Arachis paraguariensis Chodat and Hassl. is a wild peanut with very high morphogenic potential but
limited rooting ability in vitro has hampered production of large number of surviving plantlets. The
present study evaluated in vitro rooting and ex vitro survival of micro-shoots utilizing a 6 × 3 × 3 × 2
factorial experiment. Six A. paraguariensis genotypes were treated with three auxins at three concentration
levels inside either 11.4 cm × 8.6 cm × 10.2 cm polyethylene terephthalate glycol (PETG) vessel or
2.5 cm × 15 cm glass tube. The auxin treatments included indole-3-acetic acid (IAA), indole-3-butyric acid
(IBA), and -naphthaleneacetic acid (NAA) at 0.2, 0.6 and 1 mg L−1. Across all auxin treatments, PETG vessel
was superior to glass tube in terms of rooting percentage (68% vs 17%), and survival at acclimatization
(38% vs 6%) respectively. Further analysis of plantlets cultured inside PETG vessel showed that rooting
and post acclimatization survival were high on MS medium without any auxin (90% and 80%) and on
0.2 mg L−1 IBA (92% and 76%) or IAA (86% and 74%) respectively. Contrariwise, none of the shoots treated
with 1 mg L−1 NAA survived acclimatization. The 0.2 mg L−1 of IAA and IBA treatments also exhibited high
number of roots per plantlet (7 and 5), and lowest number of days to root initiation (11 and 12 days)
respectively. Since rooting was delayed (26 days) on MS medium without auxin, we recommend supplementation
of nutrient medium with 0.2 mg L−1 IBA or IAA whenever rapid rooting is desired. Overall, the
results showed that genotype, auxin type, and auxin concentration significantly altered in vitro rooting
of A. paraguariensis plantlets.