Mutant crtRB1 gene negates the unfavourable effects of opaque2 gene on germination and seed vigour among shrunken2-based biofortified sweet corn genotypes
Brijesh K. Mehta A B , Hema S. Chauhan A , Sudipta Basu A , Anjali Anand A , Aanchal Baveja A , Rajkumar U. Zunjare A , Vignesh Muthusamy A , Ashok K. Singh A and Firoz Hossain
A *
A
B Present address:
Abstract
Sweet corn is one of the most popular vegetables worldwide. However, traditional shrunken2 (sh2)-based sweet corn varieties are poor in nutritional quality. Here, we analysed the effect of (1) β-carotene hydroxylase1 (crtRB1), (2) opaque2 (o2) and (3) o2 + crtRB1 genes on nutritional quality, germination, seed vigour and physico-biochemical traits in a set of 27 biofortified sh2-based sweet corn inbreds. The biofortified sweet corn inbreds recorded significantly higher concentrations of proA (16.47 μg g−1), lysine (0.36%) and tryptophan (0.09%) over original inbreds (proA: 3.14 μg g−1, lysine: 0.18%, tryptophan: 0.04%). The crtRB1-based inbreds had the lowest electrical conductivity (EC), whereas o2-based inbreds possessed the highest EC. The o2 + crtRB1-based inbreds showed similar EC to the original inbreds. Interestingly, o2-based inbreds also had the lowest germination and seed vigour compared to original inbreds, whereas crtRB1 and o2 + crtRB1 introgressed sweet corn inbreds showed similar germination and seed vigour traits to their original versions. This suggested that the negative effect of o2 on germination, seed vigour and EC is nullified by crtRB1 in the double mutant sweet corn. Overall, o2 + crtRB1-based sweet corn inbreds were found the most desirable over crtRB1- and o2-based inbreds alone.
Keywords: crtB1, enzymes activity, germination, nutrition, opaque2, seed vigour, sweet corn.
References
Abdul-Baki AA, Anderson JD (1973) Vigor determination in soybean seed by multiple criteria. Crop Science 13, 630-633.
| Crossref | Google Scholar |
Angelovici R, Fait A, Fernie AR, Galili G (2011) A seed high-lysine trait is negatively associated with the TCA cycle and slows down Arabidopsis seed germination. New Phytologist 189, 148-159.
| Crossref | Google Scholar | PubMed |
Aung UT, McDonald MB (1995) Changes in esterase activity associated with peanut (Archis hypogaea L.) seed deterioration. Seed Science and Technology 23, 101-111.
| Google Scholar |
Babu R, Rojas NP, Gao S, Yan J, Pixley K (2013) Validation of the effects of molecular marker polymorphisms in LcyE and CrtRB1 on provitamin A concentrations for 26 tropical maize populations. Theoretical and Applied Genetics 126, 389-399.
| Crossref | Google Scholar | PubMed |
Babu BK, Agrawal PK, Saha S, Gupta HS (2015) Mapping QTLs for opaque2 modifiers influencing the tryptophan content in quality protein maize using genomic and candidate gene-based SSRs of lysine and tryptophan metabolic pathway. Plant Cell Reports 34, 37-45.
| Crossref | Google Scholar | PubMed |
Baseggio M, Murray M, Magallanes-Lundback M, Kaczmar N, Chamness J, Buckler ES, Smith ME, DellaPenna D, Tracy WF, Gore MA (2020) Natural variation for carotenoids in fresh kernels is controlled by uncommon variants in sweet corn. The Plant Genome 13(1), e20008.
| Crossref | Google Scholar | PubMed |
Baveja A, Muthusamy V, Panda KK, Zunjare RU, Das AK, Chhabra R, Mishra SJ, Mehta BK, Saha S, Hossain F (2021) Development of multinutrient-rich biofortified sweet corn hybrids through genomics-assisted selection of shrunken2, opaque2, lcyE and crtRB1 genes. Journal of Applied Genetics 62(3), 419-429.
| Crossref | Google Scholar | PubMed |
Beerelli K, Balakrishnan D, Surapaneni M, Addanki K, Rao YV, Neelamraju S (2019) Evaluation of yield and seedling vigour related traits of Swarna/Oryza nivara backcross introgression lines under three environment conditions. Ecological Genetics and Genomics 11, 100036.
