ApaL1 restriction site polymorphism analyses revealed that Gh7TMpRI was derived from the ancestral A diploid genome,while Gh7TMpR2 was from the D subgenome. Northern blot hybridization indicated that both Gh7TMpR1 and GhTTMpR2 were expressed preferentially in the elongation phase of fiber *** of the Gh7TMpRl proteins were located within the membrane structure and displayed a punctuate pattern of *** of Gh7TMpR1 in fission yeast disrupted the polar growth and caused the formation of rounded cells. These results suggest that GhT7MpRl may play a critical role in cotton fiber development,perhaps as a signaling receptor that is involved in controlling fiber elongation.

关键词： Cotton Seven-transmembrane protein Fiber elongation Alloalleles

PAT: waking up a lazy sleeping beauty

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Cell research 2007年第5期17卷 387-388页

作者： Xiangdong Fu Jianru Zuo, national center for plant gene research, Institute of genetics and Developmental Biology, Chinese Academy of Sciences, beijing 100101, china. xdfu@*** national center for plant gene research Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing China

Once upon a time, there was an unusual corn; and un-like its normal siblings, its stalk grew towards the groundrather than in an upward direction. The sleeping stature ofthe corn was portrayed in early 1930s as "lazy" by Jenkinsand Gerhardt [1]. Later, lazy appeared to be a fashionin the plant kingdom, found in rice, barley, tomato andseveral other species [2]. These seemingly magic spell-chanted plants have drawn considerable interests of plantbiologists and breeders for many decades. To breeders, thelazy phenotype, viewed as tillering or branching anglesof stems (or termed as culms in crop plants), represents akey characteristic important for controlling photosynthesisefficiency and planting density, and thus a crucial factor indetermining crop production [6]. Previous physiologicalstudies suggested that the prostrate or lazy phenotype wasattributed, at least in part, to the loss of gravitropism. Forexample, the rice lazyl plants are ageotropic, whereas wildtype (WT) plants are negatively geotropic [2]. Gravity plays a major role in plant morphogenesis bydetermining the directional growth of plant organs. Upongermination, shoots grow upward, capturing light, and bycontrast, roots penetrate the soil and grow downward, tak-

关键词： waking up a lazy sleeping beauty PAT

Heterotrimeric G protein α subunit is involved in rice brassinosteroid response

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Cell research 2006年第12期16卷 916-922页

作者： Lei Wang~(1,2) Yun-Yuan Xu~1 Qi-Bin Ma~(1,2) Dan Li~(1,2) Zhi-Hong Xu~(1,3) Kang Chong~(1,3) 1 Key Laboratory of Photosynthesis and Environmental Molecular Physiology,Institute of Botany,Chinese Academy of Sciences,Nanxincun 20,Xiangshan,beijing 100093,china,2 Graduate School of the Chinese Academv of Sciences,100046 beijing,china 3 national plant gene research center (beijing),beijing 100093,china Key Laboratory of Photosynthesis and Environmental Molecular Physiology Institute of Botany Chinese Academy of Sciences Xiangshan China Graduate School of the Chinese Academy of Sciences Beijing China national plant gene research center (beijing) Beijing China

Heterotrimeric G proteins are known to function as messengers in numerous signal transduction *** nullmutation of RGA(rice heterotrimeric G protein α subunit),which encodes the α subunit of heterotrimeric G proteinin rice,causes severe dwarfism and reduced responsiveness to gibberellic acid in ***,less is known aboutheterotrimeric G protein in brassinosteroid(BR)signaling,one of the well-understood phytohormone *** thepresent study,we used root elongation inhibition assay,lamina inclination assay and coleoptile elongation analysis todemonstrated reduced sensitivity of dl mutant plants(caused by the null mutation of RGA)to 24-epibrassinolide(24-epiBL),which belongs to brassinosteroids and plays a wide variety of roles in plant growth and ***,RGA transcript level was decreased in 24-epiBL-treated seedlings in a dose-dependent *** results show thatRGA is involved in rice brassinosteroid response,which may be beneficial to elucidate the molecular mechanisms of Gprotein signaling and provide a novel perspective to understand BR signaling in higher plants.

关键词： heterotrimeric G protein α subunit dl mutant BR signaling rice

BUD2,encoding an S-adenosylmethionine decarboxylase,is required for Arabidopsis growth and development

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Cell research 2006年第5期16卷 446-456页

作者： Chunmin Ge~(1,2,*) Xia Cui~(1,2,*) Yonghong Wang~1 Yuxin Hu~3 Zhiming Fu~1 Dongfen Zhang~(1,2) Zhukuan Cheng~1 Jiayang Li~1 ~1State Key Laboratory of plant Genomics and national center for plant gene research,Institute of genetics and Developmental Biology,Chinese Academy of Sciences,beijing 100101,china ~2Graduate School,Chinese Academy of Sciences,beijing 100101,china ~3Institute of Botany,Chinese Academy of Sciences,beijing 100093,china State Key Laboratory of plant Genomics and national center for plant gene research Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing China Graduate School Chinese Academy of Sciences Beijing China Institute of Botany Chinese Academy of Sciences Beijing China

