摘要
实现源、库的协调是小麦获得高产的生理基础。TPT是光合碳代谢与胞质其它代谢沟通的桥梁,是源、库间同化物运输的关键调控部位。研究TPT的性质、TPT在同化物分配中的凋节功能及其TPT基因的表达调控,对于提高光合作用同化物利用效率有着重要意义。但是至今对重要粮食作物小麦TPT的运输性质、生理功能及其基因表达凋控的研究尚无报导。本文研究了小麦TPT蛋白的定位、运输动力学及其对同化物分配的调节功能,TPT基因表达的影响因子,为协调源、库间的同化物分配,提高光合同化物的利用效率,增加作物产量提供依据。
根据我们已克隆的小麦TPT cDNA序列(GenBank登录号AF314182)推测的蛋白氨基酸序列,经软件分析表明羧基端的13个氨基酸序列可能是TPT的抗原决定簇。以人工合成的13肽(N’-KAK IEE EKR AKA A-C’)为半抗原,与BSA偶联后,免疫兔子获得抗血清,效价为1:500。利用TPT抗体进行Western blotting分析表明,小麦TPT蛋白仅存在于叶绿体被膜上,而在液泡膜和线粒体膜上不存在。通过TPT不可逆抑制剂[H~3]_2-DIDS标记和SDS-PAGE分析表明,小麦TPT蛋白是叶绿体被膜中最主要的蛋白,约占被膜总蛋白的15%,其分子量为35kD。
用小麦完整叶绿体,采用硅油离心法进行磷酸二羟丙酮(TP)、磷酸烯醇式丙酮酸(PEP)、葡萄糖-6-磷酸(G6P)与Pi反向交换的TPT运输动力学测定。结果表明,DHAP/Pi的最大运输活性最高,PEP/Pi次之,G6P/Pi最低。这些运输底物的Km值(由小至大)表示TPT与底物的亲和性,亲和性由高到低依次为DHAP、Pi、PEP和G6P,证明TPT的最适运输底物和运输方式为DHAP/Pi的反向交换。TPT转运活性受光的调节,光可提高V_(max),但光照并不能明显改变TPT对底物的亲和性(即TPT对其运输底物的Km值仅受光的微弱影响)。黑暗条件下,DHAP的V_(max)为21.4μmol/mg chl·h,K_m为0.019mmol/L。光照处理使DHAP的V_(max),增加54.67%,K_m值下降10.5%,表明光在调节TP合成的同时也促进了TPT的运输活性。用TPT特异性抑制剂DIDS处理的叶绿体,无论黑暗还是光照,所有底物的运输均被强烈抑制。光照条件下抑制达90-95%。
叶绿体内光合同化物磷酸丙糖(TP)有3个去向,即卡尔文循环中RuBP的再生、外运到胞质中合成蔗糖送往库器官和叶绿体内合成临时淀粉。利用TPT的专一性抑制剂比较光照条件下离体叶绿体中的终端产物淀粉合成量的差异,估算出TPT外运TP的量。当TPT被DIDS抑制,失去子运输能力时,可使叶绿体内淀粉合成增加14倍,与淀粉合成途径相关的上游代谢物PGA、TP、FBP、F6P、G6P的含量均有明显增加,增幅在25%以上。证明在保证卡尔文循环正常运转的前提下,通过TPT外运到胞质中参与蔗糖合成和其它代谢活动的TP约占93.6%,而用于叶绿体内合成淀粉的TP仅占6.4%。因此,小麦是一种典型的糖叶型植物。对光合效率不同的小麦品种进行TPT的Western blotting分析,结果表明中、低等光合效率品种的TPT表达量均较低,而高光合效率小麦品种的TPT表达量较高。因此,在对更多的小麦品种进行验证后,有可能把TPT表达的丰度作为筛选高产小麦品种的参考指标之一。
Northern blotting分析表明,渗透胁迫处理使小麦幼苗TPT在转录水平上的表达增强,但具有相同渗透势的葡萄糖等溶液处理使TPT在转录水平上的表达明显降低。在葡萄糖对小麦TPT表达调节中,低浓度范围内,即0.1-100μmol/L的葡萄糖有激活效应,当浓度大于1mmol/L时
为抑制作用。用 5 mmol/L葡萄糖溶液处理小麦幼苗 id后,TPT基因的表达开始下降,第 4 d时
受到强烈的抑制。
为了验证己糖激酶(HXK)是否参与葡萄糖对小麦TPT表达的调节,用r干CR方法克隆到一
长度为 581 hp的 HXK CDNA片段,部分构建了 dSANAi真核表达载体pBI-HXK-RNAi。电激法
转化小麦原生质体,检测 HXK活性为对照的 9.62%,表明获得了 pBIHXK-RNAi瞬时表达的原
生质体。采用’H.DIDS特异标记TPT,比较野生和转化原生质体中葡萄糖、多种葡萄糖类似物以
及 HXK抑制剂(葡萄糖胺)对 TPT表达量的影响。结果表明只有葡萄糖仰 mmol/L)或者能被 HXK
磷酸化的葡萄糖类似物2-deoxyglucose与活化的HXK共存时,TPT的表达才被抑制。初步分析
认为,葡萄糖可能是TPT对糖应答反应的信号分子,HXK对葡萄糖的磷酸化为糖应答反应所必
需。
The partitioning of photosynthate between source and sink is the physiological basis for acquiring a high yield in wheat, and triose phosphate/ phosphate translocator (TPT) may be the first regulation point for this partitioning. Understanding the characteristics of TPT, its regulation on the distribution of assimilates and the expression of tpt gene may be critical for improving the utilization rate of photosynthetic assimilates. But up to now it is unknown about the transport characteristics, the physiological function of TPT, and the expression of tpt gene in wheat. In this paper, we described the localization and transport kinetics of TPT, analyzed its physiological role in the distribution of assimilates, explored the factors that effect the expression of tpt, and studied the role of hexokinase (HXK) in the expression of tpt gene regulated by glucose in wheat, which will provide clues for coordinating the partitioning of assimilates between source and sink, improving the utilization rate of photosynthat
e, and increasing crop yield.
