LAMPIRAN
1. Alur Kerja Antagonisme In Vitro
Diinokulasikan pada media MGMC Pada jarak 3,5 cm dari tempat Inokulum bakteri.
Sebanyak 10 µl suspensi bakteri kitinolitik diteteskan cakram standard Oxoid
Diinkubasi pada suhu 300C dan dilakukan pengamatan selama lima hari
Biakan Jamur Colletotrichum sp
2. Pengamatan abnormalitas miselium Colletotrichum sp. Setelah uji antagonis
Dipotong ujung miselium
Colletotrichum sp. Pada media MGMC berbentuk block square Diletakkan pada objek glass
Diamati pertumbuhan miselium
Colletotrichumsp.yang abnormal
Biakan Jamur Colletotrichum sp
3. Pembuatan Suspensi Bakteri Kitinolitik
Disubkultur dalam media NA dan Diinkubasi ± 2 hari Diambil biakan bakteri kitinolitik dengan jarum ose Dimasukkan ke dalam tabung reaksi yang berisi akuades steril sebanyak 10 ml
Dihomogenkan dengan cara divortex
Disamakan kekeruhannya
dengan Standar Mac
farland yaitu 108 CFU/ml Biakan bakteri kitinolitik
4. Pembuatan Suspensi Jamur Colletotrichum sp
Disubkultur dalam media PDA Ditetesi dengan aquadest steril Sebanyak 10 ml kemudian dikikis dengan jarum kait agar konidia Terlepas.
Disaring dengan kain kasa
Disentrifugasi
Diencerkan sehingga mencapai kerapatan 2 x 105 konidia / ml Biakan Jamur Colletotrichum sp
Campuran Miselium dan Konidia Jamur
Filtrat
Konsentrat
Suspensi Jamur Colletotrichum sp.
5. Pengujian in vivo bakteri kitinolitik terhadap tanaman kakao
Disemprot 10 ml suspensi bakteri kitinolitik.
Tanaman disungkup plastik
Disemprot konidia
Collletotrichum dengan kerapatan
2x105 konidia/ml
Dihitung intensitas serangan dan Luas Serangan
Tanaman Kakao
Tanaman Kakao Yang Tahan Terhadap Patogen Jamur Colletotrichum sp.
6. Uji Patogenisitas
Tanaman kakao
Kontrol Positif : Pemberian Colletotrichum sp Kontrol Negatif : Tanpa Pemberian Colletotrichum sp
Perlakuan : 1. Suspensi bakteri kitinolitik KR05 + Spora Colletotrichum sp 2. Suspensi bakteri kitinolitik LK 08 + Spora Colletotrichum sp 3. Suspensi bakteri kitinolitik BK 08 + Spora Colletotrichum sp 4. Suspensi bakteri kitinolitik BK 15 + Spora Colletotrichum sp 5. Suspensi bakteri kitinolitik BK 17 + Spora Colletotrichum sp
7. Intensitas Serangan Minggu 1
Perlakuan Ulangan Total
1 2 3 4 5 BK13 0.7 2.12 0.7 0.7 0.7 4.92 6.34 4.92 7.13 4.92 19.7 7.75 55.68 KRO5 0.7 2.12 2.12 0.7 0.7 BK17 2.12 0.7 0.7 0.7 0.7 BK15 0.7 2.91 2.12 0.7 0.7 LK08 0.7 2.12 0.7 0.7 0.7 Kontrol Positif 3.53 3.53 4.06 4.06 4.52 Kontrol Negatif 0.7 2.12 3.53 0.7 0.7 Total 9.15 15.62 13.93 8.26 8.72 Fk = (55.68)2 = 3100.2624 = 88.5789 35 35 Jk T = (0.7)2 + ...+ (0.70)2 - Fk = 52.4203 JK P = (4.92)2 + ...(7.75)2 - Fk = 33.78194 5 JK B = (9.15)2 + ...+ (8.72)2 - Fk = 6.5665 7 JK Galat = JKT – JKP – JKB = 12.07186 Tabel Analisis SK DB JK KT F,Hit F.Tab 5% 1% B 4 6.5665 1.6416 3.26* 2.78 4.22 P 6 33.7819 5.6303 11.195** 2.51 3.67 G 24 12.0718 0.5029
