摘要
模式植物研究发现蔗糖非发酵-1-型相关蛋白激酶-1在植物碳氮代谢过程中有关键性开关的作用,为探讨SnRK1(蔗糖非发酵-1-型相关蛋白激酶-1)在果树碳氮代谢中的分子生物学功能,以水培平邑甜茶幼苗为试材,克隆蔗糖非发酵-1-型相关蛋白激酶-1的α和βγ亚基的编码基因,同时采用实时荧光定量PCR研究α和βγ亚基的编码基因表达量的变化,并构建α亚基的编码基因MhSnRK1的正、反以表达载体,成功转化番茄,初步研究了其生理生化功能,主要结果如下:
1.根据已知苹果SnRK1的α亚基编码基因的EST序列,设计特异引物利用RT-PCR结合RACE技术获得了蔗糖非发酵-1-型相关蛋白激酶-1的α亚基的全长编码基因,命名为MhSnRK1,GenBank注册号为EF690362;并利用相同技术扩增到平邑甜茶SnRK1的βγ亚基编码基因的片段,命名为MhAKINβγ。序列分析表明,MhSnRK1基因全长2063 bp,包含一个166 bp的5′端非编码区,1548 bp的编码区共编码515个氨基酸和348 bp的3′端非编码区,预测分子量为58.7 KD。同时MhSnRK1的氨基酸序列分析显示该基因与拟南芥、黄瓜、番茄和烟草相比有82%-88%的同源性;系统进化树分析显示,MhSnRK1基因与黄瓜α亚基编码基因(SnRK1)基因的同源关系最近。βγ亚基的片段(MhAKINβγ)与已知蔗糖非发酵-1-型相关蛋白激酶-1 AKINβγ亚基编码基因具有较高同源性,故进一步确定其为平邑甜茶蔗糖非发酵-1-型相关蛋白激酶-1的AKINβγ亚基编码基因片段。
2.荧光定量PCR分析α和βγ亚基的编码基因在不同组织的表达发现,MhSnRK1和MhAKINβγ基因在平邑甜茶的根、茎、叶中均有表达,且均在叶中的表达量最大。本文对这两个亚基的编码基因在不同环境条件下在不同组织中的表达特性的研究结果显示:硝态氮处理抑制了MhSnRK1基因在叶、根中的表达;在PEG处理下,MhSnRK1的表达水平在根中先上升(24小时),但48小时后又恢复到初始状态;叶片中,MhSnRK1的表达初期下降而48小时后又上升到初始态;在硝态氮和PEG处理下,MhAKINβγ表现出与MhSnRK1相似的表达特性。外施硝态氮,根中MhAKINβγ转录水平24小时内降低3倍,48小时后叶中也较低;在PEG处理下,MhAKINβγ的表达水平在根中先上升;叶片中,MhAKINβγ的表达初期下降10倍而48小时后又上升到初始态。
3.为进一步研究平邑甜茶SnRK1的生理生化功能,将蔗糖非发酵-1-型相关蛋白激酶-1的α亚基的全长编码基因,构建了pBI121- MhSnRK1正、反义表达载体,并对番茄进行农杆菌侵染的遗传转化。对转基因番茄果实内含物进行测定后初步发现:与对照相比,正义转基因番茄果实中可溶性总糖和淀粉含量最大升高分别30%和56%;而正义转基因番茄果实中有机酸和可溶性蛋白含量最大分别下降88%和76%。转基因植株果实发育期比野生型提早10天;果实直径在发育早期明显大于野生型。本研究测定了转基因株系和野生型叶片中淀粉含量的日变化,白天结束时,转基因株系叶片淀粉含量明显高于野生型,与野生型相比,T2-5株系中淀粉含量升高105%。转反义基因的植株与野生型相比,无论在果实内含物、果实发育期或叶片淀粉含量都没有明显变化。
SnRK1 complex plays a key role in carbon and nitrogen metabolisms of modle plants. Under water culture condition, we have cloned two subunits of SnRK1 complex from pingyitiancha seedlings. The aim of this study was to obtain a better insight into the character of the SnRK1 complex subunits encoding genes in respond to environmental signals and in carbon metabolisms in fruit trees. The expression patterns of MhSnRK1 and MhAKINβγin different tissues at different conditions were analyzed by quantitative real-time PCR. The MhSnRK1 gene derived from pingyitiancha was heterologously expressed in tomato plants to study the biological function of the SnRK1.
