Production and molecular characterization of potential seedless cybrid plants between pollen sterile Satsuma mandarin and two seedy Citrus cultivars
Production and molecular characterization of potential seedless cybrid plants between pollen sterile Satsuma mandarin and two seedy Citrus cultivars
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DOI：
10.1007/s11240-007-9266-8
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发表时间：
2007-09-01
影响因子：
3
通讯作者：
Guo, Wen-Wu
中科院分区：
文献类型：
--
作者：
Cai, Xiao-Dong;Fu, Jing;Guo, Wen-Wu
在包括温州蜜柑（Citrus unshiu Marc.）在内的高等植物中，细胞质雄性不育（CMS）是由线粒体基因组控制的。柑橘对称融合实验经常产生二倍体胞质杂种，其具有来自叶肉亲本的核DNA和来自胚性愈伤组织亲本的线粒体DNA（mtDNA）。因此，有可能通过体细胞杂交将温州蜜柑的细胞质雄性不育系作为愈伤组织亲本，转移到多籽柑橘的叶片中。本研究采用对称融合技术，将温州蜜柑细胞质雄性不育系（CMS）转化为无核细胞质杂种。CV.国青1号（G1）与沙田柚（C. grandis（L）Osbeck）和冰糖橙子（C. sinensis（L）Osbeck）。流式细胞仪分析表明，从G1 + '冰糖'橙子中回收的19株植株和从G1 + '沙田柚'中再生的35株植株中有17株为二倍体。G1 +沙田柚的其余植株均为四倍体。利用SSR和CAPS分析，证实两个组合的二倍体植株为真胞质杂种，其核DNA来自其相应的叶片亲本，mtDNA来自其共同的悬浮亲本G1温州蜜柑。G1 +沙田柚的其余植株经鉴定为带有G1线粒体DNA的体细胞杂种。叶绿体简单重复序列（cp-SSR）分析表明，两个组合的体细胞杂种/胞质杂种大多数具有双亲叶绿体类型之一，G1 +沙田柚的2个植株和G1 +冰糖橙子的所有胚状体都具有双亲叶绿体DNA（cpDNA）。这些结果表明，我们成功地将G1温州蜜柑线粒体DNA通过对称融合导入到两个目标无核柑橘品种中。
Cytoplasmic male sterility (CMS) is known to be controlled by mitochondrial genome in higher plants including Satsuma mandarin (Citrus unshiu Marc.). Citrus symmetric fusion experiments often produce diploid cybrids possessing nuclear DNA from the mesophyll parent and mitochondrial DNA (mtDNA) from the embryogenic callus parent. Therefore, it is possible to transfer CMS from Satsuma mandarin as callus parent to seedy citrus cultivars as leaf one by somatic cybridization. Herein, symmetric fusion technique was adopted to create cybrids for potential seedlessness by transferring CMS from Citrus unshiu Marc. cv. Guoqing No. 1 (G1) to two traditional Chinese seedy citrus cultivars, 'Shatian' pummelo (C. grandis (L) Osbeck) and 'Bingtang' orange (C. sinensis (L) Osbeck). Flow cytometry analysis showed that 19 plants recovered from G1 + 'Bingtang' orange and 17 of 35 plants regenerated from G1 + 'Shatian' pummelo were diploid. The remaining plants from G1 + 'Shatian' pummelo were tetraploid. The diploid plants from the two combinations were confirmed as true cybrids by simple sequence repeat (SSR) and cleaved amplified polymorphic sequence (CAPS) analysis, with nuclear DNA from their corresponding leaf parent and mtDNA from their common suspension parent, G1 Satsuma mandarin. The remaining plants from G1 + 'Shatian' pummelo were identified as somatic hybrids with mtDNA from G1. The chloroplast simple sequence repeat (cp-SSR) analysis revealed somatic hybrid/cybrid plants from the two combinations in most cases possessed either of their parental chloroplast type, and two plants from G1 + 'Shatian' pummelo and all embryoids analyzed from G1 + 'Bingtang' orange possessed chloroplast DNA (cpDNA) from both parents. These results demonstrated that we succeeded in introducing mtDNA from G1 Satsuma mandarin into the two target seedy citrus cultivars for potential seedlessness through symmetric fusion.