CRISPR/Cas9-mediated knockout of PiSSK1 reveals essential role of S-locus F-box protein-containing SCF complexes in recognition of non-self S-RNases during cross-compatible pollination in self-incompatible Petunia inflata

被引：0
作者：
Sun L. [1 ]
Kao T.-H. [1 ,2 ]
机构：
[1] Intercollege Graduate Degree Program in Plant Biology, The Pennsylvania State University, University Park, 16802, PA
[2] Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, 16802, PA
来源：
Plant Reproduction | 2018年 / 31卷 / 2期
基金：
美国国家科学基金会;
关键词：
CRISPR/Cas9; Petunia inflata; PiSSK1; S-locus F-box protein; Self-incompatibility; Skp1-like protein;
D O I：
10.1007/s00497-017-0314-1
中图分类号：
学科分类号：
摘要：
Key message: Function ofPetuniaPiSSK1. Abstract: Self-incompatibility (SI), an inbreeding-preventing mechanism, is regulated in Petunia inflata by the polymorphic S-locus, which houses multiple pollen-specific S-locus F-box (SLF) genes and a single pistil-specific S-RNase gene. S2-haplotype and S3-haplotype possess the same 17 polymorphic SLF genes (named SLF1 to SLF17), and each SLF protein produced in pollen is assembled into an SCF (Skp1–Cullin1–F-box) E3 ubiquitin ligase complex. A complete suite of SLF proteins is thought to collectively interact with all non-self S-RNases to mediate their ubiquitination and degradation by the 26S proteasome, allowing cross-compatible pollination. For each SCFSLF complex, the Cullin1 subunit (named PiCUL1-P) and Skp1 subunit (named PiSSK1), like the F-box protein subunits (SLFs), are pollen-specific, raising the possibility that they also evolved specifically to function in SI. Here we used CRISPR/Cas9-meditated genome editing to generate frame-shift indel mutations in PiSSK1 and examined the SI behavior of a T0 plant (S2S3) with biallelic mutations in the pollen genome and two progeny plants (S2S2) each homozygous for one of the indel alleles and not carrying the Cas9-containing T-DNA. Their pollen was completely incompatible with pistils of seven otherwise-compatible S-genotypes, but fully compatible with pistils of an S3S3 transgenic plant in which production of S3-RNase was completely suppressed by an antisense S3-RNase gene, and with pistils of immature flower buds, which produce little S-RNase. These results suggest that PiSSK1 specifically functions in SI and support the hypothesis that SLF-containing SCF complexes are essential for compatible pollination. © 2017, Springer-Verlag GmbH Germany, part of Springer Nature.
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页码：129 / 143
页数：14
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