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H400, Lecture 12, 10/11/01

H400, Lecture 12, 10/11/01. H400, 2001 Upcoming Due Dates 10/23/01 (Tuesday, not 10/21 ) Field Trip Reports (both trips, 1 page each) 10/25/01 (Thursday, 1:30-3:00PM; not 10/19 ) Midterm Exam

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H400, Lecture 12, 10/11/01

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  1. H400, Lecture 12, 10/11/01

  2. H400, 2001 Upcoming Due Dates • 10/23/01 (Tuesday, not 10/21) Field Trip Reports (both trips, 1 page each) • 10/25/01 (Thursday, 1:30-3:00PM; not 10/19) Midterm Exam • 11/01/01 (not 10/31) Written Cuttage Lab Report (Experiment 1 – Even groups; Experiment 2 – Odd groups

  3. Midterm Exam will cover: • Lectures • Lab exercises (understand the what & why of lab exercises) • Web assignments (preparation for Geneva field trip, and for Microprop lab, see Table 2) • Textbook: (**rechk for 2001) Assigned sections of Ch 2, 9, 10, 12 & 17; see Table 2 • Field Trips • UP TO and Including Lecture on Tuesday, 10/30/01

  4. Midterm Exam: examples of Compare & Contrast questions Compare and Contrast each of the following pairs (or triplet), in the context of their relevance to plant propagation. Your answer should make clear what each term refers to and the similarities and differences between them. (4 point each) 1.      Mitosis / meiosis – the two types of cell division involved in plant propagation. Mitosisrefers to somatic cell division involved in regeneration of new organs during asexual propagation, whereas Meiosis refers to cell division involving formation of haploid gametes and segregation of alleles, from diploid “mother” cell during the process of sexual reproduction. Black = similarity White = difference 3.      Mist / fog 4.      Root initial / root primordium 5.      In vitro / ex vitro

  5. Midterm Exam: examples of Short Answer questions Short Essays (15 points each) (1/2 – 1 page) 1.      Describe how you would determine if cuttage or micropropagation was the best method for propagating a newly introduced ornamental selection for large-scale commercial production. Assume that you have at least a year before commercial production begins and that adaquate facilities are available for either method of propagation. 2.      Briefly describe each of the stages involved in plant micropropagation via shoot culture (as described by Murashige, 1974), including the objective(s) of each stage and the strategy used to achieve those objectives. 3.      What is the effect of stock plant growth phase on rooting of cuttings and how can it be managed to optimize rooting of difficult-to-root species? 19. Describe how light, temperature and humidity interact to influence the water potential of a cutting and its ability to root. How are these environmental factors managed to maximize rootability in a mist system, compared to a closed case (polyethylene) moisture management system? Also: there may be a data interpretation question

  6. Propagation via Plant Tissue Culture

  7. Propagation Cuttage Graftage Layerage Division

  8. Asexual Propagation Cuttage Graftage Layerage Division Micropropagation

  9. Welcome to the 15 Annual conference of micropropagators . . . . . .

  10. Definitions Plant Tissue Culture- in vitro, aseptic plant culture for any purpose including genetic transformation and other plant breeding objectives, secondary product production, pathogen elimination… or forasexual(micropropagation) or sexual propagation

  11. Definitions • Plant Tissue Culture • Micropropagation - use of tissue culture for clonal plant propagation (including somatic embryogenesis) • In vitro seed or embryo culture- in vitro sexual propagation

  12. Characteristics of Most • Micropropagation Systems • aseptic culture (no other organsims) • small propagule = explant • heterotrophic nutrition

  13. History of Plant • Tissue Culture Propagation at Cornell • Lewis Knudson, 1922 - 1st in vitro embryo germination (orchid) - ie the invention of in vitro embryo culture • Knudson, L, 1922, Nonsymbiotic germination of orchid seeds, Botanical Gazette LXXIII: 19 • Gavino Rotor, 1949 - 1st in vitro vegetative propagation. • Rotor, G, 1949, A method of vegetative propagation of Phalaenopsis species and hybrids, AOS Bulletin (Dec 1) pp 738. - according to Arditti, (1993, Microprop of Orchids (CH 1), Wiley) Rotor (Dr. Mac Daniels student) was 1st in vitro vegetative propagation. • FC Stewart, 1959 - 1st somatic embryogenesis (carrot) • FC Stewart, MO Mapes, and K Mears, 1959, Growth and organized development of cultured cells, Amer. Journal of Botany 45:653

  14. Explants used for micropropagation & embyo culture • shoot meristem, tip, bud • leaf or stem (internode) • root • anther / microspore • ovule • embryo (±) associated seed parts

  15. Shoot Explants Actively growing shoot tip Dormant bud Meristem tip

  16. Forcing solution(HQC, sucrose, +/- gibberellin): • allows a dormant bud to be forced under clean (indoor) conditions. • - Paul Read et al., 1990

  17. Leaf Explants

  18. Seed “explant” (propgaule)

  19. Field Trip: October 18, 2001 AnTec Laboratory Deflasking and Compotting Slipper Orchids www.ladyslipper.com/compot2.html Specialists in Paphiopedilum and Phramapedium (tropical ladyslipper orchids), species and hybrids.

