蔷薇科植物S-RNase特异性区域分析

S-RNase在配子体型自交不亲和性反应中起关键作用,HV区段被认为是雌蕊与花粉间特异识别的关键部位。应用生物信息学方法,对蔷薇科植物的S-RNase序列分析,并对HV区段一级结构邻近区和空间邻近区作物理化学性质分析,发现HV区C端的一段氨基酸序列符合蛋白质相互作用位点的特征;HVP区也是一个多态性区段,可能参与分子识别过程。因此,蔷薇科植物中,S-RNase与花粉S基因产物的作用方式可能为S-RNase的HVC区与花粉S基因产物先非特异性结合,再以HV区和HVP区进行分子间特异识别。     

基于RNA-seq技术对乌头属铁棒锤中自交不亲和S基因的挖掘与分析

以毛茛科乌头属铁棒锤(Aconitum pendulum N.Busch)2个品系‘蓝花铁棒锤’(‘WSYB1’)和‘黄花铁棒锤’(‘WSYY1’)为材料,对其进行转录组测序(RNA-seq),采用生物信息学方法鉴定其中可能存在的花柱S基因(self-incompatibility gene)和花粉S基因,并对它们的序列特征进行分析。结果显示,转录组中共鉴定出2个在雌蕊中特异或高表达的花柱S基因(ApSRNase)和2个在雄蕊中特异表达的花粉S基因(ApSLF)。与耧斗菜(Aquilegia coerulea James)相似,铁棒锤中也存在S-RNase(S locus ribonucleases)和SLF(S locus F-box)控制的S-RNase类的自交不亲和系统,而不存在sS(stigma S-determinant)和pS(pollen S-determinant)控制的罂粟科类型的自交不亲和系统。     

S-RNase-based self-incompatibility in Petunia inflata

Background

For the Solanaceae-type self-incompatibility, also possessed by Rosaceae and Plantaginaceae, the specificity of self/non-self interactions between pollen and pistil is controlled by two polymorphic genes at the S-locus: the S-locus F-box gene (SLF or SFB) controls pollen specificity and the S-RNase gene controls pistil specificity.

Scope

This review focuses on the work from the authors'' laboratory using Petunia inflata (Solanaceae) as a model. Here, recent results on the identification and functional studies of S-RNase and SLF are summarized and a protein-degradation model is proposed to explain the biochemical mechanism for specific rejection of self-pollen tubes by the pistil.

Conclusions

The protein-degradation model invokes specific degradation of non-self S-RNases in the pollen tube mediated by an SLF, and can explain compatible versus incompatible pollination and the phenomenon of competitive interaction, where SI breaks down in pollen carrying two different S-alleles. In Solanaceae, Plantaginaceae and subfamily Maloideae of Rosaceae, there also exist multiple S-locus-linked SLF/SFB-like genes that potentially function as the pollen S-gene. To date, only three such genes, all in P. inflata, have been examined, and they do not function as the pollen S-gene in the S-genotype backgrounds tested. Interestingly, subfamily Prunoideae of Rosaceae appears to possess only a single SLF/SFB gene, and competitive interaction, observed in Solanaceae, Plantaginaceae and subfamily Maloideae, has not been observed. Thus, although the cytotoxic function of S-RNase is an integral part of SI in Solanaceae, Plantaginaceae and Rosaceae, the function of SLF/SFB may have diverged. This highlights the complexity of the S-RNase-based SI mechanism. The review concludes by discussing some key experiments that will further advance our understanding of this self/non-self discrimination mechanism.     

S-RNases and sexual incompatibility: structure, functions, and evolutionary perspectives

S-RNase-based gametophytic self-incompatibility appears to be the most phylogenetically widespread form of self-incompatibility found in the angiosperms, having been reported in the Solanaceae, Scrophulariaceae, and Rosaceae. This intraspecific breeding barrier is controlled by a single genetic locus termed S. Rejection of self-pollen has been shown to be mediated in the pistil by a highly polymorphic series of ribonucleases, but as yet the pollen component of this recognition system has not been identified. Here we review our present knowledge concerning the structure, functions, and evolution of S-RNases and the S-loci in which they reside. In addition we present two new phylogenetic analyses of S-RNases which suggest that (1). sequence variability between S-alleles is spread across the whole gene and is not as clustered as is generally believed and (2). there is evidence of recombination and/or diversifying selection in two distinct regions of S-RNases. The implications of these findings are discussed.     

Expression of a fusion protein of Pyrus pyrifolia S-RNase with glutathione-S-transferase in E. coli

S-RNase is a style-specific ribonuclease which is associated with gametophytic self-incompatibility. An expression vector of a fusion protein of Pyrus pyrifolia(Japanese pear) S3-RNase with glutathione-S-transferase (GST) was constructed and transformed into E. coli. Using this system, the fusion protein, GST-S3-RNase, was expressed as an active form and can be used for screening pollen S-gene product(s).     

Identification of Self-Incompatibility Genotypes of Apricot (<Emphasis Type="Italic">Prunus armeniaca</Emphasis> L.) by S-Allele-Specific PCR Analysis

A cDNA of 417 bp encoding an S-RNase gene, named PA S3, was isolated from apricot, Prunus aremeniaca. Nine S-alleles, S₁–S₉, were recognized by S-allele-specific PCR and confirmed by Southern blot analysis using PA S3 as probe. The S-genotypes of the six cultivars were determined and the results of self- and cross-pollination tests among the six cultivars were consistent with the predicted S-haplotypes by PCR analysis.