水稻中天冬氨酸转氨酶的分子生物学研究和转基因应用
|
摘要
水稻不仅是我国主要的粮食作物,还养活着世界一半的人口。在水稻氮代谢途径中,有一些重要的酶(例如GS、GOGAT、GDH、AAT和AS),调控着水稻氮素的代谢。有研究发现,尤其是编码天冬氨酸转氨酶的AAT基因,与控制蛋白质和氨基酸含量的QTL有对应的关系。本研究在水稻品种中花11(来自与中国农业科学院作物科学研究所的商业品种)中超量表达了AAT基因(包括来源于水稻的3个AAT基因:OsAAT1、OsAAT2和OsAAT3;来源于大肠杆菌的1个AAT基因:EcAAT),同时抑制表达AAT这个基因。得到转基因植株后,分析了转基因植株的生长和代谢水平与对照植株的差异,研究了这两个基因的表达情况和作用机理。综合分析2004年和2007年数据,得出以下几个主要结果:
1、得到了超量表达AAT的转基因植株(如上所述,共试图超量表达4个片段)。
2、t测验(t-test)表明,超量表达AAT基因的转基因植株,不管在正常生长条件下,还是各种胁迫生长条件下(包括低氮胁迫,NaCl胁迫,ABA胁迫),其生长(包括株高和干重)都与对照没有显著差异。
3、t测验(t-test)表明,超量表达AAT基因(包括OsAAT1,OsAAT2和EcAAT基因)的转基因植株的米粉中,氨基酸含量和蛋白质含量明显高于野生型对照。
4、超量表达OsAAT1基因的阳性植株的种子中的氨基酸总含量比其对应的阴性植株的含量提高了16.1%,蛋白质含量提高了22.2%;超量表达OsAAT2基因的阳性植株的种子中的氨基酸总含量比其对应的阴性植株的含量提高了12.0%,蛋白质含量提高了21.1%;超量表达EcAAT基因的阳性植株的种子中的氨基酸总含量比其对应的阴性植株的含量提高了5.4%,蛋白质含量提高了11.1%。t测验(t-test)表明,超量表达AAT基因的转基因阳性植株和其对应的阴性植株的米粉中,氨基酸含量和蛋白质含量在阳性植株中显著(P≤0.05)高于其对应的阴性植株。说明氨基酸含量和蛋白质含量的升高是由转基因作用引起的。
5、验证AAT基因家族的表达情况,发现AAT基因家族的3个成员的表达受各种胁迫的影响。同时这3个成员在植株的不同部位和不同的生长时期,表达也存在着差异。
6、超量表达EcAAT基因,能使植株体内OsAAT1和OsAAT2基因的表达上升。但是水稻体内的3个OsAAT基因的表达并不能相互调控,即超量表达任何一个OsAAT基因,并不能引起其它两个OsAAT基因表达的改变。
Aspartate aminotransferase(AAT) is a key enzyme in the synthesis of amino acids.It plays an important role in regulation of carbon and nitrogen metabolism in almost all organisms.In this study,we over-expressed in rice separately all three AAT genes from rice(OsAAT1～3) and one AAT gene from Escherichia coli(EcAAT). Over-expression was driven by the CaMV 35S promoter and transformed into rice by Agrobacterium tumefaciens-mediated transformation.Compared with control plants, the genetically engineered transformant plants showed significantly increased leaf AAT activity and seed amino acid and protein contents despite the lack of observed phenotypical changes.The total leaf AAT activities in OsAAT1,OsAAT2 and EcAAT over-expressed plants were 26.6,23.6 and 19.6 A/mg/min,which were significantly higher than that in the wild type control(17.7 A/mg/min).The amino acid content in seeds of transgenic plants over-expressing OsAAT1,OsAAT2,and EcAAT was 119.36 mg/g,102.84 mg/g and 113.72 mg/g,respectively,which were 16.1%,12.0%and 5.4%higher,respectively,than that in the control plants.The transgenic plants over-expressing OsAAT1,OsAAT2,and EcAAT had significantly higher protein contents(increased 22.2%,21.1%,and 11.1%,respectively) than wild type plants.No significant changes were found in leaf AAT activity,seed amino acid content or protein content in OsAAT3 over-expressed plants.The expression patterns of the three OsAAT genes and their different functions are also discussed.
We do some RT-PCR and real-time PCR to check the expression of AAT genes. As result shows that OsAAT1 and OsAAT2's expression level are higher than OsAAT3, and when over-expressing EcAAT gene in transfer rice,it would result in higher expression of OsAAT1 and OsAAT2,but OsAAT3 expression level is lower.Except OsAAT1 gene has higher expression level in OsAAT2 transgenic rice,over-expressing one of OsAAT genes,it couldn't induce other OsAAT genes changed their expression.
1、得到了超量表达AAT的转基因植株(如上所述,共试图超量表达4个片段)。
2、t测验(t-test)表明,超量表达AAT基因的转基因植株,不管在正常生长条件下,还是各种胁迫生长条件下(包括低氮胁迫,NaCl胁迫,ABA胁迫),其生长(包括株高和干重)都与对照没有显著差异。
3、t测验(t-test)表明,超量表达AAT基因(包括OsAAT1,OsAAT2和EcAAT基因)的转基因植株的米粉中,氨基酸含量和蛋白质含量明显高于野生型对照。
4、超量表达OsAAT1基因的阳性植株的种子中的氨基酸总含量比其对应的阴性植株的含量提高了16.1%,蛋白质含量提高了22.2%;超量表达OsAAT2基因的阳性植株的种子中的氨基酸总含量比其对应的阴性植株的含量提高了12.0%,蛋白质含量提高了21.1%;超量表达EcAAT基因的阳性植株的种子中的氨基酸总含量比其对应的阴性植株的含量提高了5.4%,蛋白质含量提高了11.1%。t测验(t-test)表明,超量表达AAT基因的转基因阳性植株和其对应的阴性植株的米粉中,氨基酸含量和蛋白质含量在阳性植株中显著(P≤0.05)高于其对应的阴性植株。说明氨基酸含量和蛋白质含量的升高是由转基因作用引起的。
5、验证AAT基因家族的表达情况,发现AAT基因家族的3个成员的表达受各种胁迫的影响。同时这3个成员在植株的不同部位和不同的生长时期,表达也存在着差异。
6、超量表达EcAAT基因,能使植株体内OsAAT1和OsAAT2基因的表达上升。但是水稻体内的3个OsAAT基因的表达并不能相互调控,即超量表达任何一个OsAAT基因,并不能引起其它两个OsAAT基因表达的改变。
Aspartate aminotransferase(AAT) is a key enzyme in the synthesis of amino acids.It plays an important role in regulation of carbon and nitrogen metabolism in almost all organisms.In this study,we over-expressed in rice separately all three AAT genes from rice(OsAAT1～3) and one AAT gene from Escherichia coli(EcAAT). Over-expression was driven by the CaMV 35S promoter and transformed into rice by Agrobacterium tumefaciens-mediated transformation.Compared with control plants, the genetically engineered transformant plants showed significantly increased leaf AAT activity and seed amino acid and protein contents despite the lack of observed phenotypical changes.The total leaf AAT activities in OsAAT1,OsAAT2 and EcAAT over-expressed plants were 26.6,23.6 and 19.6 A/mg/min,which were significantly higher than that in the wild type control(17.7 A/mg/min).The amino acid content in seeds of transgenic plants over-expressing OsAAT1,OsAAT2,and EcAAT was 119.36 mg/g,102.84 mg/g and 113.72 mg/g,respectively,which were 16.1%,12.0%and 5.4%higher,respectively,than that in the control plants.The transgenic plants over-expressing OsAAT1,OsAAT2,and EcAAT had significantly higher protein contents(increased 22.2%,21.1%,and 11.1%,respectively) than wild type plants.No significant changes were found in leaf AAT activity,seed amino acid content or protein content in OsAAT3 over-expressed plants.The expression patterns of the three OsAAT genes and their different functions are also discussed.