1. In an attempt to isolate MhSnRK1 gene, two EST sequences were used to design GSPs for 5′-RACE and 3′-RACE isolation of MhSnRK1; one EST sequence was 433-bp (CN445648) and the other was 466-bp (CX024405). Analysis revealed a 2063 bp sequence obtained by RACE was the full-length cDNA of the MhSnRK1 (Malus hupehensis Rehd. SnRK1, EF690362) gene. The nucleotide sequence of MhSnRK1 contains a 166-bp 5'-untranslated region, a 1548-bp open reading frame encoding a protein of 515 amino acids with a mass of 58.7 kDa, and a 348-bp 3'-untranslated region. The deduced amino acid sequence of MhSnRK1 showed 82.34, 87.57, 86.02, 84.08 and 82.14% identity to the Arabidopsis AKIN10, tomato LeSNF1, potato StSNF1, tobacco NPK5 and Pea, but it exhibited only 66.67 and 66.22% identity to the Barley BKIN12 and Rye RKIN1. The phylogenetic tree showed MhSnRK1 belongs to the type 1 (SnRK1) subfamily of the snf1-related kinases (exemplified by Arabidopsis AKIN10) rather than the SnRK2 subfamily (exemplified by Arabidopsis ASK1) or the SnRK3 subfamily (exemplified by Wheat WPK4). We also produce a partialβγ-subunit (MhAKINβγ) with 3′-RACE. After a 100-fold dilution of the first PCR product, the product was used for a second PCR with the primer of SN32, from which a 3′-RACE 1112-bp fragment was obtained. Analysis of this fragment showed its nucleotide sequence was highly conserved with those in other plant species (including fragments of apple, maize and Medicago truncatula) Therefore, this fragment was considered a pingyitiancha AKINβγhomologue and was designated as MhAKINβγ.
2. Expression of MhSnRK1 and MhAKINβγgenes was studied in leaves, stems and roots of two-week-old pingyitiancha seedlings by quantitative real-time PCR analysis using subunit-specific probes. An r18S gene was used as an internal standard. The results revealed that transcripts of MhSnRK1 and MhAKINβγwere detected in all tissues analysed, but the two genes expression differed among root, stem and leaf tissues. Higher levels of MhSnRK1 transcripts were expressed in leaf tissue relative to the root and stem. Similar expression patterns were observed from MhAKINβγ, with the highest transcriptional level also occurring in leaf tissue. To understand the short-term effects of nitrogen and osmotic stress on expression of MhSnRK1 or MhAKINβγ, transcriptional levels in different plant tissues were monitored by quantitative real-time PCR analysis during induction by NO3- and PEG water stress. Control plants treated with KCL or grown in normal nutrients exhibited nearly consistent levels of MhSnRK1 or MhAKINβγfrom 0 to 48 h. The NO_3~- does not affect MhSnRK1 expression as early in leaf, yet it inhibits MhSnRK1 expression after 48 h. Meanwhile, NO3- feeding reduces MhSnRK1 expression 5-fold in root within 24 h, but the transcript levels raise to control levels after 48 h. Under PEG water stress, MhSnRK1 mRNA levels increased in roots within 24 h but then declined to the levels of control after 48h. In leaves, MhSnRK1 expression decreased as early, and then increased after 48 h. Interestingly, MhAKINβγexhibits a similar expression pattern as MhSnRK1 during the NO_3~- and PEG treatment. When KNO_3 was resupplied, pMhAKINβγtranscripts decrease 3-fold in root within 24 h, and low in leaf after 48 h compared with the control. A 13-fold induction of pMhAKINβγexpression was detected after PEG treatment for 24 h in root, and then the transcript levels declined to the levels of control after 48h. MhAKINβγtranscripts in leaf decreased 10-fold within 24h compared with the control .
