Skip to main content
Log in

Comprehensive analysis of differently expressed genes and proteins in albino and green plantlets from a wheat anther culture

  • Original Paper
  • Published:
Biologia Plantarum

Abstract

The anther culture technique, although it has been widely used in wheat breeding programs, is associated with a high albinism rate that limits its application. The mechanism of albinism has not been studied explicitly at the molecular level. In this study, differently expressed genes (DEGs) and proteins (DEPs) of albino and green seedlings from a wheat anther culture were analyzed and compared using transcriptomic sequencing (RNA-Seq), two-dimensional electrophoresis, and mass spectrometry. A total of 1 892 DEGs (1 115 down-regulated and 777 up-regulated) and 43 DEPs (15 down-regulated and 28 up-regulated) that are primarily involved in photosynthetic pathways and porphyrin and chlorophyll metabolism was identified when the albino group library was used as the control. Most DEGs participated in constructing components of thylakoids and the chloroplast envelope as determined via gene ontology analysis. The expression patterns of 12 selected genes were evaluated using real-time quantitative PCR, and the results were in agreement with the RNA-Seq analysis. Our results may assist the development of new methods to decrease the albino seedling rates in wheat anther cultures.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

2-DE:

two-dimensional electrophoresis

CI:

confidence interval

DEGs:

differently expressed genes

DEPs:

differently expressed proteins

GO:

gene ontology

KEGG:

Kyoto encyclopedia of genes and genomes

MS:

mass spectrometry

PS:

photosystem

RPKM:

reads per kilo base per million reads

References

  • Andersen, S.B., Due, I.K., Olesen, A.: The response of anther culture in a genetically wide material of winter wheat (Triticum aestivum L.). — Plant Breed. 99: 181–186, 1987.

    Article  Google Scholar 

  • Andrade-Souza, V., Costa, M.G., Chen, C.X., Gmitter, F.G., Jr., Costa, M.A.: Physical location of the carotenoid biosynthesis genes Psy and beta-Lcy in Capsicum annuum (Solanaceae) using heterologous probes from Citrus sinensis (Rutaceae). — Genet. mol. Res. 10: 404–409, 2011.

    Article  CAS  PubMed  Google Scholar 

  • Ashburner, M., Ball, C.A., Blake, J.A., Botstein, D., Butler, H., Cherry, J.M., Davis, A.P., Dolinski, K., Dwight, S.S., Eppig, J.T., Harris, M.A., Hill, D.P., Issel-Tarver, L., Kasarskis, A., Lewis, S., Matese, J.C., Richardson, J.E., Ringwald, M., Rubin, G.M., Sherlock, G.: Gene ontology: tool for the unification of biology. The gene ontology consortium. — Nat. Genet. 25: 25–29, 2000.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chou, K.C., Shen, H.B.: Cell-PLoc: a package of Web servers for predicting subcellular localization of proteins in various organisms. — Nat. Protocols 3: 153–162, 2008.

    Article  CAS  PubMed  Google Scholar 

  • Cistué, L., Vallés, M.P., Echávarri, B., Sanz, J.M., Castillo, A.: Barley anther culture. — In Maluszynski, M., Kasha, K.J., Forster, B.P. Szarejko, I., (ed.): Doubled Haploid Production in Crop Plants. Pp. 29–34. Springer, Dordrecht 2003.

    Chapter  Google Scholar 

  • Daie, J.: Cytosolic fructose-1,6-bisphosphatase: a key enzyme in the sucrose biosynthetic pathway. — Photosynth. Res. 38: 5–14, 1993.

    Article  CAS  PubMed  Google Scholar 

  • Day, A., Ellis, T.H.: Chloroplast DNA deletions associated with wheat plants regenerated from pollen: possible basis for maternal inheritance of chloroplasts. — Cell 39: 359–368, 1984.

    Article  CAS  PubMed  Google Scholar 

  • Day, A., Ellis, T.H.N.: Deleted forms of plastid DNA in albino plants from cereal anther culture. — Curr. Genet. 9: 671–678, 1985.

