棉花GaMYB7和GbGLU基因启动子的克隆和初步功能验证
摘要
棉花是第一大天然纤维产物,在世界纺织工业中具有重要的经济地位。随着人们生活水平的提高和纺织技术的进步,也对棉纤维品质提出了更高的要求。而阐明纤维发育的细胞和分子机理,发现改良棉纤维的靶基因,是改良纤维品质的基础。研究这些棉花纤维特异基因的启动子既有助于进一步明确该基因的功能,也为基因工程的研究打下了基础。
棉花黄萎病(Verticillium spp)是棉花生长过程中最具破坏力的病害之一,在世界范围内流行。在我国,棉花黄萎病是棉花生产的主要障碍之一。使用棉花黄萎病病原诱导的启动子来介导抗病基因的表达,不仅会获得目的产物、达到预定目标,而且也不会产生副作用。植物抗病相关启动子的调控特性研究、分离及其应用对于提高植物抗病性极其关键。
本研究选取了两个目标基因,棉花花和纤维特异表达基因GaMYB7和棉花黄萎病诱导表达基因GbGLU,克隆了这两个基因的启动子序列,具体工作如下:
(1)通过筛选江陵中棉BAC文库的方法,得到了含有GaMYB7基因的阳性单克隆,以此作为TAIL-PCR的起始模板,获得的总长度1566bp的GaMYB7基因上游序列。经过序列分析确定了转录起始位点(TSS)、TATA box和CAATbox。通过序列分析发现,该序列包含多个光调控元件,还具有生长素和赤霉素应答的基本结构。在此基础上,构建了3个5'端启动子片段驱动GUS基因的植物表达载体。通过基因枪转化棉花胚珠瞬时表达实验发现,三个启动子片段均能指导报告基因在棉花胚珠中正常表达。TM-1棉花0DPA胚珠离体诱导实验表明,GaMYB7基因的表达受GA_3和IAA浓度的影响,10uMIAA时,GaMYB7基因的表达量最高;但是过高浓度的GA_3会抑制GaMYB7基因的表达。在20001x的光照强度下诱导处理棉花胚珠发现,与暗处理相比,早期(10天之前)光照处理可以有效提高GaMYB7基因的表达量;后期(10天之后)反而会抑制GaMYB7基因的表达。因此,GaMYB7启动子可用于研究棉花纤维的发育、品质改良以及外源基因在棉花纤维中的特异表达。
(2)通过筛选陆地棉遗传标准系TM-1 BAC文库,得到了GbGLU基因的阳性单克隆,以此作为TAIL-PCR的起始模板,分离了GbGLU基因5'侧翼序列,总长度为1357bp。PLACE分析表明,该序列含有TATA box和CAAT box,还有病原菌诱发子反应元件W-box、GT-1、MYB、MYBST1等。将GbGLU基因5'侧翼序列以不同的缺失与GUS基因融合构建了3个植物表达载体。通过农杆菌介导转化棉花胚性愈伤发现,三个启动子片段均能指导报告基因在棉花愈伤中表达。GbGLU启动子能否用于棉花抗黄萎病的转基因研究还需进一步实验证明。
Promoter can control foreign gene's expression feature:where,when or how much.Constitutive promoter,which drives the foreign gene expressed in all plant organs and throughout the plant developmental period,is widely used in plant genetic engineering studies now.This may squander the plant nutrient and energy and results in increased metabolism burden of the plant.We can control foreign gene's expression if we instead the constitutive promoter by a fiber-specific promoter or a disease inducible promoter in cotton genetic engineering.
In this study,we try to isolate a cotton fiber-specific gene(GaMYB7) promoter and a Verticillium dahliea induced up-regulation expression gene(GbGLU) promoter.
The mainly result includes two sections:
The ordered array of the pooled BAC clones was screened and the BAC clones containing the GaMYB7 gene were identified.With the BAC-DNA serving as template for the first round of TAIL-PCR,we successfully obtained a sequence 1566 base pairs in length upstream of the ATG start codon of the GaMYB7 gene.Cis-acting element prediction results showed the basic sequence structure was consisted of identified putative core promoter elements and other upstream promoter elements, including light response elements,auxin and gibberellin response elements.The effects of GA_3,IAA and light on GaMYB7 expression in fiber cells were studied by in vitro cotton ovule culture and real-time fluorescent quantitative PCR.The results showed that GaMYB7 expression could be significantly induced by 10uM IAA,5uM GA_3 and 10d light treatment,respectively.In transient assays with cotton ovules cultured in vitro,both this promoter sequence and a series of tnmcations could drive theβ-glucuronidase reporter gene(GUS) specifically in ovules and fibers.Therefore, the promoter is considered to be a useful expression element for deep researches on molecular mechanisms of cotton fiber development,modification of fiber quality and expression of foreign genes in fiber.
We also identified the BAC clones containing the GbGLU gene from upland cotton genetic standard line TM-1 BAC library screening and isolated a sequence 1357 base pairs in length upstream of the ATG start codon of the GbGLU gene, successfully.Several cis-acting elements,including W-box,GT-1 box,ARE,BOX4, as-2-box,3-AF1 binding site,Root-motif elements,were recognized with the aid of PLACE program.Four different regions of the promoter sequence of the GbGLU gene were fused to theβ-glucuronidase(GUS) coding region.In Agrobacteriummediated transient expression assay,the transcriptional activations of the promoter deletions were examined in cotton embryogenic callus.