3. In order to determine the function of MhSnRK1 in growth and development of plants cell, a construct containing full-length cDNA of MhSnRK1 gene in sense and atisense orientation driven by the constitutivecauliflower mosaic virus 35S promoter was assembled and introduced into tomato. The quality of the transgenic tomato fruits was also analysis. Compared with the wild-type, the soluble sugar and starch levels of the transgenic plant fruit were increased by approximately 20%-30% and 44%-56%, respectively. Organic acid and total soluble protein were 44%-88% and 55%-76% of the control concentrations, respectively. The results indicate MhSnRK1 has obvious effects on the quality of the tomato. Our research also showed that the fruit diameter of the transgenic plants was much thicker at the early stages of fruit development than the wild type, but were similar at later stages; the transgenic fruit ripened ten days earlier than wild type fruit.The starch content of the leaf from transformants was assayed. we found that the starch content in transgenic lines was higher than wide type in leaf at the end of the day, for example, the starch content in T2-5 transgenic line was increased by 105% compared with that in the wide-type.
引文
Alderson A, Sabelli PA, Dickinson JR, Cole D, Richardson M, Kreis M, Shewry PR, Halford NG. Complementation of snf1, a mutation affecting global regulation of carbon metabolism in yeast, by a plant protein kinase cDNA. PNAS 1991; 88:8602-8605.
A. Lovas et al., Functional diversity of potato SNF1-related kinases tested in Saccharomyces cerevisiae, Gene (2003), 321: pp. 123–129.
Baena-Gonzalez E, Rolland F, Johan M, Sheen TJ. A central integrator of transcription networks in plant stress and energy signaling. Nature 2007; 448:938-943.
Baker NG and Baker I. Starch in angiosperm pollen grains and its evolutionary significance. Am. J. Bot 1979; 66:591-600.
B.E. Kemp, Bateman domains and adenosine derivatives form a binding contract, J. Clin. Invest. (2004),113: pp. 182–184.
Bhalerao, R.P., Salchert, K., Bako, L., Okresz, L., Szabados, L., Muranaka, T., Machida, Y., Schell, J., and Koncz, C. Regulatory interaction of PRL1 WD protein with Arabidopsis SNF1-like protein kinases. Proc. Natl. Acad. Sci. USA 1999. 96: 5322–5327.
Boudsocq, M., Barbier-Brygoo, H., and Lauriere, C. Identification of nine sucrose nonfermenting 1-related protein kinases 2 activated by hyperosmotic and saline stresses in Arabidopsis thaliana. J. Biol. Chem. 2004. 279: 41758–41766.
Bouly J, Gissot L, Lessard P, Kreis M, Thomas M. Arabidopsis thaliana proteinsrelated to the yeast SIP and SNF4 interact with AKINα1, an SNF1-like protein kinase. Plant J 1999; 18:541–550.
Bradford KJ, Downie AB, Gee OH, Alvarado V, Yang H, Dahal P. Abscisic acid and gibberellin differentially regulate expression of genes of the SNF1-related kinase complex in tomato seeds. Plant Physiol 2003; 132: 1560-1576.
Carlson, M. Regulation of sugar utilization in Saccharomyces species. J.Bacteriol. 1987. 169: 4873–4877.
Celenza, J.L., and Carlson, M. Structure and expression of the SNF1 gene of Saccharomyces cerevisiae. Mol. Cell. Biol. 1984. 4: 54–60.
Celenza JL, Carlson M. A yeast gene that is essential for release from glucose repression encodes a protein kinase. Science 1986; 233:1175-1180.
Clement C, Chavant L, Burrus M, Audran JC. Anther starch variations in Lilium during pollen development. Sex. Plant Reprod 1994; 7:347-356.