    Article  CAS  Google Scholar 

  • De Buyser, J., Henry, Y., Lonnet, P., Hertzog, R., Hespel, A.: ‘Florin’: a doubled haploid wheat variety developed by the anther culture method. — Plant Breed. 98: 53–56, 1987.

    Article  Google Scholar 

  • Dunford, R., Walden, R.: Plastid genome structure and plastidrelated transcript levels in albino barley plants derived from anther culture. — Curr. Genet. 20: 339–347, 1991.

    Article  CAS  PubMed  Google Scholar 

  • Ekman, D.R., Lorenz, W.W., Przybyla, A.E., Wolfe, N.L., Dean, J.F.: SAGE analysis of transcriptome responses in Arabidopsis roots exposed to 2,4,6-trinitrotoluene. — Plant Physiol. 133: 1397–1406, 2003.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fujita Y., Takagi H., Hase T.: Cloning of the gene encoding a protochlorophyllide reductase: the physiological significance of the co-existence of light-dependent and - independent protochlorophyllide reduction systems in the cyanobacterium Plectonema boryanum. — Plant Cell Physiol. 39: 177–185, 1998.

    Article  CAS  PubMed  Google Scholar 

  • Geiger, D.R., Servaites, J.C.: Diurnal regulation of photosynthetic carbon metabolism in C3 plants. — Annu. Rev. Plant. Physiol. Plant mol. Biol. 45: 235–256, 1994.

    Article  Google Scholar 

  • Guha, S., Maheshwari, S.C.: In vitro production of embryos from anthers of Datura. — Nature 204: 497–497, 1964.

    Article  Google Scholar 

  • Harada, T., Sato, T., Asaka, D., Matsukawa, I.: Large-scale deletions of rice plastid DNA in anther culture. — Theor. appl. Genet. 81: 157–161, 1991.

    Article  CAS  PubMed  Google Scholar 

  • Hershko, A., Ciechanover, A.: The ubiquitin system. — Annu. Rev. Biochem. 67: 425–479, 1998.

    Article  CAS  PubMed  Google Scholar 

  • Hou, D.Y., Xu, H., Du, G.Y., Lin, J.T., Duan, M., Guo, A.G.: Proteome analysis of chloroplast proteins in stage albinism line of winter wheat (Triticum aestivum) FA85. — BMB Rep. 42: 450–455, 2009.

    Article  CAS  PubMed  Google Scholar 

  • Hu, D.F., Yuan, Z.D., Tang, Y.L., Liu, J.P.: [Plant cell engineering: development of a new winter wheat variety Jinghua 1 by anther culture.] — Sci. China (Series B) 3: 283–292, 1986. [In Chin.]

    Google Scholar 

  • Jansson, S.: A guide to the Lhc genes and their relatives in Arabidopsis. — Trends Plant. Sci. 4: 236–240, 1999.

    Article  CAS  PubMed  Google Scholar 

  • Just, B.J., Santos, C.A., Fonseca, M.E., Boiteux, L.S., Oloizia, B.B., Simon, P.W.: Carotenoid biosynthesis structural genes in carrot (Daucus carota): isolation, sequence-characterization, single nucleotide polymorphism (SNP) markers and genome mapping. — Theor. appl. Genet. 114: 693–704, 2007.

    Article  CAS  PubMed  Google Scholar 

  • Kokubun, N., Ishida, H., Makino, A., Mae, T.: The degradation of the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase into the 44-kDa fragment in the lysates of chloroplasts incubated in darkness. — Plant Cell Physiol. 43: 1390–1395, 2002.

    Article  CAS  PubMed  Google Scholar 

  • Larsen, E.T., Tuvesson, I.K., Andersen, S.B.: Nuclear genes affecting percentage of green plants in barley (Hordeum vulgare L.) anther culture. — Theor. appl. Genet. 82: 417–420, 1991.

    Article  CAS  PubMed  Google Scholar 

  • Lee S., Kim J.H., Yoo E.S., Lee C.H., Hirochika H., An G.: Differential regulation of chlorophyll a oxygenase genes in rice. — Plant mol. Biol. 57: 805–818, 2005.