| Crossref | Google Scholar |
Bhave MR, Lawrence S, Barton C, Hannah LC (1990) Identification and molecular characterization of shrunken-2 cDNA clones of maize. The Plant Cell 2(6), 581-588.
| Google Scholar | PubMed |
Cabrera-Soto L, Pixley KV, Rosales-Nolasco A, Galicia-Flores LA, Palacios-Rojas N (2018) Carotenoid and tocochromanol profiles during kernel development make consumption of biofortified “fresh” maize an option to improve micronutrient nutrition. Journal of Agricultural and Food Chemistry 66(36), 9391-9398.
| Crossref | Google Scholar | PubMed |
Castelli S, Mascheretti I, Cosentino C, Lazzari B, Pirona R, Ceriotti A, Viotti A, Lauria M (2018) Uniparental and transgressive expression of α-zeins in maize endosperm of o2 hybrid lines. PLoS ONE 13, e0206993.
| Crossref | Google Scholar | PubMed |
Chauhan HS, Chhabra R, Rashmi T, Muthusamy V, Zunjare RU, Mishra SJ, Gain N, Mehta BK, Singh AK, Gupta HS, Hossain F (2022a) Impact of vte4 and crtRB1 genes on composition of vitamin-E and provitamin-A carotenoids during kernel-stages in sweet corn. Journal of Food Composition and Analysis 105, 104264.
| Crossref | Google Scholar |
Chauhan HS, Muthusamy V, Rashmi T, Basu S, Anand A, Mehta BK, Gain N, Zunjare RU, Singh AK, Gupta HS, Hossain F (2022b) Characterization of crtRB1- and vte4-based biofortified sweet corn inbreds for seed vigour and physico-biochemical traits. Journal of Applied Genetics 63(4), 651-662.
| Crossref | Google Scholar | PubMed |
Cheng X, Xiong F, Wang C, Xie H, He S, Geng G, Zhou Y (2018) Seed reserve utilization and hydrolytic enzyme activities in germinating seeds of sweet corn. Pakistan Journal of Botany 50(1), 111-116.
| Google Scholar |
Chhabra R, Hossain F, Muthusamy V, Baveja A, Mehta BK, Zunjare RU (2019) Development and validation of breeder-friendly functional markers of sugary1 gene encoding starch-debranching enzyme affecting kernel sweetness in maize (Zea mays). Crop and Pasture Science 70, 868-875.
| Crossref | Google Scholar |
Chhabra R, Hossain F, Muthusamy V, Baveja A, Mehta BK, Uttamrao Zunjare R (2020) Development and validation of gene-based markers for shrunken2-Reference allele and their utilization in marker-assisted sweet corn (Zea mays Sachharata) breeding programme. Plant Breeding 139, 1135-1144.
| Crossref | Google Scholar |
Deng Y, Wang J, Zhang Z, Wu Y (2020) Transactivation of Sus1 and Sus2 by Opaque2 is an essential supplement to sucrose synthase-mediated endosperm filling in maize. Plant Biotechnology Journal 18(9), 1897-1907.
| Crossref | Google Scholar | PubMed |
Dhindsa RS, Plumb-Dhindsa P, Thorpe TA (1981) Leaf senescence: correlated with increased levels of membrane permeability and lipid peroxidation, and decreased levels of superoxide dismutase and catalase. Journal of Experimental Botany 32(1), 93-101.
| Crossref | Google Scholar |
Domergue J-B, Abadie C, Limami A, Way D, Tcherkez G (2019) Seed quality and carbon primary metabolism. Plant, Cell & Environment 42(10), 2776-2788.
| Crossref | Google Scholar | PubMed |
Dutta S, Muthusamy V, Hossain F, Baveja A, Chhabra R, Jha SK, Yadava DK, Zunjare RU (2020) Analysis of genetic variability for retention of kernel carotenoids in sub-tropically adapted biofortified maize under different storage conditions. Journal of Cereal Science 93, 102987.
| Crossref | Google Scholar |
Falco SC, Guida T, Locke M, Mauvais J, Sanders C, Ward RT, Webber P (1995) Transgenic canola and soybean seeds with increased lysine. Biotechnology 13, 577-582.
