几丁聚糖对青花菜生长生理机制及品质的影响研究
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摘要
壳聚糖是甲壳素脱去乙酰基后得到的一种高分子阳离子多聚糖，具有安全无毒，抑菌、可食和生物降解等多种特性，壳聚糖几其衍生物已被广泛地应用于医药、农业、食品、化工、环保等多个领域。由于壳聚糖的分子结构与植物细胞壁结构相似，因而作为新型的植物生长调节剂在作物上应用具有重要的意义。
1、利用不同浓度几丁聚糖对青花菜种子包衣，来研究几丁聚糖对青花菜幼苗生理指标的影响。通过本试验研究表明，不同浓度的几丁聚糖包衣对青花菜幼苗的还原糖、地上部分鲜重、地下部分鲜重及对磷钾的吸收无明显的影响，而显著地增加其中的叶绿素含量、蛋白质含量、地上部分干重、地下部分干重、适当的浓度能增加幼苗叶片中总糖含量和氮的含量；几丁聚糖包衣能显著增加青花菜叶片中游离氨的含量、硝酸还原酶、谷氨酰股合成酶和谷氨酸脱氢酶的活性；几丁聚糖种子包衣能显著增强青花菜幼苗叶片的光合作用和光化学效率。
2、青花菜品种“株绿”和“山水”在不同浓度盐胁迫下施用几丁聚糖，研究几丁聚糖对青花菜耐盐胁迫的影响。研究表明，几丁聚糖明显地增加青花菜种子的发芽率、发芽指数、种子活力指数和耐盐指数，特别是对发芽率较低的种子，几丁聚糖对其作用更加明显；在盐胁迫下，施用几丁聚糖明显地降低青花菜叶片中的MDA含量和降低叶片膜的透性，施用适当的浓度则显著地增加叶片中SOD、POD、PPO的活性，减缓逆境对叶片的伤害，增强植株的抗性，叶片中的脯氨酸含量不断地提高、蛋白质含量不断降低、还原糖基本没有变化，蔗糖、可溶性总糖在盐浓度较低时有下降趋势，后不断地增高，可见植物在受到伤害时分解体内大分子物质生成小分子物质，来增加自身的抗性。
3、通过在青花菜生长期间内喷施几丁聚糖，来研究几丁聚糖对青花菜花球内的酚类物质和品质的影响。研究表明，本研究中，不同浓度几丁聚糖喷施对青花菜花球的叶绿素、类黄酮、绿原酸、多酚总量、可溶性蛋白及可溶性总糖的含
浙江大学硕卜学位论文(2004)
量有显著增加，其中以浓度为40mgL一’影响最为显著，花球中的叶绿素、类黄
酮、绿原酸、可溶性总糖、可溶性蛋白分别较对照提高64.4%、282.5%、35.6%、
16.2%、22.7%，苯丙氨酸解氨酶(队L)、多酚氧化酶(PPO)是酚类物质合成
中关键酶，花球内酚类物质合成与苯丙氨酸解氨酶的活性没有一定的相关性，而
与多酚氧化酶的活性呈正相关。
Chitosan, a nature high molecular weight cationic polysaccharide, is normally obtained by the alkaline deacetylation of chitin. Chitosan and its derivatives have a wide range of applications due to their low toxic ,antibacterial activity,edibility and biodegradability, They have been employed, for example, to solve numerous problems in medical , agriculture ,foodstuff environmental and chemical engineering. Because the molecular structure of chitosan is similar with the structure of cell wall, it has great significance to be applied in plants as a new plant growth regulator.
1.Seeds of Broccoli were film-coated with chitosan of different concentration.Our results showed that film-coating with chitosan had no significant effect on fresh weight of ground parts and roots of Broccoli seedlings.And there was no significant effect on the absorption of N and K.But film-coating significantly improved the content of chlorophyll and protein in seedling leaves. It also improved the dry weight of ground parts and roots of cauliflower seedlings. Film-coating with chitosan of appropriate concentration can increase the content of total soluble sugar and N in seedling leaves. Film-coating with chitosan significantly improved the content of free ammonium ,the activity of NR, GDH and GS, and film-coating markedly increased photosynthetic captivity and photochemical efficiency of PS II reaction center( PS II).