棉花黄萎病(Verticillium spp)是棉花生长过程中最具破坏力的病害之一,在世界范围内流行。在我国,棉花黄萎病是棉花生产的主要障碍之一。使用棉花黄萎病病原诱导的启动子来介导抗病基因的表达,不仅会获得目的产物、达到预定目标,而且也不会产生副作用。植物抗病相关启动子的调控特性研究、分离及其应用对于提高植物抗病性极其关键。
本研究选取了两个目标基因,棉花花和纤维特异表达基因GaMYB7和棉花黄萎病诱导表达基因GbGLU,克隆了这两个基因的启动子序列,具体工作如下:
(1)通过筛选江陵中棉BAC文库的方法,得到了含有GaMYB7基因的阳性单克隆,以此作为TAIL-PCR的起始模板,获得的总长度1566bp的GaMYB7基因上游序列。经过序列分析确定了转录起始位点(TSS)、TATA box和CAATbox。通过序列分析发现,该序列包含多个光调控元件,还具有生长素和赤霉素应答的基本结构。在此基础上,构建了3个5'端启动子片段驱动GUS基因的植物表达载体。通过基因枪转化棉花胚珠瞬时表达实验发现,三个启动子片段均能指导报告基因在棉花胚珠中正常表达。TM-1棉花0DPA胚珠离体诱导实验表明,GaMYB7基因的表达受GA_3和IAA浓度的影响,10uMIAA时,GaMYB7基因的表达量最高;但是过高浓度的GA_3会抑制GaMYB7基因的表达。在20001x的光照强度下诱导处理棉花胚珠发现,与暗处理相比,早期(10天之前)光照处理可以有效提高GaMYB7基因的表达量;后期(10天之后)反而会抑制GaMYB7基因的表达。因此,GaMYB7启动子可用于研究棉花纤维的发育、品质改良以及外源基因在棉花纤维中的特异表达。
(2)通过筛选陆地棉遗传标准系TM-1 BAC文库,得到了GbGLU基因的阳性单克隆,以此作为TAIL-PCR的起始模板,分离了GbGLU基因5'侧翼序列,总长度为1357bp。PLACE分析表明,该序列含有TATA box和CAAT box,还有病原菌诱发子反应元件W-box、GT-1、MYB、MYBST1等。将GbGLU基因5'侧翼序列以不同的缺失与GUS基因融合构建了3个植物表达载体。通过农杆菌介导转化棉花胚性愈伤发现,三个启动子片段均能指导报告基因在棉花愈伤中表达。GbGLU启动子能否用于棉花抗黄萎病的转基因研究还需进一步实验证明。
Promoter can control foreign gene's expression feature:where,when or how much.Constitutive promoter,which drives the foreign gene expressed in all plant organs and throughout the plant developmental period,is widely used in plant genetic engineering studies now.This may squander the plant nutrient and energy and results in increased metabolism burden of the plant.We can control foreign gene's expression if we instead the constitutive promoter by a fiber-specific promoter or a disease inducible promoter in cotton genetic engineering.
In this study,we try to isolate a cotton fiber-specific gene(GaMYB7) promoter and a Verticillium dahliea induced up-regulation expression gene(GbGLU) promoter.
The mainly result includes two sections:
The ordered array of the pooled BAC clones was screened and the BAC clones containing the GaMYB7 gene were identified.With the BAC-DNA serving as template for the first round of TAIL-PCR,we successfully obtained a sequence 1566 base pairs in length upstream of the ATG start codon of the GaMYB7 gene.Cis-acting element prediction results showed the basic sequence structure was consisted of identified putative core promoter elements and other upstream promoter elements, including light response elements,auxin and gibberellin response elements.The effects of GA_3,IAA and light on GaMYB7 expression in fiber cells were studied by in vitro cotton ovule culture and real-time fluorescent quantitative PCR.The results showed that GaMYB7 expression could be significantly induced by 10uM IAA,5uM GA_3 and 10d light treatment,respectively.In transient assays with cotton ovules cultured in vitro,both this promoter sequence and a series of tnmcations could drive theβ-glucuronidase reporter gene(GUS) specifically in ovules and fibers.Therefore, the promoter is considered to be a useful expression element for deep researches on molecular mechanisms of cotton fiber development,modification of fiber quality and expression of foreign genes in fiber.
We also identified the BAC clones containing the GbGLU gene from upland cotton genetic standard line TM-1 BAC library screening and isolated a sequence 1357 base pairs in length upstream of the ATG start codon of the GbGLU gene, successfully.Several cis-acting elements,including W-box,GT-1 box,ARE,BOX4, as-2-box,3-AF1 binding site,Root-motif elements,were recognized with the aid of PLACE program.Four different regions of the promoter sequence of the GbGLU gene were fused to theβ-glucuronidase(GUS) coding region.In Agrobacteriummediated transient expression assay,the transcriptional activations of the promoter deletions were examined in cotton embryogenic callus.
引文
1. Annadana S, Beekwilder M J, Kuipers G. Cloning of the chrysanthemum UEP1 promoter and comparative expression in florets and leaves of Dendranthema frandiflora [J]. Transgenic Res,202,11:4337-445
2. Belbahri L, Boucher C, Candresse T. A local accumulation of the Ralstonia solanacearum PopA protein in transgenic tobacco renders a compatible plant pathogen interaction incompatible [J].Plant J,2001,28:419-421
3. Bowles D J. Defense-related proteins in higher plants [J]. Annu Rev Biochem, 1990, 59: 873-907.
4. Beasley CA and Ting IP (1973). The effects of plant growth substances on in vitro fiber development from fertilized cotton ovules [J]. Am J Bot. 60:130-139
5. C Napoli, C Lemieux, R Jorgensen. Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression [J]. Plant Cell,1990, 2:279-289
6. Clarke HR, Davis JM, Wilbert SM, et al. Wound-induced and developmental activation of apoplar tree chitinase gene promoter in transgenic tobacco[J].Plant Mol Biol.,1994,25:799-802
7. ChenYu-Ting, Yi-Ru Lee, Chin-Yuan Yang, et al. A novel papaya ACC oxidase gene (CP-ACOZ) associated with latest age fruit ripening and leaf senescence [J]. Plant Science,2003,164:531-540
8. Choi S, Begum D, Koshinsky H, et al. A new approach for the identification and cloning of genes: the pBAC wich system using Cre/lox site-specific recombination [J]. Nucleic Acids Res,2000, 28:E19
9. Chuan-Yu Hsu, Johnnie N. Jenkins, Sukumar Saha, et al.Transcriptional regulation of the lipid transfer protein gene LTP3 in cotton fibers by a novel MYB protein[J]. Plant Science, 2005,168,167-181
10. Chun-Hua Chen, Ying-Lin Cao, Wei-Cheng Hu. Apolipo protein C-II promoter T→A substitution at position-190 affects on the transcription of the gene and its relationship to hyperlipernia [J]. Biochemical and Biophysical Research Communication, 2007, 354: 62-65
11. Curr SJ, Rushton PJ. Engineering plants with increased disease resistance: how are we going to express it[J]? Trends in Biotechnology, 2005, 23: 283-290
12. Deikman J, Xu R, Kneissl ML, et al. Separation of cis elements Responsive to ethylene, fruit development, and ripening in the 5'-flanking region of the ripening-related E8 gene [J]. Plant Mol Biol, 1998,37:1001-1011
13. Delmar DP, Pear JR, Andrawis A. Genes encoding small GTP-binding proteins analogous to mammalian rac are preferentially expressed in developing cotton fibers. Mol Gen Genet, 1995, 248: 43-51
14. Digeonjf, Guiderdonie, Alaryr. Cloning of a wheat puroindo, line gene promoter by IPCR and analysis of promoter regions required for tissue-specific expression in transgenic rice seeds [J] .Plant Mol Biol,1999,39:1101-1112
15. F Gubler, JV Jacobsen. Gibberellin-responsive elements in the promoter of a barley high-pI alpha-amylase gene [J]. Plant Cell, 1992,4: 1435-1441
16. Ferguson DL, Turley R B, Kloth R H. Identification of a delta-TIP cDNA clone and determination of related A and D genome subfamilies in Gossypium species. Plant Mol Biol., 1997,34: 111-118.