We do some RT-PCR and real-time PCR to check the expression of AAT genes. As result shows that OsAAT1 and OsAAT2's expression level are higher than OsAAT3, and when over-expressing EcAAT gene in transfer rice,it would result in higher expression of OsAAT1 and OsAAT2,but OsAAT3 expression level is lower.Except OsAAT1 gene has higher expression level in OsAAT2 transgenic rice,over-expressing one of OsAAT genes,it couldn't induce other OsAAT genes changed their expression.
引文
1.郭建平,高素华,刘玲.气象条件对作物品质和产量影响的试验研究.中国东北样带(NECT)与全球变化-干旱、人类活动与生态系统.北京:气象出版社,2002:223-230
2.胡红红.水稻逆境相关转录因子的分离和功能鉴定[博士学位论文]武汉:华中农业大学图书馆,2006
3.李常健,林清华,张楚富.高等植物中氨同化酶及其同功酶研究工作.零陵师范高等专科学报,2000,21:20-22
4.李林,沙国栋,陆景淮.水稻灌浆期光温因子对稻米品质的影响.中国农业气象,1989,10:33-38
5.莫惠栋.我国稻米品质的遗传.中国农业科学,1993,26:8-14
6.潘瑞炽,董愚得.植物生理学.北京:高等教育出版社,1995
7.石岗.生物活性肽研究进展.北京农业科学,2002,20:6-9
8.沈其荣.土壤肥料通论.高等教育出版社,2001
9.唐微.Bt抗虫水稻的培育[博士学位论文]武汉:华中农业大学图书馆,2006
10.王光火,张奇春,黄昌勇.提高氮肥利用率,控制氮肥污染的新途径-SSNM.浙江大学学报,2003,29:67-70
11.王文高,陈正行.低过敏大米研究进展.粮食与油脂,2001,5:32-33
12.王章存,申瑞玲,姚惠源.大米蛋白研究进展.中国粮油学报,2004,2:1-15
13.吴昌银.水稻T-DNA插入突变体库及其Enhancer trap系的创建[博士学位论文]武汉:华中农业大学图书馆,2003
14.吴平,印莉萍,胡彬.植物氮素营养分子生理.科学出版社,2001,1-98
15.肖本泽.抗旱候选基因和启动子的水稻遗传转化分析和田间抗旱性鉴定[博士学位论文]武汉:华中农业大学图书馆,2007
16.印莉萍,刘祥林,洪剑明,邱泽生,林忠平.非固氮植物初级氮同化相关的基因及其表达调控.科学出版社,2001,271-287
17.张德颐,朱治平.植物分子生物学与生物工程.北京:科学出版社,1991,46-54
18.张云桥,吴荣生,蒋宁,刘桂华.水稻的氮素利用率与品种类型的关系.植物生理通迅,1989,2:45-47
19.钟明.水稻精米蛋白质和氨基酸含量QTL定位及遗传基础研究[硕士学位论文]武汉:华中农业大学图书馆,2007
20.Bryan JK.Synthesis of the aspartate family and branched-chain amino acids.In:Sturnpf PK and Conn EE(eds) The biochemistry of plants:a comprehensive treatise.Academic Press,1980,New York,pp 403-452
21.Cao J,Zhao JZ,Tang D,Shelton M,Earle D.Broccoli plants with pyramided cry1Ac and cry1C Bt genes control diamondback moths resistant to Cry1A and Cry1 C proteins.Theor Appl Genet,2002,105:258-264
22.Chan MT,Chang HH,Ho SL,Tong WF,Yu SM.Agrobacterium-mediated production of transgenic rice plants expressing a chimeric α-amylase promoter/β-glucuronidase gene.Plant Mol Biol,1993,22:491-506
23.Chan YL,Lin KH,Sanjaya,Liao LJ,Chen WH,Chan MT.Gene stacking in Phalaenopsis orchid enhances dual tolerance to pathogen attack.Transgenic Res,2005,14:279-288
24.Chavan JK,Duggal SK.Studies on the essential amino acid composition,protein fractions and biological value(BV) of some varieties of rice.J Sci Food Agric,1978,29:225-229
25.Chen H,Tang W,Xu C,Li X,Lin Y,Zhang Q.Transgenic indica rice plants harboring a synthetic cry2A~* gene of Bacillus thuringiensis exhibit enhanced resistance against lepidopteran rice pests.Theor Appl Genet,2005,111:1330-1337
26.Chuang CF,Meyerowitz EM.Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana.Proc Natl Acad Sci,USA,2000,97:4985-4990
27.Chu ZH,Peng KM,Zhang LD,Zhou B,Wei J,Wang SP.Construction and characterization of a normalized whole-life-cycle cDNA library of rice.Chin Sci Bull,2003,48:229-235
28.Coruzzi GM,Last R.Amino acids.In:Buchanan BB,Gruissem W,Jones RL (eds) Biochemistry and Molecular Biology of Plants.American society of plant physiologists roekville,2000,Maryland,pp 358-410.
