Marchese Annalisa, Bosković Radovan I, Caruso Tiziano, Raimondo Antonio, Cutuli Marcello, Tobutt Kenneth R
East Malling Research, New Road, East Malling, Kent ME19 6BJ, UK.
J Exp Bot. 2007;58(15-16):4347-56. doi: 10.1093/jxb/erm322.
'Kronio' is a Sicilian cultivar of sweet cherry (Prunus avium), nominally with the incompatibility genotype S(5)S(6), that is reported to be naturally self-compatible. In this work the cause of its self-compatibility was investigated. Test selfing confirmed self-compatibility and provided embryos for analysis; PCR with consensus primers designed to amplify S-RNase and SFB alleles showed that the embryos were of two types, S(5)S(5) and S(5)S(6), indicating that S(6) pollen failed, but S(5) succeeded, perhaps because of a mutation in the pollen or stylar component. Stylar RNase analysis indicated active S-RNases for both S(5) and S(6). The S-RNase alleles were cloned and sequenced; and sequences encode functional proteins. Cloning and sequencing of SFB alleles showed that S(6) was normal but S(5) had a premature stop codon upstream of the variable region HVa resulting in a truncated protein. Therefore, the self-compatibility can be attributed to a pollen-part mutation of S(5), designated S(5)', the first reported case of breakdown of self-incompatibility in diploid sweet cherry caused by a natural mutation at the S-locus. The second intron of the S-RNase associated with S(5)' contained a microsatellite smaller than that associated with S(5); primers designed to amplify across this microsatellite effectively distinguished S(5) from S(5)'. Analysis of some other Sicilian cherries with these primers indicated that S(5)' is also present in the Sicilian cultivar 'Maiolina a Rappu', and this proved to be self-compatible.
“克罗尼奥”是甜樱桃(欧洲甜樱桃)的一个西西里品种,名义上具有不亲和基因型S(5)S(6),据报道它天然自交亲和。在这项研究中,对其自交亲和的原因进行了调查。自交试验证实了其自交亲和性,并提供了用于分析的胚;用设计用于扩增S-RNase和SFB等位基因的共有引物进行PCR分析表明,胚有两种类型,即S(5)S(5)和S(5)S(6),这表明S(6)花粉败育,但S(5)花粉成功,这可能是由于花粉或花柱成分发生了突变。花柱RNase分析表明,S(5)和S(6)的S-RNase均有活性。对S-RNase等位基因进行了克隆和测序;其序列编码功能性蛋白质。SFB等位基因的克隆和测序表明,S(6)正常,但S(5)在可变区HVa上游有一个提前终止密码子,导致蛋白质截短。因此,自交亲和性可归因于S(5)的花粉部分突变,命名为S(5)',这是首次报道的由S位点自然突变导致的二倍体甜樱桃自交不亲和性破坏的案例。与S(5)'相关的S-RNase的第二个内含子包含一个比与S(5)相关的微卫星更小的微卫星;设计用于扩增该微卫星的引物能够有效地区分S(5)和S(5)'。用这些引物对其他一些西西里樱桃进行分析表明,S(5)'也存在于西西里品种“马约利纳·阿·拉普”中,并且该品种被证明是自交亲和的。