采用俄罗斯橄榄叶片和茎段诱导产生不定芽的高效再生方法(英文)

An Efficient Method for Adventitious Shoot Regeneration from Leaf and Stem-segment Explants of Russian Olive Elaeagnus angustifolia L.

俄罗斯橄榄(Elaeagnus angustifolia L.)是一种具有很重要药用价值和生态意义的植物。以俄罗斯橄榄一年生幼苗的叶片和茎段为实验材料,探讨了细胞分裂素类(6-BA和Zt)和生长素类(NAA和IBA)两类激素不同组合以及不同配比对植株再生的影响,最后建立了一个高效的俄罗斯橄榄再生方法。结果表明,MS培养基+0.5 mg/L 6-BA+0.2 mg/L NAA更适合叶片的再生,平均每个外植体能产生多达4.3个不定芽;而在MS培养基+1.0 mg/L Zt+0.5 mg/L NAA的条件下,茎段外植体再生出来的不定芽最多可以达到平均3.6个;再生芽在含有0.5 mg/L NAA的1/2 MS培养基上生根率达到100%。体外再生苗移栽到装有灭菌混合土(土:泥炭:沙子=1:1:1)的花盆中锻炼驯化,最后有77%的再生植株存活下来。此结果不仅对俄罗斯橄榄种质资源保护有重要的促进作用,另外也为其将来的遗传转化奠定了基础。

The establishment of an efficient adventitious shoot regeneration system from leaf discs and stem segments of one-year-old Russian olive（Elaeagnus angustifolia L.） was established.Shoot regeneration using leaf explants and stem segments was obtained on MS medium supplemented with different cytokinins viz.,6-benzyladenine（BA） and Zeatin（Zt）,with combination of auxins viz.,naphthaleneacetic acid（NAA） andindole-3-butyric acid（IBA）.MS ＋ 0.5mg/L 6-BA ＋ 0.2mg/L NAA was found to be a better combination for leaf regeneration,producing a maximum of 4.3 shoots per explant,whereas MS ＋ 1.0mg/L Zt ＋ 0.5mg/L NAAfor stem-segment explants produced the highest number of multiple shoots（3.6） per explant.The regenerated shoots produced 100% rooting on 1 /2 strength MS medium supplied with 0.5mg/L NAA.In vitro raised plantletswere successfully transferred to pots containing sterile soil∶ peat moss∶ sand（1∶ 1∶ 1） for acclimatization,and 77% of rooted plantlets survived under ex situ conditions.The currently employed regeneration protocol through leaf explants and stem segments facilitates germplasm conservation of this medicinally and ecologically importantspecies besides providing a standardized regeneration protocol for genetic transformation studies in the future.