近年来，植物生物技术有了飞速的发展，其应用领域益加广泛。本书是Introduction to Plant Biotechnology Second Edition的影印版，对植物的基因、基因组的组成，植物组织培养、重组DNA技术进行了全面的翔实的阐述，较之原著第一版，特别增加了体外突变发生、基金组学、生物信息学、基因转移方法和作物改良转基因方法等方面的内容。作者在生物技术和遗传学领域从事了20多年的教学和研究，在植物体外培养、基因转移、分子标记方面做出了杰出的成就，有丰富的经验。在写作本书时特别注重技术的应用，也提供了比较详细的实验室技术方案。 本书知识全面，内容丰富，集合了植物生物技术的基础知识和前沿进展，实用性强，适合相关学科的高年级本科生、研究生用作教材，也适合从事相关工作的科研技术人员参考使用。
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目录
Preface to the Second Edition Preface to the First Edition Abbreviations PART I：PLANT TISSUE CULTURE 1.INTRODUCTION New technologies Origin of biotechnology History 2.LABORATORY ORGANIZATION Washing facility General laboratory and media preparation area Transfer area Culturing facilities Light units Green houses Laboratory and personal safety 3.NUTRITION MEDIUM Facilities and equipment Units for solution preparation Media composition Inorganic nutrients Carbon and energy source Vitamins Growth regulators Organic supplements Gelling agent pH Protocol: General methodology for medium preparation 4.STERILIZATION TECHNIQUES Preparation of sterile media,containers and small instruments Steam sterilization Units of energy and pressure Dry sterilization Filter sterilization Ultra violet sterilization Maintenance of aseptic conditions Alcohol sterilization Flame sterilization Sterilization of explant Chemical sterilization Protocols Sterilization of seeds Sterilization of buds,leaf,stem,roots,tubers,scales,etc. Sterilization of tissue for immature embryos,ovules,and flower buds for anther culture 5.TYPES OF CULTURE Cytodifferentiation Organogenic differentiation Types of culture Seed culture Embryo culture Mature embryo culture Immature embryo culture/embryo rescue Applications of embryo culture Callus culture Organ culture Nucellus culture Application Endosperm culture Application Cell culture Protoplast culture Protocols Protocol for seed germination（Nicotiana） Protocol for embryo culture（Cereals-wheat,maize,barley rice,etc.） Protocol for embryo culture（Legumes-green gram,black gram,french bean,soybean,etc.） Protocol for callus induction（Nicotiana tabacum） Protocol for callus induction（Cereals-wheat,rice,maize,barley,etc.） 6.MICROPROPAGATION Axillary bud proliferation approach Meristem and shoot tip culture Bud culture Single node culture Axillary bud method Organogenesis Organogenesis via callus formation Direct adventitious organ formation Embryogenesis Advantages of micropropagation Problems associated with micropropagation Protocols Meristem and node culture of potato（Solanum tuberosum） Proliferation of axillary buds（Strawberry-Fragaria chiloensis） Organogenesis-adventitious shoot formation Organogenesis via callus formation（Nicotiana） Organogenesis via callus formation（Cereals: wheat, barley,maize, rice, etc.） Embryogenesis（Carrot） 7.CELL SUSPENSION AND SECONDARY METABOLITES Types of suspension cultures Batch culture Continuous culture Open continuous culture Closed continuous culture Semi continuous culture Growth measurements Synchronization of suspension culture cells Technique for single cell culture-Bergmann cell plating technique Applications Production of secondary metabolites Morphological and chemical differentiation Medium composition for secondary product formation Growth production patterns Environmental factors Selection of cell lines producing high amounts of a useful metabolite Product analysis Application Problems associated with secondary metabolite production Immobilized cell systems Polymers for immobilization Product release Biotransformation Protocols Protocol for cell suspension culture（Nicotiana tabacum） Protocol for cell suspension culture（Cereals- wheat, rice,maize, barley,etc.） 8.IN VITRO PRODUCTION OF HAPLOIDS Androgenic methods Anther culture Microspore culture The various factors governing the success of androgenesis Genotype Physiological status of the donor plants Stage of pollen Pretreatment of anthers Culture media Process of androgenesis The ploidy level and chromosome doubling Diploidization Significance and uses of haploids Problems Gynogenic haploids Factors affecting gynogenesis Chromosome elimination technique for production of haploids in cereals（Barley and