Figure 1.
Proliferated explants after 8 weeks of culture (A) sucrose, (B) glucose, (C) fructose, (D) sorbitol, (E) mixture 1, and (F) mixture 2.
Figure 1.
Proliferated explants after 8 weeks of culture (A) sucrose, (B) glucose, (C) fructose, (D) sorbitol, (E) mixture 1, and (F) mixture 2.
Figure 2.
(A) Effect of different levels of prior autoclaving pH on medium’s post-autoclaving pH, (B) prior autoclaving and post-autoclaving pH difference, (C) effect of medium’s post-autoclaving pH on medium’s post-cultivation pH, (D) post-autoclaving and post-cultivation pH difference. Columns followed by the same letter do not differ significantly according to the Tukey HSD test (p ≤ 0.05). The absence of letters indicates a lack of any statistical difference according to the Tukey HSD test (p ≤ 0.05).
Figure 2.
(A) Effect of different levels of prior autoclaving pH on medium’s post-autoclaving pH, (B) prior autoclaving and post-autoclaving pH difference, (C) effect of medium’s post-autoclaving pH on medium’s post-cultivation pH, (D) post-autoclaving and post-cultivation pH difference. Columns followed by the same letter do not differ significantly according to the Tukey HSD test (p ≤ 0.05). The absence of letters indicates a lack of any statistical difference according to the Tukey HSD test (p ≤ 0.05).
Figure 3.
Proliferated explants after 8 weeks of culture at (A) low pH (4.5, 4.8, 5.2), (B) medium pH (5.5, 5.8), and (C) high pH (6.0, 6.2, 6.5).
Figure 3.
Proliferated explants after 8 weeks of culture at (A) low pH (4.5, 4.8, 5.2), (B) medium pH (5.5, 5.8), and (C) high pH (6.0, 6.2, 6.5).
Figure 4.
Effect of different levels of prior autoclaving pH on (A) shoot number per explants, (B) shoot length (cm), (C) node number per shoot, (D) node number per explant, and (E) node number per cm. The line represents the change level of post-autoclaving pH in relation to prior autoclaving adjusted pH. Columns followed by the same letter do not differ significantly according to the Tukey HSD test (p ≤ 0.05). The absence of letters indicates a lack of any statistical difference according to the Tukey HSD test (p ≤ 0.05).
Figure 4.
Effect of different levels of prior autoclaving pH on (A) shoot number per explants, (B) shoot length (cm), (C) node number per shoot, (D) node number per explant, and (E) node number per cm. The line represents the change level of post-autoclaving pH in relation to prior autoclaving adjusted pH. Columns followed by the same letter do not differ significantly according to the Tukey HSD test (p ≤ 0.05). The absence of letters indicates a lack of any statistical difference according to the Tukey HSD test (p ≤ 0.05).
Figure 5.
Biplot of the proliferation parameters (shoot number, shoot length, nodes per explant, nodes per shoot, nodes per cm) of shoots produced under different levels of pH (low pH (4.5, 4.8, 5.2) cycle marker, medium pH (5.5, 5.8) triangle marker, high pH (6.0, 6.2, 6.5) reverse triangle marker) on “Krymsk 5®” explants and pH grouping (Groups I, II and III).
Figure 5.
Biplot of the proliferation parameters (shoot number, shoot length, nodes per explant, nodes per shoot, nodes per cm) of shoots produced under different levels of pH (low pH (4.5, 4.8, 5.2) cycle marker, medium pH (5.5, 5.8) triangle marker, high pH (6.0, 6.2, 6.5) reverse triangle marker) on “Krymsk 5®” explants and pH grouping (Groups I, II and III).
Figure 6.
Rooted explants after 6 weeks of culture in different combinations of sugar type and sugar concentration. (A) Sucrose 1% w/v, (B) sucrose 2% w/v, (C) sucrose 3% w/v, (D) fructose 1% w/v, (E) fructose 2% w/v, (F) fructose 3% w/v, (G) glucose 1% w/v, (H) glucose 2% w/v, (I) glucose 3% w/v, (J) sorbitol 1% w/v, (K) sorbitol 2% w/v, (L) sorbitol 3% w/v.