Celenza JL, Eng FJ, Carlson M. Molecular analysis of the SNF4 gene of Saccharomyces cerevisiae: evidence for the physical association of the SNF4 protein with the SNF1 protein kinases. Mol Cell Biol 1989; 9:5045-5054.
Coruzzi, G.M., and Zhou, L. Carbon and nitrogen sensing and signaling in plants: Emerging‘matrix effects.’Curr. Opin. Plant Biol. 2001. 4: 247–253.
D.G. Hardie and K. Sakamoto, AMPK: a key sensor of fuel and energy status in skeletal muscle, Physiology (Bethesda) (2006), 21:pp. 48–60.
D. Kerk et al., A chloroplast-localized dual-specificity protein phosphatase in Arabidopsis contains a phylogenetically dispersed and ancient carbohydrate-binding domain, which binds the polysaccharide starch, Plant J. (2006),46: pp. 400–413.
D. Stapleton et al., Mammalian AMP-activated protein kinase subfamily, J. Biol. Chem. (1996), 271: pp. 611–614.
E.M. Hrabak et al., The Arabidopsis CDPK-SnRK superfamily of protein kinases, Plant Physiol. (2003), 132: pp. 666–680.
E.N. Yoshida et al., Sequence and phylogenetic analysis of the SNF4/AMPKγsubunit gene from Drosophila melanogaster, Genome (1999), 42: pp. 1077–1087.
E.R. Hudson et al., A novel domain in AMP-activated protein kinase causes glycogen storage bodies similar to those seen in hereditary cardiac arrhythmias, Curr. Biol. (2003),13: pp. 861–866.
Estruch, F., Treitel, M.A., Yang, X., and Carlson, M. N-terminal mutations modulate yeast SNF1 protein kinase function. Genetics 1992. 132: 639–650.
F. Rolland et al., Sugar sensing and signalling in plants: conserved and novel mechanisms, Annu. Rev. Plant Biol. (2006), 57: pp. 675–709
Gao Junfeng. Experimental Technique of Plant Physiology. Beijing: Word Book Press. 2000. (in Chinese) 高俊凤.植物生理学实验技术. 2000.北京:世界图书出版社
Franchi GG, Bellani L, Nepi M, Pacini E. Types of carbohydrate reserves in pollen: localization, systematic distribution and ecophysiological significance. Flora 1996; 191:143-159.
Gibson SI. Plant sugar-response pathways: part of a complex regulatory web. Plant Physiol 2000; 124:1532–1539.
Gissot L, Polge C, Bouly JP, Lemaitre T, Kreis M, Thomas M. AKINbeta3, a plant specific SnRK1 protein, is lacking domains present in yeast and mammals non-catalytic beta-subunits. Plant Mol Biol 2004; 56:747-759.
G. Polekhina et al., AMPKβsubunit targets metabolic stress sensing to glycogen, Curr. Biol. 1(2003), 3: pp. 867–871.
Halford NG, Bouly J-P, Thomas M. SNF1-related protein kinases (SnRKs): regulators at the heart of the control of carbon metabolism and partitioning. Adv Bot Res 2000;32 :405-434.
Halford NG, Hardie DG. SNF1-related protein kinases: global regulators of carbon metabolism in plants? Plant Mol Biol 1998; 37:735-748.
Halford NG, Hey S, Jhurreea D, Laurie S, McKibbin RS, Paul M, Zhang Y. Metabolic signalling and carbon partitioning: role for Snf1-related (SnRK1) protein kinase, J Exp Bot 2003; 54:467–475.
Halford NG, Hey S, Jhurreea D, Laurie S, McKibbin RS, Zhang Y, Paul MJ. Highly conserved protein kinases involved in the regulation of carbon and amino acid metabolism. J Exp Bot 2004; 394:35-42
Halford NG, Paul MJ. Carbon metabolite sensing and signaling. Plant Biotechnol J 2003; 1:381–398.
