Wolfberry or goji berry (Lycium barbarum L.) is a member of the Solanaceae family, rich in bioactive polysaccharides (Seeram, 2008). It belongs to the genus Lycium, in which there are more than 70 species, with three main species distributed in the East and South-East Asia, South Africa, and North America (Fukuda et al., 2001).
The species L. barbarum and L. chinense are very similar and have a long tradition as food and medicinal plants in China and other Asian countries. There are also some varieties and related species such as Lycium barbarum var. aurantiocarpum, L. chinense var. potaninii, L. ruthenicum, and L. truncatum, often commercialized as low-cost adulterants (Zhang et al., 2001). For these reasons, the adulteration of commercial products cannot be excluded, and morphological, histological, and molecular differentiations are necessary (Potterat 2010; Sze et al., 2008; Zhang et al., 2001).
Large-scale propagation of woody plant species through in vitro culture techniques represents a rapid way to produce uniform materials in a short time to meet agriculture needs. The application of micropropagation for commercial-scale plant production has been well demonstrated in several plant species that contain potentially useful secondary metabolites (Frabetti et al., 2009). Many of these species have shown excellent in vitro performance in terms of the number of explants produced in a short period of time and offer the opportunity to enhance the amount of these scarce compounds for extractive manipulations (Nalawade et al., 2003; Okemo et al., 2003; Osman et al., 2012, 2013a, 2013b; Osuna et al., 2006; Parveen et al., 2005).
Wolfberry plants often are reproduced through seeds and show all the problems related to sexual propagation, i.e., low germination, absence of clonal fidelity, and nonuniform agronomical performance. Despite the increasing interests in goji cultivation, there are few papers in the scientific literature regarding the micropropagation of wolfberry (Fira et al., 2016), and almost all described micropropagation protocols start from seedlings. Our previous experiments suggested that the major problems hampering the development of in vitro propagation protocols are the deleterious postexposure effect of wolfberry varieties on Murashige and Skoog (MS) medium and the nonuniformity of the explants used for rooting induction.
On the contrary, numerous papers have described regeneration by adventitious shoot organogenesis and somatic embryogenesis, and Agrobacterium-mediated transformation and transgenic plantlets have been obtained by using adventitious organogenesis from different explants (Cui et al., 1998, 1999; Hu et al., 2001, 2002, 2006).
For many fruit species that are not easy to root and acclimatize, the rooting phase has been successfully performed ex vitro and the plants then survived when planted in the field (Benmahioul et al., 2012; Saiju, 2005). The present work has been conducted to develop an efficient and reliable protocol for the micropropagation of wolfberry (Lycium barbarum L.), focusing on the effects of mineral salts, growth regulators, and in vitro and ex vitro rooting. The simplicity of the present protocol aims not only to promote the propagation of elite cultivar but also the in vitro conservation of interesting genotypes, usable for conventional and unconventional breeding and biotechnologies.
BenmahioulB.DorionN.Kaid-HarcheM.DaguinF.2012Micropropagation and ex vitro rooting of pistachio (Pistacia vera L.)Plant Cell Tissue Organ Cult.108353358
CuiK.R.PeiX.W.WangJ.J.WangY.F.1998Effects of modulation of abscisic acid during somatic embryogenesis in Lycium barbarum LActa Biologiae Experimentalis Sinica31195201
CuiK.R.XingG.S.LiuX.M.XingG.M.WangY.F.1999Effect of hydrogen peroxide on somatic embryogenesis of Lycium barbarum LPlant Sci.146916
