臺灣博碩士論文加值系統

我們自水稻懸浮培養細胞所分泌的蛋白質當中，鑑定出兩種第三型的幾丁質酶 (Osm30和Osm34)。本研究不但針對這兩種幾丁質酶進行生化特性與生理功能的探討，也就水稻基因組資料庫中的32個第三型幾丁質酶成員，進行親緣系統樹的分析，結果顯示Osm30及Osm34分別歸類為第IIIa和IIIb亞型。
幾丁質酶屬於pathogenesis-relative (PR) 蛋白質家族成員，常參與植物的抗病防禦機制。當水稻細胞受到四種病原真菌：Rhizoctonia solani、Sarocladium oryzae、Gibberella fujikuroi和Bipolaris oryzae的感染時，Osm30和Osm34的表現量，均會被誘導增加。此外，Osm30和Osm34也受到甲基茉莉酸 (MeJA) 處理的誘導，而水楊酸 (SA) 的處理則不影響兩者基因的表現，顯示此兩種第三型幾丁質酶可能是經由JA/ethylene-dependent訊息傳導路徑，而非以SA-dependent路徑，來參與植物的抗病機制。
本研究培養水稻細胞的過程中也發現，每當更換新培養液後，水稻細胞的Osm30和Osm34基因都會被短暫誘導而增加表現，隨後又降回基礎表現量。此暫時性表現的情形可能受到繼代培養的攪拌動作 (agitation)，及更換新舊培養液時產生的滲透壓差的正向調控；此外，水稻細胞在培養過程中，會新生成並分泌一些抑制幾丁質酶表現的成分，以調節Osm34基因的表現。當水稻細胞以與逆境有關的植物生長調節劑ABA處理時，結果顯示其對於Osm30為正向的促進表現，對Osm34卻為負向的抑制表現，顯示此兩個幾丁質酶在不同的逆境下，其基因表現的調控機制可能有所不同。
綜合上述資料，Osm30和Osm34幾丁質酶在水稻細胞內平常只有基礎量的表現，當細胞感受到環境的變化，包括滲透勢改變、受到機械傷害或病原菌感染時，可能經由JA/ethylene訊息路徑，和/或由其他未知的因子，來調控共同調控其基因的表現，當水稻細胞已適應環境或威脅去除後，水稻細胞有另外的機制來節制幾丁質酶基因的表現。

Rice is one of the most important agricultural crops; biotic stresses are limiting crop productivity worldwide, hence studies aiming at identification and characterization of novel pathogen related gene(s) is gaining momentum. Chitinases belong to the pathogenesis-related protein families and play significant role in plant defense mechanism. In the present study, we identified several chitinases in rice suspension-culture, among Osm30 and Osm34 (class III) are selected for further investigations including biochemical properties, physiological functions, gene expression and regulation for future application. The phylogenetic trees of Osm30 and Osm34 with other 30 putative class III chitinases in the rice genome defines the evolutionary relationship and are classified into subclasses IIIa and IIIb respectively, indicating diverse functions in vivo. The phylogenetic analysis of Osm30 and Osm34 with other 32 putative class III chitinases in the rice genome revealed subclasses IIIa and IIIb, respectively. The expression of Osm30 and Osm34 genes was up-regulated in the presence of common rice pathogens, such as Rhizoctonia solani, Sarocladium oryzae, Gibberella fujikuroi and Bipolaris oryzae, ethylene and methyl jasmonate (MeJA), but not salicylic acid (SA). These results indicate that the expressions of these chitinases are JA/ethylene dependent and may participate in SA independent signal transduction pathway in the plant defense mechanism. Upon transfer of rice cells into fresh culture medium, Osm30 and Osm34 transcripts displayed a transient expression pattern in rice cells and subsequently reduce to basal expression level. This phenomenon may be due to mechanical agitation and osmotic change in the culture medium. In addition, rice cells secreted some inhibitory factors that regulated the Osm30 and Osm34 gene expression during culturing. It was interesting to note that ABA positively regulated the expression of Osm30 gene and inhibited Osm34. Taken together, the expression of Osm30 and Osm34 was basal level under normal culture conditions, the changes in the expression level of chitinase genes was observed after transferring into fresh culture medium. Thus rice cells sense the environmental alternation such as osmotic shock, agitation and pathogen infection probably via JA/ethylene-dependent pathway and/or other unidentified factors, to augment cellular defense mechanism.