wheat） Protocol Protocol for anther culture of cereals（Rice, barley, wheat, etc.） 9.PROTOPLAST ISOLATION AND FUSION Protoplast isolation Mechanical method Enzymatic method Physiological state of tissue and cell material Enzymes Osmoticum Protoplast purification Protoplast viability and density Culture techniques Culture medium Environmental factors Protoplast development Cell wall formation Growth,division and plant regeneration Somatic hybridization Protoplast fusion Spontaneous fusion Induced fusion methods Treatment with sodium nitrate Calcium ions at high pH Polyethylene glycol method Electrofusion Mechanism of fusion Identification and selection of hybrid cells Chlorophyll deficiency complementation Auxotroph complementation Complementation of resistance markers Use of metabolic inhibitors Use of visual characteristics Compound selection system Verification & characterization of somatic hybrids Morphology Isoenzyme analysis Chromosomal constitution Molecular techniques Genetic characterization Chromosome number in somatic hybrids Cybrids Potential of somatic hybridization Problems and limitations of somatic hybridization Protocol Protocol for protoplast isolation and fusion 10.SOMACLONALVARIATION Nomenclature Schemes for obtaining somaclonal variation Without in vitro selection With in vitro selection Factors influencing somaclonal variation Application of somaclonal variation Basis of somaclonal variation Disadvantages Gametoclonal variation 11.GERMPLASM STORAGE AND CRYOPRESERVATION Cryopreservation Raising sterile tissue cultures Addition of cryoprotectants and pretreatment Freezing Storage Thawing Determination of survival/viability TTC method of staining Evan’s Blue staining Plant growth and regeneration. Slow growth method Applications PART II: GENETIC MATERIAL AND ITS ORGANIZATION 12.GENETIC MATERIAL Sugars Amino acids Nucleotides Structural formulae of nucleotides Nomenclature of nucleoside and nucleotide compounds Polynucleotides Significance of differences between DNA and RNA Shorthand notation for polynucleotide structure Genetic material Discovery of DNA Double helix is a stable structure DNA replication 13.ORGANIZATION OF DNA AND GENE EXPRESSION Different forms of DNA Supercoiling-Tertiary structure of DNA Linking number Cruciform-Tertiary structure of DNA Eukaryotic DNA organization into nucleosomes DNA content Denaturation DNA renaturation Renaturation rate and DNA sequence complexity-Cot curves Flow of genetic information: central dogma Organization of genes in DNA molecules Operon Multigene family Plant gene structure as a discontinuous gene Control sequences TATA box AGGA box Other regulatory elements Types of RNA molecules Messenger RNA and processing Ribosomal RNA Transfer RNA Small nuclear RNA Transcription Nucleotide sequences Transcription in prokaryotes Transcription in euokaryotes Genetic code and translation Genetic code Translation Post-translational modifications PART III: RECOMBINANT DNA TECHNOLOGY 14.BASIC TECHNIQUES Agarose gel electrophoresis Pulsed field gel electrophoresis Polyacrylamide gel electrophoresis 2-Dimensional gel electrophoresis Nucleic acid blotting Southern blot analysis Northern blot analysis Protein blotting Western blot analysis Dot blot technique Autoradiography E.coil transformation Protocol Agarose gel electrophoresis 15.GENE CLONING: Cutting and Joining DNA Molecules Enzymes for cutting: Restriction endonucleases Type I,II,III endonucleases Other restriction enzymes Joining DNA molecules DNA ligase DNA-modifying enzymes Kinase Alkaline phosphatase Terminal transferase DNA polymerase S1-Nuclease λ-Exonuclease Exonuclease III Bal 31 nuclease Linkers and adaptors Protocol Restrictive digestion of plasmid DNA 16.GENE CLONING: Vectors Biology of E.coil K-12 Plasmids pBR322,pACYC184,pUC vectors,pUN121 Yeast plasmid vectors Ti plasmids Cosmids Bacteriophage vectors Biology of bacteriophage λ λ phage cloning vectors（λgtl0,λgt11,EMBL3 and 4,Charon） M13 phage Phagemids Yeast artificial chromosome（YAC） Bacterial artificial chromosome（BAC） P1 phage vectors P1 derived artificial chromosome（PAC） Shuttle vectors Expression vectors Protocols Isolation of plasmid DNA：mini