17. Fu HH, Dooner HK. A gene enriched BAC library for cloning large allele2specific fragments from maize: isolation of a 240 kb contig of the bronze region [J]. Genome Research, 2000, 10 :866-873.
18. Frijters A CJ , Zhang Z , Damme M V , et al . Construction of a bacterial artificial chromosome library containing large EcoRI and HindIII genomic fragment of lettuce [J] . Theor. Appl. Genet., 1997, 94 :390-399
19. Garbarino JE, Belknap WR. Isolation of a ubiquitin-ribosomal protein gene(ubi3) from potato and expression of its promoter in transgenic plant [J].Plant Mol Biol., 1995,24:119-122
20. Gittins JR, Pellny TK, Hiles ER, et al. Transgene expression driven by heterologous ribulose-1, 5-bisphosphate carboxylase/oxygenase small-subunit gene promoters in the vegetative tissues of apple (Malus pumila Mill) [J]. Planta,2002,210:232-236
21. Gurr SJ, Rushton PJ. Engineering plantswith increased disease resistance: how are we going t o exp ress it [J]? Trends in Biotechnology, 2005, 23: 283-290
22. Han ZY, Wang XQ, S GZ, et al. Cloning of foreign gene' s flanking sequences in transgenic rice by inverse PCR [J]. Acta. Agriculture Shanghai, 2001,17: 27-31
23. Hamilton CM, Frary A , Lewis C ,et al . Stable transfer of intact high molecular weight DNA into plant chromosomes [J].Proc.Natl.Acad.Sci.USA.,1996, 93:9975 - 9979
24. Haigler CH, Ivanova-Datcheva M, Hogan PS, et al. Carbon partitioning to cellulose synthesis [J]. Plant Mol Biol., 2001,47:29-51
25. Harmer SE, Orford SJ , Timmis JN. Characterization of six a-expansin genes in Gossypium hirsutum (upland cotton) [J]. Mol Genet Genomics, 2002,268:1-9
26. Hasegawa PM, Bressan RA, Zhu, J K, et al. Plant cellular and molecular responses to high salinity. Annu Rev Plant Physiol [J]. Plant Mol Biol., 2000, 51: 463-499
27. Hasenfratz MP, Tsou CL, Wilkins TA. Expression of two related vacuolar H~+-ATPase 16-kilodalton proteolipid genes is differentially regulated in a tissue-specific manner [J]. Plant Physiol., 1995, 108: 1395-1404
28. Higgins DG, Thompson JD and Gibson TJ. Using CLUSTAL for multiple sequence alignments [J]. Methods Enzymol., 1996,266, 383-402
29. Higgins DG, Bleasby AJand Fuchs R. CLUSTAL V: improved software for multiple sequence alignment [J]. CABIOS 1992,8,189-191
30. Higo KY, Ugawa ,M Iwamoto, et al. Plant cis-acting regulatory DNA elements (PLACE) database [J]. Nucleic Acids Research, 1999,Vol.27 No.1: 297-300
31. Hirano T, Sato T, Yaegashi K, et al. Efficient transformation of the edible basidiomycete Lentinus edodes with a vector using a glyceraldehydes-3-phosphate dehydrogenase promoter to hygromycin B resistance [J]. Molecular and General Genetics, 2000 ,263 :1047-1052
32. Hirano T, Sato T, Okawa K, et al. Isolation and characterization of the Glyceraldehydes-3-phosphate dehydrogenase gene of Lentinus edodes [J]. Bioscience biotechnology and Biochemistry, 1999,63:1223-1227
33. James JT, Dubery LA. Inhibition of Polygalacturonase from Vertiecillum dahliae by a Polygalaeturonase inhibiting Protein from cotton [J]. Phytochemistry,1996, 57:149-156
34. Jeanmougin F, Thompson JD, Gouy M, et al. Multiple sequence alignment with Clustal X [J]. Trends Biochem. Sci., 1998,23, 403-5
35. Jefferson R A. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants [J]. EMBOJ. 1987,6:3901-3907
36. Jian Xun Feng, Sheng Jian Ji, Yong Hui Shi, et al.Analysis of five differentially expressed gene families in fast elongation cotton fiber. Acta. Biochim [J]. et Biophysica Sinica, 2004, 36(1): 51-57
37. Jinfeng Suo, Xiaoe Liang, Li Pub, et al. Identification of GhMYB109 encoding a R2R3 MYB transcription factor that expressed specifically in fiber initials and elongating fibers of cotton (Gossypium hirsutum L.) [J]. Biochimica et Biophysica Acta, 2003, 1630: 25- 34
38. John ME, Keller G. Characterization of mRNA for a proline-rich protein of cotton fiber [J]. Plant Physiol., 1995, 108: 669-676
39. Kim UJ, Birren B W, Slepak T, et al. Construction and characterization of a human bacterial artificial chromosome library [J]. Genomics, 1996, 34:213-218
40. Kim HJ, Triplett BA. Cotton fiber growth in planta and in vitro. Models for plant cell elongation and cell wall biogenesis [J]. Plant Physiol., 2001, 127:1361-1366
41. Kim HJ, Triplett BA. Characterization of GhRac1 GTPase expressed in developing cotton (Gossypium hirsutum L.) fibers [J]. Biochim Biophys Acta. 2004, 679: 214-21
42. Kunpatla SP, Chandrasekharan MB, Iyer LM, et al. Induction of male sterility in plants by a chimaeric ribonuclease gene [J].Trends Plant Sci.,1998,3:97-102
43. Laosinchai W, Cui X, Brown RM, et al. A full cDNA of cotton cellulose synthase has high homology with the Arabidopsis RSW1 gene and cotton CelA1 (Accession No. AF 200453) (PGR 00-002) [J]. Plant Physiol., 2000, 122: 291
44. Liu YG, Shirano Y, Fukaki H, et al. Complementation of plant mutants with large genomic DNA fragments by a trans-formation-competent artificial chromosome vector accelerates positional cloning [J]. Proc. Natl. Acad. Sci. USA., 1999, 96:6535-6540.