| Crossref | Google Scholar | PubMed |
FAOSTAT (2022) Trade data: crops and livestock products. Available at https://www.fao.org/faostat/en/#data/TCL
Feng ZL, Liu J, Fu FL, Li WC (2008) Molecular mechanism of sweet and waxy in maize. International Journal of Plant Breeding and Genetics 2, 93-100.
| Crossref | Google Scholar |
Gibbon BC, Larkins BA (2005) Molecular genetic approaches to developing quality protein maize. Trends in Genetics 21, 227-233.
| Crossref | Google Scholar | PubMed |
Guglielminetti L, Yamaguchi J, Perata P, Alpi A (1995) Amylolytic activities in cereal seeds under aerobic and anaerobic conditions. Plant Physiology 109(3), 1069-1076.
| Crossref | Google Scholar | PubMed |
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. Journal of Genetics 97, 287-298.
| Crossref | Google Scholar | PubMed |
Hossain F, Zunjare RU, Muthusamy V, Bhat JS, Mehta BK, Sharma D, Talukder ZA, Chhabra R, Katral A, Dutta S, Chand G, Bhatt V, Mishra SJ, Gain N, Kasana R, Ikkurti G, Duo H (2022) Biofortification of maize for nutritional security. In ‘Biofortification of staple crops’. (Eds S Kumar, HK Dikshit, GP Mishra, A Singh) pp. 147–174. (Springer Nature)
Howitt CA, Pogson BJ (2006) Carotenoid accumulation and function in seeds and non-green tissues. Plant, Cell & Environment 29(3), 435-445.
| Crossref | Google Scholar | PubMed |
Jia M, Wu H, Clay KL, Jung R, Larkins BA, Gibbon BC (2013) Identification and characterization of lysine-rich proteins and starch biosynthesis genes in the opaque2 mutant by transcriptional and proteomic analysis. BMC Plant Biology 13, 60.
| Crossref | Google Scholar | PubMed |
Jin J, Long W, Wang L, Liu X, Pan G, Xiang W, Li N, Li S (2018) QTL mapping of seed vigor of backcross inbred lines derived from Oryza longistaminata under artificial aging. Frontiers in Plant Science 9, 1909.
| Crossref | Google Scholar | PubMed |
Kato-Noguchi H, Macías FA (2005) Effects of 6-methoxy-2-benzoxazolinone on the germination and α-amylase activity in lettuce seeds. Journal of Plant Physiology 162, 1304-1307.
| Crossref | Google Scholar | PubMed |
Kemper EL, Neto GC, Papes F, Moraes KCM, Leite A, Arruda P (1999) The role of opaque2 in the control of lysine-degrading activities in developing maize endosperm. The Plant Cell 11, 1981-1993.
| Crossref | Google Scholar | PubMed |
Kibinza S, Vinel D, Côme D, Bailly C, Corbineau F (2006) Sunflower seed deterioration as related to moisture content during ageing, energy metabolism and active oxygen species scavenging. Physiologia Plantarum 128, 496-506.
| Crossref | Google Scholar |
Kittock DL, Law AG (1968) Relationship of seedling vigor to respiration and tetrazolium chloride reduction by germinating wheat seeds. Agronomy Journal 60, 286-288.
| Crossref | Google Scholar |
Kravić N, Anđjelković V, Hadži-Tašković-Šukalović V, Vuletić M (2009) Antioxidant activity in seeds of maize genotypes with different percentage of exotic germplasm. Genetika 41(1), 21-28.
| Crossref | Google Scholar |
Kurilich AC, Juvik JA (1999) Quantification of carotenoid and tocopherol antioxidants in Zea mays. Journal of Agricultural and Food Chemistry 47, 1948-1955.
| Crossref | Google Scholar | PubMed |
Laughnan JR (1953) The effect of the sh2 factor on carbohydrate reserves in the mature endosperm of maize. Genetics 38, 485-499.
| Crossref | Google Scholar | PubMed |
Liu H, Mao J, Yan S, Yu Y, Xie L, Hu JG, Li T, Abbasi AM, Guo X, Liu RH (2018) Evaluation of carotenoid biosynthesis, accumulation and antioxidant activities in sweetcorn (Zea mays L.) during kernel development. International Journal of Food Science & Technology 53, 381-388.