A peptide with a length of 13 amino acids (N'-KAK. IEE EKR AKA A-C') was synthesized according to the putative amino acid sequence by 3' end of wheat tpt cDNA (39 bp) cloned by us. The peptide coupled with bovine serum albumin (BSA) was used to inject rabbit to prepare antibody. The ratio of the prepared antibody against TPT was 1:500. Western blotting analysis showed that TPT only exists in the envelope membrane of wheat chloroplasts, but not in the membrane of vacuoles and mitochondrials. SDS-PAGE and labeling of TPT with [H3]2-DIDS (4, 4'-diisothiocyano-2, 2'-stilbenedisulfonate, DIDS), an irreversible specific inhibitor, indicated that TPT is a 35 kD chloroplast membrane protein and makes up about 15 % of the total membrane proteins of chloroplasts.
Silicone-oil-layer centrifugation system was employed to study the kinetic properties of TPT, which catalyzes the counter exchange of dihydroxyacetone phosphate (DHAP)/phosphoenol -pyruvate (PEP)/glucose-6-phosphate (G6P) and inorganic phosphate (Pi). The maximal transport activity of TPT was the highest for DHAP/Pi, followed by PEP/Pi and G6P/Pi. The Km of TPT was the lowest for DHAP, followed by Pi, PEP and G6P, so the most preferred substrate of TPT is DHAP. The Vmas and Km of DHAP were 21.4 umol/mg chl-h and 0.019 mmol/L respectively in the dark. In the light, Vmax of DHAP increased by 54.67 %, and Km decreased by 10.5 %. Thus, the transport of TPT appears to be regulated by light, and it can increase Vmax, but illumination does not change the affinity of TPT to substrates, that is to say that light has few effects on the Km of substrates transported by TPT. The transport of the various substrates is strongly inhibited by DIDS whether in dark or in light.
The transport activity of TPT being inhibited leads to an obvious increase of the content of many metabolites and an accumulation of starch in chloroplasts. A change of the enzyme acitvity in wheat chloroplasts also provide a support for the flux of photosynthate to starch. Wheat is a kind of sugar-leaf plants. TPT plays an important role in the regulation on the distribution of assimilates in wheat chloroplasts and in the transport of photosynthetic products from source to sink. Its function is to export
photosynthetic assimilates out of chloroplasts efficiently and participate in all kinds of metabolism in cytosol.
Northern blotting analysis demonstrated that the expression of tpt gene increased under osmotic stress treatments in seedlings of Jing 411, but decreased obviously under the treatments with glucose, sucrose and frucrose. The expression of tpt decreased clearly in wheat seedlings when the concentration of glucose was above 5 mmol/L for 4 days. Western blotting analysis showed that the expression of tpt of the cultivar Jing 411, with a middle photosynthetic rate, Zhongpin 97-37 and Zhong 924, with a low photosynthetic rate, is lower than No 6 Wenmai and No 16 Jinan, with a higher photosynthetic rate, so the abundance of expression of tpt may act as
引文
陈珈.光合产物输出.邹承鲁等主编.当代生物学前沿.中国致公出版社,1999,第126页
陈珈,王学臣,王庆梅.代谢物转运器对光合作用的调节.娄成后等主编:作物产量形成的生理学基础.北京,中国农业出版社,第7-26页
李宏潮等.冬小麦原生质体培养及植株再生.植物学报,1991,33:706-711
萨姆布鲁克 J,弗里奇 EF,曼尼阿蒂斯 T.分子克隆(第二版),科学出版社,1996
王海波等.小麦原生质体培养—高频率的细胞团形成和植株再生.中国科学(B 辑),1990,8:828-835
王庆梅,杨树德,陈珈,叶绿体内被膜上的磷酸丙糖转运器.植物学通报,2001,18:11~15
王庆梅.小麦和水稻磷酸丙糖转运器基因的克隆及其表达的组织特异性分析,中国农业大学博士学位论文,2001
夏朝晖,李晓薇,余和芬,陈珈.盐和干旱胁迫对燕子掌叶片液泡膜 H+-ATPase 活性的影响.植物生理学报,2000,26:433-436
朱美君,陈珈,朱庆鸿,王学臣.两相分配法制备玉米根质膜及其纯度鉴定.生物化学杂志,1997,13:686-690
Arenas-Huertero F, Arroyo A, Zhou L, Sheen J, Leon P. Analysis of Arabidopsis glucose insensitive mutants, gin5 and gin6, reveals a central role of the plant development by sugar. Genes Dev, 2000, 14:2085-2096
Ball K L, Barker J H A, Halford N G et al. Immunlogical evidence that HMG-CoA reductase kinase A is the cauliflower homologue of the RKIN 1 subfamily of plant protein kinase. FEBS Lett, 1995, 377:189-192
Barker J H A, Slocombe S P, Ball K L et al. Evidence that barley 3-hydroxy-3-methylglutaryl-Coenzyme A reductase kinase is a member of the sucrose nonfermenting-1-related protein kinase family. Plant Physiol, 1996, 112: 1141-1149
Barr J E, Miller A, Rodermel S. The Transformation of Arabidopsis thaliana with Agrobacterium tumefaciens: Creating and Identifying a Homozygous Line with an Altered Hexokinase Activity. Protocol from the Department of Botany, Iowa State University, Ames, Iowa 50011
