Skip to main content

Advertisement

SpringerLink
Log in

CgSL2, an S-like RNase gene in ‘Zigui shatian’ pummelo (Citrus grandis Osbeck), is involved in ovary senescence

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

‘Zigui shatian’ pummelo (Citrus grandis Osbeck) is one nature mutant from ‘Shatian’ pummelo, which showed self-compatibility, because self-pollen tubes were not arrested in the style, moreover abnormal post-zygotic development in ovary caused seed abortion in the cultivar. Herein we constructed a cDNA library from flowers of ‘Zigui shatian’ pummelo and identified one RNase gene fragment. The full length of cDNA sequence of this gene, with an open reading frame of 834 bp, was isolated by 5′-RACE method. The gene, named as CgSL2, contained five conserved regions and two histidine residues essential for RNase activity. Phylogenetic analysis indicated that CgSL2 was mostly similar to AhSL28, an S-like RNase from Antirrhinum. Southern hybridization verified CgSL2 existed in the genome as multiple copies. qRT-PCR and RT-PCR analysis showed that the expression of CgSL2 was not tissue-specific. The expression of CgSL2 was down-regulated during senescence of stem, petal, style and stamen, whereas up-regulated during ovary senescence. Further in situ hybridization of CgSL2 in the ovary during the balloon stage to anthesis stage also showed that it dramatically increased in mature flower, consistent with qRT-PCR and RT-PCR results. These findings suggested that CgSL2 might play an important role during ovary senescence.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

SI:

Self-incompatibility

qRT-PCR:

Quantitative Real-Time Polymerase Chain Reaction

References

  1. Green PJ (1994) The ribonucleases of higher plants. Annu Rev Plant Physiol Plant Mol Biol 45:421–445

    Article  CAS  Google Scholar 

  2. Dodds PN, Clarke AE, Newbigin E (1996) A molecular perspective on pollination in flowering plants. Cell 85:141–144

    Article  CAS  PubMed  Google Scholar 

  3. Kao TH, McCubbin AG (1996) How flowering plants discriminate between self and non-self pollen to prevent inbreeding. Proc Natl Acad Sci USA 93:12059–12065

    Article  CAS  PubMed  Google Scholar 

  4. Lee HS, Huang S, Kao T (1994) S proteins control rejection of incompatible pollen in Petunia inflata. Nature 367:560–563

    Article  CAS  PubMed  Google Scholar 

  5. McClure BA, Haring V, Ebert PR, Anderson MA, Simpson RJ, Sakiyama F, Clarke AE (1989) Style self-incompatibility gene products of Nicotiana alata are ribonucleases. Nature 342:955–957

    Article  CAS  PubMed  Google Scholar 

  6. Murfett J, Atherton TL, Mou B, Gasser CS, McClure BA (1994) S-RNase expressed in transgenic Nicotiana causes S-allele specific pollen rejection. Nature 367:563–566

    Article  CAS  PubMed  Google Scholar 

  7. Anderson MA, McFadden GI, Bernatzky R, Atkinson A, Orpin T, Dedman H, Tregear G, Fernley R, Clarke AE (1989) Sequence variability of three alleles of the self-incompatibility gene of Nicotiana alata. Plant Cell 1(5):483–491

    Article  CAS  PubMed  Google Scholar 

  8. Taylor CB, Bariola PA, DelCardayré SB, Raines RT, Green PJ (1993) RNS2: a senescence-associated RNase of Arabidopsis that diverged from the S-RNases before speciation. Proc Natl Acad Sci USA 90:5118–5122

    Article  CAS  PubMed  Google Scholar 

  9. Lee HS, Singh A, Kao T (1992) RNase X2, a pistil-specific ribonuclease from Petunia inflata, shares sequence similarity with solanaceous S proteins. Plant Mol Biol 20:1131–1141

    Article  CAS  PubMed  Google Scholar 

  10. Gausing K (2000) A barley gene (rsh1) encoding a ribonuclease S-like homologue specifically expressed in young light-grown leaves. Planta 210:574–579

    Article  CAS  PubMed  Google Scholar 

  11. Wei JY, Li AM, Li Y, Wang J, Liu XB, Liu LS, Xu ZF (2006) Cloning and characterization of an RNase-related protein gene preferentially expressed in rice stems. Biosci Biotechnol Biochem 70:1041–1045