目錄
中文摘要 1
英文摘要 2

第一章 前言 3
一、抗病蛋白質與植物抗病反應 3
二、幾丁質酶的分類 4
三、植物幾丁質酶的種類 4
四、植物幾丁質酶的受質 5
五、幾丁質酶的功能 6
六、植物幾丁質酶的基因表現與調節 7
七、抗病基因的應用 8
第一章之圖、表 9
表一、病原菌相關蛋白質的分類 9
表二、醣苷水解酵素第18和19家族幾丁質酶的差異 10

第二章 水稻 (Oryza sativa L.) 分泌性蛋白質Osm30的生化及功能分析 11
第一節 摘要 11
第二節 緒言 13
第三節 材料與方法 14
一、植物材料與培養 14
(一) 水稻細胞的懸浮培養方法 14
(二) 真菌感染試驗 14
(三) 各種植物生長調節劑的處理 15
二、水稻分泌性蛋白質的收集與蛋白質N端胺基酸微定序 15
(一) 水稻分泌性蛋白質的收集與定量 15
(二) 蛋白質N端胺基酸微定序 16
三、幾丁質酶的膠體活性分析 18
四、Osm30 cDNA的PCR選殖及序列分析 18
(一) λDNA之萃取 18
(二) 聚合酶鏈反應 19
(三) 限制酵素反應 20
(四) DNA的電泳分析 20
(五) 以離心管柱純化瓊脂膠體內的DNA 20
(六) DNA之接合反應 21
(七) 細菌的轉形 21
(八) 鹼性溶菌法純化細菌質體DNA 22
(九) 大腸桿菌菌體的保存 23
五、北方轉漬分析 23
(一) RNA之萃取與純化 23
(二) RNA的電泳分析 24
(三) RNA的轉漬與分析 25
六、Osm30多株抗體的製備 27
(一) 重組蛋白質的萃取與純化 27
(二) 多株抗體的製備 29
七、西方免疫轉漬分析 31
八、細菌生長曲線之測定 32
第四節 結果 32
一、水稻細胞分泌性蛋白質的收集與幾丁質酶的活性分析 32
二、Osm30基因之選殖 33
三、Osm30在水稻細胞的生理表現情形 34
四、真菌感染會增加Osm30表現 34
五、Osm30重組蛋白質會抑制細菌生長 35
六、Osm30基因的表現受到MeJA和ABA之調控 35
第五節 討論 36
第二章之圖、表 41
圖一、水稻細胞分泌性蛋白質的分析 41
圖二、Osm30的完整基因序列，與轉譯出之胺基酸序列 43
圖三、Osm30在水稻懸浮培養細胞中的表現情形 44
圖四、真菌感染後Osm30的表現情形 46
圖五、細菌轉形Osm30基因與GFP基因後的生長曲線比較 48
圖六、各種植物生長調節劑對Osm30基因表現之影響 50
圖七、Osm30在水稻植株中的表現情形 51

第三章 水稻 (Oryza sativa L.) 分泌性蛋白質Osm34的基因選殖及第三型幾丁質酶基因家族的親緣關係分析 52
第一節 摘要 52
第二節 緒言 54
第三節 材料與方法 56
一、植物材料與培養 56
二、Osm34蛋白質的收集與N端微定序 56
三、Osm34 cDNA的Nested PCR選殖與序列分析 56
四、水稻基因組DNA的分離與南方轉漬分析 56
(一) 基因組DNA之純化 56
(二) 南方轉漬分析 57
五、水稻第III型幾丁質酶的胺基酸序列之比對分析 58
第四節 結果 58
一、Osm34蛋白質的收集與N端胺基酸微定序 58
二、Osm34基因的選殖與分析 59
三、Osm34與其他植物幾丁質酶之胺基酸序列比對 59
四、水稻基因組中Osm34相似基因的數目 60
五、水稻第III型幾丁質酶基因家族的系統分析 60
第五節 討論 62
第三章之圖、表 66
圖一、 Osm34的cDNA與胺基酸序列暨其與其他植物第III型幾丁質酶的胺基酸序列比對結果 66
圖二、南方轉漬分析結果 69
圖三、水稻第III型幾丁質酶胺基酸序列的親緣系統樹 70
圖四、水稻第III型幾丁質酶的胺基酸多序列比對結果 71
表一、水稻第III型幾丁質酶的生化特性 74