45. Liu YG, Whittier RF. Thermal Asymmetric Interlaced PCR: Automatable amplificaton and sequencing of insert end fragments from P1 and YAC clones for chromosome walking [J].Genomics, 1995, 25 :674-681
46. Liu YG, Norihiro Mitsukawa, Teruko Oosumi, et al. Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR[J].The Plant Journal, 1995, 8:457-463
47. Liu YG, Huang N. Efficient amplification of insert end sequences from bacterial artificial chromosome clones by thermal asymmetric interlaced PCR [J]. Plant Molecular Biology Reporter, 1998, 16 :175-181
48. Lander ES, Linton LM, Birren B, et al. Initial sequencing and analysis of the human genome [J].Nature,2001,409:860-921
49. Liu JJ, Ekramoddoullah AK. Root-specific expression of a western white pine PR10 gene is mediated by different promoter regions in transgenic tobacco [J]. Plant Mol Biol. 2003, 52:103-20
50. Lucia J. Pablo V. Local and systemic induction of two defense related subtilisin like protease promoters in transgenic Arabidopsis plants [J]. Plant Physiol., 2000, 124:1049-1052
51. Luo MZ, Wang YH, Frisch D, et al. Dean melon bacterial artificial chromosome (BAC) library construction using improved methods and identification of clones linked to the locus conferring resistance to melon Fusarium wilt ( Fom-2) [J] .Genome , 2001 , 44 : 154-162
52. Mozo T, Fischer S, Shizuya H, et al. Construction and characterization of the IGF Arabidopsis BAC library [J]. Mol. Gen. Genet., 1998,258: 562 - 570
53. Hamilton CM, Frary A, Xu Y, et al. Construction of tomato genomic DNA libraries in a binary BAC (BIBAC) vector [J]. Plant J., 1999, 18 :223-229
54. Iyer LM, Kumpatla SP, Chandrasekharan MB et al. Transgene silencing in monocots [J]. Plant Mol Biol.,2000, 43:323 -346
55. Ma DP, Liu HC, Creech RG, et al. Cloning and characterization of a cotton lipid transfer gene specifically expressed in fiber cells [J]. Biochim Biophys Acta, 1997, 1334:111-114.
56. Ma DP, Tan H, Si Y, Creech RG, et al. Differential expression of a lipid transfer protein gene in cotton fiber [J]. Biochim Biophys Acta, 1995, 1257: 81-84
57. Mariani C, De Beuckeleer M, Truettner J, et al. Induction of male sterility in plants by a chimaeric ribonuclease gene [J]. Nature, 1990,347: 737-742
58. Mariani C, Gossele V, De Beucheleer M, et al. A chimaeric ribonuclease-inhibitor gene restores fertility to male-sterile plants [J]. Nature, 1992,357:384-389
59. Mcelroy D, Zhang W G, Cao J. Isolation of an efficient actin promoter for use in rice transformation [J]. Plant cell, 1990,2:163-171
60. Millar AJ, Kay SA. Integration of circadian and phototransduction pathways in the network controlling CAB gene transcription in Arabidopsis [J]. Proc. Natl. Acad. Sci. USA, 1996, 93: 15491-15496.
61. Molnaar, ovasa, banaflviz. Tissue-specific signal(s) activate the Promoter of a metallocarb -oxypeptidase inhibit or gene family in potato tuber and berry [J]. Plant Mol. Biol., 2001 , 46: 301-311
62. Montgomery J, Goldman S, Deikman J, et al.. Identification of an ethylene-responsive region in the Promoter of a fruit ripening gene [J]. Proc Natl Acad Sei USA. 1993,90:5939-5943
63. Nagao RT, Goekjian VH, et al. Identification of protein-binding DNA sequences in an auxin-regulated gene of soybean [J]. Plant Mol Biol.,1993,21:1147-1150
64. Naomi SS, Ichiro M , Shigeo N, et al. Constitutive promoters available for transgene expression instead of CaMV35S RNA promoter: Arabidopsis promoters of tryptophan synthase protein β subunit and phytochrome B [J]. Plant Biotech, 2002,19:19-22
65. Nitz I, Berkefeld H, Puzio PS. Pyk10, a seedling and root specific gene and promoter from Arabidopsis thaliana. Grundler FM [J]. Plant Sci. 2001, 161:337-346
66. Nagao RT, Goekjian VH, et al. Identification of protein-binding DNA sequences in an auxin-regulated gene of soybean [J]. Plant Mol Biol., 1993, 21:1147-1150
67. Orford SJ, Timmis JN. Abundant mRNAs specific to the developing cotton fiber [J]. Theor. Appl.Genet, 1997, 94: 909-918
68. Pear JR, Kawagoe Y, Schreckengost WE. Higher plant contains homologs of the bacterial celA gene encoding the catalytic subunit of cellulose synthase [J]. Proc Natl.Acad.Sci.USA, 1996, 93: 12637-12642
69. Prestridge DS. SIGNAL SCAN: A computer program that scans DNA sequences for eukaryotic transcriptional elements [J]. CABIOS, 1991, 7, 203-206
70. Preuss ML, Kovar DR, Lee YR, et al. A plant-specific kinesin binds to actin microfilaments and interacts with cortical microtubules in cotton fibers. Plant Physiol., 2004, 136: 3945-55
71. Robinson DJ. Environmental risk assessment of release of transgenic plants containing virus-derived inserts [J].Transgen. Res.,1996,5:359-341