Figure 6.
Rooted explants after 6 weeks of culture in different combinations of sugar type and sugar concentration. (A) Sucrose 1% w/v, (B) sucrose 2% w/v, (C) sucrose 3% w/v, (D) fructose 1% w/v, (E) fructose 2% w/v, (F) fructose 3% w/v, (G) glucose 1% w/v, (H) glucose 2% w/v, (I) glucose 3% w/v, (J) sorbitol 1% w/v, (K) sorbitol 2% w/v, (L) sorbitol 3% w/v.
Figure 7.
Rooted explants after 6 weeks of culture in different combinations of sugar type and pH level. (A–F) Sucrose and pH at 4.8, 5.2, 5.5, 5.8, 6.2, and 6.5, respectively, (G–L) fructose and pH at 4.8, 5.2, 5.5, 5.8, 6.2, and 6.5, respectively.
Figure 7.
Rooted explants after 6 weeks of culture in different combinations of sugar type and pH level. (A–F) Sucrose and pH at 4.8, 5.2, 5.5, 5.8, 6.2, and 6.5, respectively, (G–L) fructose and pH at 4.8, 5.2, 5.5, 5.8, 6.2, and 6.5, respectively.
Figure 8.
Biplot of the post-autoclaving and pre-adjusted pH and rooting parameters (rooted explants, root number, and root length) of explants cultivated in rooting mediums supplemented with fructose (cycle marker) or sucrose (triangle marker). Sugar grouping (Groups I and II) is also presented.
Figure 8.
Biplot of the post-autoclaving and pre-adjusted pH and rooting parameters (rooted explants, root number, and root length) of explants cultivated in rooting mediums supplemented with fructose (cycle marker) or sucrose (triangle marker). Sugar grouping (Groups I and II) is also presented.
Table 1.
Brief presentation of shοot proliferation experiments.
Table 1.
Brief presentation of shοot proliferation experiments.
| Treatment | pH | g L−1 |
---|
Sucrose | Fructose | Glucose | Sorbitol |
---|
Experiment 1 | Treat 1 | 5.8 | 2.4 | 2.4 | 5.2 | 20 |
Treat 2 | 7.5 | 7.5 | 7.5 | 7.5 |
Treat 3 | 30 | | | |
Treat 4 | | 30 | | |
Treat 5 | | | 30 | |
Treat 6 | | | | 30 |
Experiment 2 | Treat 1 | 4.5 | 2.4 | 2.4 | 5.2 | 20 |
Treat 2 | 5.0 |
Treat 3 | 5.2 |
Treat 4 | 5.5 |
Treat 5 | 5.8 |
Treat 6 | 6.0 |
Treat 7 | 6.2 |
Treat 8 | 6.5 |
Table 2.
Brief presentation of rooting experiments.
Table 2.
Brief presentation of rooting experiments.
| Treatment | pH | Sugar Type | Sugar Concentration (% w/v) | Agar (g L−1) |
---|
Experiment 3 | Treat 1 | 5.8 | Sucrose | 1 | 9 |
Treat 2 | 2 |
Treat 3 | 3 |
Treat 4 | 5.8 | Fructose | 1 | 9 |
Treat 5 | 2 |
Treat 6 | 3 |
Treat 7 | 5.8 | Glucose | 1 | 9 |
Treat 8 | 2 |
Treat 9 | 3 |
Treat 10 | 5.8 | Sorbitol | 1 | 9 |
Treat 11 | 2 |
Treat 12 | 3 |
Experiment 4 | Treat 1 | 4.8 | Sucrose | 2 | 9 |
Treat 2 | 5.2 |
Treat 3 | 5.5 |
Treat 4 | 5.8 |
Treat 5 | 6.2 |
Treat 6 | 6.5 |
Treat 7 | 4.8 | Fructose | 2 | 9 |
Treat 8 | 5.2 |
Treat 9 | 5.5 |
Treat 10 | 5.8 |
Treat 11 | 6.2 |
Treat 12 | 6.5 |
Table 3.