表二、水稻第III型幾丁質酶的胺基酸序列相似度分析結果 76

第四章 水稻 (Oryza sativa L.) 細胞分泌性第III b亞型幾丁質酶Osm34的基因表現與調節 78
第一節 摘要 78
第二節 緒言 80
第三節 材料與方法 82
一、植物材料與培養 82
二、北方轉漬分析 82
三、Osm34多株抗體的製備 82
四、西方免疫轉漬分析 83
第四節 結果 84
一、Osm34抗體的製備與效價分析 84
二、Osm34在水稻細胞懸浮培養過程中的表現情形 84
三、真菌感染對Osm34基因表現的影響情形 85
四、水稻細胞內Osm34暫時性表現的情形 85
五、組織培養的過程對Osm34表現之影響情形 86
六、培養液酸鹼值的變化情形 86
七、培養液滲透勢的變化情形 87
八、舊培養液的成分對Osm34基因表現的影響情形 87
九、植物生長調節劑對Osm34基因表現的影響情形 88
十、蛋白質生合成抑制劑對Osm34基因表現的影響情形 89
第五節 討論 90
真菌感染會誘導Osm34基因的表現 90
Osm34在水稻細胞中呈現暫時性表現的情形 91
水稻細胞培養前期，Osm34表現量增加 92
水稻細胞培養後期，Osm34表現量下降 95
總結 96
第四章之圖、表 97
圖一、水稻細胞Osm34幾丁質酶的抗體效價分析 97
圖二、Osm34在水稻細胞懸浮培養過程中的表現情形 99
圖三、真菌感染後Osm34的表現情形 101
圖四、在繼代培養過程中，水稻細胞Osm34表現情形 103
圖五、繼代培養的攪拌動作產生的機械傷害，會影響水稻細胞內Osm34表現量 105
圖六、水稻細胞培養過程中，培養液酸鹼值的變化與Osm34的表現情形 106
圖七、水稻細胞培養液滲透勢的變化與Osm34的表現情形 108
圖八、培養過水稻細胞的培養液會抑制Osm34的表現 110
圖九、各種植物生長調節劑與乙烯抑制劑對水稻細胞Osm34基因表現之影響 112
圖十、蛋白質生合成抑制劑對水稻細胞內Osm34表現的影響 114
圖十一、各種生物性和非生物性因子對Osm34基因表現可能的調控路徑 115

第五章 綜合討論 116
水稻細胞會表現多種幾丁質酶 116
幾丁質酶的應用 117
第五章之圖、表 119
表一、水稻懸浮培養細胞所分泌的幾丁質酶之生化特性 119

參考文獻 120
附錄 127
附錄一、縮寫對照表 127
附錄二、中英對照表 131
附錄三、微生物培養基配方 133
附錄四、Murashige-Skoog培養液配方 134
附錄五、RNA電泳所使用的藥品配方 135
附錄六、PCR primers 136
附錄七、水稻細胞分泌之第三型幾丁質酶Osm34的序列資料 137
附錄八、水稻細胞分泌之第一型幾丁質酶OsmChiI-34的序列資料 139
附錄九、Osm34/pBS 和Osm34/pET24a圖譜 141
附錄十、Osm30/pBS 和Osm30/pET24a圖譜 142
附錄十一、GFP的核苷酸序列與GFP/pET24b圖譜 143
附錄十二、細菌轉形Osm30、Osm34和GFP基因後的生長曲線 144
附錄十三、Osm30基因啟動子序列 145
附錄十四、Osm34基因啟動子序列 146

參考文獻
李嘉雯 1997 水稻懸浮培養細胞33-kDa分泌性蛋白質的生物功能鑑定與基因選殖之研究 國立台灣師範大學生物系碩士論文
Abe H, Yamaguchi-Shinozaki K, Urao T, Iwasaki T, Hosokawa D and Shinozaki K. 1997. Role of arabidopsis MYC and MYB homologs in drought- and abscisic acid-regulated gene expression. Plant Cell. 19: 1859-1868.