preparation Isolation of genomic DNA by SDS-Proteinase K method 17.GENE CLONING: cDNA and Genomic Cloning and Analysis of Cloned DNA Sequences Complimentary DNA（cDNA）libraries and cloning Isolation or extraction of mRNA of mRNA Synthesis of first strand of cDNA Synthesis of second strand of DNA Cloning of cDNA Introduction to host cells Clone selection Genomic cloning Isolation of DNA Partial digestion Vectors for cloning Ligation of fragment to a vector Packaging Identification and analysis of cloned genes Probes Approaches for identification of genes Colony and plaque hybridization Immunological detection Southern blot analysis Detection of nucleic acid sequences Radioactive labeling Nonradioactive labeling Horseradish peroxidase system Digoxigenin（DIG）label!ng system Biotin-streptavidin labeling system DNA sequencing Sanger-Coulson method Maxam-Gilbert method High throughput DNA sequ.encing 18.POLYMERASE CHAIN REACTION Procedure Components in a polymerase chain reaction Inverse PCR Reverse transcriptase mediated PCR（RT-PCR） RACE：Rapid amplification of cDNA ends Quantitative RT-PCR Amplification of differentially expressed genes Differential display reverse transcriptase PCR Suppression subtraction hybridization Cloning of PCR products Restrictive site addition T/A cloning Blunt end ligation Genetic engineering by PCR Applications Advantages Problems Protocol PCR for detection of transgenes 19.IN VITRO MUTAGENESIS Site directed mutagenesis Deletion mutagenesis Unidirectional deletions Gap-sealing mutagenesis Linker scanning mutagenesis Cassette mutagenesis Oligonucleotide directed mutagenesis Gapped duplex method dUTP incorporation strategy Phosphorothioate method Chemical mutagenesis PCR-mediated in vitro mutagenesis Advantages of site directed mutagenesis Random mutagenesis Insertional mutagenesis Transposon mediated insertional mutagenesis T-DNA mediated insertional mutagenesis 20.TRANSPOSON GENETIC ELEMENTS AND GENE TAGGING Transposable elements in bacteria IS elements Composite transposons Complex transposons:the Tn3 family Transposable elements in eukaryotes Classification Class I elements Retroviruses LTR retrotransposons Non-LTR retrotransposons Class II elements Ac-Ds elements En/Spm transposable element family Transposon gene tagging Isolation of transposable element Gene tagging Characterization of transposon insertion mutation Isolation of mutant gene Isolation of intact gene Transposon tagging in heterologous species Significance of transpron tagging（mutations, gene expression, vector development） 21.GENE ISOLATION General strategies for cloning genes from plants Isolation of genes coding for specific proteins Isolation of genes that are tissue specific in function Cloning methods based on DNA insertions Subtractive cloning Map-based cloning Identification of DNA markers linked to target gene and construction of genetic maps Construction of high resolution physical maps Chromosome walking Testing of gene by transformation and complementation Chromosome jumping Chromosome landing approach 22.MOLECULAR MARKERS AND MARKER-ASSISTED SELECTION Morphological markers Biochemical markers Molecular markers Non-PCR based approaches RFLP（Procedure,construction of RFLP maps,uses of RFLP PCR-based techniques Random amplified polymorphic DNA（RAPD） DNA amplification fingerprinting（DAF） Arbitrarily primed polymerase chain reaction（AP-PCR） Amplified fragment length polymorphism（AFLP） Simple sequence repeats（SSR）or Microsatellites Microsatellite directed PCR：unanchored primers Microsatellite directed PCR：anchored primers Random amplified microsatellite polymorphic loci（.RAM.P） Selective amplification of microsatellite polymorphic Ioci Targeted PCR and sequencing Sequence tagged sites Sequence tagged microsatellites（STMs） Sequence characterized amplified regions（SCARs） Cleaved amplified polymorphic sequences（CAPs） Advantages of targeted PCR and sequencing approaches Disadvantages Fingerprinting Marker-assisted selection Protocol Protocol for RAPD analysis 23.GENE TRANSFER IN PLANTS Transient and stable gene expression Marker genes Reporter genes Opine synthase Chloramphenicol acetyl transferase β glucuronidase（GUS） Bacterial