澳洲指橘与柑橘属间原生质体电融合再生二倍体体细胞杂种

电场诱导粗柠檬（CitrusjambhiriLush,2n＝2x＝18）叶肉原生质体与澳洲指橘（MicrocitruspapuanaSwingle,2n＝2x＝18）悬浮系原生质体融合，融合产物培养后再生出丛芽，经试管嫁接得到完整植株。再生植株的细胞学检查表明它们具有18条染色体，为二倍体；植株的叶片形态与叶肉亲本（粗柠檬）一样；用6个10-mer随机引物分析再生植株的杂种特性：在4个引物（OPA－07、OPAN－07、OPE－05和OPA－08）的扩增带型图中，再生植株的带型与粗柠檬完全一样，澳洲指橘的特征带未在植株中出现；在引物OPS-13和引物OPA－04的扩增带型图中，再生植株都具有澳洲指橘的特征带。细胞学和RAPD分析的结果表明，通过对称融合得到了澳洲指橘与粗柠檬的属间二倍体体细胞杂种植株。这是柑橘属间对称融合再生二倍体叶肉亲本类型植株的首例报道。

Regeneration of Diploid Intergeneric Somatic Hybrid Plants Between Microcitrus and Citrus via Electrofusion

Leaf-derived protoplasts of Rough lemon (Citrus jambhiri Lush, 2n = 2x = 18) were electrofused with embryogenic suspension protoplasts of its relative, Microcitrus papuana Swingle (2n = 2x = 18), with an intention of creating novel germplasm. Six plants were regenerated following protoplasts fusion. Cytological examination demonstrated that they were diploids with 18 chromosomes (2n = 2x = 18). RAPD (random amplified polymorphic DNA) analyses with six arbitrary 10-mer primers showed that the regenerated plants had identical band patterns to those of Rough lemon for primers OPA-07, OPAN-07, OPE-05 and OPA-08, Whereas for the other two primers, OPA-04 and OPS-13, bands specific to M. papuana could be detected in the regenerated plants. Cytological and RAPD analysis revealed that the regenerated plants were diploid somatic hybrids between M. papuana and Rough lemon. The putative hybrids were morphologically similar to Rough lemon. This is the first report on production of diploid somatic hybrid plants between citrus with its related genus via symmetric fusion.