Bairoch A, Bucher P and Hofmann K. 1997. The PROSITE database, its status in 1997. Nucleic Acids Res. 25: 217-221.
Beintema JJ. 1994. Structural features of plant chitinases and chitin-binding proteins. FEBS Lett. 350: 159-163.
Bokma E, Rozeboom HJ, Sibbald M, Dijkstra BW and Beintema JJ. 2002. Expression and characterization of active site mutants of hevamine, a chitinase from the rubber tree Hevea brasiliensis. Eur. J. Biochem. 269: 893-901.
Boter M, Ruiz-Rivero O, Abdeen A and Prat S. 2004. Conserved MYC transcription factors play a key role in jasmonate signaling both in tomato and Arabidopsis. Genes Dev. 18: 1577-1591.
Boyle B and Brisson N. 2001. Repression of the Defense Gene PR-10a by the Single-Stranded DNA Binding Protein SEBF. Plant Cell. 13: 2525–2538.
Brameld KA, Shrader WD, Imperiali B and Goddard WA 3rd. 1998. Substrate assistance in the mechanism of family 18 chitinases: theoretical studies of potential intermediates and inhibitors. J. Mol. Biol. 280: 913-923.
Brunner FSA, Fritig B and Legrand M. 1998. Substrate specificities of tobacco chitinases. Plant J. 14: 225-234.
Chan MT, Chao YC, Yu SM. 1994. Novel gene expression system for plant cells based on induction of alpha-amylase promoter by carbohydrate starvation.
J. Biol. Chem. 269: 17635-17641.
Chen C and Chen Z. 2000. Isolation and characterization of two pathogen- and salicylic acid-induced genes encoding WRKY DNA-binding proteins from tobacco. Plant Mol. Biol. 42: 387-396.
Chen W, Provart NJ, Glazebrook J, Katagiri F, Chang HS, Eulgem T, Mauch F, Luan S, Zou G, Whitham SA, Budworth PR, Tao Y, Xie Z, Chen X, Lam S, Kreps JA, Harper JF, Si-Ammour A, Mauch-Mani B, Heinlein M, Kobayashi K, Hohn T, Dangl JL, Wang X and Zhu T. 2002. Expression profile matrix of Arabidopsis transcription factor genes suggests their putative functions in response to environmental stresses. Plant Cell 14: 559-574
Clendennen SK, Lopez-Gomez R, Gomez-Lim M, Arntzen CJ and May GD. 1998. The abundant 31-kilodalton banana pulp protein is homologous to class-III acidic chitinases. Phytochemistry. 47: 613-619.
Collinge DB, Kragh KM, Mikkelsen JD, Nielsen KK, Rasmussen U and Vad K. 1993. Plant chitinases. Plant J. 3: 31-40.
Datta K, Tu J, Oliva N and Ona I. 2001. Enhanced resistance to sheath blight by constitutive expression of infection-related rice chitinase in transgenic elite indica rice cultivars. Plant Science 160: 405-414.
de Jong AJ, Cordewener J, Lo Schiavo F, Terzi M, Vandekerckhove J, van Kammen A and De Vries SC. 1992. A carrot somatic embryo mutant is rescued by chitinase. Plant Cell 4: 425-433.
Denekamp M. and Smeekens SC. 2003. Integration of wounding and osmotic stress signals determines the expression of the AtMYB102 transcription factor gene. Plant Physiol. 132:1415-1423.
Durrant W.E. and Dong X. 2004. Systemic acquired resistance. Annu. Rev. Phytopathol. 42: 185-209.
Ezcurra I, Wycliffe P, Nehlin L, Ellerstrom M and Rask L.2000. Transactivation of the Brassica napus napin promoter by ABI3 requires interaction of the conserved B2 and B3 domains of ABI3 with different cis-elements: B2 mediates activation through an ABRE, whereas B3 interacts with an RY/G-box. Plant J. 24: 57-66
Farmer EE and Ryan CA. 1990. Interplant communication: airborne methyl jasmonate induces synthesis of proteinase inhibitors in plant leaves. Proc. Natl. Acad. Sci. USA. 87: 7713-7716.