Effect of sugar type on shoot proliferation variables.
Table 3.
Effect of sugar type on shoot proliferation variables.
Sugar Type | Shoots per Explant | Shoot Length (cm) | Nodes per Explant | Nodes per Shoot | Nodes per cm |
---|
Sucrose | 1.79 c | 1.15 b | 5.3 c | 2.3 a | 2.15 a |
Glucose | 3.07 b | 1 b | 9.91 ab | 1.99 a | 2.07 a |
Fructose | 4.08 a | 1.67 a | 13.68 a | 1.99 a | 1.22 a |
Sorbitol | 2.38 bc | 1.15 b | 8.06 bc | 1.96 a | 1.77 a |
Mixture 1 * | 2.34 bc | 1.29 ab | 7.9 bc | 2.4 a | 1.9 a |
Mixture 2 ** | 1.65 c | 1.11 b | 5.18 c | 2.19 a | 1.73 a |
Table 4.
Multiple regression and correlation analysis between shoot proliferation parameters, prior autoclaving pH, and post-autoclaving pH with probabilities and correlation coefficients.
Table 4.
Multiple regression and correlation analysis between shoot proliferation parameters, prior autoclaving pH, and post-autoclaving pH with probabilities and correlation coefficients.
| | r | p Value |
---|
Post-autoclaving pH | Prior autoclaving pH | 0.96 | *** |
Shoot number | Prior autoclaving pH | −0.67 | *** |
Shoot length | Prior autoclaving pH | −0.42 | ** |
Nodes per shoot | Prior autoclaving pH | −0.41 | ** |
Nodes per explant | Prior autoclaving pH | −0.68 | *** |
Post-autoclaving pH | Shoot number | −0.65 | *** |
Post-autoclaving pH | Shoot length | −0.46 | ** |
Post-autoclaving pH | Nodes per shoot | −0.46 | ** |
Post-autoclaving pH | Node per explant | −0.68 | *** |
Difference post A/C- post culture pH | Shoot number | 0.59 | *** |
Difference post A/C- post culture pH | Shoot length | 0.44 | *** |
Difference post A/C- post culture pH | Nodes per shoot | 0.49 | *** |
Difference post A/C- post culture pH | Nodes per explant | 0.57 | *** |
Difference post A/C- post culture pH | Nodes per cm | −0.08 | ns |
Table 5.
Effect of sugars, concentrations, and their interactions in rooting parameters (rooting percentage, root number, and length) six weeks after planting in the absence of auxins.
Table 5.
Effect of sugars, concentrations, and their interactions in rooting parameters (rooting percentage, root number, and length) six weeks after planting in the absence of auxins.
| Rooted Explants (%) | Number of Roots | Root Length (cm) |
---|
Sugar | | | |
Fructose (Fruc) | 88.3 a | 7.1 a | 0.48 a |
Glucose (Gluc) | 74.3 ab | 5.4 b | 0.28 b |
Sorbitol (Sor) | 70.8 b | 5.2 b | 0.24 b |
Sucrose (Suc) | 54.9 c | 4.5 b | 0.18 b |
Concentration (% w/v) | | | |
1 | 58.2 b | 5.14 a | 0.25 b |
2 | 77.6 a | 6.07 a | 0.24 b |
3 | 80.5 a | 5.4 a | 0.40 a |
Sugar × Concentration | | | |
Fruc × 1 | 75 abc | 6.54 ab | 0.32 bc |
Fruc × 2 | 95 a | 7.97 a | 0.38 b |
Fruc × 3 | 95 a | 6.85 ab | 0.73 a |
Gluc × 1 | 69 abc | 4.3 b | 0.28 bc |
Gluc × 2 | 71 abc | 6 ab | 0.2 bc |
Gluc × 3 | 83.3 ab | 5.3 ab | 0.37 bc |
Sor × 1 | 50 bc | 3.75 b | 0.23 bc |
Sor × 2 | 81.2 abc | 5.05 ab | 0.2 bc |
Sor × 3 | 81.2 abc | 4.58 b | 0.3 bc |
Suc × 1 | 38.9 c | 6 ab | 0.17 bc |
Suc × 2 | 63.2 bc | 5.28 ab | 0.16 c |
Suc × 3 | 62.5 bc | 4.88 ab | 0.23 bc |
Table 6.
