This document summarizes a study on the effects of chitosan and chlorocholine chloride on cocoyam minituberization. The study found that applying chitosan at concentrations of 1, 2, and 3 g/L and chlorocholine chloride at 5, 10, and 15 mg/L significantly increased average plant height, number of leaves, and leaf surface area compared to the control. A two-way ANOVA and Tukey's multiple comparisons test showed highly significant interactions between the factors and differences between the means of most treatment combinations. The column factor of chitosan and chlorocholine chloride treatment had the strongest effect on the results.

1. Advanced Statistics for
Measurement Sciences
By Mesele Tilahun Belete
University of Science and Technology, Korean Research Institute
of Bioscience and Biotechnology (KRIBB) Campus
Course Instructor: 강 남구 (Ph.D.)
Data analysis presentation
Dec 13, 2021
Email: mesele21@kribb.re.kr

2. Cocoyam (Xanthosoma sagittifolium) is the most
important root and tuber crops in the world

3. Objective of Study
This research was aimed at studying the effect of chitosan and chlocholine chloride
on the minituberization of cocoyam
Originally the data were analyzed by SPSS 16.0 with Student-Newman Keul’s and Duncan’s test with LSD 5%
But now analyzed by “GraphpadPrism 9.3” with Tukey's multiple comparison post Hoc test.
Chitosan : plant growth enhancer and antifungal, reduce stress(drought), increase yield and seed quality
Chlocholine chloride: micro tuber formation , stimulates tuber initiation by recalcitrant genotypes

4. Methodology
• This study was carried out in order to evaluate the effect of
chitosan (1, 2 and 3gL-1) and chlorocholine chloride (5, 10
and 15mgL-1) on the minituberization of cocoyam
(Xanthosoma sagittifolium).
Experiment setup
Factor
1. Control
2. Chitosan (g/L)=1, 2, 3
3. Chlorocholine chloride = mg/L=5, 10, 15
Parameters
- Average Height of plants (HP) (cm)
- Average Number of leaves (NL)
- Average Leaves surface (SF) (cm 2 ) The effect of two independent variables are considered.

5. Parameters
Times
(days)
CTH (g/L) CCC (mg/L)
control 1 2 3 5 10 15
Average Height of
plants (HP) (cm)
D0 15.08 17.87 15.59 16.41 15.33 20.44 21.06
D20 14.69 17.87 15.94 16.82 15.38 20.52 21.15
D40 12.7 17.83 16.75 16.22 12.9 20.34 20.85
D60 9.29 16.45 15.64 16.48 8.43 15.12 14.49
D80 8.69 16.89 14.65 14.98 7.71 14.53 12.47
D100 9.76 14.92 15.13 15.67 8.75 12.06 13.21
Average Number of
leaves (NL)
D0 3.2 3 2.9 2.2 2.6 3.1 3
D20 3.1 3.2 3.6 2.7 3.3 3.6 3
D40 2.9 3 3.8 2.6 3 2.5 3
D60 2.8 2.05 2.8 2.2 2.4 2.4 2.4
D80 2.8 2.3 2.2 2 2.5 2.4 2.8
D100 2.7 2.2 2.3 1.9 2.1 2.3 2.6
Average Leaves
surface (SF) (cm2 )
D0 15.87 36.39 29.79 26.11 16 20.98 24.31
D20 15.2 29.42 25.91 24.56 14.98 22.04 23.96
D40 12.07 31.26 28.4 26.9 12.72 20.91 25.87
D60 14.71 26.81 24.19 26.39 11.55 21.79 18.67
D80 12.57 26.82 21.53 22.01 10.65 22.23 17.6
D100 15.54 20.72 21.79 21.92 12.37 19.17 18.74
Treatments

