RNAi mediated silencing of lipoxygenase gene to maintain rice grain quality and viability during storage

Gayen, Dipak; Ali, Nusrat; Ganguly, Moumita; Paul, Soumitra; Datta, Karabi; Datta, Swapan K.

doi:10.1007/s11240-014-0476-6

Cited by 49 publications

(31 citation statements)

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“…Lipid degradation was proposed as one cause of seed deterioration during storage (Takano 1993). Similar to the results from soybean, the negative relationships between LOX activity and seed longevity have also been observed in maize and rice (Li et al 2007;Song et al 2007;Jiang et al 2009;Long et al 2013;Gayen et al 2014;Huang et al 2014). For example, after storing seeds of three maize varieties with and without LOX-1, 2 at 42°C and 84 % relative humidity for a period of 15 days, the variety lacking LOX-1, 2 showed a slight change in germination, but for varieties with LOX-1, 2, an obvious decline in germination was observed, suggesting LOX-1, 2 may be a definitive factor which influences seed life span (Li et al 2007).…”

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confidence: 68%

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Grain-specific reduction in lipoxygenase activity improves flour color quality and seed longevity in common wheat

et al. 2015

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In higher plants, lipoxygenases (LOXs) catalyze the oxidation of polyunsaturated fatty acids and can accumulate to relatively high levels in the seeds. However, high LOX activity has often been found associated with harmful effects on the quality attributes and the longevity of harvested seeds. Here, we report the development and characterization of three homozygous T 5 transgenic lines (designated as LOX RNAi -4, 7 and 9, respectively) with LOX activity specifically reduced in the grains of common wheat. An RNAi hairpin directed under an endosperm-specific promoter was introduced into the genome of the elite wheat variety Longchun 23 using particle bombardment. Each transgenic line (LOX RNAi -4, 7 or 9) derived from separate integration event, which carried more than 20 copies of the transgene. The three LOX RNAi lines and wild-type (WT) Longchun 23 showed a similar level of LOX transcripts in the flag leaves. However, relative to WT control, LOX RNAi -4, 7 and 9 exhibited drastic decreases in the level of LOX transcripts during grain development compared with WT control. The LOX activity levels in the harvested grains of the three transgenic lines were significantly lower than that of WT control under either normal conditions or artificial aging (AA, stored at 40°C and 85 % humidity for 6 or 9 days) treatment. The whiteness values of the flour samples from LOX RNAi -4, 7 and 9 were generally and significantly higher than those of WT control regardless of the grains were stored under normal or AA conditions. Furthermore, the germination rates of LOX RNAi -4, 7 and 9 were substantially higher than those of WT control under AA treatment. Our data demonstrate that grainspecific reduction in LOX activity by RNAi may represent a generally applicable approach for improving the quality traits and longevity of crop seeds.Keywords Lipoxygenase Á RNA interference Á Germination rate Á Whiteness Á Wheat Zhenying Dong and Bo Feng have contributed equally to this work.Electronic supplementary material The online version of this article

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confidence: 68%

“…The relationships between LOX activity and seed longevity during AA treatment have been well studied in maize and rice (Li et al 2007;Song et al 2007;Jiang et al 2009;Long et al 2013;Gayen et al 2014;Huang et al 2014). However, there have been few reports about the effect of LOXs on wheat grain deterioration.…”

Section: Discussionmentioning

confidence: 99%

Grain-specific reduction in lipoxygenase activity improves flour color quality and seed longevity in common wheat

et al. 2015

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“…RNAi vector containing r9-LOX1 gene under control of Oleosin-18 promoter (pOle-lox-006) was used for plant transformation as described in our previous study by Gayen et al (2014). The constructed RNAi vector pOle-lox-006 was introduced into the genome of golden indica rice line (SKBR-244), by PDS-1000/He particle delivery system (Bio-Rad, Hercules, CA, USA) (Datta et al 1998).…”

