Marker-assisted backcross breeding for enhancing β-carotene of QPM inbreds
Maize possesses natural variations for crtRB1 (β-carotene hydroxylase) and lcyE (lycopene-ε-cyclase) which significantly enhances provitamin A (proA) concentration. Enhancement of kernel proA carotenoids in QPM genetic background is of greater value. In this study, allelic variability for proA carotenoids was analyzed to identify promising β-carotene donor to introgress into QPM inbreds employing molecular breeding. Maize inbred MGU23379 (6.31 μg/g of β-carotene) was used as β-carotene donor to cross with recurrent parents (RPs), CB6-36 (CBML6) and CB7-28 (CBML7). In conversion program, F1, BC1F1, BC2F1, BC2F2, BC2F3, and BC2F4 materials were generated. In each generation, foreground selection was carried out with crtRB1-3′TE and umc1066. The crtRB1-3′TE segregated as per the expectation. Selection for modifiers, recombinants, and background genome of RPs was carried out in BC populations. The individuals with high recovery of recurrent parent genome were retained. Tryptophan/lysine content in introgressed progenies was on par, but β-carotene content was significantly high (6.25and 6.80 μg/g) compared to original inbreds (0.71 and 1.29 μg/g). Phenotypic data recorded for different traits in the BC2F3 populations did not show any significant difference between the converted BC2F3 families and their RPs. Also, grain yield of converted inbreds (PVCBML6 and PVCBML7) was on par with their original lines. In PVCBML6, stem anthocyanin pigmentation was reduced and silk color was changed to dark pink; whereas in PVCBML7, tassel was more erect and sparer; and silk color was changed to light pink compared to original inbreds. Converted inbreds provides an ideal platform for stacking number of nutritionally important traits.
KeywordsQPM opqaue2 crtRB1-3′TE β-Carotene Provitamin A Recurrent parent genome Molecular breeding
First author is thankful to Department of Science and Technology (DST), Govt. of India, for providing INSPIRE fellowship to pursue doctoral studies [IF120799]. All authors are thankful to Institute of Biotechnology and Maize Research Centre, PJTSAU, Hyderabad, for providing lab and field facilities to conduct the research.
Conception and design of study: SSR and DBS, Development of segregating progenies and molecular analysis: DBS. Morphological characterization: DBS and PS. Phenotyping for kernel quality, biochemical analyses: DBS, MS, and PS. Statistical analyses: PS. Drafting of the manuscript: DBS and PS.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Babu R, Nair SK, Kumar A, Venkatesh S, Sekhar JC, Singh NN, Srinivasan G, Gupta HS (2005) Two-generation marker-aided backcrossing for rapid conversion of normal maize lines to quality protein maize (QPM). Theor Appl Genet 111(5):888–897. https://doi.org/10.1007/s00122-005-0011-6 CrossRefPubMedGoogle Scholar
- Bain LE, Awah PK, Geraldine N, Kindong NP, Sigal Y, Bernard N, Tanjeko AT (2013) Malnutrition in Sub-Saharan Africa: burden, causes and prospects. Pan Afr Med J 15(1). https://doi.org/10.11604/pamj.2013.15.120.2535
- Gebremeskel S, Garcia-oliveira AL, Menkir A, Adetimirin V, Gedil M (2018) Effectiveness of predictive markers for marker assisted selection of pro-vitamin A carotenoids in medium-late maturing maize (Zea mays L.) inbred lines. J Cereal Sci 79:27–34. https://doi.org/10.1016/j.jcs.2017.09.001 CrossRefGoogle Scholar
- Harjes CE, Rocheford TR, Bai L, Brutnell TP, Kandianis CB, Sowinski SG, Stapleton AE, Vallabhaneni R, Williams M, Wurtzel ET, Yan J, Buckler ES (2008) Natural genetic variation in lycopene epsilon cyclase tapped for maize biofortification. Science 319(5861):330–333. https://doi.org/10.1126/science.1150255 CrossRefPubMedPubMedCentralGoogle Scholar
- Hossain F, Muthusamy V, Pandey N, Vishwakarma AK, Baveja A, Zunjare RU, Thirunavukkarasu N, Saha S, Manjaiah KM, Prasanna BM, Gupta HS (2018) Marker-assisted introgression of opaque2 allele for rapid conversion of elite hybrids into quality protein maize. J Genet 97(1):287–298. https://doi.org/10.1007/s12041-018-0914-z CrossRefPubMedGoogle Scholar
