Micropropagation of three berry fruit species using nodal segments from field‐grown plants

Abstract

Plant micropropagation of blueberry (Vaccinium corymbosum L. cv. Berkeley), blackberry (Rubus sp. cv. Smoothstem) and raspberry (Rubus idaeus L. cv. Gradina) was carried out from nodal segments of adult field-grown plants. Hardwood and softwood cuttings were studied as explant sources. The cultures successfully established were softwood from all three species, and hardwood only from blueberry. Shoot-bud establishment from blueberry was achieved by culturing explants in WPM salts with MS vitamins for 15 days, and then 30 days in the same medium with 18 mM Zeatin. The best results of multiplication were obtained in the same medium with 25 mM 2iP. For blackberry, shoot-bud establishment was achieved by culturing explants in MS medium for 15 days, and then in the same medium with 4 mM BA and 0.25 mM IBA. This medium was also the best for blackberry multiplication. Raspberry explants (cvs Gradina and Willamette) were cultured in MS medium for 15 days and then transferred to MS medium supplemented with 4 mm BA and 0.25 mM IBA. After 30 days of culture, only Gradina explants survived, from which shoot-bud establishment was obtained in a modified MS medium (Andersons macronutrients except calcium, with Sequestrene as the iron source) with the same growth regulators. Multiplication was achieved by subculturing explants in the same medium either with 4 mM BA plus 0.25 mM IBA or with 8 mM BA plus 0.25 mM IBA. Shoots of at least 1 cm in length from all species were rooted ex vitro in a mixture of peat and Perlite (1:1, v/v) in a mist chamber, and 100% of rooting plants were acclimated. Bacterial, fungal and viral diseases were detected in stock plants, while tests carried out in both shoots and regenerated plants revealed the absence of any kind of disease.

La elongación de brotes adventicios de frambuesa (Rubus ideaus L.) es influenciada por brasinosteroides

“…A este respecto, se han probado técnicas como la organogénesis directa e indirecta utilizando hojas de diferentes variedades comerciales (Isac, 2009) y la criopreservación de brotes apicales (Wang et al, 2005;Gupta y Reed, 2006). Algunos sistemas para la micropropagación de frambuesa han utilizadoel medio de cultivo MS (Muashige y Skoog, 1692) semisólido y líquido en inmersión temporal complementado con bencil amino purina (BAP), ácido giberélico (AG 3 ) y ácido ascórbico (Jones y Flores, 2007); también se reporta el uso de N 6 -benciladenina, tidiazuron y zeatina (Reed, 1991;Popescue Isac, 1999;González et al, 2000;Vujović et al, 2009). …”

“…In this regard, techniques like direct and indirect organogenesis have been used, using leaves from different commercial varieties (Isac, 2009) and cryopreservation of apical shoots (Wang et al, 2005;Gupta and Reed, 2006). Some systems for raspberry micro propagation have used MS medium (Muashige and Skoog, 1692) semisolid and liquid in temporary immersion supplemented with benzyl amino purine (BAP), gibberellic acid (GA 3 ) and ascorbic acid (Jones and Flores, 2007); the use of N 6 -benzyladenine, thidiazuron and zeatin (Reed, 1991;Popescue Isac, 1999;González et al, 2000;Vujović et al, 2009) has also been reported.…”

Strategies to propagate Vaccinium nuclear stocks for the Canadian berry industry

“…In vitro derived shoots (> 1.5 cm long) were rooted ex vitro in shredded sphagnum moss in pots (Qu et al 2000). Although an auxinpretreatment was found unnecessary for ex vitro rooting of blueberries (Gonzalez et al 2000), Debnath (2004a) found 85 to 95% rooting in lowbush blueberry when microshoots were dipped in 4.9 mM indole-3-butyric acid (IBA) before planting in 2 peat:1 perlite (vol/vol) medium. For ex vitro rooting, the microcuttings are generally maintained in a mist chamber with very high relative humidity (RH 95%) and then transferred to a greenhouse (85% RH) for acclimatization.…”

“…The first in vitro callus formation was induced by Nickerson and Hall (1976) in lowbush blueberry, followed by Lyrene (1978) in rabbiteye (V. Ashei Reade) and Nickerson (1978a, b) in lowbush blueberry. Subsequently, tissue culture techniques for axillary shoot proliferation have been developed for lowbush blueberry (Frett and Smagula 1983;Debnath 2004a), highbush blueberry (V. corymbosum L.) (Zimmerman and Broome 1980;Gonzalez et al 2000), rabbiteye blueberry (Lyrene 1980) and cranberry (V. macrocarpon Ait.) (Marcotrigiano and McGlew 1991;Debnath and McRae 2001a;Debnath and McRae 2005), lingonberry (V. vitis-idaea L.) (Hosier et al 1985;Reed and Abdelnour-Esquivel 1991;Debnath and McRae 2001b;Jaakola et al 2001;Debnath 2005a, b) and bilberry (V. myrtillus L.) (Jaakola et al 2001) .…”

“…Eccher and Noè (1989) reported that zeatin is less phytotoxic than 2iP in highbush blueberry culture. Gonzalez et al (2000) developed a method of highbush blueberry micropropagation where nodal segments from mature field-grown plants were established on WPM with 18 µM zeatin and multiplied on WPM with 25 µM 2iP. Kaldmäe et al (2006) established lowbush blueberry cultures on modified WPM medium supplemented with 4.56 µM zeatin and observed that thidiazuron (TDZ) promotes callus development and inhibites shoot elongation.…”