Skip to main content
SpringerLink
Log in

Identification and validation of genetic locus Rfk1 for wheat fertility restoration in the presence of Aegilops kotschyi cytoplasm

  • Original Article
  • Published:
Theoretical and Applied Genetics Aims and scope Submit manuscript

Abstract

Key message

Major fertility restorer locus for Aegilops kotschyi cytoplasm in wheat, Rfk1 , was mapped to chromosome arm 1BS. Most likely candidate gene is TraesCS1B02G197400LC , which is predicted to encode a pectinesterase/pectinesterase inhibitor.

Abstract

Cytoplasmic male sterility (CMS) is widely used for heterosis and hybrid seed production in wheat. Genes related to male fertility restoration in the presence of Aegilops kotschyi cytoplasm have been reported, but the fertility restoration-associated gene loci have not been investigated systematically. In this study, a BC1F1 population derived from a backcross between KTP116A, its maintainer line TP116B, and its restorer line LK783 was employed to map fertility restoration by bulked segregant RNA-Seq (BSR-Seq). A major fertility allele restorer locus for Ae. kotschyi cytoplasm in wheat, Rfk1, was mapped to chromosome arm 1BS, and it was contributed by LK783. Morphological and cytological studies showed that male fertility restoration occurred mainly after the late uninucleate stage. Based on simple sequence repeat and single-nucleotide polymorphism genotyping, the gene locus was located between Xnwafu_6 and Xbarc137 on chromosome arm 1BS. To further isolate the specific region, six Kompetitive allele-specific polymerase chain reaction markers derived from BSR-Seq were developed to delimit Rfk1 within physical intervals of 26.0 Mb. After searching for differentially expressed genes within the candidate interval in the anthers and sequencing analysis, TraesCS1B02G197400LC was identified as a candidate gene for Rfk1 and it was predicted to encode a pectinesterase/pectinesterase inhibitor. Expression analysis also confirmed that it was specifically expressed in the anthers, and its expression level was higher in fertile lines compared with sterile lines. Thus, TraesCS1B02G197400LC was identified as the most likely candidate gene for Rfk1, thereby providing insights into the fertility restoration mechanism for K-type CMS in wheat.

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
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

CMS:

Cytoplasmic male sterility

KASP:

Kompetitive allele-specific polymerase chain reaction

SNP:

Single-nucleotide polymorphism

BSR-seq:

Bulked segregant RNA-Seq

qRT-PCR:

Quantitative real-time PCR

SSR:

Simple sequence repeat

TCMS-K:

Thermo-sensitive cytoplasmic male sterility with Aegilops kotschyi cytoplasm

References

  • Balaji Suresh P, Srikanth B, Hemanth Kishore V, Subhakara Rao I, Vemireddy LR, Dharika N, Sundaram RM, Ramesha MS, Sambasiva Rao KRS, Viraktamath BC, Neeraja CN (2012) Fine mapping of Rf3 and Rf4 fertility restorer loci of WA-CMS of rice (Oryza sativa L.) and validation of the developed marker system for identification of restorer lines. Euphytica 187:421–435

    Article  CAS  Google Scholar 

  • Bentolila S, Alfonso AA, Hanson MR (2002) A pentatricopeptide repeat-containing gene restores fertility to cytoplasmic male-sterile plants. Pro Natl Acad Sci USA 99:10887–10892

    Article  CAS  Google Scholar 

  • Börner A, Korzun V, Polley A, Malyshev S, Melz G (1998) Genetics and molecular mapping of a male fertility restoration locus (Rfg1) in rye (Secale cereale L.). Theor Appl Genet 97:99–102

    Article  Google Scholar 

  • Bosch M, Hepler P (2005) Pectin methylesterases and pectin dynamics in pollen tubes. Plant Cell 17:3219–3226

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Brown G, Formanova N, Jin H, Wargachuk R, Dendy C, Patil P, Laforest M, Zhang J, Cheung W, Landry B (2003) The radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats. Plant J 35:262–272