| Crossref | Google Scholar |
Loesch PJ, Jr, Wiser WJ, Booth GD (1978) Emergence comparisons between opaque and normal segregates in two maize synthetics. Crop Science 18(5), 802-805.
| Crossref | Google Scholar |
Mansouri-Far C, Goodarzian-Ghahfarokhi M, Saeidi M, Abdoli M (2015) Antioxidant enzyme activity and germination characteristics of different maize hybrid seeds during ageing. Environmental and Experimental Biology 13, 177-182.
| Google Scholar |
Mehta BK, Hossain F, Muthusamy V, Zunjare RU, Sekhar JC, Gupta HS (2017) Analyzing the role of sowing and harvest time as factors for selecting super sweet (-sh2sh2) corn hybrids. Indian Journal of Genetics and Plant Breeding 77, 348-356.
| Crossref | Google Scholar |
Mehta BK, Muthusamy V, Baveja A, Chauhan HS, Chhabra R, Bhatt V, Chand G, Zunjare RU, Singh AK, Hossain F (2020a) Composition analysis of lysine, tryptophan and provitamin-A during different stages of kernel development in biofortified sweet corn. Journal of Food Composition and Analysis 94, 103625.
| Crossref | Google Scholar |
Mehta BK, Muthusamy V, Zunjare RU, Baveja A, Chauhan HS, Chhabra R, Singh AK, Hossain F (2020b) Biofortification of sweet corn hybrids for provitamin-A, lysine and tryptophan using molecular breeding. Journal of Cereal Science 96, 103093.
| Crossref | Google Scholar |
Mehta BK, Chhabra R, Muthusamy V, Zunjare RU, Baveja A, Chauhan HS, Prakash NR, Chalam VC, Singh AK, Hossain F (2021) Expression analysis of β-carotene hydroxylase1 and opaque2 genes governing accumulation of provitamin-A, lysine and tryptophan during kernel development in biofortified sweet corn. 3 Biotech 11(7), 325.
| Crossref | Google Scholar |
Mertz ET, Bates LS, Nelson OE (1964) Mutant gene that changes protein composition and increases lysine content of maize endosperm. Science 145, 279-280.
| Crossref | Google Scholar | PubMed |
Mertz LM, Henning FA, Cruz HLd, Meneghello GE, Ferrari CdS, Zimmer PD (2009) Structural differences between soybean seeds coat with contrasting permeability. Revista Brasileira de Sementes 31(1), 23-29.
| Crossref | Google Scholar |
Moehn S, Pencharz PB, Ball RO (2012) Lessons learned regarding symptoms of tryptophan deficiency and excess from animal requirement studies. The Journal of Nutrition 142, 2231S-2235S.
| Crossref | Google Scholar | PubMed |
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, e113583.
| Crossref | Google Scholar | PubMed |
Nelly Sandrine AM, Zhao H, Qin Y, Sun Q, Gong D, Pan Z, Qiu F (2020) 22KD zein content coordinates transcriptional activity during starch synthesis in maize endosperm. Agronomy 10(5), 624.
| Crossref | Google Scholar |
Oliveira TLd, Pinho RGV, Santos HOd, Silva KMdJ, Pereira EdM, Souza JLD (2020) Biochemical changes and physiological quality of corn seeds subjected to different chemical treatments and storage times. Journal of Seed Science 42, e202042038.
| Crossref | Google Scholar |
Pinzino C, Capocchi A, Galleschi L, Saviozzi F, Nanni B, Zandomeneghi M (1999) Aging, free radicals, and antioxidants in wheat seeds. Journal of Agricultural and Food Chemistry 47, 1333-1339.
| Crossref | Google Scholar | PubMed |
Prasanna BM, Palacios-Rojas N, Hossain F, Muthusamy V, Menkir A, Dhliwayo T, Ndhlela T, San Vicente F, Nair SK, Vivek BS, Zhang X, Olsen M, Fan X (2020) Molecular breeding for nutritionally enriched maize: status and prospects. Frontiers in Genetics 10, 1392.
| Crossref | Google Scholar | PubMed |
Pritchard SL, Charlton WL, Baker A, Graham IA (2002) Germination and storage reserve mobilization are regulated independently in Arabidopsis. The Plant Journal 31, 639-647.