    Article  CAS  PubMed  Google Scholar 

  • Lee, S.K., Jeon, J.S., Bornke, F., Voll, L., Cho, J.I., Goh, C.H., Jeong, S.W., Park, Y.I., Kim, S.J., Choi, S.B., Miyao, A., Hirochika, H., An, G., Cho, M.H., Bhoo, S.H., Sonnewald, U., Hahn, T.R.: Loss of cytosolic fructose-1,6-bisphosphatase limits photosynthetic sucrose synthesis and causes severe growth retardations in rice (Oryza sativa). — Plant Cell Environ. 31: 1851–1863, 2008.

    Article  CAS  PubMed  Google Scholar 

  • Lefebvre, S., Lawson, T., Zakhleniuk, O.V., Lloyd, J.C., Raines, C.A., Fryer, M.: Increased sedoheptulose-1,7-bisphosphatase activity in transgenic tobacco plants stimulates photosynthesis and growth from an early stage in development. — Plant Physiol. 138: 451–460, 2005.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li, R.Q., Yu, C., Li, Y.R., Lam, T.W., Yiu, S.M., Kristiansen, K., Wang, J.: SOAP2: an improved ultrafast tool for short read alignment. — Bioinformatics 25: 1966–1967, 2009.

    Article  CAS  PubMed  Google Scholar 

  • Lindgren, L.O., Stalberg, K.G., Hoglund, A.S.: Seed-specific overexpression of an endogenous Arabidopsis phytoene synthase gene results in delayed germination and increased levels of carotenoids, chlorophyll, and abscisic acid. — Plant Physiol. 132: 779–785, 2003.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu, J.X., Chiou, C.Y., Shen, C.H., Chen, P.J., Liu, Y.C., Jian, C.D., Shen, X.L., Shen, F.Q., Yeh, K.W.: RNA interference-based gene silencing of phytoene synthase impairs growth, carotenoids, and plastid phenotype in Oncidium hybrid orchid. — SpringerPlus 3: 478, 2014b.

    Article  PubMed  PubMed Central  Google Scholar 

  • Liu, L., Shao, Z., Zhang, M., Wang, Q.: Regulation of carotenoid metabolism in tomato. — Mol. Plant. 8: 28–39, 2014a.

    Article  PubMed  Google Scholar 

  • Lopez-Casado, G., Covey, P.A., Bedinger, P.A., Mueller, L.A., Thannhauser, T.W., Zhang, S., Fei, Z., Giovannoni, J.J., Rose, J.K.C.: Enabling proteomic studies with RNA-Seq: the proteome of tomato pollen as a test case. — Proteomics 12: 761–774, 2012.

    Article  CAS  PubMed  Google Scholar 

  • Meier, S., Tzfadia, O., Vallabhaneni, R., Gehring, C., Wurtzel, E.T.: A transcriptional analysis of carotenoid, chlorophyll and plastidial isoprenoid biosynthesis genes during development and osmotic stress responses in Arabidopsis thaliana. — BMC Syst. Biol. 5: 77, 2011.

    Article  PubMed  PubMed Central  Google Scholar 

  • Mortazavi, A., Williams, B.A., McCue, K., Schaeffer, L., Wold, B.: Mapping and quantifying mammalian transcriptomes by RNA-Seq. — Nat. Methods. 5: 621–628, 2008.

    Article  CAS  PubMed  Google Scholar 

  • Nagalakshmi, U., Wang, Z., Waern, K., Shou, C., Raha, D., Gerstein, M., Snyder, M.: The transcriptional landscape of the yeast genome defined by RNA sequencing. — Science 320: 1344–1349, 2008.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nick, S., Meurer, J., Soll, J., Ankele, E.: Nucleus-encoded lightharvesting chlorophyll a/b proteins are imported normally into chlorophyll b-free chloroplasts of Arabidopsis. — Mol. Plant. 6: 860–871, 2013.

    Article  CAS  PubMed  Google Scholar 

  • Oster, U., Tanaka, R., Tanaka, A., Rudiger, W.: Cloning and functional expression of the gene encoding the key enzyme for chlorophyll b biosynthesis (CAO) from Arabidopsis thaliana. — Plant J. 21: 305–310, 2000.

