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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (04): 579-586.doi: 10.3724/SP.J.1006.2011.00579

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Detection of Interactions between SCR and SRK in Brassica oleracea L. by Yeast Two-Hybrid System

LUO Bing1,3,XUE Li-Yan1,**,ZHU Li-Quan1,*,ZHANG He-Cui1,PENG Yi-Bo1,CHEN Song1,YANG Hong1,YANG Kun1,LI Cheng-Qiong2,WANG Xiao-Jia2,*   

  1. 1 Plant Physiology and Biochemistry Laboratory of Southwest University, Chongqing 400716, China; 2 Key Laboratory in Olericulture of Chongqing, Southwest University, Chongqing 400716, China; 3 College of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, China
  • Received:2010-09-17 Revised:2011-01-06 Online:2011-04-12 Published:2011-02-24
  • Contact: 朱利泉, E-mail: zhuliquan@swu.edu.cn, Tel: 023-68250794, Fax:023-68251914; 王小佳, E-mail: wxj@swu.edu.cn

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Abstract: For further study on mechanism of the mutual recognition between S-locus receptor kinase (SRK) and S-locus Cysteine-rich protein (SCR) in Brassica, the extracellular domains of SRK (named as eSRK) and SCR were amplified by nested PCR using Brassica oleracea L. ‘D3’ with the trait of typical self-incompatibility (SI), and the interactions between eSRK and SCR was detected by the yeast two-hybrid system. Then the full-length eSRK and the SCR were fused to the Gal4 DNA activation domain (designated pGADT7eSRK) and the Gal4 DNA binding domain (designated pGBKT7SCR) by the gene homologous recombination technique respectively. Then the pGADT7eSRK plasmid and the pGBKT7SCR were transformed into the Y187 yeast strain and the Y2HGold yeast strain respectively. The two transformed yeast strains did not exhibit autoactivation. The diploids, which were fused by the transformed Y2HGold yeast strain and the transformed Y187 yeast strain, grew on selective agar plates (SD/–Ade/–His/– Leu/–Trp/X-a-Gal/AbA), and the resulting colonies were blue, which strongly indicated that eSRK and SCR could combine with each other.

Key words: Brassica oleracea L., Self-incompatibility, S-locus receptor kinase (SRK), S-locus cysteine-rich protein (SCR), Yeast two-hybrid system