Baskin T, Remillong E, Wilson J. The impact of mannose and other carton sources on the elongation and diameter of the primary root of Arabidopsis thaliana. Aust J Plant Physiol, 2001, 38:481-488
Batz O, Scheibe R, Neuhaus H E. Transport processes and corresponding changes in metabolite levels in relation to starch synthesis in barley (Hordeum vulgare L.) etioplasts. Plant Physiol, 1992, 100: 184-190
Bhalerao R P, Salchert K, Bako L, Okresz L, Szabados L, Muranaka T, Machida Y, Schell J, Koncz C. Regulatory interaction of PRL1 WD protein with Arabidopsis SNF1-like protein kinases. Proc. Natl. Acad Sci, 1999, USA 96:5322-5327
Borchert S, GroBe H, Heldt H W. Specific transport of inorganic phosphate, glucose 6-phosphate,
dihydroxyacetone phosphate and 3-phosphoglycerate into amyloplasts from pea roots.FEBS LETTERS,1989,253:183-186
Boxall S,Martin T,Graham I.A new class of Arabidopsis mutants that is carbohydrate insensitive. Seventh International Conference on Arabidopsis Research,Norwich.1996,UK.Abstract S96
Bradford M H.Rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Ana.Biochem,1976,72:248-254
Brodzik R,Koprowski H,Yusibov V,Sirko A.Production of urease from Helicobacter pylori in transgenic tobacco plants.Cellular & Molecular Biology Letters,2000,5:357-366
Brouquisse R,Evrard A,Rolin D,Raymond P,Roby,C.Regulation of protein degradation and protease expression by mannose in maize root tips:Pi sequestration by mannose may hinder the study of its signaling properties.Plant Physiol,2001,125:1485-1498.
Burke L,Hibberd J M,Quick W P et al.The H+/sucrose cotransporter NtSUT1 is essential for sugar export from tobacco leaves.Plant Physiol,1998,118:59-68
Celenza J L,Carlson M.A yeast gene that is essential for release from glucose repression encodes a protein kinase.Science,1986,233:1175-1180
Chen J(陈珈),Wu Z-Y(吴忠义),Zhu M-J(朱美君).Solubilization & characteristics of ABA binding proteins from plasma membrane of maize roots.植物学报(Acta Bot Sin),1997,39:1015-1021 (In Chinese with English Abstract)
Copeland L,Tanner G J.Physiol Plant,1988,74:531-536
Corbesier L,Lejeune P,Bernier G.The role of carbohydrates in the induction of flowering in Arabidopsis thaliana:comparison between the wild type and a starchless mutant.Planta,1998,206: 131-137
Coruzzi,G,Bush D R.Nitrogen and carbon nutrient and metabolite signaling in plants.Plant Physiol, 2001,125:61-64
Coruzzi G M,Zhou L.Carbon and nitrogen sensing and signaling in plants:Emerging 'matrix effects.' Curr.Opin.Plant Biol,2001,4:247-253
Dai N,Schaffer A,Petreikov M et al.Overexpression of Arabidopsis hexokinase in tomato plants inhabits growth,reduces photosythsis,and induces rapid senescence.Plant Cell,1999,1:737-749
Dai N,Schaffer A,Petreikov M,Shahak Y,Levine A,Giller Y,Ratner K,Granot D.Overexpression of Arabidopsis hexokinase in tomato plants inhibits growth,reduces photosynthesis and induces rapid senescence.Plant Cell,1999,11:1253-1266
Dijkewl P P,Koch P,Bezemer R et al.Sucrose represses the developmentally controlled transient activation of the plastocyanin gene in Arabidopsis thaliana seeding.Plant Physiol,1996,110: 455-463
Dijkwel P P,Huijser C,Weisbeeek P J et al.Sucrose control of phytochrome A signaling in Arabidopsis.Plant cell,1997,9:583-595
Douce R,Holtz R B,Benson A A.Isolation and properties of the envelope of the spinach chloroplasts.J Biol Chem,1973,248:7215-7222
Douglas P,Pigaglio E,Ferrer A et al.Three spinach leaf nitrate reductase/3-hydroxy-3-methylglutary-CoA reductase kinases that are regulated by reversible phosphorylation and/or Ca2+ ions.Biochem J,1997,325:101-109
Ehness R,Ecker M,Roitsch T.Glucose and stress independently regulate source and sink metabolism and defense mechanism via signal transduction pathways involving protein phosphorylation.Plant Cell,1997,9:1825-1841
Entwistle G,Rees T.Enzymic capacities of amyloplast from wheat (Triticum aestuvum) endosperm. Biochem J,1988,255:291-396
Finkelstein R R,Gibson S I.ABA and sugar interactions regulating development:Cross-talk or voices in a crowd? Curr.Opin.Plant Biol.1999,5:26-32
Finnic C,Borch J,Collinge D B.14-3-3 proteins:Eukaryotic regulatory proteins with many functions. Plant Mol.Biol,1999,40:545-554
