Component Analysis of SCFSLF Complex in Diploid Potato

doi:10.13560/j.cnki.biotech.bull.1985.2021-0904

Abstract

Abstract:

Self-incompatibility (SI) exists widely in flowering plants,which is a way to prevent inbreeding and promote hybridization,and may effectively maintain the genotypic diversity of species. The type of SI in Plantaginaceae,Solanaceae,Rosaceae and other plants belongs to the typical gametophytic self-incompatibility (GSI). In the cross-pollination of GSI plants,the SCF^SLF complex (Skp1/Cullin1/F-box) in the pollen tube,as the E3 ubiquitin ligase,is involved in the degradation of S-RNase,which is the key factor to maintain the crossing compatibility. This study summarized the research progress of SCF^SLF complex in GSI plants,and preliminarily analyzed the components of SCF^SLF complex in diploid potato by homologous alignment,phylogenetic analysis and multiple protein sequence alignments. The gene expression and protein interaction of each component were analyzed,and it was preliminarily determined that the SCF^SLF complex in potato pollen tube may be composed of Soltu.DM.06G001040,Soltu.DM.11G004330,Soltu.DM.06G022320 and the corresponding SLF proteins.

Key words: SCF^SLF complex, gametophytic self-incompatibility, Solanum tuberosum

GAO Meng, LI Fu-ting, WEI Zhan-lin, ZHANG Sai-hang, BAI Ru-qian, SHANG Yi, MA Ling. Component Analysis of SCF^SLF Complex in Diploid Potato[J]. Biotechnology Bulletin, 2022, 38(4): 117-125.

Figures/Tables 7

Fig.1 Typical structural pattern of SCF complex

Table 1 Fluorescent quantitative primers for SSK1 and Cullin1 in potato（S. tuberosum）

引物名称 Primer	引物序列 Sequence（5'-3'）
4330-SSK1-F	ATGGAGTTGTGTTGCCAATC
4330-SSK1-R	AATCTCAGGACCTTCAAATGC
1040-CLU1-F	GTGTCATGTGCCAAGTACAA
1040-CLU1-R	CATAACGTCTATCCTTGCCG
QEF1a-F	GATGGTCAGACCCGTGAACATGC
QEF1a-R	ACGATTTCATCATACCTAGCCTTGGAGT

Table 2 Homologous comparison of SCFSLF complex components in diploid potato DM

	基因编号Gene ID	基因位点Location	E值E value
PiSSK1同源基因 PiSSK1-homologous gene	Soltu.DM.11G004330	chr11：4294163..4295355 reverse	1.71E-102
	Soltu.DM.11G016290	chr11：30776770..30778841 reverse	2.83E-44
	Soltu.DM.11G004300	chr11：4286144..4286630 forward	9.28E-43
	Soltu.DM.10G017580	chr10：47907512..47910644 forward	9.30E-42
	Soltu.DM.01G051200	chr01：87717008..87719187 forward	2.54E-41
	Soltu.DM.11G004500	chr11：4499656..4500133 reverse	1.32E-40
PiCUL1-P同源基因 PiCUL1-P-homologous genes	Soltu.DM.06G001040	chr06：1465504..1471494 reverse	0
	Soltu.DM.06G035050	chr06：58938032..58944924 reverse	0
	Soltu.DM.09G023090	chr09：58921771..58929345 reverse	0
	Soltu.DM.01G022080	chr01：60076281..60087621 forward	0
	Soltu.DM.09G003030	chr09：2388523..2390740 reverse	0
	Soltu.DM.09G002940	chr09：2349588..2351796 forward	0
	Soltu.DM.01G022100	chr01：60106453..60114114 forward	0
	Soltu.DM.01G022150	chr01：60164455..60171592 forward	0
	Soltu.DM.09G003010	chr09：2377567..2379367 reverse	0
	Soltu.DM.01G022120	chr01：60134989..60155075 forward	0
PiRbx1同源基因 PiRbx1-homologous genes	Soltu.DM.06G022320	chr06：48782308..48785375 reverse	1.82E-71

Fig. 2 SSK1 protein sequence and expression analysis A：Phylogenetic tree of SSK genes in GSI plants（The branch marked by the red triangle is the branch where Rosaceae and Solanaceae separated）. B：Sequence alignment of SSK protein in GSI plants. C：The expression of Soltu.DM.11G004330 in different organs of E172 is calculated by 2-∆∆Ct with stamen as control

Fig. 3 Cullin1 protein evolution and expression analysis A：Phylogenetic tree of CUL genes in GSI plants（The branch marked by the red triangle is the branch where Rosaceae and Solanaceae separated）. B：The expression of Soltu.DM.06G001040 in different organs of E172 is calculated by 2-∆∆Ct with stamen as control

