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LAMPIRAN
Tabel Lampiran 1.
Sifat Kimia Tanah yang Digunakan untuk Penelitian di Rumah Kaca
Sifat Kimia Tanah Pasir
(%)
Debu
(%)
Liat
(%)
PH H20 pH KC1 N (%I
c
(%)
S (mg/lOOg)
P Bray 11 (mg/lOOg) C/N ratio
KTK (me/100g) Jumlah Kation (me/100g) Kejenuhan Basa
(%)
AI - tukar (me/100g) H-tukar (me/lOOg)
Hasil Analisis
Sifat Kimia Tanah yang Digunakan untuk Penelitian di Lapang
Tabel Lampiran 2.
Hasil Analisis Sifat Kimia Tanah Kelompok Ulangan
I
I1
I11
Pasir
(%)
13/84
19/48
19/73
Debu
(%)
30180
33/53
32/84
Liat
(%)
55,36
46,99
47,43
pH H20
5,15
5/30
5/11
pH KC1
4/08
3/95
4/10
12/62
11/11
N (%)
0,14
0,13
0114
c (%I
1/73
1/42
1/56
P Bray I1 (mg/100g) 0,57
0,40
0,48
Ca (me/lOOg)
1/69
2/23
1/66
~g (me/100g)
1/25
K
(me/lOOg)
0,27
Na (me/lOOg)
0,29
C/N ratio
KTK (me/100g)
18,87
Jumlah kation (me/lOOg)
3,51
Kejenuhan basa ( % ) 18/58 Al-tukar (me/100g)
1/60
H-tukar (me/100g)
0,64
Fe (ppm)
6,67
Mn (ppm)
20~19
Cu (ppm)
1,50
Zn (ppm)
3/56
11,33
Tabel Lampiran 3.
Metode Isolasi Spora MVA (Gerdemann dan Nicolson, 1963; Daniels dan Skipper, 1982 dimodifikasi)
Sampel tanah sebanyak 50 g berat kering udara dicampur dengan 200 ml air dalam gelas piala, kemudian diaduk hingga larut homogen. Larutan tersebut dibiarkan beberapa detik agar partikel-partikel besar mengendap. Setelah mengendap, suspensi disaring melalui 4 saringan yang disusun dari
atas kebawah masing-masing ber-
ukuran 1000 pm, 425 pm,
149 pm
dan 45 pm. Saringan
1000 pm dan 425 pm untuk memisahkan partikel- partikel
besar
.
Partikel-partikel halus berikut spora yang
tertampung
pada saringan 149 pm dan 45 pm dituang kedalam tabungtabung sentrifusi masing-masing sebanyak 25 ml. Larutan sukrosa 60 % sebanyak 25 ml
ditambahkan keda-
lam tabung-tabung sentrifusi tersebut, kemudian disentrifusi selama 3 menit dengan kecepatan 2000 rpm. Supernatan disaring dengan saringan 45 pm dan dicuci dengan air mengalir. Spora yang tertahan ditampung kedalam cawan petri
di-
lengkapi dengan cawan petri plastik bergaris. Pengamatan spora dan penghitungan populasi spora MVA dengan menggunakan mikroskop dissecting. Identifikasi dan merekam
spesies spora MVA
dengan
menggunakan mikroskop compound yang dilengkapi kamera Nikon dan film ASA 100.
Tabel
1. Akar
Lampiran 4.
Metode Pewarnaan Akar Centro dan Puero (Kormanik dan McGraw, 1982 dimodif ikasi)
dicuci dengan air hingga bersih dan masukkan
dalam botol berisi FAA. 2. Akar dikeluarkan dari dalam botol
kemudian dicuci
dengan air yang mengalir untuk menghilangkan FAA. 3.
Akar dimasukkan ke dalam tabung berisi KOH 10% dan di panaskan
90°c
dalam bak air di
ruang asam
(fwe
hood) selama satu jam. 4.
Akar
dicuci dengan air yang mengalir untuk menghi-
langkan KOH hingga tampak jernih. 5. Apabila akar belum tampak jernih, ditambahkan H202
50% direndam selama 1 6.
-
5 jam.
Larutan H202 dibuang, akar dicuci dengan air yang mengalir hingga bersih dari larutan tersebut.
7.
Akar
direndam dalam HC1 0,l N
selama 10 menit kemu-
dian larutan tersebut dibuang. 8. Akar
direndam dalam asam
fuhsin-asam laktat 0,01%
atau 0.05% trypan blue dan dipanaskan 90'~ dalam bak air di ruang asam (fume hood) selama 10 -
60 menit
atau sampai terjadi penetrasi warna. 9. Larutan .trypanblue dibuang, akar diambil dan ditempatkan pada cawan petri kemudian ditambahkan larutan campuran gliserin dan aquadest (1
:
1).
10. Kolonisasi MVA pada akar siap diamati dengan menggunakan mikroskop d i s s e c t i n g .
Tabel Lampiran 5 .
Penghitungan Persentase Kolonisasi MVA pada Akar dengan Metode Gridline (Giovannetti dan Mosse, 1980)
1. Contoh akar setelah dilakukan pewarnaan, diambil seca-
2.
3.
4. 5.