Fischer K,Arbinger B,Kammerer B,Busch C,Brink S,Wallmeier H,Sauer N,Eckerskorn C,Flugge U I.Cloning and in vivo expression of functional triose phosphate/phosphate translocarors from C3-and C4-plants:evidence for the putative participation of specific amino acid residues in the recognition of phosphoenopyruvate.Plant J,1994,5:215-226
Fischer K,Kammerer B,Gutensohn M,Arbinger,Weber A.A new class of plastidic phosphate translocators:a putative link between primary and secondary metabolism by the phosphoenolpuruvate/phosphate antiporter.Plant Cell,1997,9:453-462
Fliege R,Flugge U I,Werdan K.Specific transport of inorganic phosphate,3-phosphoglycerate and triose phosphates across the inner membrane of the envelope in spinach chloroplasts.Biochim Biophys Act,1978,502:232-247
Flugge U I,Fischer K,Gross A,Sebald W,Lottspeich F,Eckerskorn C.The triose phosphate-3-phosphoglycerate-phosphate translocator from spinach chloroplasts:nucleotide sequence of a full-length cDNA clone and import of the in vitro synthesized precursor protein into chloroplasts.The EMBO Journal,1989,8:39-46
Flugge U I,Gerber J,Heldt H W.Regulation of the reconstituted chloroplast phosphate translocator by an H+ gradient.Biochim Biophys Acta,1983,638:296-304
Flugge U I,Heldt H W.The phosphate translocators of the chloroplast envelope.Isolation of the carrier protein and reconstitution of transport.Biochim Biophys Acta,1981,638:296-304
Flugge U I.Reaction mechanism and asymmetric orintation of the reconstituted chloroplast phosphate translocator.Biochimica Biophysica Acta,1992,1110:112-118
Flugge U I,Weber A.A rapid method for measuring organelle-specific substrate transport in homogenates from plant tissues.Planta,1994,194:181-185
Fu H,Park W D.Sink and vascular associated sucrose synthase functions are encoded by different gene classes in potato.Plant Cell,1995,7:1369-1385
Fujiki Y,Ito M,Nishida I,Watanabe A.Multiple S200 The Plant Cell signaling pathways in gene expression during sugar starvation:Pharmacological analysis of din gene expression in
suspensioncultured cells of Arabidopsis.Plant Physiol,2000,124:1139-1148
Gazzarrini S,McCourt P.Genetic interactions between ABA,ethylene and sugar signaling pathways. Curr.Opin.Plant Biol,2001,4:387-391
Gibson S I.Plant sugar-response pathways:Part of a complex regulatory web.Plnat Physiol,2000, 124:1532-1539
Goddijn O,Smeekens S.Sensing trehalose biosynthesis in plants.Plant J,2001,14:143-146
Godt D E,Riegel A,Roitsch T.Regulation of sucrose synthase expression in Chenopodium rubrum: characterization of sugar induced expression in photoautotrophics suspension cultures and sink tissue specific expression in plants.J Plant Physiol,1995,146:231-236
Graham I A,Denby K J,Leaver C J.Carbon catabolite repression regulates glyoxylate cycle gene expression in cucumber.Plant cell,1994,6:761-772
Gray J C.Manipulation of phosphoribulokinase and phosphate translocator activities in transgenic tobacco plants.Journal of Experimental Botany,1995,46:1309-1315
Grierson C,Du J S,de Torres Zabala M,Beggs K,Smith C,Holdsworth M,Bevan M.Separate cis sequences and trans factors direct metabolic and developmental regulation of a potato tuber storage protein gene.Plant J,1994,5:815-826
Gross A,Rucker G,Heldt H W,Flugge U I.Comparison of the kinetic properties,inhibition and labelling of the phosphate translocators from maize and spinach mesophyll chloroplasts.Planta, 1990,180:262-271
Halford N G,Hardie D G.SNF1 related protein kinases:global regulators of carbon metabolism in plants? Plant Mol Biol,1998,37:735-748
Halford N G,Vicente-Carbajosa J,Sabelli P A,Shewry P R,Hannappel U,Kreis M.Molecular analyses of a barley multigene family homologous to the yeast protein kinase gene SNF1. Plant J,1992,2: 791-797
Halford N G,Hardie D G.SNF1-related protein kinases:Global regulators of carbon metabolism in plants? Plant Mol.Biol,1998,37:735-748
Hanson J,Johannesson H,Engstrom P.Sugar-dependent alterations in cotyledon and leaf development in transgenic plants expressing the HDZhdip gene ATHB13. Plant Molecular Biology,2001,45: 247-262
Hanson R W.Nutrient control of gene transcription.J.Biol.Chem,2000,275:307-347
Hardie D G,Carling D,Carlson M.The AMP-activated/SNF1 protein kinase subfamily:Metabolic sensors of the eukaryotic cell? Annu.Rev.Biochem,1998,67:821-855.