Hannappel U, Vicente-Carbajosa J, Barker JHA, Shewry PR, Halford NG. Differential expression of two barley SNF1-related protein kinase genes. Plant Mol Biol 1995;271235-1240.
Hao L, Wang H, Sunter G, Bisaro DM. Geminivirus AL2 and L2 proteins interact with and inactivate SNF1 kinase. Plant Cell 2003; 15:1034-1048 .
Hardie DG. The AMP-activated protein kinase pathway: new players upsyream and downstream. J Cell Sci 2004; 117:5479-5487.
Hey SJ, Powers SJ, Beale M, Hawkins ND, Ward J and Halford NG. Enhanced seed phytosterol accumulation through expression of a modified HMG-CoA reductase. Plant Biotechnol J 2006; 4:219-229.
Hoekma A, hirsch PR, Hooykass PJJ, Schilperoort R. A binary plant vector strategy based on separation of the vri- and T-region of Agrobacterium tumefaciens Ti plasmid. Nature 1983; 303:179-180.
Ho, S., Chao, Y., Tong, W., and Yu, S. Sugar coordinately and differentially regulates growth- and stress- related gene expression via a complex signal transduction network and multiple control mechanisms. Plant Physiol. 2001. 125: 877–890.
Hong, S.P., Leiper, F.C., Woods, A., Carling, D., and Carlson, M. Activation of yeast Snf1 and mammalian AMP-activated protein kinase by upstream kinases. Proc. Natl. Acad. Sci. USA 2003. 100: 8839–8843.
Hong, S.P., Momcilovic, M., and Carlson, M. Function of mammalian LKB1 and Ca~(2+)/calmodulin-dependent protein kinase kinase alpha as Snf1-activating kinases in yeast. J. Biol. Chem. 2005. 280: 21804–21809.
Jiang R, Carlson M. Glucose regulates protein interactions within the yeast SNF1 protein kinase complex. Genes & Development 1996; 10 :3105-3115.
Jiang R, Carlson M. The SNF1 protein kinase and its activating subunit, SNF4, interact with distinct domains of the SIP1/SIP2/GAL83 component in the kinase complex. Mol Cell Biol 1997; 17:2099-2106.
J.W. Scott et al., CBS domains form energy-sensing modules whose binding of adenosine ligands is disrupted by disease mutations, J. Clin. Invest. (2004), 113: pp. 274–284.
K.I. Mitchelhill et al., Mammalian AMP-activated protein kinase shares structural and functional homology with the catalytic domain of yeast Snf1 protein kinase, J. Biol. Chem. (1994), 269: pp. 2361–2364.
Kleinow T, Bhalerao R, Breuer F, Umeda M, Salchert K, Koncz C. Functional identification of an Arabidopsis snf4 ortholog by screening for heterologous multicopy suppressors of snf4 deficiency in yeast. Plant J 2000; 23:115–122.
Koch, K.E. Carbohydrate-modulated gene expression in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 1996. 47: 509–540.
Kuchin, S., Treich, I., and Carlson, M. A regulatory shortcut between the Snf1 protein kinase and RNA polymerase II holoenzyme. Proc. Natl. Acad. Sci. USA 2000. 97: 7916–7920.
Lakatos L, Klein M, Hofgen R, Banfalvi Z. Potato StubSNF1 interacts with StubGAL83: a plant protein kinase complex with yeast and mammalian counterparts. Plant J 1999; 17:569-574.
Laurie S, Mckibbin RS, Halford NG. Antisense SNF-related (SnRK1) protein kinase gene represses transient activity of an a-amylase (a-Amy2) gene promoter in cultured wheat embryos. J Exp Bot 2003; 54 :739-747.
L. Gissot et al., AKINβ3, a plant specific SnRK1 protein, is lacking domains present in yeast and mammals non-catalyticβsubunits, Plant Mol. Biol. (2004), 56: pp. 747–759.