    Article  CAS  PubMed  Google Scholar 

  • Ouyang, J.W., Hu, H., Zhang, J.J., Zeng, J.Z.: [Induction and investigation of pollen plants and their offspring in wheat.] - Sci. China (Series A). 16: 72–82, 1973. [In Chin]

    Google Scholar 

  • Ouyang, J.W., Jia, S.E., Zhang, C., Chen, X., and G., F.: [A new synthetic medium (W14) for wheat anther culture.] - Annu. Rep. Inst. Genet. Acad. sin. 91–92, 1989. [In Chin.]

    Google Scholar 

  • Paulsen, H., Dockter, C., Volkov, A., Jeschke, G.: Folding and pigment binding of light-harvesting chlorophyll a/b protein (LHCIIb). - In: Rebeiz, C., Benning, C., Bohnert, H., Daniell, H., Hoober, J.K., Lichtenthaler, H., Portis, A., Tripathy, B. (ed.): The Chloroplast. Vol. 16. Pp. 231–244. Springer, Dordrecht 2010.

    Chapter  Google Scholar 

  • Peng, Y., Zhang, Y., Lv, J., Zhang, J.H., Li, P., Shi, X.L., Wang, Y.F., Zhang, H.L., He, Z.H., Teng, S.: Characterization and fine mapping of a novel rice albino mutant low temperature albino 1. — J. Genet. Genomics 39: 385–396, 2012.

    Article  CAS  PubMed  Google Scholar 

  • Plumley, G.F., Schmidt, G.W.: Light-harvesting chlorophyll a/b complexes: interdependent pigment synthesis and protein assembly. — Plant Cell 7: 689–704, 1995.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Portis, A.R., Jr.: Rubisco activase - Rubisco's catalytic chaperone. — Photosynth. Res. 75: 11–27, 2003.

    Article  CAS  PubMed  Google Scholar 

  • Qin, G., Gu, H., Ma, L., Peng, Y., Deng, X.W., Chen, Z., Qu, L.J.: Disruption of phytoene desaturase gene results in albino and dwarf phenotypes in Arabidopsis by impairing chlorophyll, carotenoid, and gibberellin biosynthesis. — Cell Res. 17: 471–482, 2007.

    Article  CAS  PubMed  Google Scholar 

  • Rampino, P., Spano, G., Pataleo, S., Mita, G., Napier, J.A., Di Fonzo, N., Shewry, P.R., Perrotta, C.: Molecular analysis of a durum wheat 'stay green' mutant: expression pattern of photosynthesis-related genes. — J. Cereal Sci. 43: 160–168, 2006.

    Article  CAS  Google Scholar 

  • Ren, X., Ye, X., Xu, H., Dong, J., Du, L., Zhang X.: [Development of new spring wheat variety Ningchun42 by anther culture.] — Crops 2: 19, 2006. [In Chin.]

    Google Scholar 

  • Schmittgen, T.D., Livak, K.J.: Analyzing real-time PCR data by the comparative C(T) method. — Nat. Protocols 3: 1101–1108, 2008.

    Article  CAS  PubMed  Google Scholar 

  • Schmitz-Linneweber, C., Williams-Carrier, R.E., Williams-Voelker, P.M., Kroeger, T.S., Vichas, A., Barkan, A.: A pentatricopeptide repeat protein facilitates the trans-splicing of the maize chloroplast rps12 pre-mRNA. — Plant Cell 18: 2650–2663, 2006.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Spreitzer, R.J.: Role of the small subunit in ribulose-1,5-bisphosphate carboxylase/oxygenase. — Arch. Biochem. Biophys. 414: 141–149, 2003.

    Article  CAS  PubMed  Google Scholar 

  • Tang, M.H., Hai, Y., Da, L.Z., Zhou X.B., Zhao Y.Y.: [The breeding and characteristics of new wheat variety Huapei 5.] — Chin. Agr. Sci. Bull. 25: 98–101, 2009. [In Chin.]

    Google Scholar 

  • Teramoto, H., Ono, T., Minagawa, J.: Identification of Lhcb gene family encoding the light-harvesting chlorophyll-a/b proteins of photosystem II in Chlamydomonas reinhardtii. — Plant Cell. Physiol. 42: 849–856, 2001.

