Ⅰ.Development and Morphology 1
1.Fertilization and Embryogeny in Maize&WILLIAM F.SHERIDAN AND JANICE K.CLARK 3
2.The Maize Shoot&R.S.POETHIG 11
3.The Maize Leaf&MICHAEL FREELING AND BARBARA LANE 17
4.The Maize Root&L.FELDMAN 29
5.Morphology and Development of the Tassel and Ear&PING-CHIN CHENG AND DAYAKER R.PAREDDY 37
6.Gametogenesis in Maize&PATRICIA BEDINGER AND SCOTT D.RUSSELL 48
7.The Patterns of Plant Structures in Maize&WALTON C.GALINAT 61
8.Genetics and the Morphological Evolution of Maize&JOHN DOEBLEY 66
9.Overview of Key Steps in Aleurone Development&VIRGINIA WALBOT 78
Ⅱ.Cell Biology 81
10.Light Microscopy Ⅰ：Dissection and Microtechnique&ANNE W.SYLVESTER AND STEVEN E.RUZIN 83
11.Light Microscopy Ⅱ：Observation，Photomicrography，and Image Analysis&STEVEN E.RUZIN AND ANNE W.SYLVESTER 95
12.Scanning Electron Microscopy&MICHELLE H.WILLIAMS AND ANNE W.SYLVESTER 108
13.Transmission Electron Microscopy：Chemical Fixation，Freezing Methods，and Immunolocalization&M.V.PARTHASARATHY 118
14.Techniques for Histology of Maize Megaspores and Embryo Sacs&S.D.RUSSELL AND D.P.WEST 135
15.Indirect Immunofluorescence：Localization of the Cytoskeleton&C.J.STAIGER 140
16.Immunocytochemistry for Light and Electron Microscopy&M.J.VARAGONA AND N.V.RAIKHEL 149
17.Immunolocalization of Nuclear Proteins&LAURIE G.SMITH 158
18.In situ Hybridization&JANE A.LANGDALE 165
19.Photography of Maize&PHILIP W.BECRAFT 180
Ⅲ.Genetics Protocols 187
20.Genetic Experiments and Mapping&E.H.COE 189
21.Growing Maize for Genetic Studies&M.G.NEUFFER 197
22.A Nine-Step Way to Characterize a Morphological Mutant&MICHAEL FREELING AND JOHN FOWLER 209
23.Mutagenesis&M.G.NEUFFER 212
24.Gene Tagging with Ac/Ds Elements in Maize&STEPHEN L.DELLAPORTA AND MARIA A.MORENO 219
25.Using Cytogenetics to Enhance Transposon Tagging with Ac Throughout the Maize Genome&DONALD AUGER AND WILLIAM F.SHERIDAN 234
26.Transposon Tagging with Spm&KAREN CONE 240
27.Transposon Tagging with Mutator&PAUL S.CHOMET 243
28.Mapping Genes with Recombinant Inbreds&BENJAMIN BURR，FRANCES A.BURR AND EILEEN C.MATZ 249
29.The Placement of Genes Using waxy-Marked Reciprocal Translocations&JOHN R.LAUGHNAN AND SUSAN GABAY-LAUGHNAN 255
30.Chimeras for Genetic Analysis&M.G.NEUFFER 258
31.The Use of Clonal Sectors for Lineage and Mutant Analysis&SARAH HAKE AND NEELIMA SINHA 262
32.Use of Segmental Aneuploids for Mutant Analysis&BEN GREENE AND SARAH HAKE 270
33.Biased Transmission of Genes and Chromosomes&WAYNE R.CARLSON 274
34.Anthocyanin Genetics&E.H.COE 279
35.Cloned Anthocyanin Genes and Their Regulation&KAREN CONE 282
36.Maize and Puccinia sorghi：A System for the Study of the Genetic and Molecular Basis of Host-Pathogen Interactions&TONY PRYOR 286
37.Disease Lesion Mutants&M.G.NEUFFER 291
38.Classification of Pollen Abortion in the Field&R.L.PHILLIPS 297
39.Genetic Fine Structure as Revealed in Pollen Assays&OLIVER E.NELSON 298
40.Genetic Fine Structure from Testcross Progeny Analysis&HUGO K.DOONER 303
41.Trisomic Manipulation&JAMES A.BIRCHLER 307
42.B-A Translocation Manipulation&WAYNE R.CARLSON 308
43.Locating Recessive Genes to Chromosome Arm with B-A Translocations&J.B.BECKETT 315
44.Dosage Analysis Using B-A Translocations&JAMES A.BIRCHLER 328
45.Marker Systems for B-A Translocations&JAMES A.BIRCHLER 330
46.Construction of Compound B-A Translocations&JAMES A.BIRCHLER 332
47.A-B-A Compound Chromosomes&JAMES A.BIRCHLER 334
48.Comprehensive List of B-A Translocations in Maize&J.B.BECKETT 336
49.Chromosomal Translocations Involving the Nucleolus Organizer Region or Satellite of Chromosome 6&R.L.PHILLIPS 342