Effect of sugars, prior autoclaving pH, and their interactions on post-autoclaving pH and rooting parameters (rooting percentage, root number, and length) six weeks after planting in the absence of auxins.
Table 6.
Effect of sugars, prior autoclaving pH, and their interactions on post-autoclaving pH and rooting parameters (rooting percentage, root number, and length) six weeks after planting in the absence of auxins.
| Rooted Explants (%) | Number of Roots | Root Length (cm) | pH Post |
---|
Sugar | | | | |
Sucrose (Suc) | 35.6 b | 4.2 b | 0.25 b | 5.24 a |
Fructose (Fruc) | 86.0 a | 5.8 a | 0.41 a | 4.73 b |
pH pre | | | | |
4.8 | 66.2 a | 6.6 a | 0.35 ab | 4.78 e |
5.2 | 69.8 a | 5.8 ab | 0.49 ab | 4.71 f |
5.5 | 67.5 a | 4.9 ab | 0.28 ab | 4.90 d |
5.8 | 57.5 a | 5.2 ab | 0.47 a | 5.00 c |
6.2 | 55.0 a | 3.4 b | 0.22 b | 5.21 b |
6.5 | 49.1 a | 4.0 ab | 0.29 ab | 5.32 a |
Sugar × pH pre | | | | |
Fruc × 4.8 | 95.0 a | 7.3 a | 0.36 a | 4.62 i |
Fruc × 5.2 | 83.3 ab | 5.5 ab | 0.57 a | 4.62 i |
Fruc × 5.5 | 85.0 ab | 5.3 ab | 0.57 a | 4.63 i |
Fruc × 5.8 | 90.0 ab | 7.0 a | 0.54 a | 4.70 h |
Fruc × 6.2 | 75.0 abc | 4.0 ab | 0.24 a | 4.9 f |
Fruc × 6.5 | 88.2 ab | 5.6 ab | 0.44 a | 4.92 ef |
Suc × 4.8 | 37.5 cde | 5.9 ab | 0.32 a | 4.94 e |
Suc × 5.2 | 56.3 bcd | 6.2 ab | 0.22 a | 4.79 g |
Suc × 5.5 | 50.0 b–e | 4.5 ab | 0.23 a | 5.16 d |
Suc × 5.8 | 25.0 de | 3.5 ab | 0.40 a | 5.31 c |
Suc × 6.2 | 35.0 de | 2.7 b | 0.20 a | 5.51 b |
Suc × 6.5 | 10.0 e | 2.5 b | 0.15 a | 5.71 a |
Table 7.
Multiple regression and correlation analysis between rooting parameters, prior autoclaving pH, and post-autoclaving pH with probabilities and correlation coefficients.
Table 7.
Multiple regression and correlation analysis between rooting parameters, prior autoclaving pH, and post-autoclaving pH with probabilities and correlation coefficients.
| | r | p Value |
---|
Post-autoclaving pH | Prior autoclaving pH | 0.59 | *** |
Rooted explants % | Prior autoclaving pH | −0.23 | ns |
Rooted explants % | Post-autoclaving pH | −0.79 | *** |
Rooted explants % | Root number | 0.52 | *** |
Rooted explants % | Root length | 0.31 | * |
Root length | Prior autoclaving pH | −0.08 | ns |
Root length | Post-autoclaving pH | −0.34 | * |
Root length | Root number | 0.14 | ns |
Root number | Prior autoclaving pH | −0.42 | *** |
Root number | Post-autoclaving pH | −0.59 | *** |