72. Rojas A, Almoguera C, Carranco R, et al. Selective activation of the developmentally regulated hahsp17G1 promoter by heat stress transcription factors [J]. Plant Physiology,2002,129:1207-1211
73. Sa Q, Wang Y, LiW, et al. The promoter of an antifungal protein gene from Gastrodia elata confers tissue s pecific and fungus-in-ducible expression patterns and responds to both salicylic acid and jasmonic acid [J]. Plant Cell Rep, 2003, 22: 79-84
74. Shizuya H, Birren B, Kim U J, et al. Cloning and stable maintenance of 300 kilo base-pair fragments of human DNA in Escherchia coli using an F-factor-based vector [J]. Proc. Natl. Acad. Sic. USA., 1992, 89: 8794 - 8797
75. Song P, Allen RD. Identification of a cotton fiber-specific acyl carrier protein cDNA by differential display [J]. Biochim Biophy Acta, 1997, 1351: 305-312
76. Streatfield SJ, Beifuss KK, Brooks CA, et al. Analysis of the maize polyubiquitin-1 promoter heat shock elements and generation of promoter variants with modified expression characteristics [J].Transgenic Res,2004,13:299-312
77. T Kasrga, kR. Mitehelson. Expression of Potential defense response genes in cotton Physiological and Molecular [J]. Plant Pathology, 2000, 56:25-31
78. T Triglia, MG Peterson and DJ Kemp. A procedure for in vitro amplification of DNA segments that lie outside the boundaries of known sequences [J]. Nucl. Acids Res. 1988, 16, 81-86
79. Tada Y, Sakamoto M, Matsuoka M, et al. Expression of a monocot LHCP promoter in transgenic rice [J]. EMBO J, 1991, 10:1803-1808
80. Tan H, Creech RG, Jenkins JN, et al. Cloning and expression analysis of two cotton (Gossypium hirsutum L.) genes encoding cell wall proline-rich proteins. DNA, 2001, 12: 367-380
81. Thompson JD, Gibson TJ, Plewniak F, et al. The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools [J]. Nucleic Acids Research, 1997,24: 4876-4882
82. Wan CY, Wilkins TA. Isolation of multiple cDNA encoding the vacuolar H~+-ATPase subunit B from developing cotton (Gossypium hirsutum L.) [J]. Plant Physiol., 1994, 106:393-394
83. Wilkins TA. Vacuolar H+-ATPase 69-kilodalton catalytic subunit cDNA from developing cotton (Gossypium hirsutum L.) ovules [J]. Plant Physiol., 1993, 102: 679-680
84. Yoon IS, Park DH, Mori H, et al. Characterization of an auxin-inducible 1-Aminocyclopropane-1- carboxylate synthase gene,VR-ACS6,of mung bean(Vigna radiate(L)Wilczek)and hormonal interactions on the promoter activity in transgenic tobacco[J].Plant Cell Physiol,1999,40:431-435
85.Yun-Kuan Liang,et al.AtMYB61,an R2R3-MYB Transcription Factor Controlling Stomatal Aperture in Arabidopsis thaliana[J].Current Biology,2005,15:1201-1206
86.ZHAO Chang-ling,GUO Wei-ming,YANG Qing,et al.Cloning of Full-length F3H of Nanjinghongxu(Prunus mume) by gDNA-based TAIL-PCR[J].2005,12,159-165
87.樊荣,万小荣,张文涛.LE-ACS6启动子在LE-ACS6:GUS转基因拟南芥中的特异性[J].植物生理与分子生物学学报,2004,30(3):351-354
88.甘莉,吕金殿,汪沛洪,等.棉花黄萎病菌分泌的糖蛋白毒素与其致病力的关[J].中国农业科学,1995,28(2):58-65
89.高玉龙.棉花防卫反应相关基因类似物的克隆与特征分析和受黄萎病诱导表达的两个全长cDNA的获得[J].博士论文,2005
90.简桂良,马存,孙文姬.1995年华北棉区棉花黄萎病大发生及防治措施[J].植物保护.1996,(3):31-35
91.冯洁,孙文姬.中国棉花枯萎病生理小种的RAPD分析[J].菌物系统,2000,19:45-50
92.冯洁,孙文姬.棉花枯萎病菌生理小种的分子指纹分析[J].棉花学报,1999,11(5):230-234
93.冯洁.棉花与黄萎病菌互作机制研究现状及展望[J].棉花学报,1998,10(6):281-284
94.李桂兰,王文颇,臧少先,等。拟南芥黑芥子酶基因根特异性表达启动子的克隆[J].河北农业大学学报.2005,28(1)49-52
95.李娜,王世明,陈军,等.SEFA-PCR法克隆灵芝鲨烯合酶基因启动子及其序列分析[J].菌物学报2006,4,245-253
96.李姗姗,迟彦,李凌飞,等.启动子克隆方法研究进展[J].中国生物工程杂志.2005,25(7):9-16
97.李胜国,刘玉乐,朱锋.基因工程雄性不育烟草的获得[J].植物学报,1995,7(8):659-660
98.刘学堂,宋晓轩.棉花黄萎病菌的研究及最新进展[J].棉花学报,1998,10(1):6-13
99.刘学堂,张元恩.我国棉花黄萎病菌群体遗传分析研究初报[J].棉花学报,1997,24-31
100.刘巧泉,于恒秀,张文娟.水稻rbcS启动子控制的外源基因在转基因水稻中的特异性表达[J].植物生理与分子生物学学报,2005,31(3):247-251
101.刘大文,谢友菊,王守才.拟南芥菜花药绒毡层启动子的克隆和序列分析[J].作物学报,2000,26(4):406-409
102.路静,赵华燕,何奕等.高等植物启动子及其应用研究进展[J].自然科学进展,2004,14(8):856-862
103.廖飞雄,潘瑞炽.植物根特异性基因表达研究[J].植物学通报,1998,15(2):8-13
104.毛自朝,于秋菊,甄伟,等.果实专一性启动子驱动iPt基因在番茄中的表达及其对番茄果实发育的影响[J].科学通报,2002,47(6):444-448
105.任茂智.棉花nodunli-like和arfl基因及其启动子的分离和功能分析[D].2004年中国农科院博士学位论文
106.石磊岩,简桂良.我国棉花黄萎病研究现状与方向[J].植物保护学报,1998,25(2):103-107
107.石磊岩,李长兴,姚大瑞,等.棉花铃病发生关键因子及预防的研究[J].棉花学报,1995,7(3):175-178
108.孙晓红,陈明杰,潘迎捷.启动子克隆概述[J].食用菌学报,2002,9(3):57-62
109.吴梅花,张丽,牛一丁,等.拟南芥逆境胁迫诱导表达基因rd29A启动子的克隆与序列分析[J].华北农学报,2005,20(2):5-7
110.王雅琴.嗜麦芽假单胞菌热休克基因dnaK启动子的亚克隆及其在大肠杆菌中功能的研究[J].工业微生物,2001,31(3):29-33
111.张天真,何金龙,闵留芳,等.棉花对黄萎病的抗性遗传模式及抗(耐)病品种的选育技术[J].作物学报.2000,26(6):673-680
112.张晓国,刘玉乐,康良仪.水稻花药特异表达基因启动子的扩增及克隆[J].武汉大学学报(自然科学版),1997,43(4):480-484
113.周兆华,郭旺珍,潘家驹,等.我国棉花黄萎病菌群体的遗传变异分析[J]中国农业科学,1999,32(2):60-65
114.周雪荣,彭仁旺,方荣祥.表达核糖核酸酶基因的雄性不育油菜的获得[J].遗传学报,1997,24(6):531-534
2. Belbahri L, Boucher C, Candresse T. A local accumulation of the Ralstonia solanacearum PopA protein in transgenic tobacco renders a compatible plant pathogen interaction incompatible [J].Plant J,2001,28:419-421
