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Table 1 Experimental and coded levels used in box-behnken design for studying the effects of cellulase (x1), macerozyme (x2), mannitol (x3) and hydrolysis time (x4) on yield of isolated protoplasts along with the predicted mean and observed responses, and FDA results

From: Optimization of isolation and transfection conditions of maize endosperm protoplasts

Experimental factor Coded symbol Coded variable levels        
Low Centre High        
−1 0 +1        
Cellulase (%) X1 1.00 1.50 2.00        
Macerozyme (%) X2 0.50 0.75 1.00        
Mannitol X3 0.40 0.60 0.80        
Hydrolysis time (h) X4 4 6 8        
Test Number Coded levels Actual levels Response
Yield, Y (×106 cells/ml) FDA
X1 X2 X3 X4 X1 X2 X3 X4 Observed Y Predicted Y (%)
Coded levels for the four factors selected for protoplast isolation
1 +1 +1 0 0 2.00 1.00 0.60 6 0.95 1.52 92.3
2 +1 −1 0 0 2.00 0.50 0.60 6 1.45 1.68 93.9
3 −1 +1 0 0 1.00 1.00 0.60 6 1.25 1.49 90.9
4 −1 −1 0 0 1.00 0.50 0.60 6 1.10 1.00 95.8
5 +1 0 +1 0 2.00 0.75 0.80 6 1.55 2.22 95.1
6 +1 0 −1 0 2.00 0.75 0.40 6 2.70 2.56 97.2
7 −1 0 +1 0 1.00 0.75 0.80 6 1.70 1.65 94.6
8 −1 0 −1 0 1.00 0.75 0.40 6 3.30 2.43 96.9
9 +1 0 0 +1 2.00 0.75 0.60 8 1.70 1.02 90.3
10 +1 0 0 −1 2.00 0.75 0.60 4 2.85 2.21 95.1
11 −1 0 0 +1 1.00 0.75 0.60 8 0.85 1.22 90.5
12 −1 0 0 −1 1.00 0.75 0.60 4 0.90 1.31 92.0
13 0 +1 +1 0 1.50 1.00 0.80 6 3.00 2.02 96.0
14 0 + −1 0 1.50 1.00 0.40 6 1.70 1.75 91.8
15 0 −1 +1 0 1.50 0.50 0.80 6 1.35 1.03 94.1
16 0 −1 −1 0 1.50 0.50 0.40 6 1.70 2.41 91.5
17 0 +1 0 +1 1.50 1.00 0.60 8 0.95 0.79 96.4
18 0 +1 0 −1 1.50 1.00 0.60 4 1.15 1.43 92.3
19 0 −1 0 +1 1.50 0.50 0.60 8 1.10 0.62 91.7
20 0 −1 0 −1 1.50 0.50 0.60 4 1.30 1.26 93.8
21 0 0 +1 +1 1.50 0.75 0.80 8 1.30 1.88 95.8
22 0 0 +1 −1 1.50 0.75 0.80 4 1.10 1.20 97.3
23 0 0 −1 +1 1.50 0.75 0.40 8 0.75 1.12 93.3
24 0 0 −1 −1 1.50 0.75 0.40 4 3.20 3.08 93.8
25 0 0 0 0 1.50 0.75 0.60 6 7.75 8.03 93.3
26 0 0 0 0 1.50 0.75 0.60 6 8.50 8.03 93.9
27 0 0 0 0 1.50 0.75 0.60 6 7.60 8.03 91.8
28 0 0 0 0 1.50 0.75 0.60 6 8.25 8.03 96.7
29 0 0 0 0 1.50 0.75 0.60 6 8.05 8.03 94.1