  20. Stem Explant: Scrophularia sp., Wansang Lim

  21. Plant tissue culture systems used for micropropagation • shoot organ culture (axillary or nodal) • root organ culture • callus culture • cell suspension culture • somatic embryogenesis • anther (microspore) culture

  22. Plant tissue culture systems used for micropropagation • shoot organ culture (axillary or nodal)

  23. Plant tissue culture systems used for micropropagation • root organ culture

  24. Plant tissue culture systems used for micropropagation • callus culture Http://www.cas.muohio.edu/~wilsonkg/biotech/html/greentc_.htm Http://www.zeta.org.au/ ~brianc/pcc.htm

  25. Plant tissue culture systems used for micropropagation cell (liquid) suspension culture Cells and clumps of cells in suspension (liquid medium, agitation) Http://www.zeta.org.au/ ~brianc/pcc.htm http://www.bio.purdue.edu/nscort/image.html

  26. Plant tissue culture systems used for micropropagation somatic embryo culture http://mars.cropsoil.uga.edu/homesoybean/somprot.htm

  27. Plant Regeneration Pathways • Organogenesis via organ culture • Direct organogenesis • Organogenesis from callus • Somatic embryogenesis

  28. Plant Regeneration Pathways • Organogenesis via shoot organ culture + auxin

  29. Organogenesis via shoot organ culture

  30. Plant Regeneration Pathways Organogenesis via root organ culture

  31. Organogenesis via root organ culture

  32. Plant Regeneration Pathways • Direct organogenesis African violet leaf culture by Michael H. Renfroe Http://www.jmu.edu/biology/biofac/facfro/cloning/cloning.html

  33. Plant Regeneration Pathways • Organogenesis from Callus: • African Violet Use of a Protoplast Regeneration System for African Violet Improvement Traud Winkelmann, Institute for Breeding of Ornamental Species Ahrensburg, Germany http://aggie-horticulture.tamu.edu/tisscult/proto/wink/wink.html

  34. Plant Regeneration Pathways • Direct Organogenesis: Scrophularia sp., Wansang Lim

  35. Plant Regeneration Pathways • Somatic embryogenesis http://mars.cropsoil.uga.edu/homesoybean/somprot.htm

  36. http://mars.cropsoil.uga.edu/homesoybean/somprot.htm

  37. Plant Regeneration Pathways • Organogenesis via organ culture • Direct organogenesis • Organogenesis from callus • Somatic embryogenesis

  38. Revised to here 10/09/01

  39. Micropropagation via Shoot Culture

  40. Applications of Shoot Culture : • propagation per se • bulking plant breeding selections • production and maintenance of disease free plants • restoration of juvenility • stock plant management

  41. Shoot culture for propagation per se Theoretical: Eg. Mum Conventional prop. -- 30K plants per year Microprop: (based on 5 fold increase every 4 wks, assuming unlimited labor, space, etc. and no losses): 1 > 5 > 25 > 125 > 625 > 3125 > 15,625 > 78,125 > 390,625 > 1,953,125 > 9,765,625 > 48,824,125 > 244,140,625

  42. Shoot culture for propagation per se • Practical Considerations: economic niches • French hybrid lilacs • Raspberries • Rhododendrons, Kalmia

  43. Applications of Shoot Culture : • propagation per se • bulking plant breeding selections • production and maintenance of disease free plants • restoration of juvenility • stock plant management

  44. Applications of Shoot Culture : • propagation per se • bulking plant breeding selections • production and maintenance of disease free plants • restoration of juvenility • stock plant management

  45. Shoot culture for production and maintenance of disease free plants • assumption: because plant tissue cultures are aspetic they are disease free • - contributed to the spread of orchid viruses

  46. Shoot culture for production and maintenance of bacterial and fungal disease-free plants propagate Or X destroy save Culture indexing: for detection ofBacteria & fungi (not virus)

  47. Shoot culture for production and maintenance of disease free plants Virus

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