Abstract:Phosphatidylinositol signaling system plays an important role in plant development. Phosphatidylinositol 4-phosphate 5-kinase (PIP5K) is one of the most crucial enzyme that catalyze PIP2 synthesis in this signaling system. The present study identified 19 CaPIP5K family genes in pepper (Capsicum annuum) genome database, among which 14 CaPIP5Ks were dispersedly distributed on 8 out of the 12 chromosomes and 5 CaPIP5Ks located on chromosome 0. Except 1 PIP5K domain was confirmed in every CaPIP5K proteins, 5~8 repeated MORN (membrane occupation and recognition nexus) domains were also detected in 8 CaPIP5Ks. All CaPIP5K proteins were predicted to locate in the cytoplasm. Moreover, cis-acting elements in CaPIP5K promoter region mainly responded to hormones, defense and stress, and low temperature. In addition, the expressional analysis indicated that CaPIP5K4-1 not only specifically expressed in flower instead of other tissues, but also gradually improved with the development of flower buds in fertile accessions as compared to sterile accession. This study provides a reference basis for further exploring the mechanism of CaPIP5K gene family in the development of anther and pollen in pepper.
[1] Akagi H, Nakamura A, Yokozeki-Misono Y, et al.2004. Positional cloning of the rice Rf-1 gene, a restorer of BT-type cytoplasmic male sterility that encodes a mitochondria-targeting PPR protein[J]. Theoretical and Applied Genetics, 108(8): 1449-1457. [2] Alves-Ferreira M, Wellmer F, Banhara A, et al.2007. Global expression profiling applied to the analysis of Arabidopsis stamen development[J]. Plant Physiology, 145(3): 747-762. [3] Bailey T L, Boden M, Buske F A, et al.2009. MEME Suite: Tools for motif discovery and searching[J]. Nucleic Acids Research, 37(2): W202-W208. [4] Barchenger D W, Said J I, Zhang Y, et al.2018. Genome-wide identification of chile pepper pentatricopeptide repeat domains provides insight into fertility restoration[J]. Journal of the Americam Society for Horticultural Science, 143(6): 418-429. [5] Bentolila S, Alfonso A A, Hanson M R.2002. A pentatricopeptide repeat-containing gene restores fertility to cytoplasmic male-sterile plants[J]. Proceedings of the National Academy of Sciences of the USA, 99(16): 10887-10892. [6] Bosland P W, Votava E J.2012. Peppers. In: Vegetable and Spice Capsicums. 2nd ed.[M]. CAB International, Wallingford, UK, pp. 230. [7] Brill J A, Yildirim S, Fabian L.2016. Phosphoinositide signaling in sperm development[J]. Seminars in Cell & Developmental Biology, 59: 2-9. [8] Brown G G, Formanová N, Jin H, et al.2003. The radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats[J]. The Plant Journal, 35(2): 262-272. [9] Chapman L A, Goring D R.2011. Misregulation of phosphoinositides in Arabidopsis thaliana decreases pollen hydration and maternal fertility[J]. Sex Plant Reproduction, 24(4): 319-326. [10] Chase C D.2007. Cytoplasmic male sterility: A window to the world of plant mitochondrial-nuclear interactions[J]. Trends in Genetics, 23(2): 81-90. [11] Cheng J W, Chen Y J, Hu Y F, et al.2020. Fine mapping of restorer-of-fertility gene based on high-density genetic mapping and collinearity analysis in pepper (Capsicum annuum L.)