1) Clonal propagation is the multiplication of genetically identical copies of a plant cultivar through asexual reproduction. This can be done in vivo through methods like cutting, layering, and grafting, or in vitro through tissue culture techniques. 2) Micropropagation through tissue culture involves taking explants like shoot tips or meristems and culturing them on nutrient media to induce multiplication. Common methods are meristem culture, organogenesis, and somatic embryogenesis. 3) Clonal propagation has advantages like producing true-to-type plants, allowing off-season production, reducing life cycles, enabling large-scale production, and facilitating germplasm conservation and genetic transformation. However, it can be costly and labor intensive

1. A Term Paper Presentation On….“Clonal Propagation In Ornamental Plants”Course teacherDr. R.GnanamProfessorPersented byBhor Sachin AshokI.D.No.-09-607-001

2. What is clone & clonal propagation? A population derived from a single individual by asexual reproduction constitutes a clone.

3. Multiplication of genetically identical copies of a cultivar by asexual reproduction is called a clonal propagation.

4. Applied in crops like potato ,apple, pear, Horticultural crops, ornamental bulbs and in tuberous plants also.

5. This method can be done well under in vitro conditions.Contd...Clonal propagation through tissue culture popularly called micro propagation.

6. Use of tissue culture for micro propagation was initiated by G. Morel(1960) who found this approach in orchid propagation.

7. Axillary or apical shoot meristems in in vitro are most widely used methods in clonal propagation.Methods of Propagation

8. METHODS OF CLONAL PROPAGATIONCLONAL PROPAGATIONIN VIVOIN VITROCUTTINGBUD PROLIFERATIONBUDDINGORGANOGENESISLAYERINGSOMATIC EMBRYOGENESISGRAFTINGDIVISION & SEPARATION

9. METHODS OF IN VITRO CLONAL PROPAGATIONBud proliferationMeristem culture

10. Shoot tip culture

11. Single node culture

12. Axillary bud cultureOrganogenesis Organogenesis via callus formation

13. Adventitious organ formationEmbryogenesis Direct embryogenesis

14. Indirect embryogenesisMAJOR STAGES OF IN VITRO CLONAL PROPAGATION

15. MAJOR STAGES OF INVITRO CLONAL PROPAGATION

16. INVITRO MULTIPLICATION TAKES PLACE THROUGH….Multiplication via Meristem culture

17. Multiplication via somatic embryogenesis.

18. Multiplication via thin cell layer

19. Multiplication by adventitious shoots

20. Multiplication through callus cultureMULTIPLICATION VIA MERISTEM CULTUREIn vitro propagation through Meristem culture is the best possible means of virus elimination and produces a large numbers of plants in a short span of time.

21. The Meristem-tip culture is an aseptic culture on artificial medium of the apical dome without leaf primordia. It measures 0.2 to 0.3 mm.

22. Pelargonium, Chrysanthemum etc. are produced from mother plants who were cleaned up by culture of meristems

23. The pathway of regeneration undergoes several steps.

24. Starting with an isolated explants, with de-differentiation followed by re-differentiation and organization into meristematic centre.MULTIPLICATION VIA SOMATIC EMBRYOGENESISSomatic embryos ,which are bipolar structures, arise from individual cells and have no vascular connection with the maternal tissue of the explant.

25. Embryos develop through direct embryogenesis or through indirect embryogenesis.

26. Succeeded in ornamental pot plants like chrysanthemum (Dendrathemagrandiflorum)

27. Commercial application of somatic embryogenesis will be accomplished only when the germination rate of somatic embryos is high up to 80–85%.

28. Fig.1.Invitro somatic embryogenesis of Euphorbia pulcherrima. (A) Isolated somatic embryos of E. Pulcherrima(B) Germination of Somatic embryo.(C) Somatic embryos derived plantlets acclimatized in the greenhouse(D) Flowering of somatic embryo-derived plants

29. MULTIPLICATION VIA THIN CELL LAYERTCLs can be excised from stem, leaf, vein, floral stalk, petiole, pedicel, bulb-scale, etc

30. Reduced cell number in TCL is important developmental process or the morphogenetic programme.

31. Applications in higher plant tissue and organ culture and genetic transformation MULTIPLICATION VIA CALLUS CULTUREDifferentiation of plants from cultured cells via shoot -root formation or somatic embryogenesis is the fastest method of cloning of plant species.

