Interaction Effects of IBA and Immersion Time on In Vivo Rooting Responses of In Vitro Raised Ex-Vitro Mass Propagated Sugarcane Plants

Gezahegn Terefe1 and Belay Tolera2

1Dilla University, School of Natural and Computational Sciences, Department of Biology

2Ethiopian Sugar Corporation Research and Development Center, Wonji Research Center, Ethiopia

Corresponding Author:
Belay Tolera
Ethiopian Sugar Corporation Research and Development Center
Wonji Research Center, Ethiopia
Tel: +251-910-18-16-44
Fax: +251-222-20-01-44
E-mail: [email protected]

Received Date: January 24, 2017; Accepted Date: February 23, 2017; Published Date: March 01, 2017

Citation: Terefe G, Tolera B. Interaction Effects of IBA and Immersion Time on In Vivo Rooting Responses of In Vitro Raised Ex-Vitro Mass Propagated Sugarcane Plants. J Plant Sci Agric Res. 2017, 1:1.

Copyright: © 2017 Terefe G, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 
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Abstract

A study was conducted at Metahara Sugar Estate of Ethiopia Sugar Industry with the objective to determine the effects of different concentrations of IBA and immersion time on ex vitro root induction of detached sugarcane plantlets. Five levels of IBA (0, 0.5, 1, 1.5 and 2 mgL-1) and four levels of immersion time (0, 1, 5 and 10 minutes) with three sugarcane genotypes were combined in factorial treatment combination arrangements with completely randomized design in pot experiment under nursery. The basal end of the detached shoots was dipped in IBA solution for different durations before the shoots were transferred into a polyethylene bag containing a mixed growing medium. The results showed that the interaction effect of IBA, immersion time and the sugarcane genotypes was very highly significant (p<0.0001) on number of roots per shoot, average root length and average survival rate of the sugarcane plantlets. Sugarcane plantlets of SP7-1284, C132-81 and C86-56 dipped in 0.5 mgL-1 IBA for 10 minutes gave the optimum rooting responses with highest survival rate in all the sugarcane genotypes studied.

Keywords

In vivo rooting; Sugarcane genotypes; IBA; Immersion time; SP70-1284; C132-81; C86-56

Introduction

For commercial planting, sugarcane is propagated by vegetative means through stem cuttings. Besides its slow rate of propagation [1-3], production of sufficient quantity disease-free planting material in such large numbers during the planting season is laborious and time consuming. In addition, it requires a substantial quantity of crushable cane that otherwise could be used for sugar and oth er by-products production. Proper use of tissue culture technology offers an opportunity to mass produce disease-free planting material and is now used to supplement commercial sugarcane propagation in many countries including Ethiopia. Recently, tissue culture technology plays a leading role in rapid multiplication of disease-free and quality planting material and thus, reported to give better cane and sugar yield as compared to the conventional seed sources [4-11].

To utilize this advantage, besides building tissue culture facilities at Metahara, Kuraz, Tendaho and Fincha sugar factories/projects, the Ethiopian Sugar Corporation has been procuring micropropagated sugarcane plantlets from other biotechnology laboratories and will going to require about five million plantlets annually. However, the high tech nature of the technology, limited skills of the supplier organizations and delay of import inputs make the plantlets supply erratic and challenging the sustainable supply. In addition, the cost of procurement is high and increasing from time to time.

The high cost involved in microproagation technology is a major constraint to its popular use in sugarcane [12]. Thus, rapid and low cost ex-vitro propagation protocols have been developed to complement microproagation technology for few sugarcane genotypes while reports on in vivo rooting are limited. During in-vitro propagation, detaching the tillers from the main shoot having common fibrous root system resulted in absence or few roots and resulted in low survival rate of the detached plantlets/tillers.

Most reports of ex vitro rooting of plant species have involved treatment with exogenous auxin such as indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and 1-naphthalene-acetic acid (NAA) with dipping the basal end for different durations for better root induction. Therefore, this study was aimed to determine the effect of different concentrations of IBA and immersion time on ex vitro rooting of three sugarcane genotypes (SP70-1284, C132-81 and C86-56).

