梅花品种“美人”叶片离体再生体系建立

The Establishment of in vitro Regeneration System of 'Meiren' (Prunus mume) Cultivar by Using the Leaves as Explants

“以美人”梅离体培养的叶片为材料进行愈伤组织诱导再生研究,诱导过程分两步,第一步是愈伤组织的诱导,诱导过程中生长素2,4-D起了决定性的作用,并且以液体培养基为佳,而且暗培养的愈伤组织再分化率明显高于光培养。具体培养基为1/2WPM+5mg/L2,4-D;第二步是在愈伤组织诱导出之后,进一步诱导愈伤组织再生出植物,在经过7d液体培养基的诱导之后将离体叶片转入1/2WPM+1.0mg/L6-BA+0.1mg/LNAA+8mg/LAgNO3+100mg/L水解乳蛋白培养,再分化率可以达到24.25%。愈伤组织分化出苗之后,进一步诱导生根“,美人”梅最佳生根培养基是1/2WPM+0.5mg/LNAA。

Prunus mume is an important flowering woody species widely used in the gardens of China.‘Meiren＇,one of the most common Prunus mume cultivars, was chosen as material to establish the in vitro regeneration system with the leaves as explants. A complete regeneration protocol was developed for this cultivar. There were two steps for the regeneration of leaves. The first step was the callus inducement of leaves. The optimum medium was modified 1/2 woody plant medium （1/2WPM） liquid medium supplemented with 5mg/L 2,4-diehlorophenoxyacetie acid （2,4-D） which plays a key role in the callus inducement processing .The callus re-differentiation ratio was higher in dark culture condition than that in light culture condition. The second step was the seedling regeneration from the callus induced at the first step. In vitro Prunus mume leaves were transferred to the regeneration medium 1/2WPM supplemented with 1.0mg/L 6-benzyladenine （6-BA）, 0.1 mg/L naphthalene-acetic acid （NAA）, 8mg/L AgNO3 and 100mg/L lactoalbumin hydrolysate （LH） after seven days inducing culture on the liquid medium. The regeneration rate can be achieve to 24.25%. After the shoot were regenerated from the callus, roots were induced from the regenerated plants. The optimum rooting medium for ‘Meiren＇ is improved 1/2 woody plant medium （WPM） supplemented with 0.5mg/L NAA.