Gametophytic self-incompatibility （GSI） is controlled by a highly polymorphic locus called the S-locus, which is an important factor that can result in seedless fruit in Citrus. The S 1 self-incompatibility locus-li...Gametophytic self-incompatibility （GSI） is controlled by a highly polymorphic locus called the S-locus, which is an important factor that can result in seedless fruit in Citrus. The S 1 self-incompatibility locus-linked pollen 3.15 gene （S1-3.15） belongs to a type of S locus gene. The role of S1-3.15 in the SI reaction of Citrus has not yet been reported. In this study, full-length sequences of cDNA and DNA encoding the S1-3.15 gene, referred to as CrS1-3.15 , were isolated from ‘Wuzishatangju’ （Self-incompatibility, SI） and ‘Shatangju’ （Self-compatibility, SC） . The predicted amino acid sequences of CrS1-3.15 between ‘Wuzishatangju’ and ‘Shatangju’ differ by only three amino acids. Compared to ‘Wuzishatangju’, three bases were substituted in the genomic DNA of CrS1-3.15 from ‘Shatangju’. Southern blot results showed that one copy of CrS1-3.15 existed in the genomic DNA of both ‘Wuzishatangju’ and ‘Shatangju’. The expression level of the CrS1-3.15 gene in the ovaries of ‘Shatangju’ was approximately 60-fold higher than that in the ovaries of ‘Wuzishatangju’. When ‘Wuzishatangju’ was cross-pollinated, the expression of CrS1-3.15 was upregulated in the ovaries at 3d, and the highest expression levels were detected in the ovaries at 6d after cross-pollination of ‘Wuzishatangju’ × ‘Shatangju’. To obtain the CrS1-3.15 protein, the full-length cDNA of CrS1-3.15 genes from ‘Wuzishatangju’ and ‘Shatangju’ was successfully expressed in Pichia pastoris. Pollen germination frequency of ‘Wuzishatangju’ was inhibited significantly with increasing CrS1-3.15 protein concentrations from SI ‘Wuzishatangju’.展开更多
基金supported by the National Natural Science Foundation of China(31000899)the Research Fund for the Doctoral Program of Higher Education of China(20104404120015 and 20114404110018)+4 种基金the Guangdong Province Science Foundation of China(06025843)the Science and Technology Planning Project of Guangzhou(2010r1-C771)the open foundation of the State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,South China Agricultural University(KSL-CUSAb-2012-09)the Key Laboratory of Innovation and Utilization for Germplasm Resources in Horticultural Crops in Southern China of Guangdong Higher Education Institutes,South China Agricultural University(No.KBL11008)the "211" Construction Fund for Key Subjects of College of Horticulture,South China Agricultural University
文摘Gametophytic self-incompatibility （GSI） is controlled by a highly polymorphic locus called the S-locus, which is an important factor that can result in seedless fruit in Citrus. The S 1 self-incompatibility locus-linked pollen 3.15 gene （S1-3.15） belongs to a type of S locus gene. The role of S1-3.15 in the SI reaction of Citrus has not yet been reported. In this study, full-length sequences of cDNA and DNA encoding the S1-3.15 gene, referred to as CrS1-3.15 , were isolated from ‘Wuzishatangju’ （Self-incompatibility, SI） and ‘Shatangju’ （Self-compatibility, SC） . The predicted amino acid sequences of CrS1-3.15 between ‘Wuzishatangju’ and ‘Shatangju’ differ by only three amino acids. Compared to ‘Wuzishatangju’, three bases were substituted in the genomic DNA of CrS1-3.15 from ‘Shatangju’. Southern blot results showed that one copy of CrS1-3.15 existed in the genomic DNA of both ‘Wuzishatangju’ and ‘Shatangju’. The expression level of the CrS1-3.15 gene in the ovaries of ‘Shatangju’ was approximately 60-fold higher than that in the ovaries of ‘Wuzishatangju’. When ‘Wuzishatangju’ was cross-pollinated, the expression of CrS1-3.15 was upregulated in the ovaries at 3d, and the highest expression levels were detected in the ovaries at 6d after cross-pollination of ‘Wuzishatangju’ × ‘Shatangju’. To obtain the CrS1-3.15 protein, the full-length cDNA of CrS1-3.15 genes from ‘Wuzishatangju’ and ‘Shatangju’ was successfully expressed in Pichia pastoris. Pollen germination frequency of ‘Wuzishatangju’ was inhibited significantly with increasing CrS1-3.15 protein concentrations from SI ‘Wuzishatangju’.