3. Bowles D J. Defense-related proteins in higher plants [J]. Annu Rev Biochem, 1990, 59: 873-907.
4. Beasley CA and Ting IP (1973). The effects of plant growth substances on in vitro fiber development from fertilized cotton ovules [J]. Am J Bot. 60:130-139
5. C Napoli, C Lemieux, R Jorgensen. Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression [J]. Plant Cell,1990, 2:279-289
6. Clarke HR, Davis JM, Wilbert SM, et al. Wound-induced and developmental activation of apoplar tree chitinase gene promoter in transgenic tobacco[J].Plant Mol Biol.,1994,25:799-802
7. ChenYu-Ting, Yi-Ru Lee, Chin-Yuan Yang, et al. A novel papaya ACC oxidase gene (CP-ACOZ) associated with latest age fruit ripening and leaf senescence [J]. Plant Science,2003,164:531-540
8. Choi S, Begum D, Koshinsky H, et al. A new approach for the identification and cloning of genes: the pBAC wich system using Cre/lox site-specific recombination [J]. Nucleic Acids Res,2000, 28:E19
9. Chuan-Yu Hsu, Johnnie N. Jenkins, Sukumar Saha, et al.Transcriptional regulation of the lipid transfer protein gene LTP3 in cotton fibers by a novel MYB protein[J]. Plant Science, 2005,168,167-181
10. Chun-Hua Chen, Ying-Lin Cao, Wei-Cheng Hu. Apolipo protein C-II promoter T→A substitution at position-190 affects on the transcription of the gene and its relationship to hyperlipernia [J]. Biochemical and Biophysical Research Communication, 2007, 354: 62-65
11. Curr SJ, Rushton PJ. Engineering plants with increased disease resistance: how are we going to express it[J]? Trends in Biotechnology, 2005, 23: 283-290
12. Deikman J, Xu R, Kneissl ML, et al. Separation of cis elements Responsive to ethylene, fruit development, and ripening in the 5'-flanking region of the ripening-related E8 gene [J]. Plant Mol Biol, 1998,37:1001-1011
13. Delmar DP, Pear JR, Andrawis A. Genes encoding small GTP-binding proteins analogous to mammalian rac are preferentially expressed in developing cotton fibers. Mol Gen Genet, 1995, 248: 43-51
14. Digeonjf, Guiderdonie, Alaryr. Cloning of a wheat puroindo, line gene promoter by IPCR and analysis of promoter regions required for tissue-specific expression in transgenic rice seeds [J] .Plant Mol Biol,1999,39:1101-1112
15. F Gubler, JV Jacobsen. Gibberellin-responsive elements in the promoter of a barley high-pI alpha-amylase gene [J]. Plant Cell, 1992,4: 1435-1441
16. Ferguson DL, Turley R B, Kloth R H. Identification of a delta-TIP cDNA clone and determination of related A and D genome subfamilies in Gossypium species. Plant Mol Biol., 1997,34: 111-118.
17. Fu HH, Dooner HK. A gene enriched BAC library for cloning large allele2specific fragments from maize: isolation of a 240 kb contig of the bronze region [J]. Genome Research, 2000, 10 :866-873.
18. Frijters A CJ , Zhang Z , Damme M V , et al . Construction of a bacterial artificial chromosome library containing large EcoRI and HindIII genomic fragment of lettuce [J] . Theor. Appl. Genet., 1997, 94 :390-399
19. Garbarino JE, Belknap WR. Isolation of a ubiquitin-ribosomal protein gene(ubi3) from potato and expression of its promoter in transgenic plant [J].Plant Mol Biol., 1995,24:119-122
20. Gittins JR, Pellny TK, Hiles ER, et al. Transgene expression driven by heterologous ribulose-1, 5-bisphosphate carboxylase/oxygenase small-subunit gene promoters in the vegetative tissues of apple (Malus pumila Mill) [J]. Planta,2002,210:232-236
21. Gurr SJ, Rushton PJ. Engineering plantswith increased disease resistance: how are we going t o exp ress it [J]? Trends in Biotechnology, 2005, 23: 283-290
22. Han ZY, Wang XQ, S GZ, et al. Cloning of foreign gene' s flanking sequences in transgenic rice by inverse PCR [J]. Acta. Agriculture Shanghai, 2001,17: 27-31
23. Hamilton CM, Frary A , Lewis C ,et al . Stable transfer of intact high molecular weight DNA into plant chromosomes [J].Proc.Natl.Acad.Sci.USA.,1996, 93:9975 - 9979
24. Haigler CH, Ivanova-Datcheva M, Hogan PS, et al. Carbon partitioning to cellulose synthesis [J]. Plant Mol Biol., 2001,47:29-51
25. Harmer SE, Orford SJ , Timmis JN. Characterization of six a-expansin genes in Gossypium hirsutum (upland cotton) [J]. Mol Genet Genomics, 2002,268:1-9
26. Hasegawa PM, Bressan RA, Zhu, J K, et al. Plant cellular and molecular responses to high salinity. Annu Rev Plant Physiol [J]. Plant Mol Biol., 2000, 51: 463-499
27. Hasenfratz MP, Tsou CL, Wilkins TA. Expression of two related vacuolar H~+-ATPase 16-kilodalton proteolipid genes is differentially regulated in a tissue-specific manner [J]. Plant Physiol., 1995, 108: 1395-1404
28. Higgins DG, Thompson JD and Gibson TJ. Using CLUSTAL for multiple sequence alignments [J]. Methods Enzymol., 1996,266, 383-402
29. Higgins DG, Bleasby AJand Fuchs R. CLUSTAL V: improved software for multiple sequence alignment [J]. CABIOS 1992,8,189-191
30. Higo KY, Ugawa ,M Iwamoto, et al. Plant cis-acting regulatory DNA elements (PLACE) database [J]. Nucleic Acids Research, 1999,Vol.27 No.1: 297-300
31. Hirano T, Sato T, Yaegashi K, et al. Efficient transformation of the edible basidiomycete Lentinus edodes with a vector using a glyceraldehydes-3-phosphate dehydrogenase promoter to hygromycin B resistance [J]. Molecular and General Genetics, 2000 ,263 :1047-1052
32. Hirano T, Sato T, Okawa K, et al. Isolation and characterization of the Glyceraldehydes-3-phosphate dehydrogenase gene of Lentinus edodes [J]. Bioscience biotechnology and Biochemistry, 1999,63:1223-1227
33. James JT, Dubery LA. Inhibition of Polygalacturonase from Vertiecillum dahliae by a Polygalaeturonase inhibiting Protein from cotton [J]. Phytochemistry,1996, 57:149-156
34. Jeanmougin F, Thompson JD, Gouy M, et al. Multiple sequence alignment with Clustal X [J]. Trends Biochem. Sci., 1998,23, 403-5
35. Jefferson R A. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants [J]. EMBOJ. 1987,6:3901-3907
36. Jian Xun Feng, Sheng Jian Ji, Yong Hui Shi, et al.Analysis of five differentially expressed gene families in fast elongation cotton fiber. Acta. Biochim [J]. et Biophysica Sinica, 2004, 36(1): 51-57
37. Jinfeng Suo, Xiaoe Liang, Li Pub, et al. Identification of GhMYB109 encoding a R2R3 MYB transcription factor that expressed specifically in fiber initials and elongating fibers of cotton (Gossypium hirsutum L.) [J]. Biochimica et Biophysica Acta, 2003, 1630: 25- 34
38. John ME, Keller G. Characterization of mRNA for a proline-rich protein of cotton fiber [J]. Plant Physiol., 1995, 108: 669-676
39. Kim UJ, Birren B W, Slepak T, et al. Construction and characterization of a human bacterial artificial chromosome library [J]. Genomics, 1996, 34:213-218
40. Kim HJ, Triplett BA. Cotton fiber growth in planta and in vitro. Models for plant cell elongation and cell wall biogenesis [J]. Plant Physiol., 2001, 127:1361-1366
41. Kim HJ, Triplett BA. Characterization of GhRac1 GTPase expressed in developing cotton (Gossypium hirsutum L.) fibers [J]. Biochim Biophys Acta. 2004, 679: 214-21
42. Kunpatla SP, Chandrasekharan MB, Iyer LM, et al. Induction of male sterility in plants by a chimaeric ribonuclease gene [J].Trends Plant Sci.,1998,3:97-102
43. Laosinchai W, Cui X, Brown RM, et al. A full cDNA of cotton cellulose synthase has high homology with the Arabidopsis RSW1 gene and cotton CelA1 (Accession No. AF 200453) (PGR 00-002) [J]. Plant Physiol., 2000, 122: 291
44. Liu YG, Shirano Y, Fukaki H, et al. Complementation of plant mutants with large genomic DNA fragments by a trans-formation-competent artificial chromosome vector accelerates positional cloning [J]. Proc. Natl. Acad. Sci. USA., 1999, 96:6535-6540.
