Development of an efficient regeneration protocol for pear rootstock Pyrodwarf and assessment of SSR variability in regenerating shoots
Development of an efficient regeneration protocol for pear rootstock Pyrodwarf and assessment of SSR variability in regenerating shoots
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DOI：
10.1080/00087114.2004.10589667
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发表时间：
2009-01
期刊：
影响因子：
2.1
通讯作者：
Martinelli Federico;M. Busconi;C. Fogher;L. Sebastiani
中科院分区：
文献类型：
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作者：
Martinelli Federico;M. Busconi;C. Fogher;L. Sebastiani
摘要在果树作物中，生物技术方法的使用尤其令人感兴趣，因为传统的育种计划在产生商业化影响之前需要很长的时间和大量的资源。为了建立梨砧木火矮病的高效植株再生体系，研究了1-萘乙酸(NAA)和噻二唑隆(TDZ)两种激素的不同组合对再生过程的影响。其他因素也进行了研究，如黑暗时间和在诱导和表达阶段使用不同的激素浓度。我们的实验表明，黑暗的长度不会影响体外再生。不同浓度的TDZ对再生的影响是不同的：6μM的效果优于2.5μM。诱导期较高的TDZ水平是诱导再生的唯一重要因素，而不依赖于表达阶段激素的存在。这两种激素不能有效地诱导另一个梨砧木Fox11的叶片外植体产生足够数量的再生芽。利用SSR标记对再生过程中的变异性进行评估。在外植体和再生芽之间观察到相同的遗传图谱。这些结果有助于将火矮菌纳入用于育种目的的基因工程实验。相反，必须评估其他激素在诱导Fox11叶片外植体体外再生方面的有效性。
Abstract The use of biotechnological methods is of particular interest in fruit tree crops since conventional breeding programs require long time and high resources before having impact upon commercialization. To develop an efficient plant regeneration protocol for pear rootstock Pyrodwarf, different combinations of two hormones, naphthalene-1-acetic acid (NAA) and Thidiazuron (TDZ) were studied on regeneration process. Other factors were also studied, such as darkness period and the use of different hormone concentrations during induction and expression phase. Our experiments showed that the length of darkness did not affect in vitro regeneration. Different concentrations of TDZ have different effects on regeneration: 6 μM has been shown to be more effective than 2.5 μM. A higher level of TDZ during induction period was the only important factor to induce regeneration independently to the presence of hormones during expression phase. The two hormones showed to be uneffective to induce a sufficient number of regenerating shoots from leaf explants of another pear rootstock, Fox11. Variability during regeneration was assessed using SSR markers. An identical genetic profile was observed between in vitro explants and regenerating shoots. These results are useful to include Pyrodwarf in genetic engineering experiments for breeding purposes. On the contrary, other hormones have to be evaluated for their effectiveness to induce in vitro regeneration in Fox11 leaf explants.