Felix G, Regenass M and Boller T. 2000. Sensing of osmotic pressure changes in tomato cells. Plant Physiol. 124: 1169-1179.
Fujimoto SY, Ohta M, Usui A, Shinshi H and Ohme-Takagi M. 2000. Arabidopsis ethylene-responsive element binding factors act as transcriptional activators or repressors of GCC box-mediated gene expression. Plant Cell. 12: 393-404.
Gal SW, Choi JY, Kim CY, Cheong YH, Choi YJ, Bahk JD, Lee SY and Cho MJ. 1998. Cloning of the 52-kDa chitinase gene from Serratia marcescens KCTC2172 and its proteolytic cleavage into an active 35-kDa enzyme. FEMS Microbiol. Lett. 160: 151-158.
Grover A and Gowthaman. 2003. Strategies for development of fungus-resistant transgenic plants. Current Science. 84: 330-340.
Hamel F, Boivin R, Tremblay C and Bellemare G. 1997. Structural and evolutionary relationships among chitinases of flowering plants. J. Mol. Evol. 44: 614–624.
Henrissat B. 1991. A classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem. J. 280: 309-316.
Hong JK and Hwang BK. 2002. Induction by pathogen, salt and drought of a basic class II chitinase mRNA and its in situ localization in pepper (Capsicum annuum). Physiol. Plant. 114: 549-558.
Itzhaki H, Maxson JM and Woodson WR. 1994. An ethylene-responsive enhancer element is involved in the senescence-related expression of the carnation glutathione-S-transferase (GSTI) gene. Proc. Natl. Acad. Sci. USA. 91: 8925-8929.
Kasprzewska A. 2003. Plant chitinases--regulation and function. Cell Mol. Biol. Lett. 8: 809-824.
Kim YK, Baek JM, Park HY, Choi YD and Kim SI. 1994. Isolation and characterization of cDNA clones encoding class I chitinase in suspension cultures of rice cell. Biosci. Biotechnol. Biochem. 58: 1164-1166.
Kim CY, Gal SW, Choe MS, Jeong SY, Lee SI, Cheong YH, Lee SH, Choi YJ, Han C, Kang YH and Cho MJ. 1998. A new class II rice chitinase, Rch2, whose induction by fungal elicitor is abolished by protein phosphatase 1 and 2A inhibitor. Plant Mol. Biol. 37: 523-534.
Kunkel BN and Brooks DM. 2002. Cross talk between signaling pathways in pathogen defense. Current Opinion in Plant Biology. 5: 325-331.
Laemmli UK. 1970. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature. 227: 680-685.
Lee SC, Kim YJ and Hwang BK. 2001. A pathogen-induced chitin-binding protein gene from pepper: its isolation and differential expression in pepper tissues treated with pathogens, ethephon, methyl jasmonate or wounding. Plant Cell Physiol. 42: 1321-1330.
Lee SC and Hwang BK. 2005. Induction of some defense-related genes and oxidative burst is required for the establishment of systemic acquired resistance in Capsicum annuum. Planta. 221: 790-800.
Li C-W and Wang Y-C. 2006. Biochemical and functional characterization of rice (Oryza sativa L.) Osm30 secretory protein — A Class III Chitinase. Bioformosa 41 (1): 31-43.
Li C-W and Wang Y-C. 2006. Gene cloning of rice (Oryza sativa L.) Osm34 secretory protein and phylogenetic analysis of the class III chitinase gene family. Bioformosa 41 (1): 44-57.
Mauch F, Mauch-mani FB and Boller T. 1988. Antifungal hydrolases in pea tissue. II. Inhibition of fungal growth by combination of chitinase and -1,3-glucanase. Plant Physiol. 88: 936-942.
Meins F, Fritig B, Linthorst HJM, Mikkelsen JD, Neuhaus J-M and Ryals J. 1994. Plant chitinase genes. Plant Mol. Biol. Rep. 12: 22–28.