29.Coruzzi GM.Primary N-assimilation into amino acids in Arabidopsis.In: Somerville CR, Meyerowitz EM, Rockville, MD (eds) The Arabidopsis book. American society of plant biologists, 2003, Maryland New York, pp 1-17
30. Crawley MJ, Brown SL, Hails RS, Kohn DD, Rees M. Transgenic crops in natural habitats. Nature, 2001,409: 682-683
31. Daniell H, Muthukumar B, Lee SB. Marker free transgenic plants: engineering the chloroplast genome without the use of antibiotic selection. Curr Genet, 2001,39:109-116
32. Datta S K, Obcemea W M, and Jana R K. Protein content of rice grain as affected by nitrogen fertilizer and some triazines and substituted ureas. Agron J, 1972, 64: 785-788
33. Ellstrand NC. When transgenes wander, should we worry? Plant Physiol, 2001, 125: 1543-1545
34. Falco SC, Guida T, Locke M, Mauvais J, Sanders C, Ward RT, Webber P. Transgenic canola and soybean seeds with increased lysine. Bio/Technology, 1995,13:577-582
35. Ferrario MS, Suzuki A, Kunz C, Valadier MH, Roux Y, Hirel B, Foyer CH. Modulation of amino acid metabolism in transformed tobacco plants deficient in Fd-GOGAT. Plant Soil, 2000,221: 67-79
36. Ferre J, Van Rie J. Biochemistry and genetics of insect resistance to Bacillus thuringiensis. Annu Rev Entomol, 2002, 47: 501-533
37. Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 1998,391:806-811
38. Gebhardt J S, Wadsworth G J, Matthews B F. Characterization of a single soybean cDNA encoding cytosolic and glyoxysomal isozymes of aspartate aminotransferase. Plant Mol Biol, 1998,37: 99-108.
39. Gelfand DH, Rudo N. Mapping of the aspartate and aromatic amino acid aminotransferase genes tyrB and aspC. J Bacteriol, 1977,133: 441-444
40. Givan CV. Aminotransferases in higher plants. In: Stumpf PK, Conn EE (eds) The biochemistry of plants. Academic Press, 1980, New York, pp 329-357
41. Gomez K A, and De Datta S K. Influence of environment on protein content of rice. Agron. J, 1975, 67: 565-568
42. Gordon SA, Fleck A, Bell J. Optimal conditions for the estimation of ammonium by the Berthelot reaction.Ann Clin Biochem,1978,15:270-275
43.Gould J,Devey M,Hasegawa O,Ulian EC,Peterson G,Smith RH.Transformation of Zea mays L.using Agrobacterium tumefaciens and the shoot apex.Plant Physiol,1991,95:426-434
44.Greco R,Ouwerkerk PB,Sallaud C,Kohli A,Colombo L,Puigdomenech P,Guiderdoni E,Christou P,Hoge JH,Pereira A.Transposon insertional mutagenesis in rice.Plant Physiol,2001,125:1175-1177
45.Gregerson R G,Miller S S,Petrowski M,Gantt J S,Vance C P.Genomic structure,expression and evolution of the alfalfa aspartate aminotransferase genes.Plant Mol Biol,1994,25,387-99
46.Haldrup A,Petersen SG,Okkels FT.The xylose isomerase gene from Thermoanaerobacterium thermosulfurogenes allows effective selection of transgenic plant cells using D-xylose as the selection agent.Plant Mol Biol,1998,37:287-296
47.Hamilton AJ,Baulcombe DC.A species of small antisense RNA in posttranscdptional gene silencing in plants.Science,1999,286:950-952
48.Hatch MD,Osmond CB.Activity,location,and role of aspartate aminotransferase isoenzymes in leaves with C4 pathway photosynthesis.Arch Biochem Biophys,1973,156:195-206
49.Heber U,Heldt HW.The chloroplast envelope:structure,function and role in leaf metabolism.Annu Rev Plant Physiol,1981,32:139-168
50.Hiei Y,Ohta S,Komari T,Kumashiro T.Efficient transformation office(Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA.Plant J,1994,6:271-282
51.Hirochlka H.Contribution of the Tos17 retrotransposon to rice functional genomics.Curr Opin Plant Biol,2001,4:118-122
52.Holmstrom K,Mantyla E,Welin B,Mandal A,Palva E.Drought tolerance in tobacco.Nature,1996,379:683-684
53.Huang J,Rozelle S,Pray C,Wang Q.Plant biotechnology in China.Science,2002,295:674-676
54.IRRI.Annual Report for 1976.Rice Res.Los Banos,Laguna,Philippines,1977
55.Jeon JS,Lee S,Jung KH,Jun SH,Jeong DH,Lee J,Kim C,Jang S,Yang K,Nam J,An K,Han MJ,Sung RJ,Choi HS,Yu JH,Choi JH,Cho SY,Cha SS,Kim SI,An G T-DNA insertional mutagenesis for functional genomics in rice.Plant J, 2000,22:561-570
56.Joersbo M.Advances in the selection of transgenic plants using non-antibiotic marker genes.Physiol Plant,2001,111:269-272
57.Jorgensen R.Altered gene expression in plants due to trans interactions between homologous genes.Trends Biotechnol,1990,8:340-344
58.Juliano B O.Rice chemistry and technology,2nd edn.American Association of Cereal Chemists,Incorporated Saint Paul,Minnesota,USA,1985
59.Karcicio M,Izbirak A.Isozyme variations in some Aegilops L.and Triticum L.species collected from Central Anatolia.Turk J Bot,2003,27:433-440
60.Krupp H K,Abilay W P,and Alvarez E I.Some water stress effects on rice.in:Rice Breeding.Rice Res.Los Banos,Laguna,Philippines.1972,663-675
61.Lam H M,Coschigano K T,Oliveira I C,Melo-Oliveira R,Coruzzi G M.The molecular-genetics of nitrogen assimilation into amino acids in higher plants.Annu.Rev.Plant Physiol.Plant Mol Bio,1996,47:569-593
62.Lin YJ,Zhang QF.Optimising the tissue culture conditions for high efficiency transformation of indica rice.Plant Cell Rep,2005,23:540-547
63.Liu Q,Kasuga M,Sakuma Y,Abe H,Miura S,Yamaguchi-Shinozaki K,Shinozaki K.Two transcription factors,DREB1 and DREB2,with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought- and low-temperature-responsive gene expression,respectively,in Arabidopsis.Plant Cell,1998,10:1391-1406
64.Lohaus G,B(u|¨)ker M,Huβmann M,Soave C,Heldt H W.Transport of amino acids with special emphasis on the synthesis and transport of asparagine in the Illinois Low Protein and Illinois High Protein strains of maize.Planta,1998,205:181-188
65.Lohaus G,Moellers C.Phloem transport of amino acids in two Brassica napus L.genotypes and one B.carinata genotype in relation to their seed protein content.Planta,2000,211:833-840
66.Marcin M,Andrzej P.Genetic control of aspartate aminotransferase isoenzymes in Aegilops and Triticum species.J Appl Genet,2004,45:411-417
67.Martins M L,Mourato M P,de Varennes e Mendonca AP.Characterization of aspartate aminotransferase isoenzymes from leaves of Lupinus albus L.cv Estoril.J Biochem Mol Biol,2002,35:220-7
68.Matzke M,Matzke AJ,Kooter JM.RNA:guiding gene silencing.Science,2001,293:1080-1083
69.Mett VL,Podivinsky E,Tennant AM,Lochhead LP,Jones WT,Reynolds PHS.A system for tissue-specific copper controllable gene expression in transgenic plants:nodule-specific antisense of aspartate aminotransferase-P2.Transgenic Res,1996,5:105-113.