    Article  CAS  PubMed  Google Scholar 

  12. Taylor CB, Green PJ (1991) Genes with homology to fungal and S-gene RNases are expressed in Arabidopsis thaliana. Plant Physiol 96:980–984

    Article  CAS  PubMed  Google Scholar 

  13. Jost W, Bak H, Glund K, Terpstra P, Beintema JJ (1991) Amino acid sequence of an extracellular, phosphate-starvation induced ribonuclease from cultured tomato (Lycopersicon esculentum) cells. Eur J Biochem 198:1–6

    Article  CAS  PubMed  Google Scholar 

  14. Dodds PN, Clarke AE, Newbigin E (1996) Molecular characterization of an S-like RNase of Nicotiana alata that is induced by phosphate starvation. Plant Mol Biol 31:227–238

    Article  CAS  PubMed  Google Scholar 

  15. Van Nerum I, Certal AC, Oliveira MM, Keulemans J, Broothaerts W (2000) PD1, an S-like RNase gene from a self-incompatible cultivar of almond. Plant Cell Rep 19:1108–1114

    Article  Google Scholar 

  16. Liang L, Lai Z, Ma W, Zhang Y, Xue Y (2002) AhSL28, a senescence- and phosphate starvation-induced S-like RNase gene in Antirrhinum. Biochim Biophys Acta 1579:64–71

    CAS  PubMed  Google Scholar 

  17. Shimizu T, Inoue T, Shiraishi H (2001) A senescence-associated S-like RNase in the multicellular green alga Volvox carteri. Gene 274:227–235

    Article  CAS  PubMed  Google Scholar 

  18. Perez-Amador MA, Abler ML, De Rocher EJ, Thompson DM, van Hoof A, LeBrasseur ND, Lers A, Green PJ (2000) Identification of BFN1, a bifunctional nuclease induced during leaf and stem senescence in Arabidopsis. Plant Physiol 122:169–179

    Article  CAS  PubMed  Google Scholar 

  19. Lers A, Khalchitski A, Lomaniec E, Burd S, Green PJ (1998) Senescence-induced RNases in tomato. Plant Mol Biol 36:439–449

    Article  CAS  PubMed  Google Scholar 

  20. Lers A, Lomaniec E, Burd S, Khalchitski A (2001) The characterization of LeNUC1, a nuclease associated with leaf senescence in tomato. Physiol Plant 112:176–182

    Article  CAS  PubMed  Google Scholar 

  21. Lers A, Sonego L, Green PJ, Burd S (2006) Suppression of LX ribonuclease in tomato results in a delay in leaf senescence and abscission. Plant Physiol 142:710–721

    Article  CAS  PubMed  Google Scholar 

  22. Langston BL, Bai S, Jones ML (2005) Increases in DNA fragmentation and induction of a senescence-specific nuclease are delayed during the senescence of ethylene-insensitive (etr1-1) transgenic petunias. J Exp Bot 56:15–23

    Article  CAS  PubMed  Google Scholar 

  23. Xu Y, Hanson MR (2000) Programmed cell death during pollination induced petal senescence in petunia. Plant Physiol 122:1323–1333

    Article  CAS  PubMed  Google Scholar 

  24. Panavas T, LeVangie R, Mistler J, Reid PD, Rubinstein B (2000) Activities of nucleases in senescing daylily petals. Plant Physiol Biochem 38:837–843

    Article  CAS  Google Scholar 

  25. Kock M, Loffier A, Abel S, Glund K (1995) Structural and regulatory properties of a family of phosphate starvation induced ribonucleases from tomato. Plant Mol Biol 27:477–485

    Article  CAS  PubMed  Google Scholar 

  26. Certal AC, Almeida BR, Boskovic R, Oliveira MM, Feijó JA (2002) Structural and molecular analysis of self-incompatibility almond (Prunus dulcis). Sex Plant Reprod 15:13–20

    Article  CAS  Google Scholar 

  27. Wiersma PA, Wu Z, Zhou L, Hampson C, Kappel F (2001) Identification of new self-incompatibility alleles in sweet cherry (Prunus avium L.) and clarification of incompatibility groups by PCR and sequencing analysis. Theor Appl Genet 102:700–708