32. But yet the cultures produced from calli are not stable genetically, which will decline plant regeneration capacity.

33. In some crops genetically stable calli also have been derived from explants of Lilium, Chrysanthemum and tomato.Basic parameters for in vitro Clonal propagation The Explant

34. The Nutrient medium

35. The culture EnvironmentEXAMPLES. . .

36. Advantages of Clonal propagation True to type plants

37. Off season production of plants

38. Reduction in life cycle of plant

39. Germplasm conservation

40. Genetic transformation

41. Large scale production of plants

42. Minimum growing space requiredLimitations of clonal propagationThis is not a simple technique

43. This technique is costly than sexual propagation

44. Woody plants does not gives response to clonal propagation

45. Plants regenerated through clonal propagation are not vigorous in growthApplication of In vitro clonal propagationIn vitro mutagenesis

46. Somaclonal variation

47. Cryopreservation

48. Genetic transformationCASE STUDIES

49. CHRYSANTHEMUM

51. Material & MethodStem nodal Explant selected

52. Surface sterilization

53. Inoculation on MS+BAP(0.5mg/l)+NAA(0.1mg/l),pH-5.6ResultAfter 2 weeks Callus Formation

54. Dark green colur & granular appearance.

55. After 5-6 weeks Sub culturingEmbryogenesisCells obtained in callus isolated from medium &Transferred to regeneration medium .

56. Subsequent subculturing on media containing BAP.

57. After 5-6 weeks small green leaf Appearance

58. Green embryonic mass transferred to MS media containing BAP(0.2-3.0mg/l)RootingTransfer of Seedlings from embryonic callus to basal MS (1/2)medium

59. Regeneration of complete plant Within 3 weeks of InoculationHardeningMedium-2-3 ml of MS Media+1% sugar CONTD. . .

60. GERBERA

62. Material & MethodSeeds soaked in Colchicine (20mg/l for 6 hours)Surface sterilization of ExplantGermination of seeds on MS basal mediumCrushed germinated seeds are used as ExplantInoculation on MS + Kn,BA+,2,4-D (1-5 mg/l) + BA,NAA,IBA (1-5 mg/l) + 5 mg/l Glutamine,pH-5.8Incubation for 10 days Transfer of callus on media containing BA,NAA,IBA (1-5 mg/l each)Hardening Media supplemented with AuxinsPlantlets are transferred to greenhouse conditionPlants shows 98%Survival rate.

63. Orchid

65. Explants available for orchid propagationShoot tip

66. Leaf Segment

67. Root segment

68. Inflorescence axis & flower bud

69. Rhizome Segment

70. Thin cell layerMaterial & method1-2 nodes as a Explant from 6 month old plantlet

71. Cross section (0.3-0.5mm thickness)excised from stem

72. Inoculation on MS media +sucrose(28gm/l)+Different conc. Of BA,Kn &IAA, pH-%.8

73. 20 ml media taken ion to glass bottle &Explant inoculation

74. Transfer of regenerated shoots to GR free media+0-20%sucrose +coconut water (for rooting)

75. Transfer in to pots under G.H. condition

76. Covering of plantlets by polythene bag

77. Survival rate is 92% GLADIOLOUS

79. ROSE

80. REFERENCE ARTICLE . . .

81. H. S. Chawla 2007 ; Introduction to plant biotechnology ; oxford & IBH publisher ; page no: 39-53.

82. S.S.Bohojwani and M.K. Razdan ; Plant tissue culture : theory and practice ; ELSEVIER Amsterdam-Oxford-New York-Tokyo (1983); page no; 313-343.

83. V K Srivastava , S Chadrasekhar; Commercialization of biotechnology for agriculture and aquaculture ; oxford & IBH publisher ; page no:215-221.

84. www.springerlink.comREFERENCES . . .

85. CONCLUSION. . .

86. DISCUSSION . . .

87. THANK U . . .