2.We sprayed chitosan on "zhulv"and "shanshui"-two variation of Broccoli under salt stress to investigate the effect of chitosan on salt tolerant of Broccoli.The results showed chitosan significantly increased the germination percentage, germination index and vigor index of Broccoli seeds and increased the salt tolerance index of Broccoli seedings. The effect was more significant on seeds of low germination percentage.Under salt stress ,chitosan significantly decreased the MDA content and membrane permeability of Broccoli leaves.Activities of SOD,POD and PPO in Broccoli leaves was significantly promoted treated with chitosan . Proline content in leaves continuously increased ,while protein content continuously decreased. Reducing sugar content changed little .Content of sucrose and total
soluble sugar decreased a little under low salt concentration and increased late with the increase of salt concentration.It indicated that plants decompose macromolecular substances to small molecule substance to improve their tolerance captivity under stress.
3.Spraying with chitosan of different concentration significantly increased the content of chlorophyll ,anthoxanthin ,chlorogentic acid, polyphenal ,soluble protein and total soluble sugar of Broccoli flowers. 40 mg L-l chitosan showed the most remarkable effet,compared to control, the content of
chlorophyll ,anthoxanthin ,chlorogentic acid, total soluble sugar and soluble protein of treated Broccoli flowers increased by 64.4%,282.5%,35.6%,16.2% 22.7% respectively.PAL and PPO are key enzymes in synthesize of phenolics.No relation was found between phenolics synthesize and PAL activity in Broccoli flowers,but it showed positive relation between phenolics synthesize and PPO activity.
1、利用不同浓度几丁聚糖对青花菜种子包衣，来研究几丁聚糖对青花菜幼苗生理指标的影响。通过本试验研究表明，不同浓度的几丁聚糖包衣对青花菜幼苗的还原糖、地上部分鲜重、地下部分鲜重及对磷钾的吸收无明显的影响，而显著地增加其中的叶绿素含量、蛋白质含量、地上部分干重、地下部分干重、适当的浓度能增加幼苗叶片中总糖含量和氮的含量；几丁聚糖包衣能显著增加青花菜叶片中游离氨的含量、硝酸还原酶、谷氨酰股合成酶和谷氨酸脱氢酶的活性；几丁聚糖种子包衣能显著增强青花菜幼苗叶片的光合作用和光化学效率。
2、青花菜品种“株绿”和“山水”在不同浓度盐胁迫下施用几丁聚糖，研究几丁聚糖对青花菜耐盐胁迫的影响。研究表明，几丁聚糖明显地增加青花菜种子的发芽率、发芽指数、种子活力指数和耐盐指数，特别是对发芽率较低的种子，几丁聚糖对其作用更加明显；在盐胁迫下，施用几丁聚糖明显地降低青花菜叶片中的MDA含量和降低叶片膜的透性，施用适当的浓度则显著地增加叶片中SOD、POD、PPO的活性，减缓逆境对叶片的伤害，增强植株的抗性，叶片中的脯氨酸含量不断地提高、蛋白质含量不断降低、还原糖基本没有变化，蔗糖、可溶性总糖在盐浓度较低时有下降趋势，后不断地增高，可见植物在受到伤害时分解体内大分子物质生成小分子物质，来增加自身的抗性。
3、通过在青花菜生长期间内喷施几丁聚糖，来研究几丁聚糖对青花菜花球内的酚类物质和品质的影响。研究表明，本研究中，不同浓度几丁聚糖喷施对青花菜花球的叶绿素、类黄酮、绿原酸、多酚总量、可溶性蛋白及可溶性总糖的含
浙江大学硕卜学位论文(2004)
量有显著增加，其中以浓度为40mgL一’影响最为显著，花球中的叶绿素、类黄
酮、绿原酸、可溶性总糖、可溶性蛋白分别较对照提高64.4%、282.5%、35.6%、
16.2%、22.7%，苯丙氨酸解氨酶(队L)、多酚氧化酶(PPO)是酚类物质合成
中关键酶，花球内酚类物质合成与苯丙氨酸解氨酶的活性没有一定的相关性，而
与多酚氧化酶的活性呈正相关。
Chitosan, a nature high molecular weight cationic polysaccharide, is normally obtained by the alkaline deacetylation of chitin. Chitosan and its derivatives have a wide range of applications due to their low toxic ,antibacterial activity,edibility and biodegradability, They have been employed, for example, to solve numerous problems in medical , agriculture ,foodstuff environmental and chemical engineering. Because the molecular structure of chitosan is similar with the structure of cell wall, it has great significance to be applied in plants as a new plant growth regulator.