50.Inversions and List of Inversions Available&G.G.DOYLE 346
51.Use of Maize Monosomics for Gene Localization and Dosage Studies&DAVID F.WEBER 350
52.Marker Systems for r-x1&JAMES A.BIRCHLER AND E.H.COE 359
53.Translocations as Genetic Markers&E.B.PATTERSON 361
54.A-A Translocations：Breakpoints and Stocks&E.H.COE 364
55.Segmental Aneuploid Analysis&JAMES A.BIRCHLER 377
56.Directed Synthesis of Segmental Transpositions&JAMES A.BIRCHLER 383
57.Practical Aspects of Haploid Production&JAMES A.BIRCHLER 386
58.Indeterminate Gametophyte（ig）：Biology and Use&JERRY L.KERMICLE 388
59.Production of a Ploidy Series&JAMES A.BIRCHLER 394
60.Absorption Cytophotometry of Nuclear DNA&RICHARD V.KOWLES，GEORGIA L.YERK AND RONALD L.PHILLIPS 396
61.Flow Cytometry for Endosperm Nuclear DNA&RICHARD V.KOWLES，GEORGIA L.YERK，LIANG SCHWEITZER，FREIDRICH SRIENC AND RONALD L.PHILLIPS 400
62.Allelism Testing of Lethal Mutations&JANICE K.CLARK AND WILLIAM K.SHERIDAN 407
63.Analysis of Cytoplasmically Inherited Mutants&KATHLEEN J.NEWTON 413
64.Male Sterility and Restorer Genes in Maize&SUSAN GABAY-LAUGHNAN AND JOHN R.LAUGHNAN 418
65.Inbred Lines of Maize and Their Molecular Markers&MICHAEL LEE 423
66.Traditional Analysis of Maize Pachytene Chromosomes&ELLEN DEMPSEY 432
67.Techniques for Preparing Whole-Mount Spreads of Maize Pachytene Chromosome Complements for Electron-Microscopic Visualization of Synaptonemal Complex Structures&MARJORIE P.MAGUIRE 442
68.A Smear Technique for the Study of Meiosis in Pollen Mother Cells of Maize&INNA GOLUBOVSKAYA 447
69.Protocol for Preparing Maize Macrospore Mother Cells for the Study of Female Meiosis and Embryo-sac Development&INNA GOLUBOVSKAYA AND N.A.AVALKINA 450
70.Preparing a Suspension of Microsporocytes for Spreading and Electron Microscopy&INNA GOLUBOVSKAYA AND Z.K.GZEBENNIKOVA 454
71.Three-dimensional Fluorescence Microscopy of Maize Chromosomes&R.KELLY DAWE 457
72.Chromosomal Behavior During Microsporogenesis&MING T.CHANG AND M.GERALD NEUFFER 460
73.A Staining Procedure for Pollen Grain Chromosomes of Maize&BRYAN KINDIGER 476
74.A Technique for the Preparation of Somatic Chromosomes of Maize&BRYAN KINDIGER 481
75.A Technique for Somatic Chromosome Preparation and C-banding of Maize&DAVID C.JEWELL AND NURUL ISLAM-FARIDI 484
76.Duplications&JAMES A.BIRCHLER 493
77.Deficiency Analysis&JAMES A.BIRCHLER 494
78.The Gametophyte Factors of Maize&OLIVER E.NELSON 496
79.Ring Chromosomes&JAMES A.BIRCHLER 503
80.In situ Hybridization of DNA and RNA Probes to Maize Chromosomes&S.M.LIVINGSTON AND R.L.PHILLIPS 504
81.Analysis of Traits with Complex Inheritance in Maize Using Molecular Markers&TIM HELENTJARIS AND MANFRED HEUN 509
82.Heterofertilization&JAMES A.BIRCHLER 514
Ⅳ.Molecular Biology Protocols 517
83.Isolation of Small Nuclear RNAs&TAMAS KISS AND WITOLD FILIPOWICZ 519
84.Plant DNA Miniprep and Microprep：Versions 2.1-2.3&STEPHEN L.DELLAPORTA 522
85.Urea-based Plant DNA Miniprep&JYCHIAN CHEN AND STEPHEN L.DELLAPORTA 526
86.High-Molecular-Weight Plant DNA Preparation for CHEF Gel Analysis&ELSBETH WALKER 528
87.Preparation of High-Molecular-Weight Maize DNA and Analysis by Pulsed-Field Gel Electrophoresis&AVRAHAM A.LEVY 530
88.Isolation of Genomic DNA from Calli&CHRISTINE A.WARREN 534
89.Isolation of DNA from Immature Cobs&CHRISTINE A.WARREN 536
90.Preparation of Nucleic Acids from Maize Microspores and Pollen&ANNE H.BROADWATER AND PATRICIA BEDINGER 538
91.Preparation of DNA and RNA from Leaves：Expanded Blades and Separated Bundle Sheath and Mesophyll Cells&TIMOTHY NELSON 541