45. Liu YG, Whittier RF. Thermal Asymmetric Interlaced PCR: Automatable amplificaton and sequencing of insert end fragments from P1 and YAC clones for chromosome walking [J].Genomics, 1995, 25 :674-681
46. Liu YG, Norihiro Mitsukawa, Teruko Oosumi, et al. Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR[J].The Plant Journal, 1995, 8:457-463
47. Liu YG, Huang N. Efficient amplification of insert end sequences from bacterial artificial chromosome clones by thermal asymmetric interlaced PCR [J]. Plant Molecular Biology Reporter, 1998, 16 :175-181
48. Lander ES, Linton LM, Birren B, et al. Initial sequencing and analysis of the human genome [J].Nature,2001,409:860-921
49. Liu JJ, Ekramoddoullah AK. Root-specific expression of a western white pine PR10 gene is mediated by different promoter regions in transgenic tobacco [J]. Plant Mol Biol. 2003, 52:103-20
50. Lucia J. Pablo V. Local and systemic induction of two defense related subtilisin like protease promoters in transgenic Arabidopsis plants [J]. Plant Physiol., 2000, 124:1049-1052
51. Luo MZ, Wang YH, Frisch D, et al. Dean melon bacterial artificial chromosome (BAC) library construction using improved methods and identification of clones linked to the locus conferring resistance to melon Fusarium wilt ( Fom-2) [J] .Genome , 2001 , 44 : 154-162
52. Mozo T, Fischer S, Shizuya H, et al. Construction and characterization of the IGF Arabidopsis BAC library [J]. Mol. Gen. Genet., 1998,258: 562 - 570
53. Hamilton CM, Frary A, Xu Y, et al. Construction of tomato genomic DNA libraries in a binary BAC (BIBAC) vector [J]. Plant J., 1999, 18 :223-229
54. Iyer LM, Kumpatla SP, Chandrasekharan MB et al. Transgene silencing in monocots [J]. Plant Mol Biol.,2000, 43:323 -346
55. Ma DP, Liu HC, Creech RG, et al. Cloning and characterization of a cotton lipid transfer gene specifically expressed in fiber cells [J]. Biochim Biophys Acta, 1997, 1334:111-114.
56. Ma DP, Tan H, Si Y, Creech RG, et al. Differential expression of a lipid transfer protein gene in cotton fiber [J]. Biochim Biophys Acta, 1995, 1257: 81-84
57. Mariani C, De Beuckeleer M, Truettner J, et al. Induction of male sterility in plants by a chimaeric ribonuclease gene [J]. Nature, 1990,347: 737-742
58. Mariani C, Gossele V, De Beucheleer M, et al. A chimaeric ribonuclease-inhibitor gene restores fertility to male-sterile plants [J]. Nature, 1992,357:384-389
59. Mcelroy D, Zhang W G, Cao J. Isolation of an efficient actin promoter for use in rice transformation [J]. Plant cell, 1990,2:163-171
60. Millar AJ, Kay SA. Integration of circadian and phototransduction pathways in the network controlling CAB gene transcription in Arabidopsis [J]. Proc. Natl. Acad. Sci. USA, 1996, 93: 15491-15496.
61. Molnaar, ovasa, banaflviz. Tissue-specific signal(s) activate the Promoter of a metallocarb -oxypeptidase inhibit or gene family in potato tuber and berry [J]. Plant Mol. Biol., 2001 , 46: 301-311
62. Montgomery J, Goldman S, Deikman J, et al.. Identification of an ethylene-responsive region in the Promoter of a fruit ripening gene [J]. Proc Natl Acad Sei USA. 1993,90:5939-5943
63. Nagao RT, Goekjian VH, et al. Identification of protein-binding DNA sequences in an auxin-regulated gene of soybean [J]. Plant Mol Biol.,1993,21:1147-1150
64. Naomi SS, Ichiro M , Shigeo N, et al. Constitutive promoters available for transgene expression instead of CaMV35S RNA promoter: Arabidopsis promoters of tryptophan synthase protein β subunit and phytochrome B [J]. Plant Biotech, 2002,19:19-22
65. Nitz I, Berkefeld H, Puzio PS. Pyk10, a seedling and root specific gene and promoter from Arabidopsis thaliana. Grundler FM [J]. Plant Sci. 2001, 161:337-346
66. Nagao RT, Goekjian VH, et al. Identification of protein-binding DNA sequences in an auxin-regulated gene of soybean [J]. Plant Mol Biol., 1993, 21:1147-1150
67. Orford SJ, Timmis JN. Abundant mRNAs specific to the developing cotton fiber [J]. Theor. Appl.Genet, 1997, 94: 909-918
68. Pear JR, Kawagoe Y, Schreckengost WE. Higher plant contains homologs of the bacterial celA gene encoding the catalytic subunit of cellulose synthase [J]. Proc Natl.Acad.Sci.USA, 1996, 93: 12637-12642
69. Prestridge DS. SIGNAL SCAN: A computer program that scans DNA sequences for eukaryotic transcriptional elements [J]. CABIOS, 1991, 7, 203-206
70. Preuss ML, Kovar DR, Lee YR, et al. A plant-specific kinesin binds to actin microfilaments and interacts with cortical microtubules in cotton fibers. Plant Physiol., 2004, 136: 3945-55
71. Robinson DJ. Environmental risk assessment of release of transgenic plants containing virus-derived inserts [J].Transgen. Res.,1996,5:359-341