1.Seeds of Broccoli were film-coated with chitosan of different concentration.Our results showed that film-coating with chitosan had no significant effect on fresh weight of ground parts and roots of Broccoli seedlings.And there was no significant effect on the absorption of N and K.But film-coating significantly improved the content of chlorophyll and protein in seedling leaves. It also improved the dry weight of ground parts and roots of cauliflower seedlings. Film-coating with chitosan of appropriate concentration can increase the content of total soluble sugar and N in seedling leaves. Film-coating with chitosan significantly improved the content of free ammonium ,the activity of NR, GDH and GS, and film-coating markedly increased photosynthetic captivity and photochemical efficiency of PS II reaction center( PS II).
2.We sprayed chitosan on "zhulv"and "shanshui"-two variation of Broccoli under salt stress to investigate the effect of chitosan on salt tolerant of Broccoli.The results showed chitosan significantly increased the germination percentage, germination index and vigor index of Broccoli seeds and increased the salt tolerance index of Broccoli seedings. The effect was more significant on seeds of low germination percentage.Under salt stress ,chitosan significantly decreased the MDA content and membrane permeability of Broccoli leaves.Activities of SOD,POD and PPO in Broccoli leaves was significantly promoted treated with chitosan . Proline content in leaves continuously increased ,while protein content continuously decreased. Reducing sugar content changed little .Content of sucrose and total
soluble sugar decreased a little under low salt concentration and increased late with the increase of salt concentration.It indicated that plants decompose macromolecular substances to small molecule substance to improve their tolerance captivity under stress.
3.Spraying with chitosan of different concentration significantly increased the content of chlorophyll ,anthoxanthin ,chlorogentic acid, polyphenal ,soluble protein and total soluble sugar of Broccoli flowers. 40 mg L-l chitosan showed the most remarkable effet,compared to control, the content of
chlorophyll ,anthoxanthin ,chlorogentic acid, total soluble sugar and soluble protein of treated Broccoli flowers increased by 64.4%,282.5%,35.6%,16.2% 22.7% respectively.PAL and PPO are key enzymes in synthesize of phenolics.No relation was found between phenolics synthesize and PAL activity in Broccoli flowers,but it showed positive relation between phenolics synthesize and PPO activity.
引文
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63. Hadwiger LA. Inducyion of phenylalanine ammonia lyase and pisatin by photo-sensitive psoralen compounds Plant Physiol, 1972,49:779-782
64. Helander, IM, Nurmiaho-Lassila EL, Ahvenainen R, et al. Chitosan disrupts the barrier properties of the outer membrane of Gram-negative bacteria
International Journal of Food Microbiology, 2001,71:235-244
65. Hadwiger LA. Loschke DC. Molecular communication in host-parasite interactions:hexosamine polymers(chitosan) as regulator compounds in race-specific and other interactions Phytopathology, 1981a, 71:756-782
66. Helander, IM, Nurmiaho-Lassila EL, hhvenainen R, et al. Chitosan disrupts the barrier properties of the outer membrane of Gram-negative bacteria International Journal of the Food Microbiology, 2001, 71:235-244
67. Hirano S, Hayashi M, Nagao N, et al. Chitinase activity of some seeds during their germination process and its inducetion by treating with chitosan and derivatives. In: Chitin and Chitosan: Sources, Chemistry, Biochemistry, Physical, Properties, and Application. Edited by Skjak B, Gudmund, London UK 1998, p743-747
68. Hirano S, Nagao N Effects of chitosan, pectic acid, lysozyme, and chitosan on the growth of several phytopathogens[J] Agric Biol Chem, 1989, 53 (11): 3065-3066
