Abstract
Nitrite reductase (NiR) is a critical enzyme involved in nitrogen metabolism in plants. A full-length cDNA sequence of 1,764 bp for the NiR gene was isolated from mulberry (Morus L.) using reverse transcription polymerase chain reaction (RT-PCR). This NiR cDNA encodes a protein of 587 amino acids with a theoretical molecular weight of 65.4 kDa. The predicted Morus alba NiR (MaNiR) protein includes a repeated hemoprotein ferredoxin-like beta-component together with a 4Fe-4S-binding domain and a siroheme-binding site, which could be expressed in a recombinant prokaryotic expression system. Nitrite reductase activity was exhibited only when both of the repeat functional domains were expressed together. Both glutamic acid and gibberellins promoted MaNiR expression in mulberry leaves, but inhibited its expression in roots and stems. Following dedifferentiation of mulberry hypocotyls and cotyledons, MaNiR expression decreased strongly. This suggests that NiR could play an important role in the in vitro regeneration of mulberry hypocotyls and cotyledons.
Similar content being viewed by others
References
Asano N, Tomioka E, Kizu H, Matsui K (1994) Sugars with nitrogen in the ring isolated from the leaves of Morus bombycis. Carbohydr Res 253:235–245
Beevers L, Hageman RH (1983) Uptake and reduction of nitrate: bacteria and higher plants. In: Läuchli A, Bieleski RL (eds) Inorganic plant nutrition, vol 15. Springer, Berlin, pp 351–375 Enc Plant Physiol (New Series)
Bremer B, Bremer K, Chase MW et al (2009) An update of the angiosperm phylogeny group classification for the orders and families of flowering plants: APG III. Bot J Linn Soc 161:105–121
Crepet WL, Nixon KC, Gandolfo MA (2004) Fossil evidence and phylogeny: the age of major angiosperm clades based on mesofossil and macrofossil evidence from Cretaceous deposits. Am J Bot 91:1666–1682
Crété P, Caboche M, Meyer C (1997) Nitrite reductase expression is regulated at the post-transcriptional level by the nitrogen source in Nicotiana plumbaginifolia and Arabidopsis thaliana. Plant J 11:625–634
Du ZH, Liu JF, Zuo YC, Zhang JF, Liu BB (2011) The nutrient components and utilization of mulberry as animal forage. Acta Prata Sin 20(5):192–200
Falcão VR, Oliveira MC, Colepicolo P (2010) Molecular characterization of nitrate reductase gene and its expression in the marine red alga Gracilaria tenuistipitata (Rhodophyta). J Appl Phycol 22:613–622
Faure JD, Vincentz M, Kronenberger J, Caboche M (1991) Co-regulated expression of nitrate and nitrite reductases. Plant J 1:107–113
Flores E, Herrero A, Guerrero MG (1987) Nitrite uptake and its regulation in the cyanobacterium Anacystis nidulans. Biochim Biophys Acta 896:103–108
Gerard J, Lepoivre P (1991) Uptake kinetics of medium constituents during batch growth of potato cell cultures. Biotechnol Lett 13:381–384
He NJ, Zhang C, Qi XW et al (2013) Draft genome sequence of the mulberry tree Morus notabilis. Nat Commun 3445:1–5
Hellens R, Mullineaux P, Klee H (2000) A guide to Agrobacterium binary Ti vectors. Trends Plant Sci 5:446–451
Huang T, Dong GF, Li GG (2002) Effects of gibberellic acid and its synthetic inhibitors on shoot regeneration from epicotyl of Citrus grandis. Plant Physiol Commun 6:559–561
Kato C, Takahashl M, Sakamoto A, Morikawa H (2004) Differential expression of the nitrite reductase gene family in tobacco as revealed by quantitative competitive RT-PCR. J Exp Bot 403:1761–1763
Kim SY, Gao JJ, Lee WC, Ryu KS, Lee KR, Kim YC (1999) Antioxidative flavonoids from the leaves of Morus alba. Arch Pharm Res 22:81–85
Li FJ, Yu YX (2006) The main factors influencing alfalfa protein content. China Feed 5:34–38
Marushige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
May SK, GU LJ, Cheng HM (2011) The role of nitrate reductase and nitrite reductase in plants. Curr Biotechnol 3:159–164
Morgan DR, Soltis DE, Robertson KR (1994) Systematic and evolutionary implications of rbcL sequence variation in Rosaceae. Am J Bot 81:890–903
Nishimura A, Ashikari M, Lin S, Takashi T, Angeles ER, Yamamoto L, Matsuoka M (2005) Isolation of a rice regeneration quantitative trait loci gene and its application to transformation systems. Proc Natl Acad Sci USA 102:11940–11944
Ogawa T, Fukuoka H, Yano H, Ohkawa Y (1999) Relationships between nitrite reductase activity and genotype-dependent callus growth in rice cell cultures. Plant Cell Rep 18:576–581
Ozawa K, Kawahigashi H (2006) Positional cloning of the nitrite reductase gene associated with good growth and regeneration ability of calli and establishment of a new selection system for Agrobacterium-mediated transformation in rice (Oryza sativa L). Plant Sci 70:384–393
Qiu YC, Duan ZA, Zhang JF (2007) Study on the factors affecting the regeneration of in vitro leaves of Sophora japonica f. ‘Liao Hong’. For Res 6:854–858
Su HY, Wu YM, Liu JX (2002) Nutrients mulberry leaves and its application in ruminant rearing. Chin Cow 1:26–28
Tsai CB, Kaiser WM, Kaldenhoff R (2003) Molecular cloning and characterization of nitrate reductase from Ricinus communis L. heterologously expressed in Pichia pastoris. Planta 217:962–970
Vijaya Chitra DS, Padmaja G (1999) Clonal propagation of mulberry (Morus indica L. cultivar M-5) through in vitro culture of nodal explants. Sci Hortic-Amsterdam 80:289–298
Vincentz M, Moureaux T, Leydecker MT, Vaucheret H, Caboche M (1993) Regulation of nitrate and nitrite reductase expression in Nicotiana plumbaginifolia leaves by nitrogen and carbon metabolites. Plant J 3:315–324
Wang XL, Yu YS, Yang Y et al (2014) Clone and expression analysis of nitrate reductase gene (MaNR) of Morus L. Sci Agric Sin 12:2465–2475
Wu XP, Liu GS, Zhu K, Peng S, Guo QY (2004) Effects of extraneous source amino acid on the contents of amino acid of tobacco leaves. Sci Agric Sin 37:357–361
Ye XG, She MY, Wang K, Du LP, Xu HJ (2012) Identification, cloning, and potential application of genes related to somatic embryogenesis in plant tissue culture. Acta Agron Sin 2:191–201
Yu MD, Xiang ZH, Feng LC, Ke YF, Zhang XY, Jing CJ (1996) The discovery and study on a natural haploid Morus notabilis Schneid. Sci Seric 22:67–71
Acknowledgments
This work was supported by the National Natural Science Foundation of China (31101769), Fundamental Research Funds for the Central Universities (XDJK2014C065).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wang, Xl., Yu, Ys., Yang, Y. et al. Molecular cloning and expression of a nitrite reductase gene from mulberry (Morus L.). Plant Cell Tiss Organ Cult 121, 301–309 (2015). https://doi.org/10.1007/s11240-014-0701-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-014-0701-3