Result and Discussion

Analysis of variance proved that the Effects of IBA, Immersion Time and sugarcane genotypes have a highly significant (IBA*time* genotype=p<0.001) effect on the number of Roots per shoot and average root length (cm) of all the sugarcane genotypes tested: C132-81, C86-56 and Sp70-1284 (Table 1).

Source of Variation DF Mean Squares
Number of Roots per shoot Average root length(cm)
IBA 3 113.61ns 16.80*
Time 2 960.41** 42.43***
IBA*Time 6 699.46*** 25.77***
Genotype 2 337.13* 33.72***
IBA*Genotype 6 39.05ns 8.31ns
Time*Genotype 4 164.92ns 22.94***
IBA*Time*Genotype 12 291.62** 25.86***

Table 1: ANOVA Summary for the interaction effects of IBA and immersion time on in vivo Rooting of sugarcane plantlets.

All the sugarcane genotypes showed statistically significant variation both in the number of roots per shoot and average root length (cm) (Table 2). In sugarcane genotype SP70-1284, the lowest number of roots per shoot(15.0) and the lowest average root length (9.67 cm) were recorded at 0 mg/l IBA and 0 minute immersion time while the lowest average survival count (20 plants per plot) was recorded at 0.5 mg/l IBA+5 minutes immersion time. Similarly, in C86-56, the lowest number of roots per shoot(19.0) and lowest average survival count(24 plants per plot) were obtained on the control treatment(0 mg/l IBA+0 minute immersion time) while the lowest average root length (11.83 cm) was obtained at 0.5 mg/l IBA+5 minutes immersion time. In sugarcane genotype C132-81, the lowest number of roots per shoot (16.67) and lowest average survival count (21 plants per plot) were found on the control treatment (0 mg/l IBA +0 minute immersion time) while the lowest average root length(11.8 cm) was found at 2 mg/l IBA with 1 minute immersion time (Table 2). In sugarcane genotype SP70-1284, the maximum number of roots per shoot (51.67) was recorded at 1.5 mg/l IBA+1 minute immersion time while the highest average root lengths were recorded at 0.5 mg/l IBA+10 minutes immersion time; and 2 mg/l IBA+5 minutes immersion time. However, in this genotype (SP70-1284), the optimum number of roots per shoot (48.33), average root length (19.66 cm) and average survival count (30 plants per plot) were obtained at 0.5 mg/l IBA with 10 minutes immersion time.

Sugarcane genotype Pooled value of the response variables
Number of roots per shoot Average root length(cm)
C86 - 56 41.38a 13.90c
C132 - 81 39.14b 17.01a
SP70 -1284 34.97c 15.46b

Table 2: Comparison of the sugarcane genotypes by their rooting responses.

In sugarcane genotype C86-56, at 0.5 mg/l IBA, increasing the duration of immersion time from 0 to 1 minute, from 1 to 5 minute, from 5 to 10 minute increased the average number of roots per shoot from 19.0 to 33.67 cm, from 33.67 to 48.33 cm and from 48.33 to 51.33 cm, respectively. In addition, the maximum number of roots per shoot (65.33), highest average root length (20 cm) and maximum survival count (30 plants per plot) were recorded 1 mg/l IBA+5 minutes immersion duration, 1 mg/l IBA+10 minutes immersion time and 0.5 mg/l IBA+10 minutes immersion time respectively. However, the optimum treatment combination selected was 0.5 mg/l IBA with 10 minutes immersion time that produced 51.33 roots per shoot with 19.66 cm root length and an average survival count of 30 plants per plot (Table 3).