Lin Y,Hwang C F,Brown J, Cheng C,L.5' proximal regions of Arabidopsis nitrate reductase genes direct nitrateinduced transcription in transgenic tobacco.Plant Physiol,1994,106:477–484
Linkohr B L,Williamson L C,Fitter A,H ,Leyser H M.Nitrate and phosphate availability and distribution have different effects on root system architecture of Arabidopsis.Plant J,2002,29:751-760
Lohrmann J,Harter K.Plant two-component signaling systems and the role of response regulators.Plant Physiol,2002,128:363–369
Lovas, A., Sos-Hegedus, A., Bimbo, A., and Banfalvi, Z. Functional diversity of potato SNF1-related kinases tested in Saccharomyces cerevisiae. Gene 2003. 321: 123–129.
Lu CA, Lin CC, Lee KW, Chen JL, Huang LF, Ho SL, Liu HJ, Hsing YI, Yu SM. The SnRK1A protein kinase plays a key role in sugar signaling during germination and seedling growth of rice. Plant Cell 2007;19(8) :2484–2499.
Mckibbin RS, Muttucumaru N, Paul MJ, Powers SJ, Burrell MM, Coates S, Purcell PC, Tiessen A, Geigenberger P, Halford NG. Production of high-starch, low-glucose potatoes through over-expression of the metabolic regulator SnRK1. Plant Biotechnol J 2006; 4:409-418.
Man AL, Purcell PC, Hannappel U, Halford NG. Potato SNF1-related protein kinase: molecular cloning, expression analysis and peptide kinase activity measurements. Plant Mol Biol 1997; 34:31–43.
McCormac AC, Cherry JR, Hershey HP, Vierstra RD, Smith H. Photoresponses of transgenic tobacco plants expressing an oat phytochrome gene. Planta 1991; 185:162–170.
Muranaka T, Banno H, Machida Y. Characterization of the tobacco protein kinase, NPK5, a homologue of Saccharomyces cerevisiae SNF1 that constitutively activates expression of the glucose–repressible SUC2 gene for a secreted invertase of S. cerevisiae. Mol Cell Biol 1994; 14:2958–2965.
Murashig T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissues. Plant Physiol 1962; 15 :473–497.
Neininger A,Back E,Bichler A,Schneiderbauer H et al.Deletion analysisof a nitrite-reductase promoter from spinach in transgenic tobacco.Planta,1994,194:186–192
N.G. Halford et al., SNF1-related protein kinases (SnRKs)– regulators at the heart of the control of carbon metabolism and partitioning. In: M. Kreis and J.C. Walker, Editors, Advances in Botanical Research, Academic Press (2000), pp. 405–434
O. Vincent and M. Carlson, Gal83 mediates the interaction of the Snf1 kinase complex with the transcription activator Sip4, EMBO J. (1999), 18: pp. 6672–6681.
Palmer S J,Berridge D M,McDonald et al.Control of leaf expansion in sunflower (Helianthus annuus L.) by nitrogen nutrition.Jour of Exp Bot,1996,47 (296):359-368
Peng J, Peng F, Zhu C, Wei S. Molecular cloning of a putative gene encoding isopentenyltransferase from pingyitiancha (Malus hupenensis) and characterization of its response to nitrate. Tree Physiol 2008; 28:899-904.
Perata, P., Matsukura, C., Vernieri, P., and Yamaguchi, J. Sugar repression of gibberellin-dependent signalling pathway in barley embryos. Plant Cell 1997. 9: 2197–2208.
Polge C, Thomas M. SNF1/AMPK/SnRK1 kinases, global regulators at the heart of energy control? Trends Plant Sci 2007; 12 :20–28.
Purcell PC, Smith AM, Halford NG. Antisense expression of a sucrose non-fermenting-1-related protein kinase sequence in potato results in decreased expression of sucrose synthase in tubers and loss of sucrose-inducibility of sucrose synthase transcripts in leaves. Plant J 1998; 14:195–202.
Radchuk R, Rodchuk V, Weschke W, Borisjur L, Weber H. Repressing the expression of the sucrose nonfermenting-1-related protein kinase gene in pea embryo causes pleiotropic defects of maturation similar to an abscisic acid-insensitive phenotype. Plant Physiol 2006; 140:263-278.