    Article  CAS  Google Scholar 

  28. Yaegaki H, Shimada T, Moriguchi T, Hayama H, Haji T, Yamaguchi M (2001) Molecular characterization of S-RNase genes and S-genotypes in the Japanese apricot (Prunus mume Sieb. et Zucc.). Sex Plant Reprod 13:251–257

    Article  CAS  Google Scholar 

  29. Certal AC, Sanchez AM, Kokko H, Broothaerts W, Oliveira MM, Feijó JA (1999) S-RNases in apple are expressed in the pistil along the pollen tube growth path. Sex Plant Reprod 12:94–98

    Article  CAS  Google Scholar 

  30. Feng JR, Chen XS, Yuan ZH, Zhang LJ, Ci ZJ, Liu XL, Zhang CY (2009) Primary molecular features of self-incompatible and self-compatible F(1) seedling from apricot (Prunus armeniaca L.) Katy x Xinshiji. Mol Biol Rep 36(2):263–272

    Article  CAS  PubMed  Google Scholar 

  31. Feng JR, Chen XS, Wu Y, Liu W, Liang Q, Zhang LJ (2006) Detection and transcript expression of S-RNase gene associated with self-incompatibility in apricot (Prunus armeniaca L.). Mol Biol Rep 33(3):215–221

    Article  CAS  PubMed  Google Scholar 

  32. Soost RK (1969) The incompatibility gene system in citrus. Proc First Int Citrus Symp 1:189–190

    Google Scholar 

  33. Liu YZ, Liu Q, Tao NG, Deng XX (2006) Efficient isolation of RNA from fruit peel and pulp of ripening navel orange (Citrus sinesis Osbeck). J Huazhong Agric Univ 25(3):300–304

    CAS  Google Scholar 

  34. Bendtsen JD, Nielsen H, Heijne GV, Brunak S (2004) Improve prediction of signal peptides: signalP 3.0. J Mol Biol 340:783–795

    Article  PubMed  Google Scholar 

  35. Cheng YJ, Guo WW, Yi HL, Pang XM, Deng XX (2003) An efficient protocol for genomic DNA extraction from Citrus species. Plant Mol Biol Rep 21:177a–177g

    Article  Google Scholar 

  36. Hu ZY, Tong Z, Wei H, Yi HL, Deng XX (2006) Mitochondrial gene expression in stamens is differentially regulated during male gametogenesis in Citrus unshiu. J Hortic Sci Biotechnol 81:565–569

    CAS  Google Scholar 

  37. Wang P, Xue YB, Lv LX (2002) Cloning and analysis of an S-like RNase gene CgSL1 from self-incompatibility cultivars in Citrus grandis Osbeck. J Wuhan Bot Res 20(2):83–88 (in Chinese with English summary)

    Google Scholar 

  38. Van Damme EJM, Hao Q, Barre A, Rouge P, Van Leuven F, Peumans WJ (2000) Major protein of resting rhizomes of Calystegia sepium (hedge bindweed) closely resembles plant RNases but has no enzymatic activity. Plant Physiol 122:433–445

    Article  PubMed  Google Scholar 

  39. Ioerger TR, Gohlke JR, Xu B, Kao TH (1991) Primary structural features of the self-incompatibility protein in Solanaceae. Sex Plant Reprod 4:81–87

    Article  Google Scholar 

  40. Huang S, Lee HS, Karunanandaa B, Kao TH (1994) Ribonuclease activity of Petunia inflata S proteins is essential for rejection of self-pollen. Plant Cell 6:1021–1028

    Article  CAS  PubMed  Google Scholar 

  41. Royo J, Kunz C, Kowyama Y, Anderson M, Clarke AE, Newbigin E (1994) Loss of a histidine residue at the active site of S-locus ribonuclease is associated with self-compatibility in Lycopersicon peruvianum. Proc Natl Acad Sci USA 91:6511–6514

    Article  CAS  PubMed  Google Scholar 

  42. Irie M, Ohgi K, Iwama M, Koizumi M, Sasayama E, Harada K, Yano Y, Udagawa J, Kawasaki M (1997) Role of histidine 46 in the hydrolysis and the reverse transphosphorylation reaction of RNase Rh from Rhizopus niveus. J Biochem Tokyo 121:849–853

    CAS  PubMed  Google Scholar 

  43. Richman AD, Broothaerts W, Kohn JR (1997) Self-incompatibility RNases from three plant families: homology or convergence? Am J Bot 84:912–917