69. Hoffman, G.J. and Jobes, J.A. Growth and water relations of cereal crops as influenced by salinity and relative humidity Agron. J. 1978,70:765-769
70. Huang and Redman, R.E. Solute adjustment to salinity and calcium supply in cultivated and wild Barley. J. of Plant Nutrition, 1995,18(7):1371-1389
71. Jia ZS, Shen DF, Xu WL. Synthesis and antibacterial activities of quatemary ammonium salt of chitosan. Carbohydrate Research, 2001,333:1-6
72. Kendra DF, Christian D, Hadwiger LA. Chtosan oligomers from Fusarium solani/pea interactions, chitinase/β-glucanase digestion of sporelings and from fungal wall chitin activitely inhibit fungalgrowth and enhance disease resistance[J].Physiological and Molecular Plant Pathology, 1989,35:215-230
73. Koukol. J. Conn EE The metabaolism of aromatic compounds in higher plant [J], io. chem 1991.236:2692-2695
74. Lafontaine PJ, Baenhamou N. Chtosan treament: an emerging straegy for enhancing sistance of greenhouse tomato plants to infection by Fusarium oxysporum f. Sp. Radicis-lycoperic[J]i. Biocontrol Sci rechnol, 1996, 6:111-124
75. Lee KY; Kwon IC; Kim YH, et. al. Jeong SY. Preparation of chitosan self-aggregates as a gene delivery system. Journal of Controlled Release, 1998, 51:213-220
76. Leslie Ch;Romani RJ.Inhibition of ethylene biosynthesis bysalicylic acid[J].Plant Physiol, 1988,88:833-837
77. Majalhaes JR and Huber DM(1991)Reponse of ammnium assimilation enzymes to nitrogen form treatments in different plant species J. Plant Nutr. 14:175-185
78. Martinez N, Giselle MA, Madrid EA, Bottini, R, et al. Indole acetic acid attenuates disease severity in potato-Phytophthora infeatans interaction and inhibits the pathogen growth in vitro. Plant Physiology and Biochemistry, 2001,39:815-823
79. Matsuhashi S, Kume T. Enhancement of antimicrobical activity of chitosan by irrdiation J Sci Food Agric, 1997, 73: 237-241
80. Mauch F, Hadwiger LA, Boller T. Antifungal hydrolases in pea tissue Plant physiol, 1988a, 87:325-333
81. Mauch F, Mauch-Mani B, boller T. antifungal hydrolases in pea tissue Ⅱ inhibition of fungal growth by combinations of chitinase and β -1,3-glucanase Plant Plant Physiol, 1988b, 88:936-942
82. Murr DP Morris Infuence of O2 and CO on o_diphenol oxidase activity in mushrooms [J] Amer. Soc hort Sci 1974.99:155-158
83. Nielsen KK, Jorgensen P, Mikkelsen JD. Antifungal activity of sugar beet chiinase against Cercospora beticola can autoradiographic study on cell wall degration Plant Patho,1994,43:979-986
84. NOHK, Na, YP, Shin HL, et al. Antibacterial activity of chitosans and
chitosan oligomers with different molecular weights, Intermational Journal of Food Microbiology, 2002, 74:65-72
85. Perez-Alfocea, F, Estan, M,T, Effects of salinity on nitrate, total nitrogen, soluble protein and free amino acid levels in tomato plants J, of Horticultural Science, 1994,68(6):1021-1027
86. Pospieszny H, Antiviroid activity of chitosan. Crop Protection, 1997, (16)2: 105-106
87. Rasmussen PH, Knudsen IMB, Elmholt S, et al. Relationship between soil cellulolytic activity and suppression of seeding blight of barley in arable soils[J]Applied Soil Ecology, 2002.19:91-96
88. Reddy MVB, Arul J, Angers P, et al. Chitosan treatment of wheat seeds induced resistance to Fusarium graminearum and improves seed quality. J. Agric. Food Chem, 1999, 47:1208-1216
89. Rejikumar JL, Surekha D, Devi S.Hydrolysis of lactose and milk when using afixed bed reator containing beta-galactosidase covalently bound onto chitosan and cross-linked poly (vinyl alcohol) .Food science and techno; ogy, 2001, 36(1): 91-98
90. Rhoades J, Rolier S. Antimicrobial actions of degraded and native chitosan agaist spoilage organisms in laboratory media and foods. Applied and Evironmental Microbiology, 2000, 66 (1): 80-86
91. Ryder T, Cramer CL, Bell Jn, et al. Elicitor rapidly induceds chalconc synthase Mrna in Phaseolus Vulgaris cell at the onset of the phytoalexi defense response[j].Proc Natl Acad Sci USA, 1984,81:5724-5728