Section: Cloning Of Rnai Vector and Plant Transformationmentioning

confidence: 99%

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

Gayen

Ali

Sarkar

et al. 2015

Planta

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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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“…Finally, the DNA constructs are relatively easy to make and can be used in the latest generation of transgene vectors. Practical RNAi technology only dates from about 2003, but it has already been demonstrated to be useful in generating variation for important lipid-related traits in oil crops (Cheng et al, 2013;Gayen et al, 2014). However, it is worth pointing out that RNAi is not a radically novel technology in terms of creating genetic variation.…”

Section: Transgenic Breeding Technologiesmentioning

confidence: 99%

“…Peroxidation of unsaturated fatty acids due to enhancement of LOX activity which directly leads to reduction in seed vigour and deterioration of grain nutritional quality. In a recent study in rice it was found that attenuating LOX activity using RNAi technology had a positive effect on seed oil stability and seed viability -in particular GC-MS analysis revealed that reduction of free fatty acid levels in non-transgenic seeds during storage was higher when compared with that of transgenic rice seeds, which might be relevant to the situation in ripe palm fruits (Gayen et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Using modern plant breeding to improve the nutritional and technological qualities of oil crops

Murphy

2014

OCL

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-The last few decades have seen huge advances in our understanding of plant biology and in the development of new technologies for the manipulation of crop plants. The application of relatively straightforward breeding and selection methods made possible the "Green Revolution" of the 1960s and 1970s that effectively doubled or trebled cereal production in much of the world and averted mass famine in Asia. During the 2000s, much attention has been focused on genomic approaches to plant breeding with the deployment of a new generation of technologies, such as marker-assisted selection, next-generation sequencing, transgenesis (genetic engineering or GM) and automatic mutagenesis/selection (TILLING, TargetIng Local Lesions IN Genomes). These methods are now being applied to a wide range of crops and have particularly good potential for oil crop improvement in terms of both overall food and non-food yield and nutritional and technical quality of the oils. Key targets include increasing overall oil yield and stability on a per seed or per fruit basis and very high oleic acid content in seed and fruit oils for both premium edible and oleochemical applications. Other more specialised targets include oils enriched in nutritionally desirable "fish oil"-like fatty acids, especially very long chain ω-3 acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), or increased levels of lipidic vitamins such as carotenoids, tocopherols and tocotrienes. Progress in producing such oils in commercial crops has been good in recent years with several varieties being released or at advanced stages of development.Résumé -Utilisation de la sélection variétale moderne pour améliorer les qualités nutritionnelles et technologiques des cultures oléagineuses. Les dernières décennies ont connu de grandes avancées dans la compréhension de la biologie des plantes et dans le développement de nouvelles technologies de manipulation des semences. L'application de méthodes de croisements variétal et les méthodes de sélection ont rendu possible la révolution verte des années 1960 et 1970 qui ont efficacement doublé voire triplé la production céréalière dans une grande partie du monde et évité des famines de masse en Asie. Durant les années 2000, davantage d'attention a été portée à des approches génomiques en sélection des plantes avec le déploiement d'une nouvelle génération de technologies, comme la sélection assistée par des marqueurs, le séquençage de la génération suivante, la transgénèse (ingénierie génétique ou OGM) et la mutagé-nèse/sélection automatique (ILLI NG, TargetIng Local Lesions IN Genomes). Ces méthodes sont désormais appliquées à un large éventail de semences et offrent, pour les oléagineux, un potentiel particulièrement intéressant à la fois en termes de rendement alimentaire et non-alimentaire, en qualité nutritionnelle et technique des huiles. Les objectifs clés incluent une amélioration du rendement global en huile et de la stabilité par graine ou par fruit, et un contenu très élevé en acide oléique da...

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RNAi mediated silencing of lipoxygenase gene to maintain rice grain quality and viability during storage

Cited by 49 publications

References 38 publications

Grain-specific reduction in lipoxygenase activity improves flour color quality and seed longevity in common wheat

Grain-specific reduction in lipoxygenase activity improves flour color quality and seed longevity in common wheat

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

Using modern plant breeding to improve the nutritional and technological qualities of oil crops

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