| Crossref | Google Scholar | PubMed |
Rao AV, Rao LG (2007) Carotenoids and human health. Pharmacological Research 55, 207-216.
| Crossref | Google Scholar | PubMed |
Rao MV, Paliyath G, Ormrod DP (1996) Ultraviolet-B- and ozone-induced biochemical changes in antioxidant enzymes of Arabidopsis thaliana. Plant Physiology 110, 125-136.
| Crossref | Google Scholar | PubMed |
Sarika K, Hossain F, Muthusamy V, Zunjare RU, Baveja A, Goswami R, Bhat JS, Saha S, Gupta HS (2018) Marker-assisted pyramiding of opaque2 and novel opaque16 genes for further enrichment of lysine and tryptophan in sub-tropical maize. Plant Science 272, 142-152.
| Crossref | Google Scholar | PubMed |
Shi Y, Li G, Tian Z, Wang Z, Wang X, Zhu Y, Chen Y, Guo S, Qi J, Zhang X, Ku L (2016) Genetic dissection of seed vigour traits in maize (Zea mays L.) under low-temperature conditions. Journal of Genetics 95(4), 1017-1022.
| Crossref | Google Scholar | PubMed |
Singh B, Zunjare RU, Shrivastava S, Chand G, Gain N, Bhatt V, Muthusamy V, Hossain F (2023) Provitamin A, lysine and tryptophan enrichment in shrunken2-based sweet corn genotypes through genomics-assisted breeding for crtRB1 and opaque2 genes. Molecular Biology Reports
| Crossref | Google Scholar |
Sun T, Zhu Q, Wei Z, Owens LA, Fish T, Kim H, Thannhauser TW, Cahoon EB, Li L (2021) Multi-strategy engineering greatly enhances provitamin A carotenoid accumulation and stability in Arabidopsis seeds. aBIOTECH 2(3), 191-214.
| Crossref | Google Scholar | PubMed |
Tome D, Bos C (2007) Lysine requirement through the human life cycle. The Journal of Nutrition 137, 1642S-1645S.
| Crossref | Google Scholar | PubMed |
Tsai C-Y, Nelson OE (1966) Starch-deficient maize mutant lacking adenosine diphosphate glucose pyrophosphorylase activity. Science 151(3708), 341-343.
| Crossref | Google Scholar | PubMed |
Wakasa K, Hasegawa H, Nemoto H, Matsuda F, Miyazawa H, Tozawa Y, Morino K, Komatsu A, Yamada T, Terakawa T, Miyagawa H (2006) High-level tryptophan accumulation in seeds of transgenic rice and its limited effects on agronomic traits and seed metabolite profile. Journal of Experimental Botany 57, 3069-3078.
| Crossref | Google Scholar | PubMed |
Wang W, Xu M, Wang G, Galili G (2018) New insights into the metabolism of aspartate-family amino acids in plant seeds. Plant Reproduction 31, 203-211.
| Crossref | Google Scholar | PubMed |
Wang H, Huang Y, Xiao Q, Huang X, Li C, Gao X, Wang Q, Xiang X, Zhu Y, Wang J, Wang W, Larkins BA, Wu Y (2020) Carotenoids modulate kernel texture in maize by influencing amyloplast envelope integrity. Nature Communications 11(1), 5346.
| Crossref | Google Scholar |
Yan J, Kandianis CB, Harjes CE, Bai L, Kim E-H, 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 β-carotene in maize grain. Nature Genetics 42, 322-327.
| Crossref | Google Scholar | PubMed |
Zhao GW, Sun Q, Wang JH (2007) Improving seed vigour assessment of super sweet and sugar-enhanced sweet corn (Zea mays saccharata). New Zealand Journal of Crop and Horticultural Science 35(3), 349-356.
| Crossref | Google Scholar |
Zhu X, Galili G (2003) Increased lysine synthesis coupled with a knockout of its catabolism synergistically boosts lysine content and also transregulates the metabolism of other amino acids in Arabidopsis seeds. The Plant Cell 15, 845-853.
| Crossref | Google Scholar | PubMed |
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. Frontiers in Plant Science 9, 178.
| Crossref | Google Scholar | PubMed |
