植物生理学实验实训综合教程(中英双语版)

本书在介绍“植物生理学实验基础”和“植物生理学综合设计性实验概述”的基础上以专题形式安排了植物种子的萌发、水分代谢、光合性能的评价、逆境生理等8个综合设计性项目实验，将植物生理学中的55个基础性实验通过不同的项目专题串联成综合设计性实验。这些实验方法，既可以独立开设基础性实验，也可以围绕相关主题开设综合设计性实验。使学生在加深植物生理学基本理论的同时，通过独立分析问题、设计实验、完成实验、解决问题，把知识转化为能力，为毕业设计、学术论文、开展科学实验研究以及完成科技成果报告等作基础训练。本教材除供高校学生使用外，也可以供从事植物生理学研究的技术人员参考。

姜倩倩，1983年11月生，女，山东省泰安人，研究生学历，农学博士学位，副教授。现为潍坊学院教师，主讲《植物生理学》、《植物生理学实验》和《无土栽培原理与技术》等课程；完成山东省本科教学改革研究项目3项，获省级教学成果二等奖2项，发表教研论文5篇。现承担教育部新农科研究与改革实践项目1项、山东省本科教学改革研究重点项目和面上项目各1项。主要从事园艺植物生理生态研究，近年来承担和主持国家自然科学基金项目2项、山东省高校科研发展计划项目1项、市级科技发展计划项目2项；获山东省科技进步三等奖1项，山东省农牧渔业丰收二等奖1项，潍坊市科技进步二等奖1项，发表科研论文近20篇，出版专著1部，获批发明专利2项，实用新型专利5项。

\"中文篇 第1章 植物生理学实验基础 ·························································3 1.1 植物生理学实验室规则................................................................................3 1.2 化学试剂的分类和配制方法........................................................................4 1.3 实验材料的采集、处理和保存....................................................................5 1.4 植物生理学实验研究的基本方法..............................................................10 第2章 植物种子的萌发 ······························································14 2.1 种子发芽率、发芽势及发芽指数的测定 .................................................15 2.2 种子生活力的快速测定..............................................................................16 2.3 种子活力的测定..........................................................................................17 2.4 种子萌发时淀粉酶的形成和活力检测......................................................19 2.5 油类种子萌发时脂肪酸含量的变化..........................................................20 2.6 种子萌发时氨基酸含量的变化..................................................................20 第3章 植物水分代谢 ·································································22 3.1 植物叶片含水量、相对含水量及水分饱和亏测定 .................................22 3.2 植物叶片自由水和束缚水含量测定（马林契克法） .............................24 3.3 植物组织水势的测定（小液流法）..........................................................25 3.4 植物组织渗透势的测定（质壁分离法） .................................................27 3.5 蒸腾速率的测定..........................................................................................28 第4章 矿质元素缺乏对植物生命活动的影响 ······································30 4.1 植物的元素缺乏症（溶液培养）..............................................................30 4.2 硝酸还原酶活性的测定..............................................................................34 4.3 根系活力的测定（α - 萘胺氧化法）.......................................................35 4.4 植株中硝态氮的测定..................................................................................38 4.5 植株磷素的测定（钼蓝法）......................................................................40 第5章 植物光合性能的评价 ························································42 5.1 光合色素的提取和理化性质......................................................................43 5.2 光合色素分离及吸收光谱的测定..............................................................44 5.3 叶绿体色素含量的测定..............................................................................46 5.4 植物叶片光合速率及气体交换参数的测定 .............................................48 5.5 叶绿素荧光参数的测定..............................................................................51 5.6 核酮糖二磷酸羧化酶（RuBPCase）活性的测定 ....................................55 5.7 磷酸烯醇式丙酮酸羧化酶（PEPCase）活性的测定 ..............................57 第6章 植物碳氮代谢 ·································································59 6.1 葡萄糖、果糖、蔗糖含量的测定..............................................................60 6.2 游离氨基酸含量的测定..............................................................................61 6.3 蔗糖合成酶和蔗糖磷酸合成酶活性的测定 .............................................63 6.4 α - 淀粉酶与β - 淀粉酶活性的测定.......................................................65 6.5 谷氨酰胺合成酶活性的测定......................................................................68 6.6 可溶性蛋白含量的测定..............................................................................69 第7章 植物生长物质的生理效应及其对植物生长发育的影响 ··············72 7.1 生长素对小麦根、芽生长的影响..............................................................72 7.2 赤霉素对小麦幼苗生长的影响..................................................................74 7.3 乙烯利对果实的催熟作用..........................................................................75 7.4 脱落酸对植物叶柄的脱落效应..................................................................75 7.5 赤霉素和脱落酸对种子萌发的影响..........................................................76 7.6 赤霉素诱导种子α - 淀粉酶的合成..........................................................78 7.7 脱落酸对气孔运动的影响..........................................................................80 7.8 液相色谱法测定植物激素含量..................................................................81 第8章 植物组织培养 ·································································83 8.1 培养基的配制...............................................................................................83 8.2 灭菌、消毒与接种......................................................................................87 8.3 继代培养与扩繁..........................................................................................90 8.4 试管苗的驯化、移栽和管理......................................................................93 第9章 植物逆境生理 ·································································95 9.1 