In Vitro Propagation and Genetic Diversity Study of Korarima (Aframomum corrorima (Braun) P.C.M. Jansen) from Southern and Northwestern Ethiopia
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Date
2017-05
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Addis Ababa Universty
Abstract
Korarima [Aframomum corrorima (Braun) P.C.M. Jansen]is perennial rhizomatous plant which is indigenous to Ethiopia and widely used for culinary and medicinal purposes.Traditionally, korarima is propagated by stem cutting which is labor intensive and time consuming process. In vitro propagation of plant has played a very important role in rapid multiplication of cultivars with desirable traits and production of healthy and disease-free plants.Surface sterilization of seeds in 25% Clorox for 30 minutes followed by dipping in 70% ethanol for one minute showed the maximum aseptic cultures and high percentage of germination (70%).The maximum shoot initiation percentage (90%) was obtained on MS medium supplemented with 1.0 mg/l BAP in combination with 0.1mg/l NAA.Shoot multiplication using 1.5 mg/l BAP in combination with 2.0 mg/l KNresulted in the highest mean shoot number per explants (5.13 ± 0.64) within four weeks time. Table sugar and laboratory grade sucrose have been found to play substantial roles in micropropagation of A. corrorima. The highest shoot number (6.67 ± 0.55) per explant was observed in explants cultured on MS medium supplemented with 3% laboratory grade sucrose followed by 4.10 ± 0.44 shoot number per explant using 3% table sugar.Full strength MS medium resulted in the highest number of shoots per explant (5.27 ± 0.53) and mean shoot length (3.08 ± 0.20 cm). The highest number of shoot per explant observed on MS liquid and solid media was 4.41 ± 0.65 and 3.00 ± 0.25 respectively. The maximum callus induction frequency (80 ± 5.77%) was obtained on MS medium containing 0.5 mg/l 2,4-D alone and 5.0 mg/l 2,4-D in combination with 0.25 mg/l BAP. The highest frequency of shoot regeneration (25.33 ± 1.99%) was obtained upon transfer of calli onto regeneration medium containing 0.5 mg/l BAP combined with 0.25 mg/l IAA. Healthy shoots were rooted on half strength MS medium containing different concentration of auxin with highest root number per shoot (9.77 ± 0.78 and 18.50 ± 1.15) obtained using solid and liquid MS media respectively. The plantlet with well developed shoot and root system were transferred to greenhouse and 78% and above 86% rate of survival has been recorded in liquid and solid media respectively.The somatic chromosome number of A. corrorima was found to be 2n = 48. The amplification of genomic DNA with six ISSR primersamong 102 individuals from 11 populations of A. corrorimayielded 83 scorable loci of which 100% were found to be polymorphic. High levels of genetic diversity and significantly high genetic differentiation (P < 0.001) involving within and among populations were revealed by this marker in all studied populations. Nei‟s gene diversity and Shannon‟s information index were 0.3041 and 1.1441 respectively. The analysis of molecular variance showed that the maximum value of genetic variation was found within populations (76%), whereas low value of genetic variance was observed among populations (24%). Both PCoA and UPGMA cluster analysis showed the clustering of all 11 populations into three groups, corresponding to the geographic distribution patterns of the population. Proper identification and characterization of the plant‟s germplasm is central in genetic improvement of the plant and for developing appropriate conservation strategies.
Key Words/ Phrases: Aframomum corrorima, callus induction, genetic diversity, in vitro regeneration, ISSR marker, micropropagation, MS medium, Plant Growth Regulator
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Keywords
Phrases, Aframomum corrorima, Callus induction, Genetic diversity, In vitro regeneration, ISSR marker, Micropropagation, MS medium, Plant Growth Regulator