6.
ra acak kemudian dipotong-potong sepanjang 1 cm dan disebarkan merata pada cawan petri. Kisi-kisi (sama sisi) dibuat pada lembar plastik (kertas putih) dengan ukuran masing-masing sisi 1,27 cm atau dengan menggunakan cawan plastik berkisi-kisi. Plastik/kertas putih berkisi tersebut No. 2 kemudian diletakkan pada cawan petri yang lebih besar, sehingga cawan petri tempat contoh akar dapat diletakkan diatasnya. Letakkan cawan-cawan petri tersebut No. 3 dibawah mikroskop d i s s e c t i n g dengan pembesaran 10 - 40 kali. Cara pengamatan dilakukan dengan menghitung akar yang terkoloni maupun yang tidak terkoloni MVA, mengikuti garis horisontal dan garis vertikal kemudian dicatat pada tabel pengamatan. Tabel pengamatan : --
No. Kisi
Total akar
Jumlah 7.
xn
Akar yang terinfeksi
yn
Persentase kolonisasi MVA pada akar dihitung dengan rumus :
Yn -
x 100 %
xn 8. Pelaksanaan praktis penghitungan persentase kolonisasi MVA tersebut digambarkan oleh Bougher et dl., (1994) sebagai berikut :
1
. A k u disebarkan dalam porti./pirim
'
p l a s t i k bergaris Akar dalam l a k t o g l i s e r
Jumlah akar
3.
Akar bermikorisa
Eituzig mengikuti g a r i s h o r i s o d t a l dan v e r t i k a l
Garis h o r i s o n t a l 111
Tabel Lampiran 6. Analisis Kecernaan Bahan Kering Secara In-Vitro (Metode PepsinSelulase) (Terry and Tilley, 1963) ~ ~ ~ 1. Contoh hijauan dikeringkan pada suhu 1 0 0 setelah kering contoh kemudian digiling dengan ukuran 1 mm dan disimpan dalam botol-botol tertutup. 2. Timbang
contoh
0,5 gram dan masukkan dalam tabung
sentrifusi 100 ml. 3. Tambahkan 50 ml
pepsin
0,2 %
kedalam larutan HC1
0,l N. 4.
Diinkubasikan pada suhu 3g°C selama 48 jam.
5. Disaring dengan sinter glass
dan dicuci dengan air
destilasi satu kali, kemudian residunya dituang kembali kedalam tabung sentrifusi. Cuci sinter glass dengan selulase 2 , 5 % dalam larutan buffer. 6. Tambahkan 50 m l larutan selulase. 7.
Diinkubasikan selama 48 jam pada suhu 39'~.
8. Disaring dengan
sinter glass
kemudian dicuci dengan
air destilasi dan diulang tiga kali. 9. Dikeringkan dalam oven satu malam kemudian ditimbang. 10. Dipanaskan pada suhu 550'~ selama 3 jam kemudian diditimbang. 11. Kecernaan bahan kering secara in-vitro dihitung dengan rumus : BK contoh KCBK
-
(BK residu - BK blangko)
=
X
BK contoh Keterangan : KCBK
=
kecernaan bahan kering hijauan
100 %
Tabel Lampiran 7.
Hasil Analisis Keragaman Produksi dan Nilai Nutrisi Legum pada Penelitian di Rumah Kaca
Kadar SK
DB
Model
Hasil BK
79
W 1 1 S*W M*W 1 S*M*W 1 P*W 4 S*P*W 4 M*P*W 4 S*M*P*W 4 Galat (2) 40 Total 119
KeteranQan:
SK DB S M P W KK
= = =
=
= = =
sumber keragaman derajat bebas spesies legum inokulasi MVA pupuk BF periode pemotongan koefisien keragaman
N
Serapan P
N
P
Tabel Lampiran 8.
Hasil Analisis Keragaman Produksi Spora dan Kolonisasi MVA pada Penelitian di Rumah Kaca Pr>F
SK
DB Jumlah Spora
Model
9
S P S*P Galat Total
1 4
4
20 29
KK. ( % )
-:
SK DB S P KK
= = =
= =
sumber keragaman derajat bebas spesies legum pupuk BF koefisien keragaman
Kolonisasi MVA
Tabel Lampiran 9. Hasil Analisis Keragaman Produksi dan Nilai Nutrisi Legum pada Penelitian di Lapang
SK
DB Hasil BK
Model
Kadar N
99
s M S*M P S*P M*P S*M*P Galat W 2 S*W 2 M*W 2 S*M*W 2 P*W 8 S*P*W 8 M*P*W 8 S*M*P*W 8 Galat (2) 80 Total 179
Keteranaan: SK = sumber keragaman DB = derajat bebas S = spesies legum M = inokulasi MVA P = pupuk BE' W = periode pernotongan KK = koefisien keragaman
P
Zn
Serapan Cu
N
P
Tabel Lampiran 10.
Hasil Analisis Keragaman Kecernaan Bahan Kering Hijauan Legum pada Penelitian di Lapang Pr>F
Sumber Keragaman
Derajat Bebas
Model
W S*W M*W S*M*W P*W S*P*W M*P*W S*M*P*W Galat (2) Total
S M P W KK
= =
= =
=
spesies inokulasi MVA pupuk BF periode pemotongan koefisien keragaman
Kecernaan Bahan Kering
Tabel Lampiran 11.
Hasil Analisis Keragaman Produksi Spora dan Kolonisasi MVA pada Penelitian di Lapang Pr>F
SK Model
DB
Jumlah Spora
Kolonisasi MVA
0,0001
0,0001
19
S
M S*M
P S*P M*P S*M*P
Galat Total KK ( % I
-:
SK DB S M P KK
= = =
= = =
sumber keragaman derajat bebas spesies legum inokulasi MVA pupuk BF koefisien keragaman