7. Table Analyzed Grouped: Two-way ANOVA (two data sets)
Two-way ANOVA Ordinary
Alpha 0.05
Source of Variation % of total variation P value P value summary Significant?
Interaction 6.941 <.001 *** Yes
Row Factor 7.917 <.001 *** Yes
Column Factor 78.79 <.001 *** Yes
ANOVA table SS DF MS F -value P value
Interaction 660.0 12 55.00 9.567 P<.001
Row Factor 752.8 6 125.5 21.82 P<.001
Column Factor 7493 2 3746 651.7 P<.001
Residual (Error) 603.6 105 5.749
Data summary
Number of columns (Column Factor) = 3
Number of rows (Row Factor) =7
Number of values =126
Column factor = (control, CTH, CCC)=3
Row factor (No. of treatment in each factor)=7
Interaction Dof = (R-1)(C-1)
Residual Dof= N-Dof* (I+R+C) -1
F-ratio=correspondingMS/MS error

8. Number of families 1
Number of comparisons per family 21
Alpha 0.05
Tukey's multiple comparisons test Mean Diff. 95.00% CI of diff. Below threshold? Summary Adjusted P Value
Control vs. CTH [1g/L] -6.407 -8.810 to -4.005 Yes *** <.001
Control vs. CTH [2g/L] -4.958 -7.360 to -2.555 Yes *** <.001
Control vs. CTH [3g/L] -4.689 -7.092 to -2.286 Yes *** <.001
Control vs. CCC[5mg/L] 0.6111 -1.792 to 3.014 No ns .988
Control vs. CCC[10mg/L] -4.042 -6.445 to -1.640 Yes *** <.001
Control vs. CCC[15mg/L] -4.195 -6.598 to -1.792 Yes *** <.001
CTH [1g/L] vs. CTH [2g/L] 1.449 -0.9533 to 3.852 No ns .542
CTH [1g/L] vs. CTH [3g/L] 1.718 -0.6844 to 4.121 No ns .332
CTH [1g/L] vs. CCC[5mg/L] 7.018 4.616 to 9.421 Yes *** <.001
CTH [1g/L] vs. CCC[10mg/L] 2.365 -0.03772 to 4.768 No ns .057
CTH [1g/L] vs. CCC[15mg/L] 2.212 -0.1905 to 4.615 No ns .092
CTH [2g/L] vs. CTH [3g/L] 0.2689 -2.134 to 2.672 No ns >.999
CTH [2g/L] vs. CCC[5mg/L] 5.569 3.166 to 7.972 Yes *** <.001
CTH [2g/L] vs. CCC[10mg/L] 0.9156 -1.487 to 3.318 No ns .912
CTH [2g/L] vs. CCC[15mg/L] 0.7628 -1.640 to 3.165 No ns .962
CTH [3g/L] vs. CCC[5mg/L] 5.300 2.897 to 7.703 Yes *** <.001
CTH [3g/L] vs. CCC[10mg/L] 0.6467 -1.756 to 3.049 No ns .984
CTH [3g/L] vs. CCC[15mg/L] 0.4939 -1.909 to 2.897 No ns .996
CCC[5mg/L] vs. CCC[10mg/L] -4.653 -7.056 to -2.251 Yes *** <.001
CCC[5mg/L] vs. CCC[15mg/L] -4.806 -7.209 to -2.403 Yes *** <.001
CCC[10mg/L] vs. CCC[15mg/L] -0.1528 -2.555 to 2.250 No ns >.999
Compare row means (main row effect)
post Hocs test