Haulser R E,Fisher K L,Flugge U I.Determination of low-abundant metabolites in plant extracts by NAD(P)H Fluorescence with a microtiter plater reader.Analytical Biochenistry,2000,281:1-8
Hausler R E,Schlieben N H,Nicolay P,Fischer K,Fischer K L,Flugge U I.Control of carbon partitioning and photosynthesis by the triose phosphate/phosphate translocator in transgenic tobacco plants (Nicotiana tabacum L) I.Comparative physiological analysis of tobacco plants with antisense repression and overexpression of th etriose phosphate/phosphate translocator.Planta,2000,210:
371-382
Hausler R E,Schlieben N H,Schulz B,Flugge U I.Compensation of decreased triose phosphate/phosphate translocator activity by accelerated starch turnover and glucose transport in transgenic in tobacco.Planta,1998,204:366-376
Heijine G von.Membrane Protein structure prediction.Hydrophobicity analysis and the positive-inside rule.J Mol Biol,1992,225:487-494
Heineke D,Kruse A,Flugge U I,Frommer W B,Riesmeier H W,Willmitzer L,Heldt H W.Effect of antisense repression of the chloroplast triose-phosphate translocator on photosynthetic metabolism in transgenic potato plants.Planta,1994,193:174-180
Heldt H W,Flugge U I.Plant biochemistry & Molecular Biology.Oxiford university press,2000,3-38
Heldt H W,Flugge U I.Metabolite transport in plant cells,in Plant Organelles.Tobin A K ed., University Press,1992,21-47
Heldt H W.Plant Biochemistry & Molecular Biology.Oxford university press,1997,3-38
Heldt H W.Measurement of metabolite movement across the envelope and of the pH in the stroma and the thylakoid space in intact chloroplasts.Methods Enzymol,1980,69:604-613
Herbers K,Meuwly P,Frommer W B et al.Systemic acquired resistance mediated by the ectopic expression of invertase:possible hexose sensing in the secretory pathway.Plant Cell,1996,8: 793-803
Herrero P,Martinez-Campa C,Moreno F.The hexokinase 2 protein participates in regulatory DNA-protein complexes necessary for glucose repression of the SUC2 gene in Saccharomyces cerevisiae.FEBS Lett,1998,434:71-76
Hilgarth C,Sauer N,Tanner W.Glucose increases the expression of the ATP/ADP translocator and the glyceraldehyde-3-phosphate dehydrogenase genes in Chlorella.J Biol Chem,1991,266: 24044-24047
Ho S L,Chao Y C,Tong W F,Yu S M.Sugar coordinately and differentialluy regulates growth-and stress-related Gene expression via a complex signal transduction network and multiple control mechanisms.Plant Physiol,2001,125:877-890
Hohmann S,Winderickx J,Winde JHd et al.Novel alleles of yeast hexokinase Pll with distinct effects on catalytic activity and catabolite repression of SUC2. Microbiology,1999,145:703-714
Hopp T P,Woods K R.Prediction of protein anti-genie determinants from amino acid sequences.Proc NatlAcadSci USA,1981,78:3824
Howitz K T,McCarty R E.Kinetic characteristics of the chloroplast envelope glycolate transporter. Biochemistry,1985,24:2645-2652
Ishiguro S,Nakamura K.Characterization of a cDNA encoding a novel DNA-binding protein,SPF1, that recognizes SP8 sequences in the 5' upstream regions of genes coding for sporamin and beta-amylase from sweet potato.Mol Gen Genet,1994,28:563-571
Jane J C,Leon P,Zhou L et al.Hexokinase as a sugar sensor in higher plants.Plant Cell,1997,9:5-19
Jang J C,Sheen J.Sugar sensing in higher plants.Plant Cell,1994,6:1665-1679
Jang J C,Leon P,Zhou L et al.Hexokinase as a sugar sensor in higher plants.Plant cell,1997,9:5-19
Jang J C,Sheen J.Sugar sensing in higher plants.Trends Plant Science,1997,2:208-214
Jang,J C,Leon P,Zhou L,Sheen J.Hexokinase as a sugar sensor in higher plants.Plant Cell,1997,9: 5-19
Janin J.Surface and inside volumes in globular proteins.Nature,1979,277:491
Jefferson R A,Kavanagh T A,Bevan M W.GUS fusions:β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.EMBO J,1987,6:3901-3907
Jefferson R,Goldsbrough A,Bevan M.Transcriptional regulation of a patatin-1 gene in potato.Plant Mol Biol,1990,14:995-1006
Karplus P A,Schulz G E.Prediction of chain flexbility in protein.N aturwissencharaften,1985,72:212
Klein D,Stitt M.Effects of 2-deoxyglucose on the expression of RBCS and the metabolism of Chenopodium rubrum cell cultures.Planta,1998,205:223-234
Knight J S,Grat J C.Expression of genes encoding the tobacco chloroplast phosphate translocator is not light-regulated and is repressed by sucrose.Molecular and General Genetics,1994,242:586-594
Koch K E.Carbohydrate modulated gene expression in plants.Annu Rev Plant Physiol Plant Mol Biol, 1996,47:509-540
Krapp A,Hofmann B,Schafer C et al.Regulation of the expression of rcbS and other photosynthetic genes by carbihydrates:a mechanism for the sink regulation of photosynthsis.Plant J,1993,3: 817-828
Kuhn C,Franceschi V R,SchulzA et al.Localization and turnover of sucrose transporters in enucleate sieve elements indicate macromolecular trafficking.Science,1997,275:1298-1300
Laby R J,Kincaid M S,Kim D,Gibson S I.The Arabidopsis sugar-insensitive mutants sis4 and sis5 are defective in abscisic acid synthesis and response.Plant J,2000,23:587-596
Laemmli U K.Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature,1970,222:680-685
Lalond S,Boles E,Hellmann H et al.The dual function of sugar carriers:transport and sugar sensing. Plant Cell,1999,11:707-726
Leegood R C,Sharkey T D,Caemmerer S (eds),Kluwer Academic,Dordrecht,Belgium,2000,53-83
Lemoine R,Barker L,Frommer W T et al.Identification of a pollen NtSUT3 from tobacco.FEBS Lett, 1999,454:325-330
Lemoine R,Gallet O,Gaillard C et al.Plasma membrane vesicles from source and sink leaves.Plant Physiol,1992,100:1150-1156
Lilly R M,Fitzgerakd M P,Reints K G,Walker D A.Criteria of intactness and the photosynthetic activity of spinach chloroplast preparations.New Phytol,1975,75:1-10