luciferase（Lux F2） Firefly luciferase（Luc） Green fluorescent protein（GFP） Anthocyanins Selectable markers Antibiotic resistance markers Antimetabolite resistance marker Herbicide resistance markers Chimeric gene vectors Gene transfer methods Vector mediated gene transfer Agrobacterium mediated DNA transformation Classification of Agrobacteria Tumor inducing principle and the Ti plasmid Organization of Ti plasmid T-DNA transfer Agrobacterium vectors Cointegrate vectors Binary vectors Transformation techniques using Agrobacterium Agrobacterium mediated virus infection-Agroinfection Viruses-mediated gene transfer Caulimoviruses Gemini viruses RNA viruses Vectorless or direct DNA transfer Physical gene transfer methods Electroporation Particle bombardment/microprojectile/biolistics Macroinjection Microinjection Liposome-mediated transformation Silicon carbide fiber mediated transformation Ultrasound-mediated DNA transformation DNA transfer via pollen Chemical gene transfer methods PEG-mediated gene transfer Calcium phosphate coprecipitation The polycation DMSO technique DEAE dextran procedure Status and expression of transferred genes Gene silencing Detection of intrusive DNA Causes of gene silencing Strategies for avoiding gene silencing Protocols Agrobacterium mediated transformation Gene transfer by biolistic method：transient expression 24.TRANSGENICS IN CROP IMPROVEMENT Resistance to biotic stresses Insect resistance Resistance genes from microorganisms Resistance genes from higher plants Resistance genes from animals Virus resistance Coat-protein-mediated cross protection Non-structural protein-mediated resistance Antisense and sense mediated resistance Satellite RNA protection Pathogen targeted resistance Disease resistance Pathogenesis related proteins Anti-microbial proteins Engineering toxin insensitivity Phytoalexins Manipulation of disease resistance genes Resistance to abiotic stresses Herbicide resistance Transgenics for quality Transgenics for improved storage Longer life transgenic flowers Transgenics for flower color and shape Transgenics for male sterility Transgenics for terminator seed Mechanism in pure line seed production Terminator technology for use in hybrid seed,production Transgenic plants as bioreactors Carbohydrates Lipids Protein quality Vitamin and mineral Biodegradable plastic Proteins,peptides and vaccines Production of edible vaccines Commercial transgenic crops Impact of recombinant DNA technology 25.GENOMICS Mapping of prokaryotic genomes E.coil genome Mapping of eukaryotic genomes Construction of linkage maps with molecular markers Gene libraries Screening of libraries and constructing contigs Sequencing New vectors BACs and PACs and the shot gun approach Gene location in DNA sequences Sequence inspection Experimental techniques Hybridization tests cDNA sequencing Yeast (S.cerevisiae) genome Human genome Rice genome Functional genomics Computer analysis Experimental analysis Gene inactivation Gene overexpression Phage display Yeast two hybrid system Patterns of gene expression Gene expression assays by measuring levels of RNA transcripts SAGE DNA chip technology Oligonucleotide based chips cDNA based chips Hybridization and detection methods Double stranded DNA chips Characteristic features of DNA chips/microarrays Applications of DNA chips Proteomics 26.BIOINFORMATICS Development of bioinformatics Databases Online education Access to databases Applicatinos Challanges 27.INTELLECTUAL PROPERTY RIGHTS Protection of intellectual property World organizations Forms of protection Copyright Trademark Patent Patent application International patenting and patent cooperation treaty Uses of technical information in patent documents Revocation of patent Patenting of biological material Geographical indications Trade secret Designs Know-how Plant breeders rights Functions of UPOV Farmer’s rights Convention on biodiversity Plant variety protection Some case studies on plant patents Patenting of basmati rice in USA Revocation of turmeric patent Revocation of neem patent Appendix References Glossary Author index Subject index
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