Saran

BAB V. PENUTUP

5.2 Saran

Dari hasil penelitian yang telah diperoleh maka disarankan untuk melakukan penelitian lebih lanjut mengenai identifikasi dan cara ekstraksi isolat bakteri endofit, sehingga senyawa metabolit sekunder yang terkandung didalamnya dapat terekspresi.

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LAMPIRAN

Lampiran 2. Skema Kerja Penelitian

Daun Rambutan Segar

Sterilisasi Permukaan Daun Rambutan

Isolasi Bakteri Endofit

Pemurnian dan Peremajaan Isolat - Pembuatan Stock Culture dan

Working Culture

Fermentasi

Skrining Fitokimia dan Uji Aktivitas Antibakteri

Identifikasi Bakteri Endofit - Makroskopis

- Mikroskopis

Lampiran 3. Sterilisasi Permukaan Daun Alkohol 70% 1 menit NaOCl 5.25 % 5 menit Alkohol 70% 30 detik Akudes steril 1 menit Akudes steril 1 menit Daun Rambutan Cuci Bersih Kontrol Daun

Lampiran 4. Pemurnian dan Identifikasi Isolat

Metode Streak Stock

Makroskopis - Bentuk koloni - Bentuk tepi koloni - Warna koloni

Mikroskopis

- Pewarnaan Gram - Uji katalase

Lampiran 5. Fermentasi Bakteri Endofit Media NB Isolat digojog 170 rpm, 27^oC, 48 jam Sentrifugasi 3000 rpm, 20 menit, 4^oC Supernatan Uji Aktivitas Antibakteri Skrining Fitokimia KLT Reagen

Lampiran 6. Uji Aktivitas Antibakteri Supernatan Bakteri uji NaCl 0.9% 5 mL 0.1 % suspensi bakteri digojog 120 rpm, 27^oC E. coli 4 jam S. thypimurium 10 jam S. aureus 3 jam B. subtilis 13 jam 1 ml suspensi bakteri Metode Pour Plate Media NB

Lampiran 7. Hasil Isolasi Bakteri Endofit Pada Daun Rambutan

Lampiran 8. Hasil Pemurnian Isolat Bakteri Endofit

DR1 DR2

Lampiran 9. Skrining Fitokimia Daun Segar

Kontrol Saponin Alkaloid Tanin

Hasil : Positif Hasil : Negatif Hasil : Positif Busa Stabil (a) Dragendorff Hitam-hijau

(b) Meyer

Fenolik Terpenoid/Steroid Flavonoid

Hasil : Positif Hasil : Positif Hasil : Positif Ungu tua (a) Merah (bawah) (a) Abu-abu

(b) Kuning-kehijauan (b) Warna hilang

(a) (b)

(b) (a)

(b) (a)

Lampiran 10. Uji Katalase Isolat Gambar Keterangan

+

+

+

+

DR1

DR2

DR3

DR4

Lampiran 11. Uji Aktivitas Antibakteri Uji Keterangan Escherichia coli (a) DR1 (b) DR2 (c) DR3 (d) DR4 (-) Media steril (+) Kloramfenikol Salmonella thypimurium (a) DR1 (b) DR2 (c) DR3 (d) DR4 (-) Media steril (+) Kloramfenikol Staphylococcus aureus (a) DR1 (b) DR2 (c) DR3 (d) DR4 (-) Media steril (+) Kloramfenikol (a) (a) (b) (a) (b) (b) (c) (c) (c) (d) (d) (d) (+) (+) (+) (-) (-) (-)

Bacillus subtilis (a) DR1 (b) DR2 (c) DR3 (d) DR4 (-) Media steril (+) Kloramfenikol (b) (c) (d) (-) (+) (a)

Lampiran 12. Identifikasi Isolat Bakteri Endofit

Isolat Pewarnaan Gram Keterangan

DR1 Pewarnaan Gram : Gram negatif Bentuk Sel : Kokus

Skala : Perbesaran 1000x

DR2 Pewarnaan Gram : Gram negatif Bentuk Sel : Batang

Skala : Perbesaran 1000x

DR3 Pewarnaan Gram : Gram negatif Bentuk Sel : Kokus

Skala : Perbesaran 1000x

DR4 Pewarnaan Gram : Gram positif Bentuk Sel : Kokus

Lampiran 13. Identifikasi Bakteri Uji

No. Bakteri Keterangan

1. Bakteri : Staphylococcus aureus

Pewarnaan Gram : Gram positif Bentuk Sel : kokus

2. Bakteri : Bacillus subtilis

Pewarnaan Gram : Gram positif Bentuk Sel : basil

3. Bakteri : Escherichia coli

Pewarnaan Gram : Gram negatif Bentuk Sel : kokus

4. Bakteri : Salmonella thypimurium

Pewarnaan Gram : Gram negatif Bentuk Sel : basil

Lampiran 14. Karakteristik Koloni Bakteri Pada Media Agar