Specific functions of single pistil S-RNases in S -gene homozygous Pyrus germplasm

Gametophytic self-incompatibility (SI) is regulated by S -allele recognition; that is, pollen in a style with the same S -genotype will undergo programmed cell death and stop growing so that it is unable to complete double fertilization, ultimately resulting in the SI response. S-RNase is the female determinant of SI in pear ( Pyrus ). In the Pyrus genome, there are two different S-RNase alleles at the S -locus, which generate two different S-RNase products in the pistil. The extracted S-glycoprotein is actually a protein complex. In this study, artificial self-pollination was conducted at the bud stage to overcome SI in ‘Huanghua’ ( S 1 S 2 ) pear. Seven plants homozygous for S 1 -RNase and four homozygous for S 2 -RNase were selected from the selfed progeny of ‘Huanghua’ by S -gene molecular identification biotechnology. We investigated the function of single S-RNases isolated from the pistils of S -gene homozygous Pyrus germplasm. The pollen of ‘Huanghua’ could smoothly pass through the style of the S -gene homozygous germplasm and complete fertilization. S-RNases were extracted from flower styles of different genotypes and used to treat different types of pollen. The S-RNase from ‘Huanghua’ completely inhibited the growth of S 1 S 2, S 1 S 1 , and S 2 S 2 pollen, while the S-RNase from homozygous germplasm allowed some S 1 S 2 pollen and different single genotypes of pollen to continue growing. These results further validate the core events of SI including cytoskeleton depolymerization and programmed cell death. By iTRAQ-based proteomic analysis of style proteins, a total of 13 S-RNase-related proteins were identified. In summary, we have created reliable S-RNase gene homozygous germplasm, which will play a crucial role in further research on SI in pear and in the development of the pear industry.