脯氨酸含量的测定......................................................................................96 9.2 总黄酮含量的测定......................................................................................97 9.3 抗坏血酸（AsA）含量的测定..................................................................98 9.4 谷胱甘肽（GsH）含量的测定.................................................................102 9.5 细胞质膜透性的检测——电导率法........................................................103 9.6 生物膜过氧化程度的鉴定——丙二醛含量的测定 ...............................104 9.7 植株组织细胞死亡的鉴定（台盼蓝染色法） .......................................106 9.8 过氧化氢（H2O2）含量的测定................................................................107 9.9 超氧阴离子产生速率的测定....................................................................108 9.10 超氧化物歧化酶（SOD）活性的测定...................................................110 9.11 过氧化物酶（POD）活性的测定..........................................................111 9.12 过氧化氢酶（CAT）活性的测定...........................................................113 附　录 ····················································································115 附录1 相关理论知识巩固与思考..................................................................115 附录2 化学品标志符号..................................................................................202 附录3 常用酸碱的浓度..................................................................................203 附录4 常用固态酸、碱、盐的物质的量浓度配制参考 .............................203 附录5 常用缓冲液的配制..............................................................................204 附录6 常用酸碱指示剂..................................................................................209 附录7 标准计量单位......................................................................................210 附录8 常用植物生长物质的一些化学特性..................................................212 参考文献 ·················································································213 英文篇 Chapter 1 Experimental Basis of Plant Physiology ...................................217 1.1 Plant physiology laboratory rules.................................................................217 1.2 Classification and preparation methods of chemical reagent.......................218 1.3 Collection, processing and preservation of experimental material...............220 1.4 Basic methods of experimental studies on plant physiology.........................228 Chapter 2 Plant Seed Germination ..............................................................232 2.1　 Determination of seed germination rate, germination potential and germination index........................................................................................233 2.2　 Rapid determination of seed viability..........................................................234 2.3　 Determination of seed vigor.........................................................................236 2.4　 The formation and determination of the amylase activity during seed germination..................................................................................................238 2.5　 Changes in fatty acid content of oil seeds during germination.....................239 2.6　 Changes in amino acid content during seed germination.............................241 Chapter 3　 Plant Water Metabolism ..............................................................243 3.1　 Determination of plant leaf water content, relative water content and water saturation deficit.................................................................................244 3.2　 Determination of free and bound water content in plant leaves (Malincic method) .......................................................................................245 3.3　 Determination of plant tissue water potential (Small flow method) .............247 3.4　 Determination of plant tissue osmotic potential (Plasma wall separation method) ......................................................................................249 3.5　 Determination of transpiration rate..............................................................251 Chapter 4　 Effects of Mineral Element Deficiency on Plant Life Activities ........................................................................................254 4.1　 Element deficiency symptoms in plants (solution culture) ..........................255 4.2　 Determination of nitrate reductase activity..................................................259 4.3　 Determination of root activity (α - naphthylamine oxidation method) .......261 4.4　 Determination of nitrate nitrogen in plants..................................................263 4.5　 Determination of plant phosphorus (molybdenum blue method) .................265 Chapter 5　 Evaluation of Plant Photosynthetic Performance ....................268 5.1　 Extraction and physicochemical properties of photosynthetic pigments......269 5.2　 Separation of photosynthetic pigments and determination of absorption spectra.........................................................................................................271 5.3　 Determination of chloroplast pigment content..............................................273 5.4　 Determination of plant leaf