FiraA.JosheeN.CristeaV.SimuM.HartaM.PamfilD.ClapaD.2016Optimization of micropropagation protocol for Goji berry (Lycium barbarum L.)Bull. Univ. Agr. Sci. Veterinary Med. Cluj-Napoca. Hort.73141150
FrabettiM.Gutiérrez-PesceP.Mendoza-De GyvesE.RuginiE.2009Micropropagation of Teucrium fruticans L., an ornamental and medicinal plantIn Vitro Cell. Dev. Biol. Plant45129134
FukudaT.YokoyamaJ.OhashiH.2001Phylogeny and biogeography of the genus Lycium (Solanaceae): Inferences from chloroplast DNA sequencesMol. Phylogenet. Evol.192246258
HuZ.GuoG.Q.ZhaoD.L.LiL.H.ZhengG.C.2001Shoot regeneration from cultured leaf explants of Lycium barbarum and Agrobacterium-mediated transformationRuss. J. Plant Physiol.48529535
HuZ.HuY.GaoH.H.ZhengG.C.2002High-efficiency transformation of Lycium barbarum mediated by Agrobacterium tumefaciens and transgenic plant regeneration via somatic embryogenesisPlant Cell Rep.21233237
HuZ.WuY.R.LiW.GaoH.H.2006Factors affecting Agrobacterium tumefaciens – mediated genetic transformation of Lycium barbarum LIn Vitro Cell. Dev. Biol. Plant42461466
IvanovaM.Van StadenJ.2009Nitrogen source, concentration, and NH4 (+): NO3 (−) ratio influence shoot regeneration and hyperhydricity in tissue cultured Aloe polyphyllaPlant Cell Tissue Organ99167174
LloydG.McCownB.1980Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot tip cultureIntl. Plant Prop. Soc. Proc.30421427
Mendoza de GyvesE.RoyaniJ. I.RuginiE.2007Efficient method of micropropagation and in vitro rooting of teak (Tectona grandis L.) focusing on large-scale industrial plantationsAnn. For. Sci.6417378
NalawadeS.M.SagareA.P.LeeC.Y.KaoC.L.TsayH.S.2003Studies on tissue culture of Chinese medicinal plant resources in Taiwan and their sustainable utilizationBot. Bull. Acad. Sin.447998
OsmanN.I.AwalA.SidikN.J.AbdullahS.2012Antioxidant activities of in vitro seedlings of Lycium barbarum (Goji) by diphenyl picrylhydrazyl (DPPH) assayInt. J. Pharm. Pharm. Sci.4Suppl.137141
OsmanN.I.AwalA.SidikN.J.AbdullahS.2013aCallus induction and somatic embryogenesis from leaf and nodal explants of Lycium barbarum L. (Goji)Biotechnology (Faisalabad)1213645
OsmanN.I.AwalA.SidikN.J.AbdullahS.2013bIn vitro regeneration and antioxidant properties of Lycium barbarum L. (goji)J. Teknologi623538
OsunaL.Tapia-PérezM.E.FigueroaO.Jiménez-FerrerE.Garduño-RamírezM.L.González-GarzaM.T.Carranza RosalesP.Cruz-VegaD.E.2006Micropropagation of Lepidium virginicum (Brassicaceae), a plant with antiprotozoal activityIn Vitro Cell. Dev. Biol. Plant42596600
ParveenV.SharmaV.K.MandalA.K.2005A protocol for micropropagation of an important medicinal plant—Oroxylum indicum Vent. through cotyledonary nodal explantsAnn. For. (Dehra Dun)135358
PotteratO.2010Goji (Lycium barbarum and L. chinense): Phytochemistry, pharmacology and safety in the perspective of traditional uses and recent popularityPlanta Med.761719
RuginiE.1992Involvement of polyamines in auxin and Agrobacterium rhizogenes-iduced rooting of fruit trees in vitroJ. Amer. Soc. Hort. Sci.117532553
SaijuH.K.2005Tree tissue culture and ex vitro sand rooting for reforestation p. 151–154. In: K. Suzuki K. Ishii S. Sakurai and S. Sasaki (eds.). Plantation technology in tropical forest science. Springer Berlin
SarasketaA.Gonzalez-MoroM.B.Gonzalez-MuruaC.MarinoD.2016Nitrogen source and external medium pH interaction differentially affects root and shoot metabolism in ArabidopsisFront. Plant Sci.729
SeeramN.P.2008Berry fruits: Compositional elements, biochemical activities, and the impact of their intake on human health, performance, and diseaseJ. Agr. Food Chem.563627629
SzeS.C.W.SongJ.X.WongR.N.S.FengY.B.NgT.B.TongY.ZhangK.Y.B.2008Application of SCAR (sequence characterized amplified region) analysis to authenticate Lycium barbarum (wolfberry) and its adulterantsBiotechnol. Appl. Biochem.51521
ZhangK.Y.B.LeungH.W.YeungH.W.WongR.N.S.2001Differentiation of Lycium barbarum from its related Lycium species using random amplified polymorphic DNAPlanta Med.67379381