Liu X J,Prat S,Willmitzer L,Formmer W B.Cis regulatory elements directing tube-specific and sucrose-inducible expression of a chimeric class I patatin promoter/Gus-gus fusion.Mol Gen Genet, 1994,223:401-406
Loddcnkotter B,Kammerer B,Fischer K,Flugge U I.Expression of the functional mature chloroplast
triose phosphate translocator in yeast and purification of the histidin-tagged protein by a single step chromatography step.Proc Natl Acad Sci USA,1993,90:2155-2159
Lue M Y,Lee H.Protein phosphatase inhibitors enhance the expression of an a-amylase gene,a-Amys, in cultured rice cells.Biochem Biophs Res Commun,1994,205:807-816
Maloney P C.A consensus structure for membrane transport.Res Microbiol,1990,141:374-383
Martin T,Hellman H,Schmidt R et al.Identification of mutants in metabolically regulated gene expression.Plant J,1997,11:53-62
Menu T,Rothan C,Dai N,Petreikov M et al.Cloning and characterization of a cDNA encoding hexohinase from tomato.Plant Science,2001,160:209-218
Menu T,Rothan C,Dai N,Petreikov M,Etienne C,Destrac-Irvine A,Schaffer A,Granot D,Richard B. Cloning and characterization of a cDNA encoding hexokinase from tomato.Plant sci,2001,160: 209-218
Mita S,Hirano H,Nakamura K.Negative regulation in the expression of a sugar-inducible gene in Arabidopsis thaliana.A recessive mutation causing enhanced expression of a gene for beta-amylase. Plant Physiol,1997,114:575-582
Mita S,Murano N,Akaike M et al.Mutants of Arabidopsis thaliana with pleiotropic effects on the expression of the gene for β-amylase and on the accumulation of anthocyanin that are inducible by sugars.Plant J,1997,11:841-851
Moore B D,Sheen J.Plant sugar sensing and signaling:A complex reality.Trends Plant Sci,1999,4: 250.
Muranaka T,Banno H,Machida Y.Charaterization of tobacco protein kinase NPK5,a homolog of Saccharomyces cerevisiae SNF1 that consititutively actives expression of the glucose-repressible SUC2 gene for a secreted invertase of S.Cerevisiae.Mol Cell Biol,1994,14:2958-2965
Naohiro A,Tatsuro H,Sakiko T et al.Molecular cloning and expression analysis of a gene for a sucrose transporter in maize.Plnat Cell Physiol,1999,40:1072-1078
Nathalie N,Serge D,Remi L.The sucrose tranporter of celery identification and expression during salt stress.Plant Physiol,2000,122:1447-1456
Nemeth K,Putnoky P,Bhalerao R et al.Pleiotropic control of gllucose and hormone responese by PRL1,a nuclear WD protein,in Arabidopsis.Genes Dev,1998,12:3059-3073
Ohto M,Hayashi K,Isobe M,Nakamura K.Involvenent of Ca2+ signalling in the sugar-inducible expression of genes coding for sporamin and β-amylase of sweet potato.Plant J,1995,7:297-307
Ohto M,Onai K,Furukawa Y,Aoki E,Araki T,Nakamura K.Effects of sugar on vegetative development and floral transition in Arabidopsis.Plant Physiol,2001,127:252-261
Ohto M A,Nakamura K.Sugar-induced increase of calcium dependent protein kinase associated with the plasma membrane in leaf tissue of tobacco.Plant Physiol,1995,109:973-981
Ozcan S,Dover J,Johnston M.Glucose sensing and signaling by two glucose receptors in the yeast Saccharomyces cerevisiate.EMBO J,1998,17:2566-2573
Ozcan S,Dover J,Rosenwald A G et al.Two glucose transporters in Saccharomyces cereviside are
glucose sensors that generate a signal for induction of gene expression.Proc Natl Acad Sci USA, 1996,93:1-5
Pakers J M R,Guo D,Hodges R S.New hydrophilicity scale derived from high-performance liquid chromatography peptide retention data:correlation of predicted surface residues with antigenicity and X-ray derived accessible sites.Biochemistry,1986,25:5425
Paul M,Foyer C H.Sink regulation of photosynthesis.J Experinental Botany,2001,52:1383-1400
Paul M,Pellny T,Goddijn O.Enhancing photosynthesis with sugar signals.Trends Plant Science,2001, 6:197-200
Pego J V,Weisbeek P J,Smeekens S C M.Mannose inhibits Arabidopsis germination via a hexokinase-mediated step.Plant Physiol,1999,119:1017-1023
Production of Antipeptide Antisera.Current Protocols,1991,supplement 1,Unit 9. 4
Purcell P C,Smith A M,Haifond N G.Antisense expression of a sucrose nonfermenting related protein kinease sequence in potato results in decreased expression of sucrose syntase in tubers and lose of sucrose-inducibility of sucrose synthase transcripts in leaves.Plant J,1998,14:195-202
Qingmei Wang,Jia Chen,Xuechen Wang,Wei Sha,Jinyue Sun.Cloning and expression analysis of triose phosphate/phosphate translocator gene from wheat.Acta Botanica Sinica,2002,44(1) :67-71
Quick W P,Scheibe R,Neuhaus H E.Induction of hexosephosphate translocator activity in Spinach chloroplasts.Plant Physiol,1995,109:113-121
Randez-Gil F,Herrero P,Sanz P,Prieto J,Moreno F.Hexokinase PⅡ has a double cytosolic-nuclear localization in Saccharomyces cerevisiae.FEBS Lett,1998,425:475-478
Riesmeier J W,Hirner B,Frommer W B.Potato sucrose transporter expression in minor veins indicates a role in phloem loading.Plant cell,1993,5:1591-1598
Roitsch T,Bittner M,Godt D E.Induction of apoplastic in vertase of Chenopodium rubrum by D-glucose and a glucose analog and tissue-specific expression suggest a role in sink-regulation. Plant Physiol,1995,108:285-294
Roitsch T.Source-sink regulation by sugar and stress.Curr.Opin.Plant Biol,1999,2:198-206
Rolland F,Moore B,Sheen J.Sugar Sensing and Signaling in Plants.The Plant Cell,Supplement 2002, S185-S205
Rolland F,Winderickx J,Thevelein J M.Glucose-sensing mechanisms in eukaryotic cells.Trends in Biochemical Sciences,2001,26:310-317
Rumpo M E,Edwards G E,Yousif A E,Keegstra K.Specific labeling of the phosphate translocator in C3 and C4 mesophyll chloroplasts by tritiated dihydro-DIDS (1,2-ditritio-1,2-[2,2'-disulfo-4, 4'-diisothiocyano] diphenylethane).Plant Physiol,1988,86:1193-1198
Sambrook J,Fritsch E F,Maniatis T.Molecular cloning:A laboratory Mannual,2nd.New York:Cold Spring Harbor Laboratory Press.1989.
