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Down-regulation of lipoxygenase gene reduces degradation of carotenoids of golden rice during storage

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Down-regulation of lipoxygenase enzyme activity reduces degradation of carotenoids of bio-fortified rice seeds which would be an effective tool to reduce huge post-harvest and economic losses of bio-fortified rice seeds during storage.

Bio-fortified provitamin A-enriched rice line (golden rice) expressing higher amounts of β-carotene in the rice endosperm provides vitamin A for human health. However, it is already reported that degradation of carotenoids during storage is a major problem. The gene responsible for degradation of carotenoids during storage has remained largely unexplored till now. In our previous study, it has been shown that r9-LOX1 gene is responsible for rice seed quality deterioration. In the present study, we attempted to investigate if r9-LOX1 gene has any role in degradation of carotenoids in rice seeds during storage. To establish our hypothesis, the endogenous lipoxygenase (LOX) activity of high-carotenoid golden indica rice seed was silenced by RNAi technology using aleurone layer and embryo-specific Oleosin-18 promoter. To check the storage stability, LOX enzyme down-regulated high-carotenoid T3 transgenic rice seeds were subjected to artificial aging treatment. The results obtained from biochemical assays (MDA, ROS) also indicated that after artificial aging, the deterioration of LOX-RNAi lines was considerably lower compared to β-carotene-enriched transgenic rice which had higher LOX activity in comparison to LOX-RNAi lines. Furthermore, it was also observed by HPLC analysis that down-regulation of LOX gene activity decreases co-oxidation of β-carotene in LOX-RNAi golden rice seeds as compared to the β-carotene-enriched transgenic rice, after artificial aging treatment. Therefore, our study substantially establishes and verifies that LOX is a key enzyme for catalyzing co-oxidation of β-carotene and has a significant role in deterioration of β-carotene levels in the carotenoid-enriched golden rice.

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Abbreviations

LOX:

Lipoxygenase

MDA:

Malondialdehyde

ROS:

Reactive oxygen species

VAD:

Vit-A deficiency

DCFDA:

2′, 7′-Dichlorofluorescin diacetate

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Acknowledgments

We thankfully acknowledge Department of Biotechnology (DBT), Government of India (Sanction no. BT/01/COE/06/05) and DST-PURSE, University of Calcutta for financial support. We are also grateful to the Leibniz Institute of Plant Genetics and Crop Plant Research, D–06466 Gatersleben, Germany for providing the pIPKb-006 destination vector.

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  1. Nusrat Ali

    Present address: Department of Forest Genetics and Plant Physiology, Umeå Plant Science Centre, Swedish University of Agricultural Sciences, 90183, Umeå, Sweden

Authors and Affiliations

  1. Laboratory of Translational Research on Transgenic Crops, Department of Botany, University of Calcutta, 35 Ballygunge Circular Road, Kolkata, 700019, West Bengal, India

    Dipak Gayen, Nusrat Ali, Sailendra Nath Sarkar, Swapan K. Datta & Karabi Datta

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  1. Dipak Gayen
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Correspondence to Karabi Datta.

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Gayen, D., Ali, N., Sarkar, S.N. et al. Down-regulation of lipoxygenase gene reduces degradation of carotenoids of golden rice during storage. Planta 242, 353–363 (2015). https://doi.org/10.1007/s00425-015-2314-4

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