Long-term preservation of Lotus tenuis adventitious buds

Abstract

Encapsulation-dehydration, encapsulation-vitrification, and vitrification were tested for cryopreservation of Lotus tenuis (Fagaceae) adventitious buds clusters (ABCs) obtained by a direct regeneration system from leaves cultures. Among them, the PVS3-based vitrification procedure was found to be useful for survival and regrowth of the preserved explants. For vitrification, the ABCs were dehydrated in a solution containing 2 M glycerol + 0.4 M sucrose for 25 min at room temperature, submerged in PVS3 solution for 1 h at 0 °C, then immersed in liquid nitrogen for 48 h and rapidly rewarmed. Afterword, the explants were unloaded in MS liquid medium with 1.2 M sucrose for 30 min. The washed tissues were dried superficially on filter paper and cultured in semisolid hormone-free MS medium containing 0.1 M sucrose. All cultures were maintained at 25 °C in the dark for 10 days and transferred to the light conditions. With this procedure, 79 ± 5.3% survival and more than 80% of the plantlets displaying a phenotype similar to the non-treated control after acclimatization. The data settled from ISSR showed no genetic dissimilarities between in vitro regenerants derived from cryopreserved tissues and the non-treated plants. Thus, our results indicate that the use of vitrification-based PVS3 solution offers a simple, accurate, and appropriate procedure for the cryopreservation of L. tenuis adventitious buds.

Droplet-vitrification methods for apical bud cryopreservation of yacon [Smallanthus sonchifolius (Poepp. and Endl.) H. Rob.]

“…The PVS3 method, on the other hand, has been optimized for species such as Allium cepa var. Aggregatum at 3 hours treatment duration, where this method resulted in 58% recovery rate (Wang et al 2020), and Lotus tenuis INTA Pampa with at PVS3 treatment time duration of 1 hour and recovery rate post cryopreservation, reaching 79% (Espasandin et al 2019). In the case of yacon, we found that both solutions proved to be effective at the same treatment time duration (60 min) with up to a 73% recovery rate post cryopreservation.…”

Cryopreservation and Cryotolerance Mechanism in Zygotic Embryo and Embryogenic Callus of Oil Palm

“…ISSR markers detected no genetic difference between regenerated plantlets [32]. ISSR markers have been used to detect the genetic stability of citrus [33], apple [34], pineapple [35], and lotus tenuis [36]. There is a need to develop an improved cryopreservation method for oil palm conservation.…”

Impact of Water Level on Species Quantity and Composition Grown from the Soil Seed Bank of the Inland Salt Marsh: An Ex-Situ Experiment

“…It is naturalized in other parts of the world, where it is known as an important forage crop of otherwise useless saline soils [68]. It is flood tolerant [69] and well established on restored saline wetlands [39].…”

“…Trifolium fragiferum has many commonalities with the previously mentioned species. It is flood-tolerant species of alkali and salty wet meadows [62,68,69]. It is also an important forage crop [70].…”