Cryopreservation of mat rush lateral buds by air dehydration using aluminum cryo-plate

Abstract

A cryopreservation protocol based on air dehydration of explants placed on aluminium cryo-plates, termed D cryo-plate, was successfully developed for in vitro mat rush (Juncus decipiens Nakai) lateral buds. The buds of line 'Hiroshima 4gou(1)' with basal stems were dissected from multiple shoots and precultured overnight at 25°C on solid MS medium containing 0.3 M sucrose. Precultured buds were placed on aluminium cryo-plates, each one with 10 elliptical wells (2.5 mm long, 1.5 mm wide and 0.75 mm deep) and embedded in calcium alginate gel with 0.4 M sucrose and 1 M glycerol. Osmoprotection was performed by immersing the cryo-plates with buds for 30 min at 25°C in loading solution (2 M glycerol+1.0 M sucrose). Buds were dehydrated to 26% moisture content (fresh weight) by placing the cryo-plates for 2.5 h either in the air current of a laminar flow cabinet or in Petri dishes containing silica gel. Cooling was performed by placing the cryo-plates in uncapped cryotubes, which were immersed in liquid nitrogen. For rewarming, cryo-plates were immersed in liquid medium with 1.0 M sucrose for 15 min at room temperature. Under these conditions, regrowth of cryopreserved buds of line, 'Hiroshima 4gou (1)' was 93% after four weeks culture. The average regrowth of 20 mat rush lines was 88%. The D cryo-plate procedure will facilitate cryostorage of mat rush germplasm.

Cryopreservation of blueberry shoot tips derived from in vitro and current shoots using D cryo-plate technique

“…The D cryo-plate technique developed by Niino et al (2013Niino et al ( , 2014 was performed with some modifications (Matsumoto et al 2015). The cryo-plates ) used were obtained from the Genetic Resources Center, National Agriculture and Food Research Organization (Tsukuba, Japan).…”

“…Recently, two novel cryogenic protocols using aluminum cryo-plates were developed, the osmo-dehydration method (V cryo-plate; Sekizawa et al 2011;Yamamoto et al 2011Yamamoto et al , 2012aYamamoto et al , 2012b and the air-dehydration method (D cryoplate; Niino et al 2013). The major advantages of the cryo-plate methods are their user-friendly procedures, minimization of shoot injury, and rapid cooling and warming, which results in higher recovery (Matsumoto et al 2015;Niino et al 2013Niino et al , 2014Salma et al 2014). Furthermore, the D cryo-plate method can be used with larger samples that are sensitive to physical damage and toxicity from certain cryoprotectants, such as PVS2 .…”

Improvement to the D Cryo-plate Protocol Applied to Practical Cryopreservation of <i>In Vitro</i> Grown Potato Shoot Tips

“…Niino et al (2014) indicated the need to avoid some shoot tips detaching themselves from the cryoplates during manipulations. The addition of 1 M glycerol and 0.4 M sucrose to the sodium alginate gel prevented the dropping of mat rush buds from the cryoplates .…”

“…1A1). The base D cryo-plate protocol was derived from the protocol developed by Niino et al (2014) and Yamamoto et al (2015) as follows: 1.…”

Cryopreservation of Persimmon Shoot Tips from Dormant Buds Using the D Cryo-plate Technique

“…Excised shoot tips were cryopreserved using the D cryo-plate technique developed by Niino et al (2013Niino et al ( , 2014 with some modifications (Matsumoto et al, 2014a, b). The cryo-plates No.…”