Sauer N,Stadler R.A sink-specific H+/monosaccharide cotransporter from Nicotiana tabacum:cloning and heterologous expression in bakers yeast.Plant J,1993,4:601-610
Sauer N,Tanner W.The hexose carrier from chorella.cDNA cloning of a eukaryotic H+-cotransporter.
FEBS Lett,1989,259:43-46
Schneider A,Hausler R E,Kolukisaoglu O,Kunze R,van der Graaff E,Schwacke R,Catoni E, Desimone M,Flugge U I.An Arabidopsis thaliana knock-out mutant of the chloroplast triose phosphate/phosphate translocator is severely compromised only when starch synthesis,but not starch mobilisation is abolished.Plant J,2002,32:685-699
Schulz B,Frommer W B,Flugge U I,Hummel S,Fischer K,Willmitzer L.Expression of the triose phospate translocators gene from potato is light dependent and restricted to green tissues.Mol Gen Genet,1993,238:357-361
Sehnarrenberger C.Planta,1990,181:249-255
Sehnke P C,DeLille J M,Ferl R J.Consummating signal transduction:The role of 14-3-3 proteins in the completion of signal-induced transitions in protein activity.Plant Cell,2002,14 (suppl.), S339-S354.
Sheen J.Metabolic repression of transcription in higher plants.Plant Cell,1990,2:1027-1038
Sheen J.Metabolic repression of transcription in higher plants.Plant Cell,1990,2:1027-1038
Sheen J.Protein phosphatase activity is required for lightinducible gene expression in maize.EMBO J, 1993,12:3497-3505
Sheen J.C4 gene expression.Annu.Rev.Plant Physiol.Plant Mol.Biol,1999,50:187-217
Sil A K,Alam S,Xin P,Ma L,Morgan M,Lebo C M,Woods M P,Hopper J E.The Gal3p-Gal80p-Gal4p transcription switch of yeast:Gal3p destabilizes the Gal80p-Gal4p complex in response to galactose and ATP.Mol.Cell.Biol,1999,19:7828-7840
Smeekens S.Sugar induced signal transduction in plants.Annu Rev Plants Physiol Plant Mol Biol,2000, 51:49-82
Sonnewald U,Lerchl J,Zrenner R,and Frommer W.Manipulation of sink-source relations in transgenic plants.Plant,cell & environment,1994,17:649-658
Stein J C,Hansen G.Mannose induces an endonuclease responsible for DNA laddering in plant cells. Plant Physiol,1999,121:71-80
Stitt M,McC R,Lilley,Gerhardt R,Heldt H W.Metabolite levels in specific cells and subcellar compartments of plant leaves.Methods in Enzymology,1989,174:518-552
Stitt M,Krapp A.The interaction between elevated carbon dioxide and nitrogen nutrition:The physiological and molecular background.Plant Cell Environ,1999,22:583-621
Stulke J,Hillen W.Carbon catabolite repression in bacteria.Curr.Opin.Microbiol,1999,2:195-201
Sugden C,Donaghy P,Halford N G et al.Two SNF1 related protein kinases from spinach leaf phosphorylate and inactivate HMG-CoA reductase,nitrate reductase and sucrose phosphate synthase in vitro.Plant Physiol,1999,120:257-274
Sugden C,Crawford R M,Halford N G,Hardie D G.Regulation of spinach SNF1-related (SnRK1) kinases by protein kinases and phosphatases is associated with phosphorylation of the T loop and is regulated by 5_-AMP.Plant J,1999,19:433-439
Sun Y(孙玉),Chen J(陈珈),Sun D-Y(孙大业).Extra cellular calmodulin stimulates the transplasma
membrane redox reaction of root protoplasts in Zea Mays.植物学报(Acta Bot Sin),1998,40: 437-441 (in Chinese with English Abstract)
Sylvie L,Eckhard B,Hanjo H et al.The dual function of sugar carriers:transport and sugar sensing. Plant Cell,1999,11:707-726
Takeda T,Toyofuku K,Matsukura C,Yamaguchi J.Sugar transporters involved in flowering and grain development of rice.J Plant Physiol,2001,158:465-470
Takedas S,Mano S,Ohto M,et al.Inhibitors of protein in phosphatase 1 and 2A block the sugar-inducible gene expression in plants.Plant Physiol,1994,106:567-574
Tang G-Q,Luscher M,Sturm A.Antisense repression of vacuolar and cell wall intertase in transgenic carrot alters early plant development and sucrose partitioning.Plant Cell 1999,11:177-189
Thomas B R,Rodriguez R L.Metabolite signals regulate gene expression and source/sink relations in cereal seedlings.Plant Physiol,1994,106:1235-1239