photosynthetic rate and gas exchange parameters..276 5.5　 Determination of chlorophyll fluorescence parameters................................280 5.6　 Determination of ribulose diphosphate carboxylase (RuBPCase) activity.........................................................................................................286 5.7　 Determination of phosphoenolpyruvate carboxylase (PEPCase) activity.........................................................................................................289 Chapter 6　 Plant Carbon and Nitrogen Metabolism ...................................291 6.1　 Determination of sucrose, glucose and fructose content...............................292 6.2　 Determination of free amino acid content....................................................294 6.3　 Determination of sucrose synthase and sucrose phosphatase activities.......296 6.4　 Determination of α - amylase and β - amylase activities..........................298 6.5　 Determination of glutamine synthetase activity............................................302 6.6　 Determination of soluble protein content.....................................................304 Chapter 7　 The Physiological Effects of Plant Growth Substances and Their Effects on Plant Development .........................................307 7.1　 Effects of auxin on the growth of wheat root and shoot.................................308 7.2　 Effects of gibberellin on the growth of wheat seedlings................................310 7.3　 Effect of ethephon on the fruit ripening.......................................................310 7.4　 Abscissive effects of abscisic acid (ABA) on plant petioles.........................311 7.5　 Effects of gibberellin (GA3) and abscisic acid (ABA) on seed germination..................................................................................................313 7.6　 Synthesis of α - amylase in seed induced by gibberellin (GA3) .................314 7.7　 Effects of abscisic acid (ABA) on stomatal movement.................................317 7.8　 Determination of phytohormones by liquid chromatography........................318 Chapter 8　 Plant Tissue Culture ....................................................................320 8.1　 Preparation of culture medium.....................................................................321 8.2　 Sterilization, disinfection and inoculation....................................................325 8.3　 Subculture and propagation.........................................................................330 8.4　 Acclimation, transplanting and management of tube seedlings...................334 Chapter 9　 Plant Stress Physiology ..............................................................336 9.1　 Determination of proline content..................................................................337 9.2　 Determination of total flavonoid content......................................................339 9.3　 Determination of ascorbic acid (AsA, AA) ..................................................341 9.4　 Determination of glutathione (GsH) ............................................................345 9.5　 Determination of cytoplasmic membrane permeability - conductance method.........................................................................................................347 9.6　 Identification of degree of membrance peroxidation - determination of malondialdehyde content (MDA).................................................................349 9.7　 Identification of cell death in plant tissues (Trypan blue staining method) .351 9.8　 Determination of hydrogen peroxide (H2O2) content....................................352 9.9　 Determination of superoxide anion production rate.....................................354 9.10　 Determination of superoxide dismutase (SOD) activity..............................356 9.11　 Determination of peroxidase (POD) activity...............................................358 9.12　 Determination of catalase (CAT) activity...................................................360 The Appendix .....................................................................................................362 Appendix 1　 Consolidation and reflection of relative theoretical knowledge ......362 Appendix 2　 Chemical symbol ...........................................................................477 Appendix 3　 Concentrations of commonly usedacids and bases ........................477 Appendix 4　 Reference table of molarity preparation of commonly used solid acids, bases and salts ............................................................478 Appendix 5　 Preparation of commonly used buffers ...........................................478 Appendix 6　 Commonly used acid - base indicators ..........................................484 Appendix 7　 Standard unit of measurement .......................................................485 Appendix 8　 Some chemical properties of common plant growth substances .....487 Bibliography ........................................................................................................488\"