92.Isolation of RNA from Wx and wx Endosperms&SUSAN R.WESSLER 545
93.RNA Isolation from Electroporated Protoplasts&KENNETH R.LUEHRSEN AND JANE HERSHBERGER 547
94.Procedures for Isolating Mitochondria and Mitochondrial DNA and RNA&KATHLEEN J.NEWTON 549
95.Isolation of Maize Chloroplasts and Chloroplast DNA&STEVEN RODERMEL 556
96.In vitro Capping of Maize Mitochondrial RNA and Transcription Initiation Site Characterization by RNase Protection&R.MICHAEL MULLIGAN 559
97.Editors’ Note&VIRGINIA WALBOT AND MICHAEL FREELING 565
98.Southern Blots of Maize Genomic DNA&CHRISTINE A.WARREN AND JANE HERSHBERGER 566
99.Southern Blot Hybridization&STEPHEN L.DELLAPORTA AND MARIA A.MORENO 569
100.Northern Blotting&KENNETH B.LUEHRSEN 572
101.RNase Protection Assay&KENNETH R.LUEHRSEN 575
102.Genomic Sequencing in Maize&ANNA-LISA PAUL AND ROBERT J.FERL 579
103.The Polymerase Chain Reaction：Applications to Maize Transposable Elements&ANNE B.BRITT AND DAVID J.EARP 586
104.In vitro Synthesis of Capped and Polyadenylated mRNA for Translation Studies in vitro and in vivo&DANIEL R.GALLIE 592
105.Construction of a Genomic Library in Lambda Phage&CLIFFORD F.WEIL AND THOMAS E.BUREAU 595
106.Agroinfection&JESUS ESCUDERO AND BARBARA HOHN 599
107.Polyethylene Glycol-mediated DNA Uptake into Maize Protoplasts&LESZEK A.LYZNIK AND THOMAS K.HODGES 603
108.DNA Delivery into Maize Cell Cultures Using Silicon Carbide Fibers&H.F.KAEPPLER AND D.A.SOMERS 610
109.Transient Gene Expression Assay by Electroporation of Maize Protoplasts&KENNETH R.LUEHRSEN AND JEFFREY R.DE WET 613
110.Assay of Bacterial Chloramphenicol Acetyl Transferase in Transformed Maize Tissues&STEVE GOFF 616
111.Assay of Firefly Luciferase in Transformed Maize Tissues&STEVE GOFF 619
112.Storage of Frozen Maize Tissue&SUSAN R.WESSLER 622
113.Maize Methods—Starch Biosynthetic Genes&L.CURTIS HANNAH，EDWIN R.DUKE，KAREN E.KOCH AND B.GREGORY COBB 624
114.Identifying and Characterizing the TATA Box Promoter Sequence Element in a Maize Nuclear Gene&JULIE M.VOGEL 630
115.Promoters&KENNETH R.LUEHRSEN 633
116.Introns&KENNETH R.LUEHRSEN 636
117.Characterization of Zein Genes and Their Regulation in Maize Endosperms&GARY A.THOMPSON AND BRIAN A.LARKINS 639
118.Overview of Cloning Genes Using Transposon Tagging&VICKI CHANDLER 647
119.How RFLP Loci Can Be Used to Assist Transposon-Tagging Efforts&STEVEN P.BRIGGS AND WILLIAM D.BEAVIS 653
Ⅴ.Cell Culture Protocols 661
120.Regeneration of Plants from Somatic Cell Cultures：Applications for in vitro Genetic Manipulation&CHARLES L.ARMSTRONG 663
121.Initiation，Maintenance，and Plant Regeneration of Type Ⅱ Callus and Suspension Cells&J.C.SELLMER，S.W.RITCHIE，I.S.KIM AND T.K.HODGES 671
122.Production of Transgenic Maize Plants via Microprojectile-Mediated Gene Transfer&MICHAEL FROMM 677
123.In vitro Selection&D.A.SOMERS AND K.A.HIBBERD 685
124.Selection of Stable Transformants from Black Mexican Sweet Maize Suspension Cultures&JULIE ANDERSON KIRIHARA 690
125.Maize Protoplast Culture&R.D.SHILLITO，G.K.CARSWELL AND C.KRAMER 695
126.Anther and Microspore Culture&J.F.PETOLINO AND A.D.GENOVESI 701
127.In vitro Culture of Maize Kernels&BURLE G.GENGENBACH AND ROBERT J.JONES 705
128.In vitro Ear Culture System&RICHARD V.KOWLES 709
129.Maize Inflorescence Culture&R.I.GREYSON 712
130.Shoot Meristem Culture&ERIN E.IRISH 715
131.Establishment and Culture of Maize Endosperm&JACK C.SHANNON 719
132.In vitro Pollen Germination&D.B.WALDEN 723
133.Axillary Bud in vitro Culture：Asexual Propagation of Maize&R.I.GREYSON AND D.B.WALDEN 725
Index 727