Melchers LS, Apotheker-de Groot M, van der Knaap JA, Ponstein AS, Sela-Buurlage MB, Bol JF, Cornelissen BJ, van den Elzen PJ and Linthorst HJ. 1994. A new class of tobacco chitinases homologous to bacterial exo-chitinases displays antifungal activity. Plant J. 5: 469-480.
Morimoto K, Karita S, Kimura T, Sakka K, and Ohmiya K. 1997. Cloning, sequencing, and expression of the gene encoding Clostridium paraputrificum chitinase ChiB and analysis of the functions of novel cadherin-like domains and a chitin-binding domain. J. Bacteriol. 179: 7306-7314.
Montgomery J, Goldman S, Deikman J, Margossian L and Fischer RL. 1993. Identification of an ethylene-responsive region in the promoter of a fruit ripening gene. Proc. Natl. Acad. Sci. USA. 90: 5939-5943
Murashige T and Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15: 473-479.
Nagasaki H, Yamamoto K, Shomura A, Koga-ban Y, Takasuga A, Yano M, Minobe Y and Sasaki T. 1997. Rice class III chitinase homologues isolated by random cloning of rice cDNAs. DNA Res. 4: 379-385.
Neuhaus J-M, Fritig B, Linthorst HJM, Meins F, Mikkelsen JD and Ryals J. 1996. A revised nomenclature for chitinase genes. Plant Mol. Biol. Rep. 14: 102-104.
Nishizawa Y, Kawakami A, Hibi T, He D-Y, Shibuya N and Minami E. 1999. Regulation of the chitinase gene expression in suspension-cultured rice cells by N-acetylchitooligosaccharides: differences in the signal transduction pathways leading to the activation of elicitor-responsive genes. Plant Mol. Biol. 39: 907-914.
Park H-Y, Pan C-H, So M-Y, Ahn J-H, Jo D-H and Kim S-I. 2001. Purification, Characterization, and cDNA Cloning of Rice Class III Chitinase. Mol. Cells. 13: 69-76.
Park CH, Kim S, Park JY, Ahn IP, Jwa NS, Im KH and Lee YH. 2004. Molecular characterization of a pathogenesis-related protein 8 gene encoding a class III chitinase in rice. Mol Cells. 17: 144-150.
Patil RS, Ghormade V and Deshpande MV. 2000. Chitinolytic enzymes: an exploration. Enzyme Microb. Technol. 26: 473-483.
Peumans WJ, Proost P, Swennen RL and Van Damme EJ. 2002. The abundant class III chitinase homolog in young developing banana fruits behaves as a transient vegetative storage protein and most probably serves as an important supply of amino acids for the synthesis of ripening-associated proteins. Plant Physiol. 130: 1063-1072.
Pieterse CM and van Loon LC. 1999. Salicylic acid-independent plant defence pathways. Trends Plant Sci. 4: 52-58.
Rakwal R, Yang G and Komatsu S. 2004. Chitinase induced by jasmonic acid, methyl jasmonate, ethylene and protein phosphatase inhibitors in rice. Mol. Biol. Rep. 31: 113-119.
Ryals JA, Neuenschwander UH, Willits MG, Molina A, Steiner HY and Hunt MD. 1996. Systemic acquired resistance. Plant Cell. 8: 1809-1819.
Sambrook J, Maniatis T and Fristsch EF. 1989. Molecular Cloning: A Library Manual, 2nd ed. Cold Spring Harbor Laboratory press, Cold Spring Harbor, New York.
Selitrennikoff CP. 2001. Antifungal proteins. Applied and Environmental Microbiology. 67: 2883-2894.
Shirzadegan M, Christie P and Seemann J. 1991. An efficient method for isolation of RNA from tissue cultured plant cells. Nucleic Acids Res 19: 6055-6056.
Suzukawa K, Yamagami T, Ohnuma T, Hirakawa H, Kuhara S, Aso Y and Ishiguro M. 2003. Mutational analysis of amino acid residues involved in catalytic activity of a family 18 chitinase from tulip bulbs. Biosci. Biotechnol. Biochem. 67: 341-346.
Takakura Y, Ito T, Saito H, Inoue T, Komari T and Kuwata S. 2000. Flower-predominant expression of a gene encoding a novel class I chitinase in rice (Oryza sativa L.). Plant Mol. Biol. 42: 883-897.