70.Meyer P.Transcriptional transgene silencing and chromatin components.Plant Mol Biol,2000,43:221-234
71.Miesak B H,Coruzzi G M.Molecular and physiological analysis of Arabidopsis mutants defective in cytosolic or chloroplastic aspartate aminotransferase.Plant Physiol,2002,129:650-60
72.Morino K,Olsen OA,Shimamoto K.Silencing of the alettrone-specific Ltp2-gus gene in transgenic rice is reversed by transgene rearrangements and loss of aberrant transcripts.Plant Cell Physiol,2004,45:1500-1508
73.Murooka Y,Mori Y,Hayashi M.Variation of the amino acid content of Arabidopsis seeds by expressing soybean aspartate aminotransferase gene.J Biosci Bioeng,2002,94:225-230
74.Napoli C,Lemieux C,Jorgensen R.Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans.Plant Cell,1990,2:279-289
75.Rawstone S,Michin FR,Summerfield RJ,Cookson C,Coombs J.Carbon and nitrogen metabolism in legume root nodules.Phytochemistry,1980,19:341-355
76.Resurrection A P,Hara T,Juliano B O,and Yoshida,S.Effect of temperature during ripening on grain quality of rice.Soil Sci Plant Nutr,1977,23:109-112
77.Romero C,Belles J,Vaya J,Serrano R,Culianez-Macia F.Expression of the yeast trehalose-6-phosphate synthasegene in transgenic tobacco plants:pleitropic phenotypes include drought tolerance.Planta,1997,201:293-297
78.Rudman D,Meister A.Transamination in Escherichia coli.J Biol,1953,200:591-604
79.Ryan E,Fottrell PF.Subcellular localization of enzymes involved in the assimilation of ammonia by soybean root nodules.Plant Mol Biol,1974,19:2647-2652
80.Sadowski I,Ma J,Triezenberg S,Ptashne M.GAL4-VP16 is an unusually potent transcriptional activator.Nature,1988,335:563-564
81.Sambrook J,Russell DW.Molecular cloning:a laboratory manual.(3rd edn). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2001, pp 1.31-1.119
82. Schwechheimer C, Smith C, Bevan MW. The activities of acidic and glutamine-rich transcriptional activation domains in plant cells: design of modular transcription factors for high-level expression. Plant Mol Biol, 1998, 36: 195-204
83. Sentoku N, Taniguchi M, Sugiyama T, Ishimaru K, Ohsugi R, Takaiwa F, Toki S. Analysis of the transgenic tobacco plants expressing Panicum miliaceum aspartate aminotransferase genes. Plant Cell Rep, 2000,19: 598-603
84. Setoyama C, Ding S, Choudhury BK, Joh T, Takeshima H, Suzuki T, Shimada K. Regulatory regions of the mitochondrial and cytosolic isoenzymes genes participating in the malate-aspartate shuttle. J Biol Chem, 1990,265: 1293-1299
85. Shaul O, Galili G. Threonine overproduction in transgenic tobacco plants expressing a mutant desensitized aspartate kinase of Escherichia coli. Plant Physiol, 1992,100:1157-1163
86. Silvente S, Camas A, Lara M. Molecular cloning of the cDNA encoding aspartate aminotransferase from bean root nodules and determination of its role in nodule nitrogen metabolism. J Exp Bot, 2003, 54: 1545-51
87. Simon-sarkadi L, Kocsy G, Varhegyi A, Galiba G, De ronde J A. Stress-induced changes in the free amino acid composition in transgenic soybean plants having increased Proline content. Biologia Plantarum, 2006, 50: 793-796
88. Song J, Yamamoto K, Shomura A, Yano M, Minobe Y, Sasaki T. Characterization and mapping of cDNA encoding aspartate aminotransferase in rice, Oryza sativa L. DNA Res, 1996, 3: 303-310
89. Sotelo A, Sousa V, Montalvo I, Hernandez M, Hernandez A L. Chemical composition of different fractions of 12 Mexican varieties of rice obtained during milling. Cereal Chem, 1990, 67: 209-212
90. Sharp PA. RNA interference-2001. Genes Dev, 2001, 15: 485-490
91. Shen Q, Zhang P, Ho TH. Modular nature of abscisic acid (ABA) response complexes: composite promoter units that are necessary and sufficient for ABA induction of gene expression in barley. Plant Cell, 1996, 8: 1107-1119
92. Springer PS. Gene traps: tools for plant development and genomics. Plant Cell, 2000,12: 1007-1020
93. Stam M, Mol JN, Kooter JM. The silence of genes in transgenic plants. Ann Bot, 1997,79:3-12
94.Stempak JM,Sohn K J,Chiang EP,Shane B,Kim YI.Cell and stage of transformation-specific effects of folate deficiency on methionine cycle intermediates and DNA methylation in an in vitro model.Carcinogenesis,2005,26:981-990
95.Takahashi M,Uematsu Y,Kashiwaba K,Yagasaki K,Hajika M,Matsunaga R,Komatsu K,Ishimoto M.Accumulation of high levels of free amino acids in soybean seeds through integration of mutations conferring seed protein deficiency.Planta,2003,217:577-586
96.Taniguchi M,Sawaki H,Sasakawa H,Hase T,Sugiyama T.Cloning and sequence analysis of cDNA encding aspartate aminotrandferase isozymes from Panicum miliaceum L.,a C4 plant.Eur j Biochem,1992,204:611-620
97.Tian YC,Zheng JB,Yu HM,Liang HY,Li CQ,Wang JM.Studies of transgenic hybrid poplar 741 carrying two insect-resistant genes.Acta Bot Sinica,2000,42:263 -268
98.Tu J,Zhang G,Datta K,Xu C,He Y,Zhang Q,Khush GS,Datta SK.Field performance of transgenic elite commercial hybrid rice expressing bacillus thuringiensis Δ-endotoxin.Nat Biotechnol,2000,18:1101-1104
99.Udvardi M K,Kahn M L.Isolation and analysis of a eDNA clone that encodes an alfalfa(Medicago sativa) aspartate aminotransferase.Mol Gen Genet,1991,231:97-105
