Regeneration and characterization of plants derived from leaf in vitro culture of two sweet orange (Citrus sinensis (L.) Osbeck) cultivars
Regeneration and characterization of plants derived from leaf in vitro culture of two sweet orange (Citrus sinensis (L.) Osbeck) cultivars
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两个甜橙（Citrus sinensis (L.) Osbeck）品种叶离体培养植物的再生和表征
DOI：
10.1016/j.scienta.2008.10.004
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发表时间：
2009-03-03
影响因子：
4.3
通讯作者：
Liu, Ji-Hong
中科院分区：
文献类型：
--
作者：
Khan, Ehsan Ullah;Fu, Xing-Zheng;Liu, Ji-Hong
对冰糖城和瓦伦西亚两个甜橙品种的叶片外植体培养进行了研究。以外植体出芽率、平均每外植体出芽数和不定芽形成能力为指标，研究了培养基数、光照条件、外植体年龄和基因型等因素对再生反应的影响。外植体在含有不同生长调节剂的MT盐诱导培养基(SIM)上诱导产生不定芽，其中以SIM_1(MT+0.5 mg L-1 BA+0.5 mg L-1激动素+0.1 mg L-1NAA+3%蔗糖+0.8%琼脂，pH 5.8)诱导叶片不定芽效果最好。两个品种对不定芽再生的反应差异极显著，冰糖城的反应比瓦伦西亚强。与未发育叶片相比，成熟叶片外植体培养具有更好的不定芽再生能力。然而，本文所用的两种光照条件对不定芽再生的影响并不显著。表型观察和随机扩增多态DNA分析证实，两个基因型的所有再生植株与供体植株在遗传上完全相同，表明再生植株中没有可检测到的遗传变异。本文提供的数据表明，已成功地实现了叶片外植体的直接植株诱导。据我们所知，这是关于柑橘叶片外植体直接再生不定芽的首次报道，这将为今后柑橘遗传操作提供一个替代来源。(C)2008爱思唯尔B.V.保留所有权利。
In the current work attempts were made to investigate culture of leaf explants derived from in vitro seedlings of two sweet orange (Citrus sinensis (L) Osbeck) cultivars, Bingtangcheng and Valencia. Effects of several factors, including culture medium, lighting condition, explant age and genotype on regeneration response were examined based on three parameters, percentage of explants producing shoots, mean number of shoots per explant and shoot forming capacity. Culture of the explants on shoot-inducing media (SIM) composed of MT salts supplemented with different growth regulators gave rise to disparate shoot regeneration, in which SIM1 (MT + 0.5 mg L-1 BA + 0.5 mg L-1 Kinetin + 0.1 mg L-1 NAA + 3% sucrose + 0.8% agar, pH 5.8) was shown to be the most effective medium for direct induction of shoots from leaf explants. Highly significant difference in the response of shoot bud regeneration was noted between the two cultivars, with Bingtangcheng being more responsive than Valencia. Culture of explants from fully developed leaves led to better shoot regeneration capacity in comparison to undeveloped ones. However, the two lighting conditions used herein did not cause significant difference in shoot regeneration. Phenotypic observation and randomly amplified polymorphic DNA (RAPD) analysis confirmed that all the regenerated plants from both genotypes were genetically identical to their donor plants, suggesting absence of detectable genetic variation in the regenerated plants. The data presented here demonstrated that direct initiation of plants from leaf explants has been successfully accomplished. To our knowledge, this is the first report on direct regeneration of shoots from leaf explants in Citrus, which will provide an alternative source for citrus genetic manipulation in the future. (C) 2008 Elsevier B.V. All rights reserved.