Polyamines are implicated in regulating various developmental processes in plants,but their exact roles and how theygovern these processes still remain *** report here an Arabidopsis bushy and dwarf mutant,bud2,which resultsfrom the complete deletion of one member of the small gene family that encodes S-adenosylmethionine decarboxylases(SAMDCs) necessary for the formation of the indispensable intermediate in the polyamine biosynthetic ***2 plant has enlarged vascular systems in inflorescences,roots,and petioles,and an altered homeostasis *** double mutant of bud2 and samdcl,a knockdown mutant of another SAMDC member,is embryo lethal,demonstrat-ing that SAMDCs are essential for plant *** results suggest that polyamines are required for the normalgrowth and development of higher plants.

关键词： BUD2 polyamines embryogenesis plant architecture Arabidopsis

LEAFY HEAD2,which encodes a putative RNA-binding protein,regulates shoot development of rice

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Cell research 2006年第3期16卷 267-276页

作者： Guo Sheng Xiong~(1,3) Xing Ming Hu~2 Yong Qing Jiao~1 Yan Chun Yu~1 Cheng Cai Chu~1 Jia Yang Li~1 Qian Qian~2 Yong Hong Wang~11 State Key Laboratory of plant Genomics and national center for plant gene research,Institute of genetics and Developmental Biology,Chinese Academy of Sciences,beijing 100101,china 2 State Key Laboratory of Rice Biology,china national Rice research Institute,Chinese Academy of Agricultural Sciences,Hangzhou 310006,china 3 Graduate University of Chinese Academy of Sciences,beijing 100049,china State Key Laboratory of plant Genomics and national center for plant gene research Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing China Graduate University of Chinese Academy of Sciences Beijing China State Key Laboratory of Rice Biology China National Rice Research Institute Chinese Academy of Agricultural Sciences Hangzhou China

During vegetative development,higher plants continuously form new leaves in regular spatial and temporal *** with abnormal leaf developmental patterns not only provide a great insight into understanding the regulatorymechanism of plant architecture,but also enrich the ways to its modification by which crop yield could be ***,we reported the characterization of the rice leafy-head2 (lhd2) mutant that exhibits shortened plastochron,dwarfism,reduced tiller number,and failure of phase transition from vegetative to reproductive *** and histologicalstudy revealed that the rapid emergence of leaves in lhd2 was resulted from the rapid initiation ofleafprimordia whereasthe reduced tiller number was a consequence of the suppression of the tiller bud *** molecular and geneticanalysis showed that LHD2 encodes a putative RNA binding protein with 67% similarity to maize *** ofgenome-scale expression profiles between wild-type and lhd2 plants suggested that LHD2 may regulate rice shoot devel-opment through KNOXand hormone-related *** similar phenotypes caused by LHD2 mutation and the conservedexpression pattern of LHD2 indicated a conserved mechanism in controlling the temporal leaf initiation in grass.

关键词： phyllotaxy plastochron LHD2 RNA-binding protein stem elongation plant architecture Oryza sativa L

Survival of Human Metallothionein-2 Transplastomic Chlamydomonas reinhardtii to Ultraviolet B Exposure

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Acta Biochimica et Biophysica Sinica 2006年第3期38卷 187-193页

作者： Yu-Kun ZHANG~1 Gui-Fang SHEN~2 Bing-Gen RU~1*1 national Laboratory of Protein Engineering and plant genetic Engineering,Peking University,beijing 100871,china 2 Biotechnology research Institute,the Chinese Academy of Agricultural Sciences,beijing 100081,china national Laboratory of Protein Engineering and plant genetic Engineering Peking UniversityBeijing 100871 China Biotechnology research Institute the Chinese Academy of Agricultural SciencesBeijing 100081 China

Solar ultraviolet (UV) radiation has a great influence on green organisms,especially planktonlike Chlamydomonas.A human metallothionein-2 gene,which is generally considered to have an anti-radiationfunction by its coding product,was transferred into the chloroplast genome of Chlamydomonas *** dynamically measure the UV effects on Chlamydomonas cells grown in liquid tris-acetate-phosphatemedium,a new method was developed based on the relationship between the chlorophyll content of an algalculture and its absorbance at 570 nm after the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium *** this experiment,both the wild-type and the transplastomic *** ceils were cultivated in 96-well microplates containing liquid tris-acetate-phosphate medium in the absence or presence of zinc,copper,cadmium and *** transgenic *** showed a higher resistance than wild-type to UV-Bexposure under all the examined *** in the medium had positive impacts on both types of cells,but had significant influence only on the transplastomic ***,the high cell viability of the trans-genie alga at the end of the 8 h UV-B treatment disappeared after a 20-h recovery *** did notprotect cells from UV-B damage,but clearly enhanced the growth of both wild-type and transgenic ***.

