Analysis if Mitochondrial Genomes in Citrus Interspecific Somatic Hybrids Produced by Protoplastfusion

原生質體融合創造的體細胞雜種對柑橘改良具有重要價值。本文選取4株6年生的朋娜臍橙(Citrus sinensis Osbeck)+粗檸檬(C. jambhiri)種間體細胞雜種植株進行遺傳分析。倍性流式細胞儀和核基因組SSR分析表明它們是真正的異源四倍體雜種。葉綠體SSR分析表明其葉綠體基因組為隨機遺傳。其線粒體基因組的特異PCR擴增表明，這些體細胞雜種除共有一條1,100bp左右的譜帶外，還出現了葉肉親本粗檸檬的特異帶。PCR產物的進一步Tas Ⅰ限制性酶切揭示，葉肉親本的特徵帶無變化，但共有帶在所有樣品中均被酶切，而且體細胞雜種的帶型與懸浮系親本朋娜臍橙相同。共有帶的DNA測序和DNA序列的Tas Ⅰ限制性酶切位點檢索很好地解釋了線粒體基因組的CAPS分析帶型結果。這是CAPS分析和DNA測序揭示柑橘體細胞雜種線粒體基因組重組的首例報導。

關鍵字

柑橘； CAPS ；胞質基因組； DNA測序；簡單序列重複；體細胞雜種

並列摘要

Somatic hybrids produced via protoplast fusion are valuable germplasm for citrus improvement. Herein, four randomly selected six-year-old interspecific somatic hybrid plants between Bonanza sweet orange (Citrus sinensis Osbeck) and rough lemon (C. jambhiri), were thoroughly analyzed. Flow cytometry and nuclear SSR analysis confirmed them as true tetraploid somatic hybrids. Chloroplast SSR analysis showed the random inheritance nature of chloroplast DNA. PCR amplification of mitochondrial genome by universal primer pair showed that the somatic hybrids had the specific band from rough lemon (the leaf parent) while all samples shared a common band. CAPS analysis by further TasI restriction endonuclease cut of the PCR products, however, revealed the band specific to Bonanza orange, the embryogenic callus parent, was also present in all these analyzed hybrids. Further sequencing of the common band and searching for restriction endonuclease recognition sites well explained the banding pattern by CAPS analysis. It was concluded that mitochondrial recombination in citrus somatic hybrids occurred as revealed by CAPS analysis and DNA sequencing.

並列關鍵字

Citrus ； Cleaved amplified polymorphic sequence CAPS ； Cytoplasmic genome ； DNA sequencing ； Simple sequence repeat SSR ； Somatic hybrids

國際替代計量

Analysis if Mitochondrial Genomes in Citrus Interspecific Somatic Hybrids Produced by Protoplastfusion

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