92. Sathiyabama M and Balasubramanian R. Chitosan induces resistance components in Arachis hypogaea against leaf rust caused by Puccinia arachidis Speg Crop Protection, 1998, 17(4):307-313
93. Singh Nk, Handa A k, Hasegawa P M et al .Electrophoretic protein patterna in cultured cells of tobacco adapted to NaCl[J] plant Physiol,
1983, 72(5):535
94. Srivastava H.S and Singh R.P(1987).Role and regulation of L-glutamate dehydrogenase activity in higher plants Phtochemistry. 23(3):597-610
95. Suntornsuk W, Pochanavanich P, Suntornsuk L. Fungal chitosan production on foodprocessing by-products. Process Biochemistry, 2002, 37:727-729
96. Teixeira MA, Patterson WT, Dunn EJ, et al. Assessment of chitosan gels for the controlled release of agrochemical .Ind Eng Chem Res, 1990 (29): 1205-1209
97. Thanou M: Florea BI: Geldof M et al.Q uaternized chitosan oligomers as novel gene delivery vectors in epithelial cell lines Biomaterials, 2002: 153-159
98. Vander P, Varum KM, Domard A, et al. Comparison of the ability of partially N-acetylated chitosans and chitooligosaccharides to elicit resistance reactions in wheat leaves Plant Physiol, 1998, 118:1353-1359
99. Vasyukova NI, ZI, Z ionov' eva SV, Il' inskaya Li, et al. Modulation of plant resistance to diseases by water-soluble chitosan .Applied Biochemistry Microbiology, 2001, 37:115-122
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101. Xuan TL, NagasawaN, Matsuhashi S, et. al. Effect of radiation-degraded chitosan on plantsstressed with vanadium. Radiation Physics and Chemistry, 2001, 61:171-175
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61. Gaudot ED, Slezack S,Dassi, et al. plant hydrolytic enzymes (chitinases and β-glucanase) in root reactions to pathogenic and symbiotic microorganisms .Plant and Soil, 1996,185:211-221
62. Hadwiger LA, Beckman JM. Chitosan as a component of Pea-Fusarium solani interactions. Plant physiol, 1980, 66:205-211
63. Hadwiger LA. Inducyion of phenylalanine ammonia lyase and pisatin by photo-sensitive psoralen compounds Plant Physiol, 1972,49:779-782
64. Helander, IM, Nurmiaho-Lassila EL, Ahvenainen R, et al. Chitosan disrupts the barrier properties of the outer membrane of Gram-negative bacteria
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65. Hadwiger LA. Loschke DC. Molecular communication in host-parasite interactions:hexosamine polymers(chitosan) as regulator compounds in race-specific and other interactions Phytopathology, 1981a, 71:756-782
66. Helander, IM, Nurmiaho-Lassila EL, hhvenainen R, et al. Chitosan disrupts the barrier properties of the outer membrane of Gram-negative bacteria International Journal of the Food Microbiology, 2001, 71:235-244
67. Hirano S, Hayashi M, Nagao N, et al. Chitinase activity of some seeds during their germination process and its inducetion by treating with chitosan and derivatives. In: Chitin and Chitosan: Sources, Chemistry, Biochemistry, Physical, Properties, and Application. Edited by Skjak B, Gudmund, London UK 1998, p743-747
68. Hirano S, Nagao N Effects of chitosan, pectic acid, lysozyme, and chitosan on the growth of several phytopathogens[J] Agric Biol Chem, 1989, 53 (11): 3065-3066
69. Hoffman, G.J. and Jobes, J.A. Growth and water relations of cereal crops as influenced by salinity and relative humidity Agron. J. 1978,70:765-769
70. Huang and Redman, R.E. Solute adjustment to salinity and calcium supply in cultivated and wild Barley. J. of Plant Nutrition, 1995,18(7):1371-1389
71. Jia ZS, Shen DF, Xu WL. Synthesis and antibacterial activities of quatemary ammonium salt of chitosan. Carbohydrate Research, 2001,333:1-6
72. Kendra DF, Christian D, Hadwiger LA. Chtosan oligomers from Fusarium solani/pea interactions, chitinase/β-glucanase digestion of sporelings and from fungal wall chitin activitely inhibit fungalgrowth and enhance disease resistance[J].Physiological and Molecular Plant Pathology, 1989,35:215-230