Robinson D . The responses of plants to non-uniform supplies of nutrients.New Phytol,1994,127:635-674
Rolland F, Moore B, Sheen J. Sugar sensing and signaling in plants. Plant Cell 2002; 14: S185–S205.
Schwachtje J, Minchin PEH, Jahnke S, van Dongen JT, Schittko U, Baldwin IT. SNF1-related kinases allow plants to tolerate herbivory by allocating carbon to roots. PNAS 2006; 103:12935-1294.
Sheen, J., Zhou, L., and Jang, J.C. Sugars as signaling molecules. Curr. Opin. Plant Biol. 1999. 2: 410–418.
Shen, W., and Hanley-Bowdoin, L. Geminivirus infection up-regulates the expression of two Arabidopsis protein kinases related to yeast SNF1- and mammalian AMPK-activating kinases. Plant Physiol. 2006.142: 1642–1655.
Slocombe SP, Laurie S, Bertini L, Beaudoin F, Dickinson JR, and Halford NG. Molecular cloning of Snlp1, a novel protein that interacts with SNF1-related protein kinase (SnRK1). Plant Mol Biol 2002; 49:31-44.
Smeekens S. Sugar-induced signal transduction in plants. Annual Review of Plant Physiology and Plant Molecular Biology 2000; 51:49–81.
S.P. Davies et al., Purification of the AMP-activated protein kinase on ATP-gamma-sepharose and analysis of its subunit structure, Eur. J. Biochem. (1994),223: pp. 351–357.
S.P. Hong et al., Activation of yeast Snf1 and mammalian AMP-activated protein kinase by upstream kinases, Proc. Natl. Acad. Sci. U. S. A. (2003),100: pp. 8839–8843.
S.P. Hong et al., Function of mammalian LKB1 and Ca~(2+)/calmodulin-dependent protein kinase kinaseαas Snf1-activating kinases in yeast, J. Biol. Chem. (2005),280: pp. 21804–21809.
S.S. Lin et al., Sip2, an N-myristoylated beta subunit of Snf1 kinase, regulates aging in Saccharomyces cerevisiae by affecting cellular histone kinase activity, recombination at rDNA loci, and silencing, J. Biol. Chem.(2003), 278: pp. 13390–13397.
Sugden C, Donaghy PG, Halford NG, Hardie DG. Two SNF1-related protein kinases from spinach leaf phosphorylate and inactivate 3-hydroxy-3-methylglutaryl-coenzyme A reductase, nitrate reductase, and sucrose phosphate synthase in vitro. Plant Physiol 1999; 120:257–274.
Sugiharto B,Sugiyama T.Effects of nitrate and ammonium on gene expression of phosphoenolpyruvate carboxylase and nitrogen metabolism in maize leaf tissue during recovery from nitrogen stress.Plant Physiol,1992,98:1403–1408
Takano M, Kajiya-Kanegae H, Funatsuki H, Kikuchis. Rice has two distinct classes of protein kinase genes related to SNF1 of Saccharomyces cerevisiae, which are differently regulated in early seed development. Mol Gen Genet 1998; 260 :388-394.
Tan Dong-mei. The physiology and biochemistry of programmed Malus siversii and M. hupehensis cell death under drought stress. Scientia Agricultura Sinica 2007; 40:980-986.
Thelander M, Olsson T, Ronne H. Snf1-related protein kinase 1 is needed for growth in a normal day–night light cycle. EMBO J 2004; 23:1900-1910.
T.J. Iseli et al., AMP-activated protein kinaseβsubunit tethersαandγsubunits via its C-terminal sequence (186-270), J. Biol. Chem. (2005), 280: pp. 13395–13400.
T. Muranaka et al., Characterization of tobacco protein kinase NPK5, a homolog of Saccharomyces cerevisiae SNF1 that constitutively activates expression of the glucose-repressible SUC2 gene for a secreted invertase of S. cerevisiae, Mol. Cell. Biol. (1994), 14: pp. 2958–2965.