    Article  CAS  Google Scholar 

  44. Ushijima K, Sassa H, Tao R, Yamane H, Dandekar AM, Gradziel TM, Hirano H (1998) Cloning and characterization of cDNAs encoding S-RNAses in almond (Prunus dulcis): primary structural features and sequence diversity of the RNAses in the Rosaceae. Mol Gen Genet 260:261–268

    Article  CAS  PubMed  Google Scholar 

  45. Lu X, Zhou W, Gao F (2009) Cloning, characterization and localization of CHS gene from blood orange, Citrus sinensis (L.) Osbeck cv. Ruby. Mol Biol Rep 36(7):1983–1990

    Article  CAS  PubMed  Google Scholar 

  46. Chai LJ, Biswas MK, Ge XX, Deng XX (2010) Isolation, characterization, and expression analysis of an SKP1-like gene from ‘Shatian’ Pummelo (Citrus grandis Osbeck). Plant Mol Biol Rep. doi:10.1007/s11105-010-0184-2

  47. Hua ZH, Kao TH (2006) Identification and characterization of components of a putative Petunia S-locus F-box-containing E3 ligase complex involved in S-RNase-based self-incompatibility. Plant Cell 18:2531–2553

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This research was supported by Natural Science Foundation of China (NSFC, No. 30830078 and NSFC, No. 30921002). We thank Pro. Wenwu Guo and Mr. Zhiyong Pan for valuable suggestions.

Author information

Authors and Affiliations

  1. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, 430070, China

    Lijun Chai, Xiaoxia Ge, Qiang Xu & Xiuxin Deng

Authors
  1. Lijun Chai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaoxia Ge
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qiang Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xiuxin Deng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xiuxin Deng.

Additional information

The novel nucleotide sequence data published here has been deposited in the EMBL/DDBJ/GenBank databases under accession number Fj917371.

Electronic supplementary material

Below are the links to the electronic supplementary material.

Supplementary material 1 (DOC 71 kb)

Fig. S1

Isolation of the full-length cDNA of CgSL2 gene in the flower of ‘Zigui shatian’ pummelo. 1. PCR product to confirm the open reading frame of CgSL2 gene. 2. PCR product of 5’ RACE. (TIFF 1012 kb)

Fig. S2

Nucleotide sequence of CgSL2 cDNA from ‘Zigui Shatian’ pummelo and its deduced amino acid sequence. The start and stop codons are in bold. Signal peptide of 27 amino acid residues are boxed by solid line. The predicted N-glycosylation site is dotted boxed. Two long-term arrows indicated primer position of Real-time PCR. (TIFF 310 kb)

Fig. S3

Sequence alignment of CgSL2 with other RNases. Multiple sequence alignment was performed using the Clustal W program. The Genbank Accession No. of aligned sequences are as follows: Arabidopsis RNS2 from Arabidopsis thaliana (NP_030524); Antirrhinum S-like RNase28 from Antirrhinum hispanicum subsp. Mollissimum (CAC50874); Maize RNase from Zea mays (ACG36234); Rice RNase from Oryza sativa Japonica Group (BAB19805); Prunus S1-RNase from Prunus avium (BAA83479). Darkest shading indicates residues that are identical in all sequences; lighter shading denotes residues that are identical in most of sequences or functionally identical in all of them. Conserved regions C1-C5 are boxed. The two histidine residues involved in active sites of RNases are indicated by arrows. Asterisk indicates conserved cysteine residues. (TIFF 2360 kb)

Fig. S4

A phylogenetic tree of plant RNase derived from the alignment of 16 S-like RNases and 28 S-RNase using the Neighbor-joining method in MEGA 4.0. The arrow indicates the position of CgSL2. The plant S-RNases are derived from Solanaceae, Scrophulariaceae and Roasaceae. Lineages with the numbers of the introns are also indicated. (TIFF 2081 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chai, L., Ge, X., Xu, Q. et al. CgSL2, an S-like RNase gene in ‘Zigui shatian’ pummelo (Citrus grandis Osbeck), is involved in ovary senescence. Mol Biol Rep 38, 1–8 (2011). https://doi.org/10.1007/s11033-010-0070-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-010-0070-x

Keywords

Search

Navigation