100.Vain P,Afolabi AS,Wodand B,Shape JW.Transgene behaviour in populations of rice plants transformed using a new dual binary vector system:pGreen/pSoup.Theor Appl Genet,2003,107:210-217
101.van der Salm T,Bosch D,Honee C,Feng L,Munsterman E,Bakker P,Stiekema W J,Visser B.Insect resistance of transgenic plants that express modified Bacillus thuringiensis cryⅠA(b) and cryⅠC genes:a resistance management strategy.Plant Mol Biol,1994,26:51-59
102.Wang W M,Zhao Q,Yu J J,Zhu D Y,Ao G M.Transfer of high Lysine gene sb401 into rice and analysis for protein and amino acid content in transgenic rice seeds.Acta Agronomica Sinica,2005,31:603-607
103.Wadsworth G J,Marmaras S M,Matthews B F.Isolation and characterization of soybean cDNA clone encoding the plastid form of aspartate aminotransferase.Plant Mol Biol,1993,21:993-1009
104.Walbot V.Saturation mutagenesis using maize transposons.Curr Opin Plant Biol,2000,3:103-107
105.Wang WM,Zhao Q,Yu JJ,Zhu DY,Ao GM.Transfer of high lysine gene sb401into rice and analysis for protein and amino acid content in transgenic rice seeds.Acta Agronomica Sinica,2005,31:603-607
106.Wang X L,Larkins B A.Genetic analysis of amino acid accumulation in opaque-2 maize endosperm,Plant Physiol,2001,125:1766-1777
107.Waterhouse PM,Graham MW,Wang MB.Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA.Proc Natl Acad Sci,USA,1998,95:13959-13964
108.Wesley SV,Helliwell CA,Smith NA,Wang MB,Rouse DT,Liu Q,Gooding PS,Singh SP,Abbott D,Stoutjesdijk PA,Robinson SP,Gleave AP,Green AG,Waterhouse PM.Construct design for efficient,effective and high-throughput gene silencing in plants.Plant J,2001,27:581-590
109.Wilkie SE,Roper JM,Smith AG,Warren MJ.Isolation,characterisation and expression of a cDNA clone encoding plastid aspartate aminotransferase from Arabidopsis thaliana.Plant Mol Biol,1995,27:1227-1233
110.Winefield C S,Farnden K J,Reynold P H,Marshall C J.Evolutionary analysis of aspartate aminotransferases.J Mol Evol,1995,40:455-63
111.Wu C,Li X,Yuan W,Chen G,Kilian A,Li J,Xu C,Zhou DX,Wang S,Zhang Q.Development of enhancer trap lines for functional analysis of the rice genome.Plant J,2003,35:418-427
112.Xiong L,Yang Y.Disease resistance and abiotic stress tolerance in rice are inversely modulated by an abscisic acid-inducible mitogen-activated protein kinase.Plant Cell,2003,15:745-759
113.Yamaya T,Obara M,Nakajima H,Sasaki S,Hayakawa T,Sato T.Genetic manipulation and quantitative-trait loci mapping for nitrogen recycling in rice.J Exp Bot,2002,53:917-92
114.Yoon DB,Kang KH,Kim HJ,Ju HG,Kwon SJ,Suh JP,Jeong OY,Ahn SN.Mapping quantitative trait loci for yield components and morphological traits in an advanced backcross population between Oryza grandiglumis and the O.sativa japonica cultivar Hwaseongbyeo.Theor Appl Genet,2006,112:1052-1062
115.Zhang HX,Blumwald E.Transgenic salt-tolerant tomato plants accumulate salt in foliage but not in fruit.Nat Biotechnol,2001,19:765-768
116.Zhang HX,Hodson JN,Williams JP,Blumwald E.Engineering salt-tolerant Brassica plants:characterization of yield and seed oil quality in transgenic plants with increased vacuolar sodium accumulation.Proc Natl Acad Sci,2001,98:12832-12836
2.胡红红.水稻逆境相关转录因子的分离和功能鉴定[博士学位论文]武汉:华中农业大学图书馆,2006
3.李常健,林清华,张楚富.高等植物中氨同化酶及其同功酶研究工作.零陵师范高等专科学报,2000,21:20-22
4.李林,沙国栋,陆景淮.水稻灌浆期光温因子对稻米品质的影响.中国农业气象,1989,10:33-38
5.莫惠栋.我国稻米品质的遗传.中国农业科学,1993,26:8-14
6.潘瑞炽,董愚得.植物生理学.北京:高等教育出版社,1995
7.石岗.生物活性肽研究进展.北京农业科学,2002,20:6-9
8.沈其荣.土壤肥料通论.高等教育出版社,2001
9.唐微.Bt抗虫水稻的培育[博士学位论文]武汉:华中农业大学图书馆,2006
10.王光火,张奇春,黄昌勇.提高氮肥利用率,控制氮肥污染的新途径-SSNM.浙江大学学报,2003,29:67-70
11.王文高,陈正行.低过敏大米研究进展.粮食与油脂,2001,5:32-33
12.王章存,申瑞玲,姚惠源.大米蛋白研究进展.中国粮油学报,2004,2:1-15
13.吴昌银.水稻T-DNA插入突变体库及其Enhancer trap系的创建[博士学位论文]武汉:华中农业大学图书馆,2003
14.吴平,印莉萍,胡彬.植物氮素营养分子生理.科学出版社,2001,1-98
15.肖本泽.抗旱候选基因和启动子的水稻遗传转化分析和田间抗旱性鉴定[博士学位论文]武汉:华中农业大学图书馆,2007
16.印莉萍,刘祥林,洪剑明,邱泽生,林忠平.非固氮植物初级氮同化相关的基因及其表达调控.科学出版社,2001,271-287
17.张德颐,朱治平.植物分子生物学与生物工程.北京:科学出版社,1991,46-54
18.张云桥,吴荣生,蒋宁,刘桂华.水稻的氮素利用率与品种类型的关系.植物生理通迅,1989,2:45-47
19.钟明.水稻精米蛋白质和氨基酸含量QTL定位及遗传基础研究[硕士学位论文]武汉:华中农业大学图书馆,2007
20.Bryan JK.Synthesis of the aspartate family and branched-chain amino acids.In:Sturnpf PK and Conn EE(eds) The biochemistry of plants:a comprehensive treatise.Academic Press,1980,New York,pp 403-452
21.Cao J,Zhao JZ,Tang D,Shelton M,Earle D.Broccoli plants with pyramided cry1Ac and cry1C Bt genes control diamondback moths resistant to Cry1A and Cry1 C proteins.Theor Appl Genet,2002,105:258-264
22.Chan MT,Chang HH,Ho SL,Tong WF,Yu SM.Agrobacterium-mediated production of transgenic rice plants expressing a chimeric α-amylase promoter/β-glucuronidase gene.Plant Mol Biol,1993,22:491-506
23.Chan YL,Lin KH,Sanjaya,Liao LJ,Chen WH,Chan MT.Gene stacking in Phalaenopsis orchid enhances dual tolerance to pathogen attack.Transgenic Res,2005,14:279-288
24.Chavan JK,Duggal SK.Studies on the essential amino acid composition,protein fractions and biological value(BV) of some varieties of rice.J Sci Food Agric,1978,29:225-229
25.Chen H,Tang W,Xu C,Li X,Lin Y,Zhang Q.Transgenic indica rice plants harboring a synthetic cry2A~* gene of Bacillus thuringiensis exhibit enhanced resistance against lepidopteran rice pests.Theor Appl Genet,2005,111:1330-1337