    Article  CAS  PubMed  Google Scholar 

  • Cao S, Guo X, Liu D, Chang X, Zhang A (2004) Preliminary gene-mapping of photoperiod-temperature sensitive genic male sterility in wheat (Triticum aestivum L.). Acta Genet Sin 31:293–298

    CAS  PubMed  Google Scholar 

  • Chai S, Yun H, Wang H, Li A, Cao P, Li D (1999) A study on fertility-restoration of CMS systems with Aegilops kotschyi or Ae. varibilis cytoplasm and 1BL-1RS chromosome in common wheat. Acta Bot Boreali-occident Sin 19:654–658

    CAS  Google Scholar 

  • Chen L, Liu Y (2014) Male sterility and fertility restoration in crops. Annu Rev Plant Biol 65:579–606

    Article  CAS  PubMed  Google Scholar 

  • Dong D, Li Z, Yuan F, Zhu S, Chen P, Yu W, Yang Q, Fu X, Yu X, Li B, Zhu D (2012) Inheritance and fine mapping of a restorer-of-fertility (Rf) gene for the cytoplasmic male sterility in soybean. Plant Sci 188–189:36–40

    Article  PubMed  Google Scholar 

  • Du H, Zhu J, Su H, Huang M, Wang H, Ding S, Zhang B, Luo A, Wei S, Tian X, Xu Y (2017) Bulked Segregant RNA-seq reveals differential expression and SNPs of candidate genes associated with waterlogging tolerance in Maize. Front Plant Sci 8:1022

    Article  PubMed  PubMed Central  Google Scholar 

  • Fei H, Sawhney VK (2001) Ultrastructural characterization of male sterile33 (ms33) mutant in Arabidopsis affected in pollen desiccation and maturation. Can J Bot 79:118–129

    Google Scholar 

  • Fujii S, Toriyam K (2009) Suppressed expression of retrograde-regulated male sterility restores pollen fertility in cytoplasmic male sterile rice plant. Proc Natl Acad Sci USA 106:9513–9518

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gupta PK, Balyan HS, Gahlaut V, Saripalli G, Pal B, Basnet BR, Joshi AK (2019) Hybrid wheat: past, present and future. Theor Appl Genet 132:2463–2483

    Article  PubMed  Google Scholar 

  • Hanson MR, Bentolila S (2004) Interactions of mitochondrial and nuclear genes that affect male gametophyte development. Plant Cell 16(Suppl):S154–S169

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hu J, Wang K, Huang W, Liu G, Gao Y, Wang J, Huang Q, Ji Y, Qin X, Wan L, Zhu R, Li S, Yang D, Zhu Y (2012) The rice pentatricopeptide repeat protein RF5 restores fertility in Hong-Lian cytoplasmic male-sterile lines via a complex with the glycine-rich protein GRP162. Plant Cell 24:109–122

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Itabashi E, Iwato N, Fujii S, Kazama T, Toriyama K (2011) The fertility restorer gene, Rf2, for Lead Rice-type cytoplasmic male sterility of rice encodes a mitochondrial glycine-rich protein. Plant J 65:359–367

    Article  CAS  PubMed  Google Scholar 

  • IWGSC (2018) Shifting the limits in wheat research and breeding using a fully annotated reference genome. Science 361:7191

    Article  Google Scholar 

  • Jordan D, Mace ES, Henzell R, Klein P, Klein R (2010) Molecular mapping and candidate gene identification of the Rf2 gene for pollen fertility restoration in sorghum [Sorghum bicolor (L.) Moench]. Theor Appl Genet 120:1279–1287

    Article  CAS  PubMed  Google Scholar 

  • Kazama T, Toriyama K (2014) A fertility restorer gene, Rf4, widely used for hybrid rice breeding encodes a pentatricopeptide repeat protein. Rice 7:28

