水浮莲(Pistia stratiotes Linn.)的体外再生与繁殖(英文)

In vitro Regeneration and Propagation of Pistia stratiotes:An Ideal Aquatic Plant for Biomanufacturing and Bioremediation

建立了水生单子叶植物水浮莲(PistiastratiotesLinn.)通过器官发生途径的体外高效再生与繁殖方法.采用叶、茎节和匍匐茎为外植体诱导愈伤组织,只有茎节能够在添加2,4-D和6-BA的MS基本培养基上形成愈伤组织,而叶和茎在含有不同组合植物激素的培养基上都不能够诱导愈伤产生.将愈伤组织转到添加6-BA和NAA的MS固体分化培养基可以在2wk内形成小苗,将小苗移至含NAA的MS固体生根培养基形成完整的植株.将生根苗转入无植物激素的不同基本液体培养基里比较其生长效果,其中含有2倍大量元素的SH培养基最适合其生长繁殖,在2wk内可以由1个小苗繁殖出10个新的植株.本研究是关于该植物体外再生的首次报道.水浮莲体外再生及繁殖系统的建立不仅可以用于在无菌条件下进行基础生理生化研究,还可以用于该植物遗传转化系统的建立.由于该植物生长迅速且为无性繁殖,生产成本低,通过基因工程方法表达外源基因将可以用于重组药用蛋白的生产及污染水体的转基因植物修复.

An efficient and rapid in vitro regeneration protocol via organogenesis has been developed for water lettuce （Pistia stratiotes）. Different explants of the plant, e.g., leaves, condensed stems and stolons, were tested for in vitro regeneration. Calli could be initiated only from the condensed stems on Murashige and Skoog （MS） agar medium supplemented with 30 g L^-1 sucrose plus 2.26 μmol/L of 2,4-dichlorophenoxyacetic acid （2,4-D） and 0.88 μmol/L of 6-benzylaminopurine （6-BA）, while calli could not be formed from the leaves and stolons on the media with different combinations of plant growth regulators. Shoots emerged when the calli were transferred onto shoot initiationg medium （MS agar medium modified with 4.44 μmol/L BA and 0.54 μmol/L α-naphthaleneacetic acid, NAA） in two weeks. The regenerated shoots were rooted spontaneously on this medium or when transferred to rooting medium （MS agar medium supplemented with 0.54 μmol/L NAA）. The rooted plants were transferred to different liquid media without plant growth regulator for comparing their multiplication, and it was found that the medium with two-fold macroelements of Schenk and Hildebrandt （SH） was the best. The regenerated plants were multiplied rapidly in this liquid medium and 10 new plants were produced from each parent plant in two weeks. This is the first report on in vitro regeneration of water lettuce. The regeneration and clonal propagation protocol described here has the potential not only to provide an axenic model system for biochemical studies of this aquatic monocotyledonous plant, but also to be widely applied in many fields, such as genetic modification of this plant, production of pharmaceutical recombinant proteins and bioremediation of contaminated water. Fig 3, Tab 1, Ref24