    Article  CAS  PubMed  Google Scholar 

  • Tucker, W.C., Du, Z.Y., Hein, R., Gromet-Elhanan, Z., Richter, M.L.: Role of the ATP synthase alpha-subunit in conferring sensitivity to tentoxin. — Biochemistry 40: 7542–7548, 2001.

    Article  CAS  PubMed  Google Scholar 

  • Tuvesson, I.K., Pedersen, S., Andersen, S.B.: Nuclear genes affecting albinism in wheat (Triticum aestivum L.) anther culture. — Theor. appl. Genet. 78: 879–883, 1989.

    Article  CAS  PubMed  Google Scholar 

  • Wang, K., Wang, S., Zhou, X., Lin, Z., Li, J., Du, L., Tang, Y., Xu, H., Yan, Y., Ye, X.: Development, identification, and genetic analysis of a quantitative dwarfing somatic variation line in wheat. — Crop Sci. 53: 1032–1041, 2013.

    Article  CAS  Google Scholar 

  • Wang, L., Feng, Z., Wang, X., Wang, X., Zhang, X.: DEGseq: an R package for identifying differentially expressed genes from RNA-seq data. — Bioinformatics 26: 136–138, 2010.

    Article  PubMed  Google Scholar 

  • Wang, Z., Gerstein, M., Snyder, M.: RNA-Seq: a revolutionary tool for transcriptomics. — Nat. Rev. Genet. 10: 57–63, 2009.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yaronskaya, E., Ziemann, V., Walter, G., Averina, N., Borner, T., Grimm, B.: Metabolic control of the tetrapyrrole biosynthetic pathway for porphyrin distribution in the barley mutant albostrians. — Plant J. 35: 512–522, 2003.

    Article  CAS  PubMed  Google Scholar 

  • Yosef, I., Irihimovitch, V., Knopf, J.A., Cohen, I., Orr-Dahan, I., Nahum, E., Keasar, C., Shapira, M.: RNA binding activity of the ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit from Chlamydomonas reinhardtii. - J. biol. Chem. 279, 10148–10156, 2004.

    Article  CAS  PubMed  Google Scholar 

  • Zhan, G.M., Li, R.J., Hu, Z.Y., Liu, J., Deng, L.B., Lu, S.Y., Hua, W.: Cosuppression of RBCS3B in Arabidopsis leads to severe photoinhibition caused by ROS accumulation. — Plant Cell Rep. 33: 1091–1108, 2014.

    Article  CAS  PubMed  Google Scholar 

  • Zhang, H., Li, J., Yoo, J.H., Yoo, S.C., Cho, S.H., Koh, H.J.: Rice Chlorine-1 and Chlorina-9 encode ChID and ChlI subunits of Mg-chelatase, a key enzyme for chlorophyll synthesis and chloroplast development. — Plant mol. Biol. 62: 325–337, 2006.

    Article  CAS  PubMed  Google Scholar 

  • Zhou, H., Konzak, C.F.: Improvement of anther culture methods for haploid production in wheat. — Crop Sci. 29: 817–821, 1989.

    Article  Google Scholar 

  • Zhou, X., Wang, K., Lv, D., Wu, C., Li, J., Zhao, P., Lin, Z., Du, L., Yan, Y., Ye, X.: Global analysis of differentially expressed genes and proteins in the wheat callus infected by Agrobacterium tumefaciens. — PloS ONE 8: e79390, 2013.

    Article  Google Scholar 

  • Ziegler, G., Dressler, K., Hess, D.: Investigations on the anther culturability of four German spring wheat cultivars and the influence of light on regeneration of green vs. albino plants. — Plant Breed. 105: 40–46, 1990.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to X. G. Ye.

Additional information

Acknowledgments: This research was financially supported by grants in part from the National Natural Science Foundation of China (31401380 and 31371621) and from the Ministry of Agriculture in China (2014ZX08010-004). The first two authors contributed equally to this work.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhao, P., Wang, K., Zhang, W. et al. Comprehensive analysis of differently expressed genes and proteins in albino and green plantlets from a wheat anther culture. Biol Plant 61, 255–265 (2017). https://doi.org/10.1007/s10535-016-0662-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10535-016-0662-y

Additional key words

Navigation