    Article  PubMed  PubMed Central  Google Scholar 

  • Kihara H (1951) Substitution of nucleus and its effects on genome manifestations. Cytologia 16:177–193

    Article  Google Scholar 

  • Kitazaki K, Arakawa T, Matsunaga M, Yui-Kurino R, Matsuhira H, Mikami T, Kubo T (2015) Post-translational mechanisms are associated with fertility restoration of cytoplasmic male sterility in sugar beet (Beta vulgaris). Plant J 83:290–299

    Article  CAS  PubMed  Google Scholar 

  • Klein RR, Klein PE, Mullet JE, Minx P, Rooney WL, Schertz KF (2005) Fertility restorer locus Rf1 of sorghum (Sorghum bicolor L.) encodes a pentatricopeptide repeat protein not present in the colinear region of rice chromosome 12. Theor Appl Genet 111:994–1012

    Article  CAS  PubMed  Google Scholar 

  • Li S, Liu Z, Jia Y, Ye J, Yang X, Zhang L, Song X (2019) Analysis of metabolic pathways related to fertility restoration and identification of fertility candidate genes associated with Aegilops kotschyi cytoplasm in wheat (Triticum aestivum L.). BMC Plant Biol 19:252

    Article  PubMed  PubMed Central  Google Scholar 

  • Li W, Liu Z, Meng C, Jia Y, Song X (2019) Identification of genes related to the regulation of anther and pollen development in mu-type cytoplasmic male sterile wheat (Triticum aestivum) by transcriptome analysis. Crop Pasture Sci 70:306–317

    Article  CAS  Google Scholar 

  • Li H, Zhang G, Zhang L, Niu N (2008) Genetics analysis and SSR markers on fertility restorer gene of male sterility of wheat with Aegilops kotschyi cytoplasm. J JNWAFU 36:65–70

    Google Scholar 

  • Liang FS, Wang B (2003) Heredity and gene mapping of male sterility in wheat. Yi Chuan 25:461–465

    CAS  PubMed  Google Scholar 

  • Liu F, Cui X, Horner HT, Weiner H, Schnable PS (2001) Mitochondrial aldehyde dehydrogenase activity is required for male fertility in maize. Plant Cell 13:1063–1078

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu Z, Li S, Li W, Liu Q, Zhang L, Song X (2020) Comparative transcriptome analysis indicates that a core transcriptional network mediates isonuclear alloplasmic male sterility in wheat (Triticum aestivum L.). BMC Plant Biol 20:1–19

    Google Scholar 

  • Liu Z, Liu P, Long F, Hong P, He Q, Yang G (2012) Fine mapping and candidate gene analysis of the nuclear restorer gene Rfp for pol CMS in rapeseed (Brassica napus L.). Theor Appl Genet 125:773–779

    Article  CAS  PubMed  Google Scholar 

  • Liu B, Sun L, Gao Q, Zhang Y, Liu S (2000) Genetic analysis of male–sterility restoration in wheat with Ae.kotschyi cytoplasm. J Shandong Agri Univ 31:11–14

    Google Scholar 

  • Liu B, Sun Q, Sun L, Gao Q, Xie C, Dou B, Ni Z, Wei Y, Zhang Y (2002) RAPD and ISSR markers of fertility restoring gene for Aegilops kotschyi cytoplasmic male sterility in wheat. Acta Bot Sin 44:446–450

    CAS  Google Scholar 

  • Liu C, Zhou Q, Dong L, Wang H, Liu F, Weng J, Li X, Xie C (2016) Genetic architecture of the maize kernel row number revealed by combining QTL mapping using a high-density genetic map and bulked segregant RNA sequencing. BMC Genom 17:915

    Article  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Mukai Y, Tsunewaki K (1979) Basic studies on hybrid wheat breeding: VIII. a new male sterility-fertility restoration system in common wheat utilizing the cytoplasms of Aegilops kotschiyi and Ae. variabilis. Theor Appl Genet 54:153–160

