Experimen of plant cell tissue and organ culture

A       IDENTITAS

          Hari/Tanggal    : Kamis/12 Juli 2012

          Waktu               : 14.30-17.30 dan 18.30-21.30

B       KEGIATAN                     

         Experimen of plant cell tissue and organ culture

C       RINGKASAN KEGIATAN

          Setelah menyelesaikan kelas pagi, kami harus kembali ke kampus yaitu ke laboratorium kultur jaringan C5006 untuk melihat cara kerja kultur jaringan. Laboratorium kultur jaringan kecil tapi peralatannya lengkap. Aku merekam beberapa bagian keterangan dari Dr. Cui. Kelas sore berakhir pukul 05.00 a.m. Pukul 18.30 kami harus kembali lagi untuk melakukan propagasi pada tanaman anggrek.

Beberapa persyaratan dalam kultur jaringan

a.    Jaringan yang sesuai (some tissues culture better than others)

b.    Media tanam yang sesuai dan mengandung bahan organik maupun anorganik yang dibutuhkan tanaman untuk tumbuh

c.    Dilakukan dalam kondisi aseptik

2 Types of cell & tissues:

a.    Many different types of cells

b.    Varying degrees of specialization (meristematic, embryonic, and reproductive)

c.    The steps culturing plant tissue:

1.      Multiplication-the explants gives rise to a callus (a mass of loosely arranged cells) which is manipulated by varying is manipulated by varying sugar concentrations and the auxin (low): cytokinin (high) ratios to form multiple shoots

2.      The callus may be subdivided a number of times

3.      Root formation – the shoots are transferred to a growth medium with relatively higher auxin: cytokinin ratios

Atau

1.        Selection of the plant tissue (explant) from a healthy vigorous ‘ mother plant ‘ – this is often the apical bud, but can be other tissue

2.    This tissue must be sterilized to remove microbial contaminants. Elimination of microbial contaminants:

·           Surface contaminants- principally microbial saprophytes that are eliminated by surface disinfectation

·           Internal contaminants- principally pathogens that are eliminated by thermotherapy (35-40 C) and culture of explants free of organisms or by antibiotics

·           Maintenance of asepsis (free from microorganisms) – during excision and culture – procedures are carried out in sterile laminar flow positive pressure hoods (0.3 µm filters)

·           Establishment of the  explants in a culture medium. The medium sustains the plant cells and encourages cell division. It can be solid or liquid

·           Each plant species has  particular medium requirements that must be establisheds by trial  and error

·           The rooted shoots are potted up (deflasked) and ‘ hardened off ‘ by gradually decreasing the humidity

·           This is necessary as many young tissue culture plants have no waxy cuticle to prevent water loss

Medium kultur jaringan mengandung:

1.      Inorganic salt formulations

2.      Source of carbohydrate sucrose or sugar

3.      Vitamins

4.      Water

5.      Plant hormones- auxins ( stimulates root development), cytokinins ( stimulates shoot development), GA’s

Generally, the ratio of these two hormones can determine plant development:

·            Auxin     Cytokinin = root development

·            Auxin     Cytokinin = shoot development

·         Auxin = cytokinin = callus development

6.      Solidifying agents