26.Chuang CF,Meyerowitz EM.Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana.Proc Natl Acad Sci,USA,2000,97:4985-4990
27.Chu ZH,Peng KM,Zhang LD,Zhou B,Wei J,Wang SP.Construction and characterization of a normalized whole-life-cycle cDNA library of rice.Chin Sci Bull,2003,48:229-235
28.Coruzzi GM,Last R.Amino acids.In:Buchanan BB,Gruissem W,Jones RL (eds) Biochemistry and Molecular Biology of Plants.American society of plant physiologists roekville,2000,Maryland,pp 358-410.
29.Coruzzi GM.Primary N-assimilation into amino acids in Arabidopsis.In: Somerville CR, Meyerowitz EM, Rockville, MD (eds) The Arabidopsis book. American society of plant biologists, 2003, Maryland New York, pp 1-17
30. Crawley MJ, Brown SL, Hails RS, Kohn DD, Rees M. Transgenic crops in natural habitats. Nature, 2001,409: 682-683
31. Daniell H, Muthukumar B, Lee SB. Marker free transgenic plants: engineering the chloroplast genome without the use of antibiotic selection. Curr Genet, 2001,39:109-116
32. Datta S K, Obcemea W M, and Jana R K. Protein content of rice grain as affected by nitrogen fertilizer and some triazines and substituted ureas. Agron J, 1972, 64: 785-788
33. Ellstrand NC. When transgenes wander, should we worry? Plant Physiol, 2001, 125: 1543-1545
34. Falco SC, Guida T, Locke M, Mauvais J, Sanders C, Ward RT, Webber P. Transgenic canola and soybean seeds with increased lysine. Bio/Technology, 1995,13:577-582
35. Ferrario MS, Suzuki A, Kunz C, Valadier MH, Roux Y, Hirel B, Foyer CH. Modulation of amino acid metabolism in transformed tobacco plants deficient in Fd-GOGAT. Plant Soil, 2000,221: 67-79
36. Ferre J, Van Rie J. Biochemistry and genetics of insect resistance to Bacillus thuringiensis. Annu Rev Entomol, 2002, 47: 501-533
37. Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 1998,391:806-811
38. Gebhardt J S, Wadsworth G J, Matthews B F. Characterization of a single soybean cDNA encoding cytosolic and glyoxysomal isozymes of aspartate aminotransferase. Plant Mol Biol, 1998,37: 99-108.
39. Gelfand DH, Rudo N. Mapping of the aspartate and aromatic amino acid aminotransferase genes tyrB and aspC. J Bacteriol, 1977,133: 441-444
40. Givan CV. Aminotransferases in higher plants. In: Stumpf PK, Conn EE (eds) The biochemistry of plants. Academic Press, 1980, New York, pp 329-357
41. Gomez K A, and De Datta S K. Influence of environment on protein content of rice. Agron. J, 1975, 67: 565-568
42. Gordon SA, Fleck A, Bell J. Optimal conditions for the estimation of ammonium by the Berthelot reaction.Ann Clin Biochem,1978,15:270-275
43.Gould J,Devey M,Hasegawa O,Ulian EC,Peterson G,Smith RH.Transformation of Zea mays L.using Agrobacterium tumefaciens and the shoot apex.Plant Physiol,1991,95:426-434
44.Greco R,Ouwerkerk PB,Sallaud C,Kohli A,Colombo L,Puigdomenech P,Guiderdoni E,Christou P,Hoge JH,Pereira A.Transposon insertional mutagenesis in rice.Plant Physiol,2001,125:1175-1177
45.Gregerson R G,Miller S S,Petrowski M,Gantt J S,Vance C P.Genomic structure,expression and evolution of the alfalfa aspartate aminotransferase genes.Plant Mol Biol,1994,25,387-99
46.Haldrup A,Petersen SG,Okkels FT.The xylose isomerase gene from Thermoanaerobacterium thermosulfurogenes allows effective selection of transgenic plant cells using D-xylose as the selection agent.Plant Mol Biol,1998,37:287-296
47.Hamilton AJ,Baulcombe DC.A species of small antisense RNA in posttranscdptional gene silencing in plants.Science,1999,286:950-952
48.Hatch MD,Osmond CB.Activity,location,and role of aspartate aminotransferase isoenzymes in leaves with C4 pathway photosynthesis.Arch Biochem Biophys,1973,156:195-206
49.Heber U,Heldt HW.The chloroplast envelope:structure,function and role in leaf metabolism.Annu Rev Plant Physiol,1981,32:139-168
50.Hiei Y,Ohta S,Komari T,Kumashiro T.Efficient transformation office(Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA.Plant J,1994,6:271-282
51.Hirochlka H.Contribution of the Tos17 retrotransposon to rice functional genomics.Curr Opin Plant Biol,2001,4:118-122
52.Holmstrom K,Mantyla E,Welin B,Mandal A,Palva E.Drought tolerance in tobacco.Nature,1996,379:683-684
53.Huang J,Rozelle S,Pray C,Wang Q.Plant biotechnology in China.Science,2002,295:674-676
54.IRRI.Annual Report for 1976.Rice Res.Los Banos,Laguna,Philippines,1977
55.Jeon JS,Lee S,Jung KH,Jun SH,Jeong DH,Lee J,Kim C,Jang S,Yang K,Nam J,An K,Han MJ,Sung RJ,Choi HS,Yu JH,Choi JH,Cho SY,Cha SS,Kim SI,An G T-DNA insertional mutagenesis for functional genomics in rice.Plant J, 2000,22:561-570
56.Joersbo M.Advances in the selection of transgenic plants using non-antibiotic marker genes.Physiol Plant,2001,111:269-272
57.Jorgensen R.Altered gene expression in plants due to trans interactions between homologous genes.Trends Biotechnol,1990,8:340-344
58.Juliano B O.Rice chemistry and technology,2nd edn.American Association of Cereal Chemists,Incorporated Saint Paul,Minnesota,USA,1985
59.Karcicio M,Izbirak A.Isozyme variations in some Aegilops L.and Triticum L.species collected from Central Anatolia.Turk J Bot,2003,27:433-440