    Article  CAS  PubMed  Google Scholar 

  • Sajer O, Schirmak U, Hamrit S, Horn R (2020) Mapping of the new fertility restorer gene Rf-PET2 close to Rf1 on linkage group 13 in sunflower (Helianthus annuus L.). Genes 11:269

    Article  CAS  PubMed Central  Google Scholar 

  • Song X, Qian H, Zhang L (2014) Cytogenetic analysis of cytoplasmic male sterility in wheat line KTP116A and molecular mapping of two thermo-sensitive restoration genes. Euphytica 196:129–136

    Article  CAS  Google Scholar 

  • Tsunewaki K (2015) Fine mapping of the first multi-fertility-restoring gene, Rfmulti, of wheat for three Aegilops plasmons, using 1BS-1RS recombinant lines. Theor Appl Genet 128:723–732

    Article  CAS  PubMed  Google Scholar 

  • Van Ooijen JW (2006) JoinMap 4, Software for the calculation of genetic linkage maps in experimental populations. Kyazma BV, Wageningen

    Google Scholar 

  • Wang N, Liu ZY, Zhang Y, Li CY, Feng H (2017) Identification and fine mapping of a stay-green gene (Brnye1) in pakchoi (Brassica campestris L. ssp. chinensis). Theor Appl Genet 131:673–684

    Article  PubMed  Google Scholar 

  • Wang Z, Zou Y, Li X, Zhang Q, Chen L, Wu H, Su D, Chen Y, Guo J, Da L, Long Y, Zhang Y, Liu Y (2006) Cytoplasmic male sterility of rice with boro II cytoplasm is caused by a cytotoxic peptide and is restored by two related PPR motif genes via distinct modes of mRNA silencing. Plant Cell 18:676–687

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xu D, Qu S, Tucker MR, Zhang D, Liang W, Shi J (2019) Ostkpr1 functions in anther cuticle development and pollen wall formation in rice. BMC Plant Biol 19:104

    Article  PubMed  PubMed Central  Google Scholar 

  • Yao Y, Zhang G, Liu H, Wang J (2002) Studies on correlation between intine and K-type cytoplasmic male sterility in wheat. Acta Bot Boreali-occident Sin 22:333–337

    CAS  Google Scholar 

  • Zhan K, Cheng J, Cui D, Fan L, Xu H, Song Y (2006) Genetic analysis of fertility restoring genes for male sterile line with Ae. kotschyi cytoplasm in wheat [Triticum aestivum (L.) Thell.]. Acta Agron Sin 32:873–877

    Google Scholar 

  • Zhang Z, Wang Y, Zheng Y (2006) AFLP and PCR-based markers linked to Rf3, a fertility restorer gene for S cytoplasmic male sterility in maize. Mol Genet Genom 276:162–169

    Article  CAS  Google Scholar 

  • Zhao C, Zhao G, Geng Z, Wang Z, Wang K, Liu S, Zhang H, Guo B, Geng J (2018) Physical mapping and candidate gene prediction of fertility restorer gene of cytoplasmic male sterility in cotton. BMC Genom 19:6

    Article  Google Scholar 

  • Zhou Y, He Z, Zhang G, Xia L, Chen X, Gao Y, Jing Z, Yu G (2004) Utilization of 1BL/1RS translocation in wheat breeding in China. Acta Agron Sin 30:531–535

    CAS  Google Scholar 

Download references

Acknowledgements

This study was funded by the National Natural Science Foundation of China (31771874). We also thank Dr. Dejun Han, State Key Laboratory of Crop Stress Biology for Arid Areas, College of Agronomy (Northwest A&F University), China, for providing the KASP genotyping platform support.