Theis T. and Stahl U. 2004. Antifungal protein: targets, mechanism and prospective applications. Cell. Mol. Life Sci. 61: 437-455.
Thomashow MF. 1999. Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Annu. Rev. Plant Physiol. Plant Mol. Biol. 50: 571–599.
Thomma BPHJ, Eggermont K, Penninckx IAMA, Mauch-Mani B, Vogelsang R, Cammue BPA, Broekaert WF. 1998. Separate jasmonate-dependent and salicylate-dependent defense-response pathways in arabidopsis are essential for resistance to distinct microbial pathogens. Proc. Natl. Acad. Sci. USA. 95: 15107-15111.
Thompson JD, Higgins DG and Gibson TJ. 1994. ClustalW: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22: 4673–4680.
Titarenko E, Rojo E, Leó J and Sánchez-Serrano JJ. 1997. Jasmonic acid-dependent and -independent signaling pathways control wound-induced gene activation in Arabidopsis thaliana. Plant Physiol. 115: 817–826.
Truong NH, Park SM, Nishizawa Y, Watanabe T, Sasaki T and Itoh Y. 2003. Structure, heterologous expression, and properties of rice (Oryza sativa L.) family 19 chitinases. Biosci. Biotechnol. Biochem. 67: 1063-1070.
Van Loon LC. and van Strien EA. 1999. The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins. Physiol. Mol. Plant Pathol. 55: 85-97.
Verwoerd TC, Dekker BM and Hoekema A. 1989. A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res. 17: 2362-2362.
Watanabe T, Kobori K, Miyashita K, Fujii T, Sakai H, Uchida M and Tanaka H. 1993. Identification of glutamic acid 204 and aspartic acid 200 in chitinase A1 of Bacillus circulans WL-12 as essential residues for chitinase activity. J. Biol. Chem. 268:18567-18572.
Wu L, Ueda T, Messing J. 1995. The formation of mRNA 3'-ends in plants. Plant J. 8: 323-329.
Xu Y, Zhu Q, Panbangred W, Shrasu K and Lamb C. 1996. Regulation, expression and function of a new basic chitinase gene in rice (Oryza sativa L.). Plant Mol. Biol. 30: 387-401.
Yamagami T, Mine Y and Ishiguro M. 1998. Complete amino acid sequence of chitinase-a from bulbs of gladiolus (Gladiolus gandavensis). Biosci. Biotechnol. Biochem. 62: 386-389.
Yamagami T, Tsutsumi K and Ishiguro M. 2000. Cloning, sequencing, and expression of the tulip bulb chitinase-1 cDNA. Biosci. Biotechnol. Biochem. 64: 1394-1401.
Yeh S, Moffatt BA, Griffith M, Xiong F, Yang DS, Wiseman SB, Sarhan F, Danyluk J, Xue YQ, Hew CL, Doherty-Kirby A and Lajoie G. 2000. Chitinase genes responsive to cold encode antifreeze proteins in winter cereals. Plant Physiol. 124: 1251-1264.
Yen SK, Chung MC, Chen PC and Yen HE. 2001. Environmental and developmental regulation of the wound-induced cell wall protein WI12 in the halophyte ice plant. Plant Physiol. 127: 517-528.
Yu D, Chen C and Chen Z. 2001. Evidence for an important role of WRKY DNA binding proteins in the regulation of NPR1 gene expression. Plant Cell 13: 1527-1540.
Yu S-M, Kuo Y-H, Sheu G, Sheu Y-J and Liu L-F. 1991. Metabolic derepression of -amylase gene expression in suspension-cultured cells of rice. J. Biol. Chem. 266: 21131-21137.
Zhong R, Kays SJ, Schroeder BP and Ye Z-H. 2002. Mutation of a Chitinase-Like Gene Causes Ectopic Deposition of Lignin, Aberrant Cell Shapes, and Overproduction of Ethylene. Plant Cell. 14: 165-179.
Zhu Q and Lamb CJ. 1991. Isolation and characterization of a rice gene encoding a basic chitinase. Mol. Gen. Genet. 226: 289-296.