Fig. 4 Multiple sequence alignment of Cullin1 protein in potato

Fig. 5 Interaction analysis of Soltu.DM.11G004330 and SLF A：Y2H of Soltu.DM.11G004330 and SLF（-WLHA,synthetic dropout media lacking tryptophan,leucine,histidine,and adenine（-Trp/W,-Leu/L,-His/H,-adenine/Ade）;100,10-1,and 10-2 indicates concentration of E.coli;pGBKT7-53 and pGADT7-T are used as positive control;pGBKT7-Lam and pGADT7-T are used as negative control;4330 is abbreviated from Soltu.DM.11G004330）. B：LUC complementation of Soltu.DM.11G004330 and SLF.（The left figure denotes how injections were carried out on tobacco leaves and 4330 was the abbreviation of Soltu.DM.11G004330）

References 21

[1]	McClure B, Cruz-García F, Romero C. Compatibility and incompatibility in S-RNase-based systems[J]. Ann Bot, 2011, 108(4):647-658. doi: 10.1093/aob/mcr179 URL
[2]	Fujii S, Kubo K, Takayama S. Non-self- and self-recognition models in plant self-incompatibility[J]. Nat Plants, 2016, 2(9):16130. doi: 10.1038/nplants.2016.130 pmid: 27595657
[3]	Matsumoto D, Tao R. Recognition of S-RNases by an S locus F-box like protein and an S haplotype-specific F-box like protein in the Prunus-specific self-incompatibility system[J]. Plant Mol Biol, 2019, 100(4/5):367-378. doi: 10.1007/s11103-019-00860-8 URL
[4]	Hua Z, Kao TH. Identification and characterization of components of a putative Petunia S-locus F-box-containing E3 ligase complex involved in S-RNase-based self-incompatibility[J]. Plant Cell, 2006, 18(10):2531-2553. doi: 10.1105/tpc.106.041061 URL
[5]	Li S, Sun P, Williams JS, et al. Identification of the self-incompatibility locus F-box protein-containing complex in Petunia inflata[J]. Plant Reprod, 2014, 27(1):31-45. doi: 10.1007/s00497-013-0238-3 URL
[6]	李红艳, 翟赛亚, 徐明磊. 植物配子体型自交不亲和性作用机理研究进展[J]. 南方园艺, 2015, 26(3):76-78.
	Li HY, Zhai SY, Xu ML. Research progress on gametophytic self-incompatibility in plants[J]. South Hortic, 2015, 26(3):76-78.
[7]	Williams JS, Wu L, Li S, et al. Insight into S-RNase-based self-incompatibility in Petunia:recent findings and future directions[J]. Front Plant Sci, 2015, 6:41. doi: 10.3389/fpls.2015.00041 pmid: 25699069
[8]	Luu DT, Qin X, Morse D, et al. S-RNase uptake by compatible pollen tubes in gametophytic self-incompatibility[J]. Nature, 2000, 407(6804):649-651. doi: 10.1038/35036623 URL
[9]	Goldraij A, Kondo K, Lee CB, et al. Compartmentalization of S-RNase and HT-B degradation in self-incompatible Nicotiana[J]. Nature, 2006, 439(7078):805-810. doi: 10.1038/nature04491 URL
[22]	Kubo K, Entani T, Takara A, et al. Collaborative non-self recognition system in S-RNase-based self-incompatibility[J]. Science, 2010, 330(6005):796-799. doi: 10.1126/science.1195243 URL
[23]	Entani T, Kubo K, Isogai S, et al. Ubiquitin-proteasome-mediated degradation of S-RNase in a solanaceous cross-compatibility reaction[J]. Plant J, 2014, 78(6):1014-1021. doi: 10.1111/tpj.12528 URL
[24]	Kubo KI, Tsukahara M, Fujii S, et al. Cullin1-P is an essential component of non-self recognition system in self-incompatibility in Petunia[J]. Plant Cell Physiol, 2016, 57(11):2403-2416. doi: 10.1093/pcp/pcw152 URL
[25]	Chang LC, Guo CL, Lin YS, et al. Pollen-specific SKP₁-like proteins are components of functional scf complexes and essential for lily pollen tube elongation[J]. Plant Cell Physiol, 2009, 50(8):1558-1572. doi: 10.1093/pcp/pcp100 URL
[26]	Ren Y, Hua QZ, Pan JY, et al. SKP₁-like protein, CrSKP₁-e, interacts with pollen-specific F-box proteins and assembles into SCF-type E3 complex in ‘Wuzishatangju’(Citrus reticulata Blanco)pollen[J]. PeerJ, 2020, 8:e10578.