[J]. Theoretical and Applied Genetics, 133(3): 889-902. [12] Chou K C, Shen H B.2010. A new method for predicting the subcellular localization of eukaryotic proteins with both single and multiple sites: Euk-mPLoc 2.0[J]. PLOS ONE, 5(4): e9931. [13] Colombo N, Galmarini C R.2017. The use of genetic, manual and chemical methods to control pollination in vegetable hybrid seed production: A review[J]. Plant Breeding, 136(4): 287-299. [14] Cui X, Wise R P, Schnable PS.1996. The rf2 nuclear restorer gene of male-sterile T-cytoplasm maize[J]. Science, 272(5226): 1334-1336. [15] Dahan J, Mireau H.2013. The Rf and Rf-like PPR in higher plants, a fast evolving subclass of PPR genes[J]. RNA Biology, 10(9): 1469-1476. [16] Deslorie S, Gherbi H, Laloui W, et al.2003. Identification of the fertility restoration locus, Rfo, in radish, as a member of the pentatricopeptide-repeat protein family[J]. EMBO Reports, 4(6): 588-594. [17] Feng Y, Zheng Q, Song H, et al.2015. Multiple loci not only Rf3 involved in the restoration ability of pollen fertility, anther exsertion and pollen shedding to S type cytoplasmic male sterile in maize[J]. Theoretical and Applied Genetics, 128(11): 2341-2350. [18] Fujii S, Toriyama K.2009. Suppressed expression of RETROGRADE_REGULATED MALE STERILITY restores pollen fertility in cytoplasmic male sterile rice plants[J]. Proceedings of the National Academy of Sciences of the USA, 106(23): 9513-9518. [19] Fujii S, Bond C S, Small I D.2011. Selection patterns on restorer-like genes reveal a conflict between nuclear and mitochondrial genomes throughout angiosperm evolution[J]. Proceedings of the National Academy of Sciences of the USA, 108(4): 1723-1728. [20] Gulyas G, Shin Y, Kim H.2010. Altered transcript reveals an orf507 sterility-related gene in chili pepper (Capsicum annuum L.)[J]. Plant Molecular Biology Reports, 28(4): 605-612. [21] Hamamura Y, Nagahara S, Higashiyama T.2012. Double fertilization on the move[J]. Current Opinion in Plant Biology, 15(1): 70-77. [22] Hasegawa H, Noguchi J, Yamashita M, et al.2012. Phosphatidylinositol 4-phosphate 5-kinase is indispensable for mouse spermatogenesis[J]. Biology of Reproduction, 86(5): 136, 1-12. [23] Havey M J.2004. The Use of Cytoplasmic Male Sterility for Hybrid Seed Production. In: Daniell H, Chase C, editors. Molecular Biology and Biotechnology of Plant Organelles[M]. Springer, Dordrecht, pp. 623-634. [24] Hu B, Jin J, Guo A Y, et al.2015. GSDS 2.0: An upgraded gene feature visualization server[J]. Bioinformatics, 31(8): 1296-1297. [25] Hu J, Huang W, Huang Q, et al.2014. Mitochondria and cytoplasmic male sterility in plants[J]. Mitochondrion, 19: 282-288. [26] Ischebeck T, Stenzel I, Heilmann I.2008. Type B phosphatidylinositol-4-phosphate 5-kinases mediate Arabidopsis and Nicotiana tabacum pollen tube growth by regulating apical pectin secretion[J]. Plant Cell, 20(12): 3312-3330. [27] Itabashi E, Iwata N, Fujii S, et al.2011. The fertility restorer gene, Rf2, for lead rice-type cytoplasmic male sterility of rice encodes a mitochondrial glycine-rich protein[J]. The Plant Journal, 65(3): 359-367. [28] Janska H, Sarria R, Woloszynska M, et al.1998. Stoichiometric shifts in the common bean mitochondrial genome leading to male sterility and spontaneous reversion to fertility[J]. The Plant Cell, 10(7): 1163-1180. [29] Ji J J, Huang W, Yin Y X, et al.2014. Development of a SCAR marker for early identification of S-cytoplasm based on mitochondrial SRAP analysis in pepper (Capsicum annuum L.)[J]. Molecular Breeding, 33(3): 679-690. [30] Jo Y D, Ha Y, Lee J H, et al.2016. Fine mapping of restorer-of-fertility in pepper (Capsicum annuum L.) identified a candidate gene encoding a pentatricopeptide repeat (PPR)-containing protein[J]. Theoretical and Applied Genetics, 129(10): 2003-2017. [31] Kaul M L H. (eds). 