原生质体电融合再生柑橘属间体细胞杂种

电场诱导酸橙(CitrusaurantitumL.)叶肉原生质体与澳洲指橘(MicrocitruspapuanaSwingle)悬浮系原生质体融合，融合产物经培养再生出具有三种叶片形态的植株，绝大部分与酸橙叶片完全相似(即叶肉亲本型植株)；有2棵植株叶片大且厚；有1棵植株出现二叶和三叶。细胞学检查表明这些植株均为二倍体。采用RAPD标记对前两种植株进行杂种特性分析，共选用4个10-mer随机引物进行扩增。在引物OPAA-17的扩增产物中，再生植株的带型与酸橙一致；在引物OPA-08的扩增产物中，再生植株的带型与酸橙或澳洲指橘相似；而在引物OPA-04和OPA-07的扩增产物中，再生植株的带型出现三种情况，4个引物的扩增结果证明所分析的植株均是体细胞杂种。综合染色体检查和RAPD标记的结果，表明获得了酸橙与澳洲指橘属间二倍体体细胞杂种植株。

PRODUCTION OF INTERGENERIC SOMATIC HYBRID PLANTS VIA PROTOPLAST ELECTROFUSION IN CITRUS

Leaf derived protoplasts of Sour orange (Citrus aurantium L. ) were fused electrically with embryogenic protoplasts of Microcitrus papuana Swingle. Plants were regenerated from the fusion products, which were characteristic with three types of leaf morphology. Most of the plants were identical to Sour orange (namely, Leaf-parent-type plant) and two plants had large and thick leaves whereas one plant had bifoliate and trifoliate leaves. Chromosome examination showed that these plants were diploid with 18 chromosomes (2n = 2x = 18). RAPD analysis was employed to verify the hybrid characteristicsof the plants in the first two types. Four 10-mer arbitrary primers with polymorphism were chosen. Band pattern of the plants was similar with the leaf parent (Sour orange) for the primer OPAA-17. Band pattern of the plants was similar with either Sour orange or M. papuana for OPA-08. As for OPA-07 and OPA-04 three kinds of band profiles were detected. Results of RAPD marker. together with chromosome determination, indicated that all of the analyzed plants were intergeneric diploid somatic hybrids between Sour orange and M. papuana.