60.Krupp H K,Abilay W P,and Alvarez E I.Some water stress effects on rice.in:Rice Breeding.Rice Res.Los Banos,Laguna,Philippines.1972,663-675
61.Lam H M,Coschigano K T,Oliveira I C,Melo-Oliveira R,Coruzzi G M.The molecular-genetics of nitrogen assimilation into amino acids in higher plants.Annu.Rev.Plant Physiol.Plant Mol Bio,1996,47:569-593
62.Lin YJ,Zhang QF.Optimising the tissue culture conditions for high efficiency transformation of indica rice.Plant Cell Rep,2005,23:540-547
63.Liu Q,Kasuga M,Sakuma Y,Abe H,Miura S,Yamaguchi-Shinozaki K,Shinozaki K.Two transcription factors,DREB1 and DREB2,with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought- and low-temperature-responsive gene expression,respectively,in Arabidopsis.Plant Cell,1998,10:1391-1406
64.Lohaus G,B(u|¨)ker M,Huβmann M,Soave C,Heldt H W.Transport of amino acids with special emphasis on the synthesis and transport of asparagine in the Illinois Low Protein and Illinois High Protein strains of maize.Planta,1998,205:181-188
65.Lohaus G,Moellers C.Phloem transport of amino acids in two Brassica napus L.genotypes and one B.carinata genotype in relation to their seed protein content.Planta,2000,211:833-840
66.Marcin M,Andrzej P.Genetic control of aspartate aminotransferase isoenzymes in Aegilops and Triticum species.J Appl Genet,2004,45:411-417
67.Martins M L,Mourato M P,de Varennes e Mendonca AP.Characterization of aspartate aminotransferase isoenzymes from leaves of Lupinus albus L.cv Estoril.J Biochem Mol Biol,2002,35:220-7
68.Matzke M,Matzke AJ,Kooter JM.RNA:guiding gene silencing.Science,2001,293:1080-1083
69.Mett VL,Podivinsky E,Tennant AM,Lochhead LP,Jones WT,Reynolds PHS.A system for tissue-specific copper controllable gene expression in transgenic plants:nodule-specific antisense of aspartate aminotransferase-P2.Transgenic Res,1996,5:105-113.
70.Meyer P.Transcriptional transgene silencing and chromatin components.Plant Mol Biol,2000,43:221-234
71.Miesak B H,Coruzzi G M.Molecular and physiological analysis of Arabidopsis mutants defective in cytosolic or chloroplastic aspartate aminotransferase.Plant Physiol,2002,129:650-60
72.Morino K,Olsen OA,Shimamoto K.Silencing of the alettrone-specific Ltp2-gus gene in transgenic rice is reversed by transgene rearrangements and loss of aberrant transcripts.Plant Cell Physiol,2004,45:1500-1508
73.Murooka Y,Mori Y,Hayashi M.Variation of the amino acid content of Arabidopsis seeds by expressing soybean aspartate aminotransferase gene.J Biosci Bioeng,2002,94:225-230
74.Napoli C,Lemieux C,Jorgensen R.Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans.Plant Cell,1990,2:279-289
75.Rawstone S,Michin FR,Summerfield RJ,Cookson C,Coombs J.Carbon and nitrogen metabolism in legume root nodules.Phytochemistry,1980,19:341-355
76.Resurrection A P,Hara T,Juliano B O,and Yoshida,S.Effect of temperature during ripening on grain quality of rice.Soil Sci Plant Nutr,1977,23:109-112
77.Romero C,Belles J,Vaya J,Serrano R,Culianez-Macia F.Expression of the yeast trehalose-6-phosphate synthasegene in transgenic tobacco plants:pleitropic phenotypes include drought tolerance.Planta,1997,201:293-297
78.Rudman D,Meister A.Transamination in Escherichia coli.J Biol,1953,200:591-604
79.Ryan E,Fottrell PF.Subcellular localization of enzymes involved in the assimilation of ammonia by soybean root nodules.Plant Mol Biol,1974,19:2647-2652
80.Sadowski I,Ma J,Triezenberg S,Ptashne M.GAL4-VP16 is an unusually potent transcriptional activator.Nature,1988,335:563-564
81.Sambrook J,Russell DW.Molecular cloning:a laboratory manual.(3rd edn). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2001, pp 1.31-1.119
82. Schwechheimer C, Smith C, Bevan MW. The activities of acidic and glutamine-rich transcriptional activation domains in plant cells: design of modular transcription factors for high-level expression. Plant Mol Biol, 1998, 36: 195-204
83. Sentoku N, Taniguchi M, Sugiyama T, Ishimaru K, Ohsugi R, Takaiwa F, Toki S. Analysis of the transgenic tobacco plants expressing Panicum miliaceum aspartate aminotransferase genes. Plant Cell Rep, 2000,19: 598-603
84. Setoyama C, Ding S, Choudhury BK, Joh T, Takeshima H, Suzuki T, Shimada K. Regulatory regions of the mitochondrial and cytosolic isoenzymes genes participating in the malate-aspartate shuttle. J Biol Chem, 1990,265: 1293-1299
85. Shaul O, Galili G. Threonine overproduction in transgenic tobacco plants expressing a mutant desensitized aspartate kinase of Escherichia coli. Plant Physiol, 1992,100:1157-1163
86. Silvente S, Camas A, Lara M. Molecular cloning of the cDNA encoding aspartate aminotransferase from bean root nodules and determination of its role in nodule nitrogen metabolism. J Exp Bot, 2003, 54: 1545-51
87. Simon-sarkadi L, Kocsy G, Varhegyi A, Galiba G, De ronde J A. Stress-induced changes in the free amino acid composition in transgenic soybean plants having increased Proline content. Biologia Plantarum, 2006, 50: 793-796
88. Song J, Yamamoto K, Shomura A, Yano M, Minobe Y, Sasaki T. Characterization and mapping of cDNA encoding aspartate aminotransferase in rice, Oryza sativa L. DNA Res, 1996, 3: 303-310