[27]	Zeng B, Wang JY, Hao Q, et al. Identification of a novel SBP₁-containing SCFSFB complex in wild dwarf almond(Prunus tenella)[J]. Front Genet, 2019, 10:1019. DOI: 10.3389/fgene.2019.01019. doi: 10.3389/fgene.2019.01019 pmid: 31708966
[28]	Yuan H, Meng D, Gu ZY, et al. A novel gene, MdSSK1, as a component of the SCF complex rather than MdSBP₁ can mediate the ubiquitination of S-RNase in apple[J]. J Exp Bot, 2014, 65(12):3121-3131. doi: 10.1093/jxb/eru164 pmid: 24759884
[29]	Huang J, Zhao L, Yang QY, et al. AhSSK1, a novel SKP₁-like protein that interacts with the S-locus F-box protein SLF[J]. Plant J, 2006, 46(5):780-793. pmid: 16709194
[30]	Zhao L, Huang J, Zhao ZH, et al. The Skp1-like protein SSK₁ is required for cross-pollen compatibility inS-RNase-based self-incompatibility[J]. Plant J, 2010, 62(1):52-63. doi: 10.1111/j.1365-313X.2010.04123.x URL
[10]	Roldán JA, Rojas HJ, Goldraij A. Disorganization of F-actin cytoskeleton precedes vacuolar disruption in pollen tubes during the in vivo self-incompatibility response in Nicotiana alata[J]. Ann Bot, 2012, 110(4):787-795. doi: 10.1093/aob/mcs153 URL
[11]	O’Brien M, Major G, Chantha SC, et al. Isolation of S-RNase binding proteins from Solanum chacoense:identification of an SBP₁(RING finger protein)orthologue[J]. Sex Plant Reprod, 2004, 17(2):81-87. doi: 10.1007/s00497-004-0218-8 URL
[12]	McClure BA, Gray JE, Anderson MA, et al. Self-incompatibility in Nicotiana alata involves degradation of pollen rRNA[J]. Nature, 1990, 347(6295):757-760. doi: 10.1038/347757a0 URL
[13]	Xu C, Li MF, Wu JK, et al. Identification of a canonical scf(slf)complex involved In S-RNase-based Self-incompatibility Of Pyrus(rosaceae)[J]. Plant Mol Biol, 2013, 81(3):245-257. doi: 10.1007/s11103-012-9995-x URL
[14]	曾斌, 余镇藩, 马鑫鑫, 等. 基于S-核酸酶的自交不亲和cullin1基因的研究进展[J]. 分子植物育种, 2019, 17(2):466-470.
	Zeng B, Yu ZF, Ma XX, et al. Research progress of self-incompatibility Cullin1 gene based on S-RNase[J]. Mol Plant Breed, 2019, 17(2):466-470.
[15]	Zheng N, Schulman BA, Song LZ, et al. Structure of the Cul1-Rbx1-Skp1-F boxSkp2 SCF ubiquitin ligase complex[J]. Nature, 2002, 416(6882):703-709. doi: 10.1038/416703a URL
[16]	Farras R. SKP₁-SnRK protein kinase interactions mediate proteasomal binding of a plant SCF ubiquitin ligase[J]. EMBO J, 2001, 20(11):2742-2756. doi: 10.1093/emboj/20.11.2742 URL
[17]	秘彩莉, 刘旭, 张学勇. F-box蛋白质在植物生长发育中的功能[J]. 遗传, 2006, 28(10):1337-1342.
	Bi CL, Liu X, Zhang XY. The function of F-box protein in plant growth and development[J]. Hereditas, 2006, 28(10):1337-1342.
[18]	Yu H, Zhang Y, Moss BL, et al. Untethering the TIR1 auxin receptor from the SCF complex increases its stability and inhibits auxin response[J]. Nat Plants, 2015, 1:14030. doi: 10.1038/nplants.2014.30 URL
[19]	Qiao H, Wang F, Zhao L, et al. The F-box protein AhSLF-S2 controls the pollen function of S-RNase-based self-incompatibility[J]. Plant Cell, 2004, 16(9):2307-2322. doi: 10.1105/tpc.104.024919 URL
[20]	Entani T, Takayama S, Iwano M, et al. Relationship between polyploidy and pollen self-incompatibility phenotype in Petunia hybrida Vilm[J]. Biosci Biotechnol Biochem, 1999, 63(11):1882-1888. doi: 10.1271/bbb.63.1882 URL
[21]	Ushijima K, Sassa H, Dandekar AM, et al. Structural and transcriptional analysis of the self-incompatibility locus of almond:identification of a pollen-expressed F-box gene with haplotype-specific polymorphism[J]. Plant Cell, 2003, 15(3):771-781. doi: 10.1105/tpc.009290 URL