1998. Male Sterility in Higher Plants[M]. Heidelberg: Springer-Verlag, Berlin, pp. 278-287. [32] Kazama T, Nakamura T, Watanbe M, et al.2008. Suppression mechanism of mitochondrial ORF79 accumulation by Rf1 protein in BT-type cytoplasmic male sterile rice[J]. The Plant Journal, 55(4): 619-628. [33] Kim D H, Kim B D.2005. Development of SCAR markers for early identification of cytoplasmic male sterility genotype in chili pepper (Capsicum annuum L.)[J]. Molecules and Cells, 20(3): 416-422. [34] Kim D H, Kang J G, Kim B D.2007. Isolation and characterization of cytoplasmic male sterility-associated orf456 gene of chili pepper (Capsicum annuum L.)[J]. Plant Molecular Biology, 63(4): 519-532. [35] Kim D S, Kim B D.2006. The organization of mitochondrial atp6 gene region in male fertile and CMS lines of pepper (Capsicum annuum L.)[J]. Current Genetics, 49(1): 59-67. [36] Kitazaki K, Arakawa T, Matsunaga M, et al.2015. Post-translational mechanisms are associated with fertility restoration of cytoplasmic male sterility in sugar beet[J]. Plant Journal, 83(2): 290-299. [37] Klein R R, Klein P E, Mullet J E, et al.2005. KF: Fertility restorer locus Rf1 of sorghum (Sorghum bicolor L.) encodes a pentatricopeptide repeat protein not present in the collinear region of rice chromosome 12[J]. Theoretical and Applied Genetics, 111(6): 994-1012. [38] Koizuka N, Imai R, Fujimoto H, et al.2003. Genetic characterization of a pentatricopeptide repeat protein gene, orf687, that restores fertility in the cytoplasmic male-sterile Kosena radish[J]. Plant Journal, 34(4): 407-415. [39] Komori T, Ohta S, Murai N, et al.2004. Map-based cloning of a fertility restorer gene, Rf-1, in rice (Oryza sativa L.)[J]. Plant Journal, 37(3): 315-325. [40] Lee J, Yoon J B, Park H G.2008. Linkage analysis between the partial restoration (pr) and the restorer-of-fertility (Rf) loci in pepper cytoplasmic male sterility[J]. Theoretical Applied Genetics, 117(3): 383-389. [41] Livak K J, Schmittgen T D.2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method[J]. Methods, 25(4): 402-408. [42] Matsuhira H, Kagami H, Kurata M, et al.2012. Unusual and typical features of a novel restorer-of-fertility gene in sugar beet (Beta vulgaris L.)[J]. Genetics, 192(4):1347-1358. [43] Min W K, Lim H, Lee Y P, et al.2008. Identification of a third haplotype of the sequence linked to the restorer-of-fertility (Rf) gene and its implications for male sterility phenotypes in peppers (Capsicum annuum L.)[J]. Molecules and Cells, 25(1): 20-29. [44] Novak F, Betlach J, Dubovsky J.1971. Cytoplasmic male sterility in sweet pepper (Capsicum annuum L.). I. Phenotype and inheritance of male sterile character[J]. Zeitschrift fur Pflanzenzuchtung, 65: 129-140. [45] Ortega F A, Barchenger D W, Wei B Q, et al.2020. Development of a genotype-specific molecular marker associated with restoration-of-fertility (Rf) in chile pepper (Capsicum annuum)[J]. Euphytica, 216(3): 43 [46] Whitley P, Hinz S, Doughty J.2009. Arabidopsis FAB1/PIKfyve proteins are essential for development of viable pollen[J]. Plant Physiology, 151(4): 1812-1822. [47] Peterson P A.1958. Cytoplasmically inherited male sterility in Capsicum[J]. The American Naturalist, 92(863): 111-119. [48] Saitou N, Nei M.1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees[J]. Molecular Biology and Evolution, 4(4): 406-425. [49] Sousa E, Kost B, Malho R.2008. Arabidopsis phosphatidylinositol-4-monophosphate 5-kinase 4 regulates pollen