89. Sotelo A, Sousa V, Montalvo I, Hernandez M, Hernandez A L. Chemical composition of different fractions of 12 Mexican varieties of rice obtained during milling. Cereal Chem, 1990, 67: 209-212
90. Sharp PA. RNA interference-2001. Genes Dev, 2001, 15: 485-490
91. Shen Q, Zhang P, Ho TH. Modular nature of abscisic acid (ABA) response complexes: composite promoter units that are necessary and sufficient for ABA induction of gene expression in barley. Plant Cell, 1996, 8: 1107-1119
92. Springer PS. Gene traps: tools for plant development and genomics. Plant Cell, 2000,12: 1007-1020
93. Stam M, Mol JN, Kooter JM. The silence of genes in transgenic plants. Ann Bot, 1997,79:3-12
94.Stempak JM,Sohn K J,Chiang EP,Shane B,Kim YI.Cell and stage of transformation-specific effects of folate deficiency on methionine cycle intermediates and DNA methylation in an in vitro model.Carcinogenesis,2005,26:981-990
95.Takahashi M,Uematsu Y,Kashiwaba K,Yagasaki K,Hajika M,Matsunaga R,Komatsu K,Ishimoto M.Accumulation of high levels of free amino acids in soybean seeds through integration of mutations conferring seed protein deficiency.Planta,2003,217:577-586
96.Taniguchi M,Sawaki H,Sasakawa H,Hase T,Sugiyama T.Cloning and sequence analysis of cDNA encding aspartate aminotrandferase isozymes from Panicum miliaceum L.,a C4 plant.Eur j Biochem,1992,204:611-620
97.Tian YC,Zheng JB,Yu HM,Liang HY,Li CQ,Wang JM.Studies of transgenic hybrid poplar 741 carrying two insect-resistant genes.Acta Bot Sinica,2000,42:263 -268
98.Tu J,Zhang G,Datta K,Xu C,He Y,Zhang Q,Khush GS,Datta SK.Field performance of transgenic elite commercial hybrid rice expressing bacillus thuringiensis Δ-endotoxin.Nat Biotechnol,2000,18:1101-1104
99.Udvardi M K,Kahn M L.Isolation and analysis of a eDNA clone that encodes an alfalfa(Medicago sativa) aspartate aminotransferase.Mol Gen Genet,1991,231:97-105
100.Vain P,Afolabi AS,Wodand B,Shape JW.Transgene behaviour in populations of rice plants transformed using a new dual binary vector system:pGreen/pSoup.Theor Appl Genet,2003,107:210-217
101.van der Salm T,Bosch D,Honee C,Feng L,Munsterman E,Bakker P,Stiekema W J,Visser B.Insect resistance of transgenic plants that express modified Bacillus thuringiensis cryⅠA(b) and cryⅠC genes:a resistance management strategy.Plant Mol Biol,1994,26:51-59
102.Wang W M,Zhao Q,Yu J J,Zhu D Y,Ao G M.Transfer of high Lysine gene sb401 into rice and analysis for protein and amino acid content in transgenic rice seeds.Acta Agronomica Sinica,2005,31:603-607
103.Wadsworth G J,Marmaras S M,Matthews B F.Isolation and characterization of soybean cDNA clone encoding the plastid form of aspartate aminotransferase.Plant Mol Biol,1993,21:993-1009
104.Walbot V.Saturation mutagenesis using maize transposons.Curr Opin Plant Biol,2000,3:103-107
105.Wang WM,Zhao Q,Yu JJ,Zhu DY,Ao GM.Transfer of high lysine gene sb401into rice and analysis for protein and amino acid content in transgenic rice seeds.Acta Agronomica Sinica,2005,31:603-607
106.Wang X L,Larkins B A.Genetic analysis of amino acid accumulation in opaque-2 maize endosperm,Plant Physiol,2001,125:1766-1777
107.Waterhouse PM,Graham MW,Wang MB.Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA.Proc Natl Acad Sci,USA,1998,95:13959-13964
108.Wesley SV,Helliwell CA,Smith NA,Wang MB,Rouse DT,Liu Q,Gooding PS,Singh SP,Abbott D,Stoutjesdijk PA,Robinson SP,Gleave AP,Green AG,Waterhouse PM.Construct design for efficient,effective and high-throughput gene silencing in plants.Plant J,2001,27:581-590
109.Wilkie SE,Roper JM,Smith AG,Warren MJ.Isolation,characterisation and expression of a cDNA clone encoding plastid aspartate aminotransferase from Arabidopsis thaliana.Plant Mol Biol,1995,27:1227-1233
110.Winefield C S,Farnden K J,Reynold P H,Marshall C J.Evolutionary analysis of aspartate aminotransferases.J Mol Evol,1995,40:455-63
111.Wu C,Li X,Yuan W,Chen G,Kilian A,Li J,Xu C,Zhou DX,Wang S,Zhang Q.Development of enhancer trap lines for functional analysis of the rice genome.Plant J,2003,35:418-427
112.Xiong L,Yang Y.Disease resistance and abiotic stress tolerance in rice are inversely modulated by an abscisic acid-inducible mitogen-activated protein kinase.Plant Cell,2003,15:745-759
113.Yamaya T,Obara M,Nakajima H,Sasaki S,Hayakawa T,Sato T.Genetic manipulation and quantitative-trait loci mapping for nitrogen recycling in rice.J Exp Bot,2002,53:917-92
114.Yoon DB,Kang KH,Kim HJ,Ju HG,Kwon SJ,Suh JP,Jeong OY,Ahn SN.Mapping quantitative trait loci for yield components and morphological traits in an advanced backcross population between Oryza grandiglumis and the O.sativa japonica cultivar Hwaseongbyeo.Theor Appl Genet,2006,112:1052-1062
115.Zhang HX,Blumwald E.Transgenic salt-tolerant tomato plants accumulate salt in foliage but not in fruit.Nat Biotechnol,2001,19:765-768
116.Zhang HX,Hodson JN,Williams JP,Blumwald E.Engineering salt-tolerant Brassica plants:characterization of yield and seed oil quality in transgenic plants with increased vacuolar sodium accumulation.Proc Natl Acad Sci,2001,98:12832-12836