73. Koukol. J. Conn EE The metabaolism of aromatic compounds in higher plant [J], io. chem 1991.236:2692-2695
74. Lafontaine PJ, Baenhamou N. Chtosan treament: an emerging straegy for enhancing sistance of greenhouse tomato plants to infection by Fusarium oxysporum f. Sp. Radicis-lycoperic[J]i. Biocontrol Sci rechnol, 1996, 6:111-124
75. Lee KY; Kwon IC; Kim YH, et. al. Jeong SY. Preparation of chitosan self-aggregates as a gene delivery system. Journal of Controlled Release, 1998, 51:213-220
76. Leslie Ch;Romani RJ.Inhibition of ethylene biosynthesis bysalicylic acid[J].Plant Physiol, 1988,88:833-837
77. Majalhaes JR and Huber DM(1991)Reponse of ammnium assimilation enzymes to nitrogen form treatments in different plant species J. Plant Nutr. 14:175-185
78. Martinez N, Giselle MA, Madrid EA, Bottini, R, et al. Indole acetic acid attenuates disease severity in potato-Phytophthora infeatans interaction and inhibits the pathogen growth in vitro. Plant Physiology and Biochemistry, 2001,39:815-823
79. Matsuhashi S, Kume T. Enhancement of antimicrobical activity of chitosan by irrdiation J Sci Food Agric, 1997, 73: 237-241
80. Mauch F, Hadwiger LA, Boller T. Antifungal hydrolases in pea tissue Plant physiol, 1988a, 87:325-333
81. Mauch F, Mauch-Mani B, boller T. antifungal hydrolases in pea tissue Ⅱ inhibition of fungal growth by combinations of chitinase and β -1,3-glucanase Plant Plant Physiol, 1988b, 88:936-942
82. Murr DP Morris Infuence of O2 and CO on o_diphenol oxidase activity in mushrooms [J] Amer. Soc hort Sci 1974.99:155-158
83. Nielsen KK, Jorgensen P, Mikkelsen JD. Antifungal activity of sugar beet chiinase against Cercospora beticola can autoradiographic study on cell wall degration Plant Patho,1994,43:979-986
84. NOHK, Na, YP, Shin HL, et al. Antibacterial activity of chitosans and
chitosan oligomers with different molecular weights, Intermational Journal of Food Microbiology, 2002, 74:65-72
85. Perez-Alfocea, F, Estan, M,T, Effects of salinity on nitrate, total nitrogen, soluble protein and free amino acid levels in tomato plants J, of Horticultural Science, 1994,68(6):1021-1027
86. Pospieszny H, Antiviroid activity of chitosan. Crop Protection, 1997, (16)2: 105-106
87. Rasmussen PH, Knudsen IMB, Elmholt S, et al. Relationship between soil cellulolytic activity and suppression of seeding blight of barley in arable soils[J]Applied Soil Ecology, 2002.19:91-96
88. Reddy MVB, Arul J, Angers P, et al. Chitosan treatment of wheat seeds induced resistance to Fusarium graminearum and improves seed quality. J. Agric. Food Chem, 1999, 47:1208-1216
89. Rejikumar JL, Surekha D, Devi S.Hydrolysis of lactose and milk when using afixed bed reator containing beta-galactosidase covalently bound onto chitosan and cross-linked poly (vinyl alcohol) .Food science and techno; ogy, 2001, 36(1): 91-98
90. Rhoades J, Rolier S. Antimicrobial actions of degraded and native chitosan agaist spoilage organisms in laboratory media and foods. Applied and Evironmental Microbiology, 2000, 66 (1): 80-86
91. Ryder T, Cramer CL, Bell Jn, et al. Elicitor rapidly induceds chalconc synthase Mrna in Phaseolus Vulgaris cell at the onset of the phytoalexi defense response[j].Proc Natl Acad Sci USA, 1984,81:5724-5728
92. Sathiyabama M and Balasubramanian R. Chitosan induces resistance components in Arachis hypogaea against leaf rust caused by Puccinia arachidis Speg Crop Protection, 1998, 17(4):307-313
93. Singh Nk, Handa A k, Hasegawa P M et al .Electrophoretic protein patterna in cultured cells of tobacco adapted to NaCl[J] plant Physiol,
1983, 72(5):535
94. Srivastava H.S and Singh R.P(1987).Role and regulation of L-glutamate dehydrogenase activity in higher plants Phtochemistry. 23(3):597-610
95. Suntornsuk W, Pochanavanich P, Suntornsuk L. Fungal chitosan production on foodprocessing by-products. Process Biochemistry, 2002, 37:727-729
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