Comparison of two in vitro radiation-induced mutagenesis protocols for the Citrus rootstock Swingle citrumelo
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Abstract
In order to establish an efficient in vitro radiation-induced mutagenesis scheme for the rootstock Swingle citrumelo, two protocols were compared: Protocol 1 (seeds were irradiated and axillary shoots from the regenerated plantlets were propagated) and Protocol 2 (epicotyl segments were irradiated and the induced adventitious shoots were propagated). Radiosensitivity against 60Co gamma rays, shoot induction and three root induction treatments (MS, MS + NAA and MS + IBA) were evaluated. The efficiency regarding the total number of plants and the time required until their acclimatization was compared. The mutated population size was estimated. Mutagenic doses (LD30) of 29 and 37 Gy were determined for seeds and epicotyl segments, respectively. Shoot induction was higher than 93 % in both protocols. Root induction was significantly higher for adventitious shoots on MS + NAA, due to a higher secondary root induction (92 %), which also enhanced their adaptation to substrate (91,8 vs. 70,4 %). Starting from 1000 seeds, 50 and 70 mutants were estimated from Protocols 1 and 2, respectively, considering the proposed LD30 and 0,5 % frequency of induced mutations. It was demonstrated that Protocol 2: radiation-induced mutagenesis based on adventitious organogenesis was more efficient than Protocol 1, based on seed irradiation and axillary shoots propagation; since half the time (seven months) is required to obtain a higher number of mutants. It was mainly due to a high root induction potential and a better root architecture, which allowed for a higher plant ex vitro survival.
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