Keterangan: ** = berbeda sangat nyata * = berbeda nyata
8. Luas Serangan Minggu 1
Perlakuan Ulangan Total
1 2 3 4 5 BK13 0.7 4.52 0.7 0.7 0.7 7.32 11.14 7.32 11.14 7.32 22.6 11.14 77.98 KRO5 0.7 4.52 4.52 0.7 0.7 BK17 4.52 0.7 0.7 0.7 0.7 BK15 0.7 4.52 4.52 0.7 0.7 LK08 0.7 4.52 0.7 0.7 0.7 Kontrol Positif 4.52 4.52 4.52 4.52 4.52 Kontrol Negatif 0.7 4.52 4.52 0.7 0.7 Total 12.54 27.82 20.18 8.72 8.72 Fk = (77.98)2 = 6080.8804 = 173.739 35 35 Jk T = (0.7)2 + ...+ (0.70)2 - Fk = 122.5766 JK P = (7.32)2 + ...(11.14)2 - Fk = 35.02 5 JK B = (12.54)2 + ...+ (8.72)2 - Fk = 39.19 7 JK Galat = JKT – JKP – JKB = 48.36 Tabel Analisis SK DB JK KT F,Hit F.Tab 5% 1% B 4 39.19 9.79 4.85** 2.78 4.22 P 6 35.02 5.83 2.89 * 2.51 3.67 G 24 48.36 2.015
Keterangan : * = Berbeda Nyata
9. Intensitas Serangan Minggu 2
Perlakuan Ulangan Total
1 2 3 4 5 BK13 0.7 2.12 0.7 0.7 0.7 4.92 6.34 4.92 7.13 4.92 19.7 7.75 55.68 KRO5 0.7 2.12 2.12 0.7 0.7 BK17 2.12 0.7 0.7 0.7 0.7 BK15 0.7 2.91 2.12 0.7 0.7 LK08 0.7 2.12 0.7 0.7 0.7 Kontrol Positif 3.53 3.53 4.06 4.06 4.52 Kontrol Negatif 0.7 2.12 3.53 0.7 0.7 Total 9.15 15.62 13.93 8.26 8.72 Fk = (77.98)2 = 6080.8804 = 173.739 35 35 Jk T = (0.7)2 + ...+ (0.70)2 - Fk = 122.5766 JK P = (7.32)2 + ...(11.14)2 - Fk = 35.02 5 JK B = (12.54)2 + ...+ (8.72)2 - Fk = 39.19 7 JK Galat = JKT – JKP – JKB = 48.36 Tabel Analisis SK DB JK KT F,Hit F.Tab 5% 1% B 4 39.19 9.79 4.85** 2.78 4.22 P 6 35.02 5.83 2.89 * 2.51 3.67 G 24 48.36 2.015
Keterangan : * = Berbeda Nyata
10. Luas Serangan Minggu 2
Perlakuan Ulangan Total
1 2 3 4 5 BK13 0.7 4.52 0.7 0.7 0.7 7.32 11.14 7.32 11.14 7.32 22.6 11.14 77.98 KRO5 0.7 4.52 4.52 0.7 0.7 BK17 4.52 0.7 0.7 0.7 0.7 BK15 0.7 4.52 4.52 0.7 0.7 LK08 0.7 4.52 0.7 0.7 0.7 Kontrol Positif 4.52 4.52 4.52 4.52 4.52 Kontrol Negatif 0.7 4.52 4.52 0.7 0.7 Total 12.54 27.82 20.18 8.72 8.72 Fk = (77.98)2 = 6080.8804 = 173.739 35 35 Jk T = (0.7)2 + ...+ (0.70)2 - Fk = 122.5766 JK P = (7.32)2 + ...(11.14)2 - Fk = 35.02 5 JK B = (12.54)2 + ...+ (8.72)2 - Fk = 39.19 7 JK Galat = JKT – JKP – JKB = 48.36 Tabel Analisis SK DB JK KT F,Hit F.Tab 5% 1% B 4 39.19 9.79 4.85** 2.78 4.22 P 6 35.02 5.83 2.89 * 2.51 3.67 G 24 48.36 2.015
Keterangan : * = Berbeda Nyata
11. Intensitas Serangan Minggu 3
Perlakuan Ulangan Total