Author information

Author notes

  1. Yanru Chen and Yulin Jia have contributed equally to this work

Authors and Affiliations

  1. College of Agronomy, Northwest A&F University, Yangling, Shaanxi, China

    Yanru Chen, Yulin Jia, Fuqiang Niu, Yongfeng Wu, Jiali Ye, Xuetong Yang, Lingli Zhang & Xiyue Song

Authors
  1. Yanru Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yulin Jia
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fuqiang Niu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yongfeng Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jiali Ye
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xuetong Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Lingli Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xiyue Song
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

XS and LZ conceived and designed the study. YC and YJ performed the experiments and analyzed the data. FN, YW, JY, and XY provided advice and technical assistance. YC and YJ wrote the article with contributions from all of the authors. All of the authors read and approved the manuscript.

Corresponding authors

Correspondence to Lingli Zhang or Xiyue Song.

Ethics declarations

Conflict of interest

The authors declare no conflicts of interest.

Additional information

Communicated by Steven S. Xu.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

122_2020_3738_MOESM1_ESM.docx

Fig. S1 Percentage of SNP distributions. The figure on the left shows the percentage of total SNPs and the number of SNP with a Delta-SNP index ≥ 0.5. The outer ring on the right shows the distribution (percentages) of the total SNPs on each wheat chromosome and the inner ring shows the distribution (percentages) on each wheat chromosome with a Delta-SNP index ≥ 0.5 (DOCX 47 kb)

122_2020_3738_MOESM2_ESM.docx

Fig. S2 Partial SSR analysis of the BC1F1 population derived from KTP116A//LK783/T116B using markers Xgwm18 and Xnwafu_5 on chromosome 1BS. M, DNA marker 2000; P1, KTP116A; P2, LK783; M1, mixed pool of 20 sterile individuals with a self-setting rate of 0; M2, mixed pool of 20 fertile individuals with a self-setting rate higher than 85%. The black line denotes sterile individuals from the mapping population with a self-setting rate of 0. (DOCX 319 kb)

122_2020_3738_MOESM3_ESM.docx

Fig. S3 Expression analysis by qRT-PCR of root, stem, leaf, and anther tissues for the four candidate genes in the target interval. The results for different tissues from LK783 and KTP116A indicated no significant differences in the relative expression levels of the four genes in the anthers. The ordinates in the two graphs represent the 2–ΔΔCT values (DOCX 109 kb)

122_2020_3738_MOESM4_ESM.docx

Fig. S4 Alignment of amino acid sequences of TraesCS1B02G197400LC from LK783, KTP116A, and reference genome sequence (IWGSC_v1.1_LC_gene). Ten non-synonymous SNPs were detected and a 7-bp deletion is shown in the red box at 1,177 bp (DOCX 162 kb)

122_2020_3738_MOESM5_ESM.docx

Fig. S5 PCR and polyacrylamide gel electrophoresis analysis results for parental lines and the extremely fertility mixed pools using Xnwafu_4, Xucr_5, and Xucr_6. Marker Xnwafu_4 has polymorphisms at 224 bp between parental lines as well as mixed pools, whereas Xucr_5 and Xucr_6 linked to Rfv1 (Tsunewaki 2015) have no polymorphisms at 500 bp and 1100 bp, respectively. The arrow refers to the PCR products size of the three markers. M, DNA marker 2000; P1, KTP116A; P2, LK783; M1, mixed pool of 20 sterile individuals with a self-setting rate of 0; M2, mixed pool of 20 fertile individuals with a self-setting rate higher than 85% (DOCX 75 kb)

Supplementary file6 (XLSX 37kb)

Supplementary file7 (XLSX 33 kb)

Supplementary file8 (XLSX 13 kb)

Supplementary file9 (XLSX 83 kb)

Supplementary file10 (XLSX 37 kb)

Supplementary file11 (XLSX 563 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, Y., Jia, Y., Niu, F. et al. Identification and validation of genetic locus Rfk1 for wheat fertility restoration in the presence of Aegilops kotschyi cytoplasm. Theor Appl Genet 134, 875–885 (2021). https://doi.org/10.1007/s00122-020-03738-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00122-020-03738-2

Search

Navigation