9. Test details Mean 1 Mean 2 Mean Diff. SE of diff. N1 N2 q DF
control vs. CTH [1g/L] 9.648 16.06 -6.407 0.7992 18 18 11.34 105.0
control vs. CTH [2g/L] 9.648 14.61 -4.958 0.7992 18 18 8.773 105.0
control vs. CTH [3g/L] 9.648 14.34 -4.689 0.7992 18 18 8.297 105.0
control vs. CCC[5mg/L] 9.648 9.037 0.6111 0.7992 18 18 1.081 105.0
control vs. CCC[10mg/L] 9.648 13.69 -4.042 0.7992 18 18 7.153 105.0
control vs. CCC[15mg/L] 9.648 13.84 -4.195 0.7992 18 18 7.423 105.0
CTH [1g/L] vs. CTH [2g/L] 16.06 14.61 1.449 0.7992 18 18 2.565 105.0
CTH [1g/L] vs. CTH [3g/L] 16.06 14.34 1.718 0.7992 18 18 3.041 105.0
CTH [1g/L] vs. CCC[5mg/L] 16.06 9.037 7.018 0.7992 18 18 12.42 105.0
CTH [1g/L] vs. CCC[10mg/L] 16.06 13.69 2.365 0.7992 18 18 4.185 105.0
CTH [1g/L] vs. CCC[15mg/L] 16.06 13.84 2.212 0.7992 18 18 3.914 105.0
CTH [2g/L] vs. CTH [3g/L] 14.61 14.34 0.2689 0.7992 18 18 0.4758 105.0
CTH [2g/L] vs. CCC[5mg/L] 14.61 9.037 5.569 0.7992 18 18 9.854 105.0
CTH [2g/L] vs. CCC[10mg/L] 14.61 13.69 0.9156 0.7992 18 18 1.620 105.0
CTH [2g/L] vs. CCC[15mg/L] 14.61 13.84 0.7628 0.7992 18 18 1.350 105.0
CTH [3g/L] vs. CCC[5mg/L] 14.34 9.037 5.300 0.7992 18 18 9.378 105.0
CTH [3g/L] vs. CCC[10mg/L] 14.34 13.69 0.6467 0.7992 18 18 1.144 105.0
CTH [3g/L] vs. CCC[15mg/L] 14.34 13.84 0.4939 0.7992 18 18 0.8739 105.0
CCC[5mg/L] vs. CCC[10mg/L] 9.037 13.69 -4.653 0.7992 18 18 8.234 105.0
CCC[5mg/L] vs. CCC[15mg/L] 9.037 13.84 -4.806 0.7992 18 18 8.504 105.0
CCC[10mg/L] vs. CCC[15mg/L] 13.69 13.84 -0.1528 0.7992 18 18 0.2703 105.0
A p-value of 5% means that 5% of all tests will result in false positives. A q-value of 5% means that 5% of significant results will result in false positives.

10. Source of Variation Degrees of Freedom Sum of Squares Mean square
Column Factor 2 7493 3746
Row Factor 6 752.8 125.5
Interaction 12 660.0 55.00
Residual (error) 105 603.6 5.749
Total 125 9509
Does Column Factor have the same effect at all values of Row Factor?
Interaction accounts for 6.941% of the total variance.
F = 9.57 DFn = 12, DFd = 105
The P value is < 0.0001
If there is no interaction overall, there is a less than 0.01% chance of randomly observing so much
interaction in an experiment of this size. The interaction is considered extremely significant.
Since the interaction is statistically significant, the P values that
follow for the row and column effects are difficult to interpret.
Does Column Factor affect the result?
Column Factor accounts for 78.79% of the total variance.
F = 651.67 DFn = 2, DFd = 105
The P value is < 0.0001
If Column Factor has no effect overall, there is a less than 0.01% chance of randomly observing an
effect this big (or bigger) in an experiment of this size. The effect is considered
extremely significant.
Does Row Factor affect the result?
Row Factor accounts for 7.917% of the total variance.
F = 21.82 DFn = 6, DFd = 105
The P value is < 0.0001
If Row Factor has no effect overall, there is a less than 0.01% chance of randomly observing an
effect this big (or bigger) in an experiment of this size. The effect is considered extremely significant.
Data Narration

11. Means
Average
Height of
plants (HP)
(cm)
Average
Number of
leaves (NL)
Average
Leaves
surface (SF)
(cm2)
Row means
Control 11.7 2.917 14.33 9.648
CTH [1g/L] 16.97 2.625 28.57 16.06
CTH [2g/L] 15.62 2.933 25.27 14.61
CTH [3g/L] 16.1 2.267 24.65 14.34
CCC[5mg/L] 11.42 2.65 13.05 9.037
CCC[10mg/L] 17.17 2.717 21.19 13.69
CCC[15mg/L] 17.21 2.8 21.53 13.84
Column means 15.17 2.701 21.22 13.03

12. 0
5
10
15
20
25
30
control CTH [1g/L] CTH [2g/L] CTH [3g/L] CCC[5mg/L] CCC[10mg/L] CCC[15mg/L]
Effect of chitosan and chlorocholine chloride on the growth of cocoyam plants
Average
Height of
plants (HP)
(cm)
Average
Number of
leaves (NL)
Average
Leaves
surface (SF)
(cm2)