关键词： ultraviolet B MTT transplastomic Chlamydomonas metallothionein

Disruption of phytoene desaturase gene results in albino and dwarf phenotypes in Arabidopsis by impairing chlorophyll, carotenoid, and gibberellin biosynthesis

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Cell research 2007年第5期17卷 471-482页

作者： Genji Qin1 Hongya Gu1,2 Ligeng Ma1,3 Yiben Peng1 Xing Wang Deng1,3 Zhangliang Chen1 Li-Jia Qu1,2 1 Peking-Yale Joint center for plant Molecular genetics and Agrobiotechnology, national Laboratory for Protein Engineering and plant genetic Engineering, College of Life Sciences, Peking University, beijing 100871, china 2 The national plant gene research center (beijing), beijing 100101, china 3 Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven Connecticut 62520-8104, USA Peking-Yale Joint center for plant Molecular genetics and Agrobiotechnology National Laboratory for Protein Engineering and Plant Genetic Engineering College of Life Sciences Peking University Beijing China The national plant gene research center (beijing) Beijing China Department of Molecular Cellular and Developmental Biology Yale University New Haven Connecticut USA

Carotenoids play an important role in many physiological processes in plants and the phytoene desaturase gene （PDS3）encodes one of the important enzymes in the carotenoid biosynthesis pathway. Here we report the identification andanalysis of a T-DNA insertion mutant of PDS3 gene. Functional complementation confirmed that both the albino anddwarf phenotypes of the pds3 mutant resulted from functional disruption of the PDS3 gene. Chloroplast development wasarrested at the proplastid stage in the pds3 mutant. Further analysis showed that high level of phytoene was accumulatedin the pds3 mutant. Addition of exogenous GA3could partially rescue the dwarf phenotype, suggesting that the dwarfphenotype of the pds3 mutant might be due to GA deficiency. Microarray and RT-PCR analysis showed that disruptingPDS3 gene resulted in gene expression changes involved in at least 20 metabolic pathways, including the inhibition ofmany genes in carotenoid, chlorophyll, and GA biosynthesis pathways. Our data suggest that the accumulated phytoenein the pds3 mutant might play an important role in certain negative feedbacks to affect gene expression of diverse cel-lular pathways.

关键词： Arabidopsis thaliana phytoene desaturase albino dwarf microarray analysis

Identification of AtENT3 as the main transporter for uridine uptake in Arabidopsis roots

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Cell research 2006年第4期16卷 377-388页

作者： Kun Ling Chen~(1,2,*) Min Xin Xu~(2,3,*) Guang Yong Li~(2,3,*) Hui Liang~2 Zong Liang Xia~(2,3) Xin Liu~2 Ji Shu Zhang~1 Ai Min Zhang~2 Dao Wen Wang~2 ~1School of Life Sciences,Northwest Agricultural and Forestry University,Yangling 712100,china ~2center for plant gene research(beijing)and the State Kev Laboratory of plant Cell and Chromosome Engineering,Institute of genetics and Developmental Biology,Chinese Academy of Sciences,Datun Road,Chaovang District,beijing 100101,china ~3Graduate School of the Chinese Academy of Sciences,Yuquan Road,Shijingshan District,beijing 100039,china School of Life Sciences Northwest Agricultural and Forestry University Yangling China center for plant gene research (beijing) and the State Key Laboratory of plant Cell and Chromosome Engineering Institute of Genetics and Developmental Biology Chinese Academy of Sciences Datun Road Chaoyang District Beijing China Graduate School of the Chinese Academy of Sciences Yuquan Road Shijingshan District Beijing China

Previous studies have shown that Arabidopsis equilibrative nucleoside transporters （AtENTs） possess transport activitieswhen produced in yeast cells and are differentially expressed in Arabidopsis ***,we report further analysis onthe nucleoside transport activities and transcriptional patterns of AtENT *** recombinant proteins of AtENTs3,6,and 7,but not those of AtENTs 1,2,4,and 8,were found to transport thymidine with high *** to previ-ous suggestion that AtENT1 may not transport uridine,this work showed that recombinant AtENT1 was a pH-dependentand high-affinity transporter of *** grown on MS plates,the AtENT3 knockout plants were more tolerant tothe cytotoxic uridine analog 5-fluorouridine than wild-type plants and the knockout plants ofAtENT 1 or ***-sistent with this observation,the AtENT3 knockout line exhibited a significantly decreased ability to take up [3H]uridinevia the roots when compared with wild-type plants and the plants with mutated AtENT 1 or *** indicates thatAtENT3,but not AtENTs 1 and 6,is the main transporter for uridine uptake in Arabidopsis *** transcription ofAtENTs 1,3,4,6,7,and 8 was regulated in a complex manner during leaf development and *** contrast,thesix AtENT members were coordinately induced during seed *** work provides new information on thetransport properties of recombinant AtENT proteins and new clues for future studies of the in vivo transport activitiesand physiological functions of the different ENT proteins in Arabidopsis plants.

关键词： Arabidopsis equilibrative nucleoside transporter expression pattern nucleoside transport functional analysis