72. Rojas A, Almoguera C, Carranco R, et al. Selective activation of the developmentally regulated hahsp17G1 promoter by heat stress transcription factors [J]. Plant Physiology,2002,129:1207-1211
73. Sa Q, Wang Y, LiW, et al. The promoter of an antifungal protein gene from Gastrodia elata confers tissue s pecific and fungus-in-ducible expression patterns and responds to both salicylic acid and jasmonic acid [J]. Plant Cell Rep, 2003, 22: 79-84
74. Shizuya H, Birren B, Kim U J, et al. Cloning and stable maintenance of 300 kilo base-pair fragments of human DNA in Escherchia coli using an F-factor-based vector [J]. Proc. Natl. Acad. Sic. USA., 1992, 89: 8794 - 8797
75. Song P, Allen RD. Identification of a cotton fiber-specific acyl carrier protein cDNA by differential display [J]. Biochim Biophy Acta, 1997, 1351: 305-312
76. Streatfield SJ, Beifuss KK, Brooks CA, et al. Analysis of the maize polyubiquitin-1 promoter heat shock elements and generation of promoter variants with modified expression characteristics [J].Transgenic Res,2004,13:299-312
77. T Kasrga, kR. Mitehelson. Expression of Potential defense response genes in cotton Physiological and Molecular [J]. Plant Pathology, 2000, 56:25-31
78. T Triglia, MG Peterson and DJ Kemp. A procedure for in vitro amplification of DNA segments that lie outside the boundaries of known sequences [J]. Nucl. Acids Res. 1988, 16, 81-86
79. Tada Y, Sakamoto M, Matsuoka M, et al. Expression of a monocot LHCP promoter in transgenic rice [J]. EMBO J, 1991, 10:1803-1808
80. Tan H, Creech RG, Jenkins JN, et al. Cloning and expression analysis of two cotton (Gossypium hirsutum L.) genes encoding cell wall proline-rich proteins. DNA, 2001, 12: 367-380
81. Thompson JD, Gibson TJ, Plewniak F, et al. The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools [J]. Nucleic Acids Research, 1997,24: 4876-4882
82. Wan CY, Wilkins TA. Isolation of multiple cDNA encoding the vacuolar H~+-ATPase subunit B from developing cotton (Gossypium hirsutum L.) [J]. Plant Physiol., 1994, 106:393-394
83. Wilkins TA. Vacuolar H+-ATPase 69-kilodalton catalytic subunit cDNA from developing cotton (Gossypium hirsutum L.) ovules [J]. Plant Physiol., 1993, 102: 679-680
84. Yoon IS, Park DH, Mori H, et al. Characterization of an auxin-inducible 1-Aminocyclopropane-1- carboxylate synthase gene,VR-ACS6,of mung bean(Vigna radiate(L)Wilczek)and hormonal interactions on the promoter activity in transgenic tobacco[J].Plant Cell Physiol,1999,40:431-435
85.Yun-Kuan Liang,et al.AtMYB61,an R2R3-MYB Transcription Factor Controlling Stomatal Aperture in Arabidopsis thaliana[J].Current Biology,2005,15:1201-1206
86.ZHAO Chang-ling,GUO Wei-ming,YANG Qing,et al.Cloning of Full-length F3H of Nanjinghongxu(Prunus mume) by gDNA-based TAIL-PCR[J].2005,12,159-165
87.樊荣,万小荣,张文涛.LE-ACS6启动子在LE-ACS6:GUS转基因拟南芥中的特异性[J].植物生理与分子生物学学报,2004,30(3):351-354
88.甘莉,吕金殿,汪沛洪,等.棉花黄萎病菌分泌的糖蛋白毒素与其致病力的关[J].中国农业科学,1995,28(2):58-65
89.高玉龙.棉花防卫反应相关基因类似物的克隆与特征分析和受黄萎病诱导表达的两个全长cDNA的获得[J].博士论文,2005
90.简桂良,马存,孙文姬.1995年华北棉区棉花黄萎病大发生及防治措施[J].植物保护.1996,(3):31-35
91.冯洁,孙文姬.中国棉花枯萎病生理小种的RAPD分析[J].菌物系统,2000,19:45-50
92.冯洁,孙文姬.棉花枯萎病菌生理小种的分子指纹分析[J].棉花学报,1999,11(5):230-234
93.冯洁.棉花与黄萎病菌互作机制研究现状及展望[J].棉花学报,1998,10(6):281-284
94.李桂兰,王文颇,臧少先,等。拟南芥黑芥子酶基因根特异性表达启动子的克隆[J].河北农业大学学报.2005,28(1)49-52
95.李娜,王世明,陈军,等.SEFA-PCR法克隆灵芝鲨烯合酶基因启动子及其序列分析[J].菌物学报2006,4,245-253
96.李姗姗,迟彦,李凌飞,等.启动子克隆方法研究进展[J].中国生物工程杂志.2005,25(7):9-16
97.李胜国,刘玉乐,朱锋.基因工程雄性不育烟草的获得[J].植物学报,1995,7(8):659-660
98.刘学堂,宋晓轩.棉花黄萎病菌的研究及最新进展[J].棉花学报,1998,10(1):6-13
99.刘学堂,张元恩.我国棉花黄萎病菌群体遗传分析研究初报[J].棉花学报,1997,24-31
100.刘巧泉,于恒秀,张文娟.水稻rbcS启动子控制的外源基因在转基因水稻中的特异性表达[J].植物生理与分子生物学学报,2005,31(3):247-251
101.刘大文,谢友菊,王守才.拟南芥菜花药绒毡层启动子的克隆和序列分析[J].作物学报,2000,26(4):406-409
102.路静,赵华燕,何奕等.高等植物启动子及其应用研究进展[J].自然科学进展,2004,14(8):856-862
103.廖飞雄,潘瑞炽.植物根特异性基因表达研究[J].植物学通报,1998,15(2):8-13
104.毛自朝,于秋菊,甄伟,等.果实专一性启动子驱动iPt基因在番茄中的表达及其对番茄果实发育的影响[J].科学通报,2002,47(6):444-448
105.任茂智.棉花nodunli-like和arfl基因及其启动子的分离和功能分析[D].2004年中国农科院博士学位论文
106.石磊岩,简桂良.我国棉花黄萎病研究现状与方向[J].植物保护学报,1998,25(2):103-107
107.石磊岩,李长兴,姚大瑞,等.棉花铃病发生关键因子及预防的研究[J].棉花学报,1995,7(3):175-178
108.孙晓红,陈明杰,潘迎捷.启动子克隆概述[J].食用菌学报,2002,9(3):57-62
109.吴梅花,张丽,牛一丁,等.拟南芥逆境胁迫诱导表达基因rd29A启动子的克隆与序列分析[J].华北农学报,2005,20(2):5-7
110.王雅琴.嗜麦芽假单胞菌热休克基因dnaK启动子的亚克隆及其在大肠杆菌中功能的研究[J].工业微生物,2001,31(3):29-33
111.张天真,何金龙,闵留芳,等.棉花对黄萎病的抗性遗传模式及抗(耐)病品种的选育技术[J].作物学报.2000,26(6):673-680
112.张晓国,刘玉乐,康良仪.水稻花药特异表达基因启动子的扩增及克隆[J].武汉大学学报(自然科学版),1997,43(4):480-484
113.周兆华,郭旺珍,潘家驹,等.我国棉花黄萎病菌群体的遗传变异分析[J]中国农业科学,1999,32(2):60-65
114.周雪荣,彭仁旺,方荣祥.表达核糖核酸酶基因的雄性不育油菜的获得[J].遗传学报,1997,24(6):531-534