1 2 3 4 5 BK13 0.7 2.12 0.7 0.7 0.7 4.92 6.34 4.92 7.75 4.92 19.7 7.75 56.3 KRO5 0.7 2.12 2.12 0.7 0.7 BK17 2.12 0.7 0.7 0.7 0.7 BK15 0.7 3.53 2.12 0.7 0.7 LK08 0.7 2.12 0.7 0.7 0.7 Kontrol Positif 3.53 3.53 4.06 4.06 4.52 Kontrol Negatif 0.7 2.12 3.53 0.7 0.7 Total 9.15 16.24 13.93 8.26 8.72 Fk = (56.3)2 = 3169.69 = 90.5625 35 35 Jk T = (0.7)2 + ...+ (0.70)2 - Fk = 54.3575 JK P = (4.92)2 + ...(7.75)2 - Fk = 33.6434 5 JK B = (9.15)2 + ...+ (8.72)2 - Fk = 7.4047 7 JK Galat = JKT – JKP – JKB = 13.30934 Tabel Analisis SK DB JK KT F,Hit F.Tab 5% 1% B 4 7.4047 1.8511 3.33* 2.78 4.22 P 6 33.6434 5.6072 10.112** 2.51 3.67 G 24 13.3093 0.5545
Keterangan : * = Berbeda Nyata
12. Luas Serangan Minggu 3
Perlakuan Ulangan Total
1 2 3 4 5 BK13 0.7 4.52 0.7 0.7 0.7 7.32 11.14 7.32 11.14 7.32 22.6 11.14 77.98 KRO5 0.7 4.52 4.52 0.7 0.7 BK17 4.52 0.7 0.7 0.7 0.7 BK15 0.7 4.52 4.52 0.7 0.7 LK08 0.7 4.52 0.7 0.7 0.7 Kontrol Positif 4.52 4.52 4.52 4.52 4.52 Kontrol Negatif 0.7 4.52 4.52 0.7 0.7 Total 12.54 27.82 20.18 8.72 8.72 Fk = (77.98)2 = 6080.8804 = 173.739 35 35 Jk T = (0.7)2 + ...+ (0.70)2 - Fk = 122.5766 JK P = (7.32)2 + ...(11.14)2 - Fk = 35.02 5 JK B = (12.54)2 + ...+ (8.72)2 - Fk = 39.19 7 JK Galat = JKT – JKP – JKB = 48.36 Tabel Analisis SK DB JK KT F,Hit F.Tab 5% 1% B 4 39.19 9.79 4.85** 2.78 4.22 P 6 35.02 5.83 2.89 * 2.51 3.67 G 24 48.36 2.015
Keterangan :** = Berbeda Sangat Nyata * = Berbeda Nyata
13. Intensitas serangan Minggu 4
Perlakuan Ulangan Total
1 2 3 4 5 BK13 0.7 2.12 0.7 0.7 2.12 6.34 7.76 6.34 7.75 4.92 20.12 7.75 60.98 KRO5 0.7 2.12 2.12 0.7 2.12 BK17 2.12 2.12 0.7 0.7 0.7 BK15 0.7 3.53 2.12 0.7 0.7 LK08 0.7 2.12 0.7 0.7 0.7 Kontrol Positif 3.53 3.53 4.06 4.06 4.94 Kontrol Negatif 0.7 2.12 3.53 0.7 0.7 Total 9.15 17.66 13.93 8.26 11.98 Fk = (60.98)2 = 3718.5604 = 106.2445 35 35 Jk T = (0.7)2 + ...+ (0.70)2 - Fk = 54.7339 JK P = (6.34)2 + ...(7.75)2 - Fk = 106.2445 5 JK B = (9.15)2 + ...+ (11.98)2 - Fk = 8.2399 7 JK Galat = JKT – JKP – JKB = 14.78758 Tabel Analisis SK DB JK KT F,Hit F.Tab 5% 1% B 4 8.2399 2.0599 3.34* 2.78 4.22 P 6 31.7064 5.2844 8.577** 2.51 3.67 G 24 14.7875 0.6161
Keterangan : * = Berbeda Nyata ** = Berbeda Sangat Nyata
14. Luas Serangan Minggu 4
Perlakuan Ulangan Total
1 2 3 4 5 BK13 0.7 4.52 0.7 0.7 4.52 11.14 14.96 11.14 11.14 7.32 22.6 11.14 89.44 KRO5 0.7 4.52 4.52 0.7 4.52 BK17 4.52 4.52 0.7 0.7 0.7 BK15 0.7 4.52 4.52 0.7 0.7 LK08 0.7 4.52 0.7 0.7 0.7 Kontrol Positif 4.52 4.52 4.52 4.52 4.52 Kontrol Negatif 0.7 4.52 4.52 0.7 0.7 Total 12.54 31.64 20.18 8.72 16.36 Fk = (89.44)2 = 7999.5136= 228.55 35 35 Jk T = (0.7)2 + ...+ (0.70)2 - Fk = 127.5868 JK P = (11.14)2 + ...(11.14)2 - Fk = 28.358 5 JK B = (12.54)2 + ...+ (16.36)2 - Fk = 44.2016 7 JK Galat = JKT – JKP – JKB = 55.0272 Tabel Analisis SK DB JK KT F,Hit F.Tab 5% 1% B 4 44.2016 11.050 4.82** 2.78 4.22 P 6 28.358 4.72 2.06 tn 2.51 3.67 G 24 55.0272 2.29
Keterangan : tn = Tidak ada perbedaan nyata ** = berbeda Sangat Nyata
FOTO PENELITIAN