- Kandianis CB, Stevens R, Liu W, Palacios N, Montgomery K, Pixley K, White WS, Rocheford T (2013) Genetic architecture controlling variation in grain carotenoid composition and concentrations in two maize populations. Theor Appl Genet 126(11):2879–2895. https://doi.org/10.1007/s00122-013-2179-5 CrossRefPubMedPubMedCentralGoogle Scholar
- Muthusamy V, Hossain F, Thirunavukkarasu N, Choudhary M, Saha S, Bhat JS, Prasanna BM, Gupta HS (2014) Development of β-carotene rich maize hybrids through marker-assisted introgression of β-carotene hydroxylase allele. PLoS One 9(12):e113583. https://doi.org/10.1371/journal.pone.0113583 CrossRefPubMedPubMedCentralGoogle Scholar
- Muthusamy V, Hossain F, Thirunavukkarasu N, Saha S, Gupta HS (2015) Allelic variations for lycopene-ɛ-cyclase and β-carotene hydroxylase genes in maize inbreds and their utilization in β-caroteneenrichment programme. Cogent Food Agric 1(1033141). https://doi.org/10.1080/23311932.2015.1033141
- Rodriguez-Amaya DB (2001) A guide to carotenoid analysis in foods. Washington, ILSI pressGoogle Scholar
- Sagare DB, Shetti P, Surender M, Reddy SS, Pradeep T, Anuradha G (2018) Maize: potential crop for provitamin A biofortification. Maydica 63(2):11 Available at: https://journals-crea.4science.it/index.php/maydica/article/view/1645 Google Scholar
- Selvi DT, Senthil N, Yuvaraj A, Joel AJ, Mahalingam A, Nagarajan P, Vellaikumar S, Srimathi P, Raveendran M, Nepolean T (2014) Assessment of crtRB1 polymorphism associated with increased β-carotene content in maize (Zea mays L.) seeds. Food Biotechnol 28(1):41–49. https://doi.org/10.1080/08905436.2013.870077 CrossRefGoogle Scholar
- Singh R, Ram L (2014) DNA Aided Introgression of opaque2 allele for development of quality protein maize. Int J Sci Res 3(11): 3006–3011 Available at: googlescholar Google Scholar
- Tanumihardjo SA, Anderson C, Kaufer-Horwitz M, Bode L, Emenaker NJ, Haqq AM, Satia JA, Silver HJ, Stadler DD (2007) Poverty, obesity, and malnutrition: an international perspective recognizing the paradox. J Am Diet Assoc 107(11):1966–1972. https://doi.org/10.1016/j.jada.2007.08.007 CrossRefPubMedGoogle Scholar
- Vallabhaneni R, Gallagher CE, Licciardello N, Cuttriss AJ, Quinlan RF, Wurtzel ET (2009) Metabolite sorting of a germplasm collection reveals the hydroxylase3 locus as a new target for maize provitamin A biofortification. Plant Physiol 151(3):1635–1645. https://doi.org/10.1104/pp.109.145177 CrossRefPubMedPubMedCentralGoogle Scholar
- Vignesh M, Hossain F, Nepolean T, Supradip S, Agrawal PK, Guleria SK, et al (2012) Genetic variability for kernel beta-carotene and utilization of crtRB1 3’TE gene for biofortification in maize (Zea mays L.). Indian J Genet Plant Breed 72: 189–194 Available at: http://hdl.handle.net/10883/3175
- Vivek BS, Krivanek AF, Palacios-Rojas N, Twumasi-Afriyie S, Diallo AO (2008) Breeding quality protein maize (QPM): protocols for developing QPM cultivars. Mexico: CIMMYT Available at: http://hdl.handle.net/10883/801
- Yan J, Kandianis CB, Harjes CE, Bai L, Kim EH, Yang X, Skinner DJ, Fu Z, Mitchell S, Li Q, Fernandez MGS, Zaharieva M, Babu R, Fu Y, Palacios N, Li J, DellaPenna D, Brutnell T, Buckler ES, Warburton ML, Rocheford T (2010) Rare genetic variation at Zea mays crtRB1 increases beta-carotene in maize grain. Nat Genet 42(4):322–327. https://doi.org/10.1038/ng.551 CrossRefPubMedGoogle Scholar
- Zunjare RU, Hossain F, Muthusamy V, Baveja A, Chauhan HS, Thirunavukkarasu N, Saha S, Gupta HS (2017) Influence of rare alleles of β-carotene hydroxylase and lycopene epsilon cyclase genes on accumulation of provitamin A carotenoids in maize kernels. Plant Breed 136(6):872–880. https://doi.org/10.1111/pbr.12548 CrossRefGoogle Scholar
- Zunjare RU, Hossain F, Muthusamy V, Baveja A, Chauhan HS, Bhat JS, Thirunavukkarasu N, Saha S, Gupta HS (2018) Development of biofortified maize hybrids through marker-assisted stacking of β-carotene hydroxylase, lycopene-ε-cyclase and opaque2 genes. Front Plant Sci 9(178). https://doi.org/10.3389/fpls.2018.00178