Proximate Composition Analysis and Establishment of an efficient in vitro Micropropagation Protocol for Anchote [Coccinia abyssinca (Lam.) Cogn]. from Eastern Wollega, Ethiopia

Abstract

Anchote (Cocinia abysinica (Lam.) Cogn) is an annual trailing vine belonging to family cucurbitaceae and is a potential major root and tuber cultivated plant produced in west Oromia (East Wollega) ,Ethiopia as food, cultural ,social and economical crop. This study aimed to: i) assess the nutritional content of Anchote leaves ,stems and roots. ii) determine the optimum sterilization procedure for Anchote explants. iii) compare effects of different cytokines and auxins concentrations in callus induction and direct regeneration from Anchote explants. Standard method of Association of Official Analytical Chemist (AOAC) was used to determine the crude protein, Crude fat, total ash, crude fiber and moisture contents of the sample. The collected parameters were analyzed using INSTA GRAPPAD software (San Diego, California ,). Analyses of variance (ANOVA) was computed for each parameter and the data were summarized as mean ± SD. The mean difference were assessed by Student-Newman-Keuls Multiple Comparisons Tests at5% level of significance. For tissue culture, young nodal and leaf explants were selected from healthy plants and subjected to sterilization using 0.19% and 0.385% of NaOCl commercial bleach JIK® at 5 ,10 15 minutes intervals. As the result showed that the highest protein was observed in leaf (18.3±1) and this indicate that the leaf is good source of protein compared to the protein found in the stem and root parts .The result of ANOVA showed that protein in leaves significantly different at α=0.05. However the lowest protein was observed in roots (8.2±0.1 ).The highest mineral element found in the stem was Magnesium (Mg) with mean average 336±0.4 Mg/100 followed by potassium (K ) which was found with mean average 323±3 Mg/100 and Calcium(Ca ) was found with mean average of 313.5±1 Mg/100.Maximum clean survival explants xvi (84.5±0.5) were obtained with 0.19%NaOCl and 15Minutes) for nodal explants Similarly, Maximum clean survival explants (82.5±0.) were obtained with 0.19%NaOCl and 10 Minutes) for leaf explants .BAP, TDZ , Kn,2,4-D and NAA were used for callus induction. The highest percentage of callus induction (80±0.2) and (78±2) were obtained with 5μM BAP and 5μM 2,4-D) for leaf and nodal explants respectively. For regeneration of micro shoots, the nodes cultured on Murashige and Skoog's medium(MS) supplemented with 2.5 μM BAP gave the best result with (75±0.1) regeneration frequency and highest shoot number (3.2±0.5)). Similarly ,the concentration of 0.025 μM TDZ in combination with 2.5μM BAP gave highest shoot number (4.2±0.1) . The highest root initiation percentage (86 ±0.3) was shown by 0.5 μM NAA on full MS Media with maximum shoot number (4.6±4.6). Finally the best hardening (acclimatization) was obtained from Soil: Manure (1:1) mixture with mean percentage (83.33±1) of survival rate.