Umemura, T., Perata, P., Futsuhara, Y., and Yamaguchi, J. Sugar sensing and alpha-amylase gene repression in rice embryos. Planta 1998. 204: 420–428.
Vos J,Putten P E L, Van der.Effects of partial shading of the potato plant on photosynthesis of treated leaves,leaf area expansion and allocation of nitrogen and dry matter in component plant parts.Euro Jour of Agronomy: the Journal of the European Society for Agronomy,2001,14 (3): 209-220
Wang F, Sanz A, Brenner ML, Smith A. Sucrose synthase, starch accumulation, and tomato fruit sink strength. Plant Physiol 1993; 101 :321-327.
Wang Guan-lin, Fang Hong-jun. Plant gene engineering. Beijing: Science Press. 2002. (in China) 王关林,方宏筠.植物基因工程. 2002.北京:科学出版社.
Wilson, W.A., Hawley, S.A., and Hardie, D.G. Glucose repression/derepression in budding yeast: SNF1 protein kinase is activated by phosphorylation under derepressing conditions, and this correlates with a high AMP:ATP ratio. Curr. Biol. 1996. 6: 1426–1434.
Woods A, Cheung PC, Smith FC, Davison MD, Scott J, BeriRK, Carling D. Characterization of AMP-activated protein kinase beta and gamma subunits. Assembly of the heterotrimeric complex in vitro. J Biol Chem 1996; 271:10282-10290.
Xie zhujie, Jiang dong, Dai tingbo, Cao weixing. Sugar siganl and its regulation on C/N metabolism gene in plant. Journal of plant physiology 2002; 38: 399-405. 谢祝捷,姜东,戴廷波,曹卫星.植物中的糖信号及其对碳氮代谢基因的调控.植物生理学通讯, 2002. (4): 399-405.
Yang Hongqiang, Jie Yuling. Studies of Individual Difference in Seedlings of Apple Rootstock. Journal of Shandong Agricultural University. 1997.28(4) :487-491. 杨洪强,接玉玲.苹果砧木实生苗根系个体差异研究.山东农业大学学报,1997.28(4) :487-491.
Yu, S.M., Kuo, Y.H., Sheu, G., Sheu, Y.J., and Liu, L.F. Metabolic derepressionof alpha-amylase gene expression in suspension-cultured cells of rice. J. Biol. Chem. 1991. 266: 21131–21137.
Yu, S.M. Cellular and genetic responses of plants to sugar starvation. Plant Physiol. 1999. 121: 687–693.
Yu, S.M., Lee, Y.C., Fang, S.C., Chan, M.T., Hwa, S.F., and Liu, L.F. Sugars act as signal molecules and osmotica to regulate the expression of alpha-amylase genes and metabolic activities in germinating cereal grains. Plant Mol. Biol. 1996. 30: 1277–1289.
Zhang Y, Shewry PR, Jones H, Barcelo, P, Lazzeri PA, Halford NG. Expression of antisense SnRK1 protein kinase sequence causes abnormal pollen development and male sterility in transgenic barley. Plant J 2001; 28:431–441.
Zhao Chunmei, Wang Li, Yi Shuying and Liu Jian. The Construction and the Chilling-Resistance Ability of Endoplasmic Reticulum Small Heat Shock Protein (ER-sHSP) Transgenic Tomato Plants. Acta Horticulture Sinica, 2006. (5): 989-994. (in Chinese) 赵春梅,王丽,伊淑莹,刘箭.番茄转ER-sHSP基因植株构建及抗冷性研究.园艺学报,2006. (5): 989-994.
Zhao shijie, Shi guoan, Dong xinchun. Experimental Technique of Plant Physiology. Science and technology of china agriculture press. 2002. 84-85. 赵世杰,史国安,董新纯.植物生理学实验指导.中国农业科学技术出版社, 2002, 84-85.