[1]Bateman A J. Self-incompatibility systems in angiosperms: III. Cruciferae Hered, 1955, 9: 53–68
[2]Wheeler M J, Graaf B H J, Hadjiosif N, Perry R M, Poulter N S, Osman K. Identification of the pollen self-incompatibility determinant in Papaver rhoeas. Nature, 2009, 459: 992–995
[3]Stein J C, Howlett B, Boyes D C, Nasrallah M E, Nasrallah J B. Molecular cloning of a putative receptor protein kinase gene encoded at the self-incomaptibility locus of Brassica oleracea. Proc Natl Acad Sci USA, 1991, 88: 8816–8820
[4]Takayama S, Shimosato H, Shiba H, Funato M, Che F S, Watanabe M, Iwano M, Isogai A. Direct ligand–receptor complex interaction controls Brassica self-incompatibility. Nature, 2001, 413: 534–538
[5]Schopfer C R, Nasrallah M E, Nasrallah J B. The male determinant of self-incompatibility in Brassica. Science, 1999, 286: 1697–1700
[6]Suzuki G, Kai N, Hirose T, Nishio T, Takayama S, Isogai A, Watanabe M, Hinata K. Genomic organization of the S locus: identification and characterization of genes in SLG/SRK region of S9 haplotype of Brassica campestris (syn. rapa). Genetics, 1999, 153: 391–400
[7]Nasrallah J B, Yu S M, Nasrallah M E. Self-incompatibility genes of Brassica oleracea: expression, isolation and structure. Proc Nat Acad Sci USA, 1988, 85: 5551–5555
[8]Shiba H, Takayama S, Iwano M, Shimosato H, Funato M, Nakagawa T, Che F S, Suzuki G, Watanabe M, Hinata K, Isogai A. A pollen coat protein, SP11/SCR, determines the pollen S-specificity in the self-incompatibility of Brassica species. Plant Physiol, 2001, 125: 2095–2103
[9]Rong X-Y(荣小营), Zhu Li-Q(朱利泉), Wang Y(王永), Gao Q-G(高启国), Chen X-D(陈晓丹), Yang Y(杨洋), Wang X-J(王小佳). Localization of MLPK and SSP genes for self-incompatibility of Brassica oleracea by fluorescence in situ hybridization. Acta Agron Sin (作物学报), 2009, 35(5): 802–808 (in Chinese with English abstract)
[10]Takasaki T, Hatakeyama K, Suzuki G, Watanabe M, Isogai A, Hinata K. The S receptor kinase determines self-incompatibility in Brassica stigma. Nature, 2000, 403: 913–916
[11]Gao Q-G(高启国), Song M(宋明), Niu Y(牛义), Yang K(杨昆), Zhu L-Q(朱利泉), Wang X-J(王小佳). Test of interaction between THL1 and SRK from Brassica oleracea L. in self-incompatibility signaling process. Acta Agron Sin (作物学报), 2008, 34(6): 1–10 (in Chinese with English abstract)
[12]Nasrallah J B, Nasrallah M E. Pollen-stigma signaling in the sporophytic self-incompatibility response. Plant Cell, 1993, 5: 1325–1335
[13]Kachroo A, Schopfer C R, Nasrallah M E, Nasrallah J B. Allele-specific receptor-ligand interactions in Brassica self-incompatibility. Science, 2001, 293: 1824–1826
[14]Shimosato H, Yokota N, Shiba H, Iwano M, Entani T, Che F S, Watanabe M, Isogai A, Takayama S. Characterization of the SP11/SCR high-affinity binding site involved in self/nonself recognition in Brassica self-incompatibility. Plant Cell, 2007, 19: 107–117
[15]Higashiyama T. Peptide Signaling in Pollen–Pistil Interactions. Plant Cell Physiol, 2010, 51: 177–189
[16]Oender K, Niedermayr P, Hintner H, Richter K, Koller L, Trost A, Bauer J W, Hundsberger H. Relative quantitation of protein–protein interaction strength within the yeast two-hybrid system via fluorescence β-galactosidase activity detection in a high-throughput and low-cost manner. ASSAY Drug Dev Technol, 2006, 4: 709–719
[17]Bower M S, Matias D D, Fernandes C E, Mazzurco M, Gu T, Rothstein S, Goring D R. Two members of the thioredoxin-h family interact with the kinase domain of a Brassica S locus receptor kinase. Plant Cell, 1996, 8: 1641–1650
[18]Gu T, Mazzurco M, Matias D D, Sulaman W, Goring D R. Binding of a novel arm repeat protein to the kinase domain of the S locus receptor kinase. Proc Natl Acad Sci USA, 1998, 95: 382–387
[19]Mazzurco M, Sulaman W, Helen E, Cock J M, Goring D R. Further analysis of the interactions between the Brassica S receptor kinase and three interacting proteins (ARC1, THL1 and THL2) in the yeast two-hybrid system. Plant Mol Biol, 2001, 45: 365–376
[20]Naithani S, Chookajorn T, Ripoll D R, Nasrallah J B. Structural modules for receptor dimerization in the S-locus receptor kinase extracellular domain. Proc Natl Acad Sci USA, 2007, 104: 12211–12216
[21]Nishio T, Kusaba M, Sakamoto K, Ockendon D J. Polymorphism of the kinase domain of the S-locus receptor kinase gene (SRK) in Brassica oleracea L. Theor Appl Genet, 1997, 95: 335–342
[22]Watanabe M, Ito A, Takada Y, Ninomiya C, Kakizaki T, Takahata Y, Hatakeyam K, Hinata K, Suzuki G, Takasaki T, SattaY, Shiba H, Takayama S, Isogai A. Highly divergent sequences of the pollen self-incompatibility (S) gene in class-I S haplotypes of Brassica campestris (syn. rapa) L. FEBS Lett, 2000, 473: 139–144
[23]Nasrallah M E, Liu P, Broyles S S, Boggs N A, Nasrallah J B. Natural variation in expression of self-incompatibility in Arabidopsis thaliana: Implications for the evolution of selfing. Proc Natl Acad Sci USA, 2004, 101: 16070–16074
[24]Kemp B P, Doughty J. S cysteine-rich (SCR) binding domain analysis of the Brassica self-incompatibility S-locus receptor kinase. New Phytol, 2007, 175: 619–629
[25]Fujimoto R, Sugimura T, Nishio T. Gene conversion from SLG to SRK resulting in self-compatibility in Brassica rapa. FEBS Lett, 2006, 580: 425–430
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