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Molecular Characterization and Expression Analysis of S1 self-incompatibility Locus-linked Pollen 3.15 Gene in Citrus reticulata

查看全文 作　　者：Hong Xia [1,2]Miao;Zi Xing [1,3]Ye;Yong Hua [1,3]Qin;Gui Bing [1,3,2]Hu 高影响力作者 机构地区：[1]College of Horticulture,South China Agricultural University;[2]State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,South China Agricultural University;[3]Key Laboratory of Biology and Genetic Improvement of Horticultural Crops-South China,Ministry of Agriculture高影响力机构 出　　处：《Journal of Integrative Plant Biology》索引2013年第55卷第5期,共10页高影响力期刊 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);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 摘　　要：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'. 关 键 词：配子体自交不亲和 花粉萌发 基因座 表达分析 柑橘 SOUTHERN杂交 特征和 CDNA序列
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