  Response of Sugarcane Genotypes
IBA(mg/l) Immersion Time (minute)
SP70-1284 C86-56 C132-81
Number of roots per shoot Average root length (cm) Average survival count per plot Number of roots per shoot Average root length (cm) Average survival count per plot Number of roots per shoot Average root length (cm) Average survival count per plot
0 0 15.00y 9.67q 21g 19.00w 12.27n 24e 16.67x 16.50h 21g
0.5 1 38.67n 13.66m 25f 33.67p 17.33g 28c 48.67g 21.06b 28c
0.5 5 36.67o 18.33f 20g 48.33g 11.83o 28c 31.33q 14.16k 29b
0.5 10 48.33g 19.66d 30a 51.33d 19.66d 30a 41.67l 21.00b 29b
1 1 43.67j 13.33m 29b 50.00e 13.83m 28c 50.00e 17.16g 29b
1 5 37.33n 15.33i 29b 65.33a 17.33g 29b 51.33d 16.66h 29b
1 10 35.33o 17.66g 28c 21.67v 20.00c 28c 26.00r 15.16i 29b
1.5 1 51.67d 15.83i 28c 59.00b 16.66h 27d 37.00n 15.16i 29b
1.5 5 25.00t 15.33i 28c 33.00p 15.43i 28c 38.33n 15.00i 27d
1.5 10 25.67s 14.00l 27d 34.67p 17.33g 27d 44.67i 22.00a 27d
2 1 49.33f 16.50h 28c 42.50k 13.97m 28c 52.00c 11.80p 26e
2 5 23.00u 19.33d 28c 34.00p 18.75f 27d 35.00o 14.00i 26e
2 10 25.00t 12.33n 27d 46.00h 12.95n 27d 39.00m 19.50d 27d
CV 10.95 8.64
SE 0.63 1.41

Table 3: Interaction effects of IBA and Immersion time on in vivo rooting of sugarcane.

In sugarcane genotype C132-81, even though the maximum number of roots per shoot (52.0) and highest average root length (22.0 cm) were found at 2 mg/l IBA+1 minute immersion time and 1.5 mg/l IBA+10 minute immersion time, respectively, the optimum treatment combination selected was 0.5mg/l IBA +5 minutes immersion time that gave 31.33 roots per shoot with 14.16 average root length and an average survival count of 29 plants per plot. The absence of IBA in the treatment (control) resulted in reduced number of roots per shoot and hence the survivals count of the plantlets per plot. However, high concentration of IBA (2 mg/l) gave the highest number of roots per shoot (52.0) with the shortest average root length (11.8 cm) with subsequent reduced survival count (26 plants per plot). Even though the response of different genotypes differs for the same treatment, use of high concentration of IBA may result in excessive rooting with short root length followed by low survival count. Thus, uses of optimum concentration of IBA with appropriate immersion duration is essential in inducing in vivo rooting in sugarcane and help improve survival rate of in vivo detached sugarcane plantlets in ex-vitro propagation system (EPS). Comparison of the current results with the previous findings was impossible due to the scarcity of research reports on similar activities.

Conclusion

The conventional method of sugarcane planting material propagation has diverse limitations while procurement of large quantity microproagation sugarcane planting materials to complement the conventional method is costly and erratic. Similarly, on ex-vitro propagation protocols developed for three sugarcane genotypes: SP70-1284,C132-81 and C86-56; to complement microproagation technology, separation of tillers from the main shoot having common fibrous root system resulted in absent or few roots with subsequent plant death and hence reduced survival rate. Thus, ex-vitro root induction protocol for ex vitro propagation three sugarcane genotypes ‘SP70-1284, C132 and C86-56’ has been developed. The result proved that ex-vitro rooting of ex vitro propagated sugarcane plants is highly dependent on the interaction effects IBA, Immersion duration and the sugarcane genotypes. Treatment combination containing 0.5 mgL-1 IBA for 10 minutes was found to give optimum ex-vitro rooting response with the highest survival rate of the plantlets in all the three sugarcane genotypes studied. On this treatment combination, sugarcane genotype SP70-1284 produced 48.33 roots with 19.66 cm root length resulting in 100% survival rate of the plantlets while C86-56 gave 51.33 roots having 19.66 cm average root length with 100% survival rate of the plantlets. Similarly, C132-81 produced 41.67 roots with 21.0 cm root length and 96.67% survival rate. Thus, the current findings will help improve the root induction response of the sugarcane genotypes and hence their survival rate.

References

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