Tian W,Huang J,Zeng W.Transient expression of β-glucuronidase gene in protoplast of Triticum aestivum L.Genetic Manipulation in Plants,1989,5:29-35
Truernit E,Sauer N.The promoter of the Arabidopsis thaliana SUC 2 sucrose-H+ symporter gene directs expression of β-glucuronidase to the phloem:evidence for phloem loading and unloading by SUC 2. Planta,1995,196:564-570
Umemura T A,Perata P,Futsuhara Y et al.Sugar sensing and α-amylase gene repression in rice embryos.Planta,1998,204:420-428
Uyeda K.Curr Top Cell Regul,1992,33:31-46
van Oosten J J,Gerbaud A,Huijser C,Dijkwel P P,ChuaN-H,Smeekens S C M.An Arabidopsis mutant showing reduced feedback inhibition of photosynthesis.Plant J,1997,12:1011-1020
Vasil V,Redway F,Vasil I K.Regeneration of plants from embryogenic suspension culture protoplasts of wheat (Triticum aestivum L.).Bio/technology,1986,8:429-434
Waddell I D,Zomerschoe A G,Voice M W et al.Cloning and expression of hepatic microsomal glucose transport protein.Biochem J,1992,286:173-177
Walker D A,Baldry C W,Cockburn W.Photosynthesis by isolated chloroplasts,simultaneous measurement of carbon assimilation and oxygen evolution.Plant Physiol,1968,43:1419-1422
Wang Q-M(王庆梅),Chen J(陈珈),Wang X-C(王学臣),Sha W(沙伟),Sun J-Y(孙金月).Cloning and expression analysis of triose phosphate/phosphate translocator gene from wheat.Ada Bot Sin (植物学报),2002,44:67-71 (In English with Chinese Abstract)
Wang Q-M (王庆梅),Yang S-D(杨树德),Chen J(陈珈).Triose phosphate translocaotr in the inner membrance of chloroplast.Chinese Bulletin of Botany(植物学通报),2001,18:11-15 (In Chinese with English Abstract)
Welling G W,Weijer W J,Van der zee R et al.Prediction of sequential antigenic regions in proteins. FEBS Lett,1985,188:215
Wiese A,Groner F,Sonnewald U,Deppner H,Lerchl J,Hebbeker U,Flugge U I,Weber A.Spinach hexokinase I is located in the outer envelope membrance of plastids.FEBS Letters,1999,461:
13-18
Willey D L,Fischer K,Wachter E.Molecular cloning and structural analysis of the phosphate translocator from pea chloroplasts and its comparison to the spinach phosphate translocator.Planta, 1991,183:451-461
Wu Z-Y(吴忠义),Chen J(陈珈),Zhu M-J(朱美君).Identification of the photoaffinity-labelled abscisic acid binding proteis from Maize microsome.生物物理与生物化学学报(Acta Biochim Biophys Sin),1996,28:694-697 (in Chinese with English Abstract)
Yokoyama R,Hirose T,Fujii N et al.The rolC promoter of Agrobacterium rhizogenes Ri-plasmid is activated by sucrose in transgenic plants.Mol Gen Genet,1994,224:15-22
Yu He-Fen,Chen Jia,Wang Xue-Chen.Characterization of cytosolic Vl-ATPase in Pea root.Acta botanica sinica,2003,45:164-168
Yu S-M:Cellular and genetic responses of plants to sugar starvation.Plant Physiol,1999,121:687-693
Zaghmout O M F,Trolinder N L.Factors affecting transient gene expression in protoplasts isolated from very slowly growing embryogenic callus cultures of wheat (Triticum aestivum L.).Theor Appl Genet,1993,86:721-730
Zenke F T,Engles R,Vollenbroich V,Meyer J,Hollenberg C P,Breunig K D.Activation of Gal4p by galactosedependent interaction of galactokinase and Ga180p.Science,1996,272:1662-1665
Zhou A-F,Xia G-M,Chen X-L,Chen H-M.Production of somatic hybrid plants between two types of wheat protoplasts and the protoplasts of Haynaldia villosa.Acta Botanica Sinica,2002,44: 1004-1008
Zhou H,Stiff C M,Konzak C F.Stably transformed callus of wheat by electroporation-induced direct gene transfer.Plant Cell Reports,1993,12:612-616
Zhou L,Jang J,Sheen J.Glucose insensitive (gin) mutants define downstream pathways for sugar signaling in Arabidopsis development.1996,Seventh International Conference on Arabidopsis Research,Norwich,UK.Abstract S94
Zou J,Qi Q,Katavic V,Marillia E F,Taylor D C.Effects of antisense repression of an Arabidopsis thaliana pyruvate dehydrogenase kinase cDNA on plant development.Plant Molecular Biology, 1999,41:837-849
Zubay G L.Metabolism of carbohydrate.In:Biochemistry 4th edition,Wm.C.Brown Publishers,1998, 265-438