tube growth and polarity by modulating membrane recycling[J]. The Plant Cell, 20(11): 3050-3064. [50] Tamura K, Stecher G, Peterson D, et al.2013. MEGA6: Molecular evolutionary genetics analysis version 6.0[J]. Molecular Biology and Evolution, 30(12): 2725-2729. [51] Tsunewaki K.2015. Fine mapping of the first multi-fertility-restoring gene, Rfmulti, of wheat for three Aegilops plasmons, using 1BS-1RS recombinant lines[J]. Theoretical and Applied Genetics, 128(4): 723-732. [52] Tuteja R, Saxena R K, Davila J, et al.2013. Cytoplasmic male sterility-associated chimeric open reading frames identified by mitochondrial genome sequencing of four cajanus genotypes[J]. DNA Research, 20(5): 485-495. [53] Ugalde J M, Rodriguez-Furlán C, De R R, et al.2016. Phosphatidylinositol 4-phosphate 5-kinases 1 and 2 are involved inthe regulation of vacuole morphology during Arabidopsis thaliana pollen development[J]. Plant Science, 250: 10-19. [54] Uyttewaal M, Arnal N, Quadrado M, et al.2008. Characterization of Raphanus sativus pentatricopeptide repeat proteins encoded by a fertility restorer locus for Ogura cytoplasmic male sterility[J]. The Plant Cell, 20(12): 3331-3345. [55] Voorrips R E.2002. MapChart: Software for the graphical presentation of linkage maps and QTLs[J]. The Journal of Heredity, 93(1): 77-78. [56] Wang Z, Zou Y, Li X, et al.2006. Cytoplasmic male sterility of rice with boroⅡcytoplasm is caused by a cytotoxic peptide and is restored by two related PPR motif genes via distinct modes of mRNA silencing[J]. The Plant Cell, 18(3): 676-687. [57] Wang Z W, Wang C D, Mei S Y, et al.2015. An insertion-deletion at a pentatricopeptide repeat locus linked to fertility transition to cytoplasmic male sterility in radish (Raphanus sativus L.)[J]. Molecular Breeding, 35(4): 1-5. [58] Wei B Q, Bosland P W, Zhang Z H, et al.2020. A predicted NEDD8 conjugating enzyme gene identified as a Capsicum candidate Rf gene using bulk segregant RNA sequencing[J]. Horticulture Research, 7(1): 210. [59] Wei B Q, Wang L L, Bosland P W, et al.2020. A joint segregation analysis of the inheritance of fertility restoration for cytoplasmic male sterility in pepper[J]. Journal of American Society for Horticultural Science, 145(1): 3-11. [60] Wei B Q, Wang L L, Bosland P W, et al.2019. Comparative transcriptional analysis of Capsicum flower buds between a sterile flower pool and a restorer flower pool provides insight into the regulation of fertility restoration[J]. BMC Genomics, 20(1): 837. [61] Xu N, Gao X Q, Zhao X Y, et al.2011. Arabidopsis AtVPS15 is essential for pollen development and germination through modulating phosphatidylinositol 3-phosphate formation[J]. Plant Molecular Biology, 77(3): 251-260. [62] Xue H W, Chen X, Mei Y.2009. Function and regulation of phospholipid signalling in plants[J]. Biochemical Journal, 421(2): 145-156. [63] Zhang H G, Che J L, Ge Y S, er al.2017a. Ability of Rf5 and Rf6 to restore fertility of chinsurah boroⅡ-type cytoplasmic male sterile Oryza sativa (ssp. japonica) lines[J]. Rice, 10(1): 2. [64] Zhang H M, Wu J Q, Dai Z H, et al.2017b. Allelism analysis of Br Rfp locus in different restorer lines and map-based cloning of a fertility restorer gene, Br Rfp1, for pol CMS in Chinese cabbage (Brassica rapa L.)[J]. Theoretical and Applied Genetics, 130(3): 539-547. [65] Zhang Z H, Zhu Y S, Cao Y C, et al.2020. Fine mapping of the male fertility restoration gene CaRf032 in Capsicum annuum L.[J]. Theoretical and Applied Genetics, 133(4): 1177-1187.