Generation of stable ‘low phytic acid’ transgenic rice through antisense repression of the 1<scp>d</scp>‐<i>myo</i>‐inositol 3‐phosphate synthase gene (<i>RINO1</i>) using the 18‐kDa oleosin promoter

Kuwano, Mio; Mimura, Tetsuro; Takaiwa, Fumio; Yoshida, Kaoru

doi:10.1111/j.1467-7652.2008.00375.x

Cited by 104 publications

(74 citation statements)

References 58 publications

(97 reference statements)

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“…Ali et al (2013) also developed low-phytate rice with no apparent developmental defects by seed-specific silencing of IPK1. However, defects in seed weight, germination frequency, and seedling emergence were observed in several other trials, reflecting a role for InsP 6 in seed development (Bilyeu et al, 2008;Kuwano et al, 2009;Li et al, 2014). In this study, we have shown that expression of InsP 6 -sensitive Gle1 variants enhanced plant growth, seed yield, and seed performance of the phytate-deficient ipk1 mutant, suggesting a novel strategy for improving lpa plants.…”

Section: Discussionmentioning

confidence: 76%

InsP6-Sensitive Variants of the Gle1 mRNA Export Factor Rescue Growth and Fertility Defects of the ipk1 Low-Phytic-Acid Mutation in Arabidopsis

Lee

Cho

et al. 2015

The Plant Cell

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Myo-inositol-1,2,3,4,5,6-hexakisphosphate (InsP6), also known as phytic acid, accumulates in large quantities in plant seeds, serving as a phosphorus reservoir, but is an animal antinutrient and an important source of water pollution. Here, we report that Gle1 (GLFG lethal 1) in conjunction with InsP6 functions as an activator of the ATPase/RNA helicase LOS4 (low expression of osmotically responsive genes 4), which is involved in mRNA export in plants, supporting the Gle1-InsP6-Dbp5 (LOS4 homolog) paradigm proposed in yeast. Interestingly, plant Gle1 proteins have modifications in several key residues of the InsP6 binding pocket, which reduce the basicity of the surface charge. Arabidopsis thaliana Gle1 variants containing mutations that increase the basic charge of the InsP6 binding surface show increased sensitivity to InsP6 concentrations for the stimulation of LOS4 ATPase activity in vitro. Expression of the Gle1 variants with enhanced InsP6 sensitivity rescues the mRNA export defect of the ipk1 (inositol 1,3,4,5,6-pentakisphosphate 2-kinase) InsP6-deficient mutant and, furthermore, significantly improves vegetative growth, seed yield, and seed performance of the mutant. These results suggest that Gle1 is an important factor responsible for mediating InsP6 functions in plant growth and reproduction and that Gle1 variants with increased InsP6 sensitivity may be useful for engineering high-yielding low-phytate crops.

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Section: Discussionmentioning

confidence: 76%

InsP6-Sensitive Variants of the Gle1 mRNA Export Factor Rescue Growth and Fertility Defects of the ipk1 Low-Phytic-Acid Mutation in Arabidopsis

Lee

Cho

et al. 2015

The Plant Cell

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“…The generation of double or triple knockout mutants for the MIPS/IPS gene family should help determine the role of these genes in Arabidopsis InsP 6 metabolism. Alternatively, MIPS/IPS genes may be suppressed using anti-sense or RNAi-mediated gene silencing approaches which have been successfully employed in rice (Kuwano et al 2009) and soybean (Nunes et al 2006). Since T-DNA insertion mutants were obtained for only two of the four AtIMP genes, it is possible that the other genes have a role in seed InsP 6 levels.…”

Section: Discussionmentioning

confidence: 99%

Identification of genes necessary for wild-type levels of seed phytic acid in Arabidopsis thaliana using a reverse genetics approach

Kim

Tai

2011

Mol Genet Genomics

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The majority of phosphorus (P) in seeds is found in phytic acid (InsP(6)) which accumulates as the mixed salt phytate. InsP(6) is generally considered to be an anti-nutrient and the development of low phytic acid (lpa) seed crops is of significant interest. We have employed a reverse genetics approach to examine the impact of disrupting genes involved in inositol phosphate metabolism on Arabidopsis seed InsP(6) levels. Our analysis revealed that knockout mutations in three genes (AtITPK1, AtITPK4, and AtMIK/At5g58730) reduced seed InsP(6) in addition to knockouts of four previously reported genes (AtIPK1, AtIPK2β, AtMRP5, and At5g60760). Seeds of these lpa mutants also exhibited reduced germination under various stress conditions. The greatest reduction in InsP(6) (>70%) was observed in atmrp5 seeds which were also among the least sensitive to the stresses examined. Expression analysis of the lpa genes revealed three distinct patterns in developing siliques consistent with their presumed roles. Disruption of each lpa gene resulted in changes in the expression in some of the other lpa genes indicating that transcription of lpa genes is modulated by other constituents of InsP(6) metabolism. While all the lpa genes represent possible targets for genetic engineering of low phytate seed crops, mutations in AtMRP5, AtMIK, and At5g60760 may be most successful for conventional approaches such as mutation breeding.

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“…In the present study, we have developed transgenic golden rice with reduced LOX activity by silencing endogenous lipoxygenase gene to reduce co-oxidation of carotenoids using RNAi technology which is very powerful tool for crop improvement (Saurabh et al 2014). The RNAi-mediated gene silencing of lox gene was driven by tissue-specific Oleosin-18 promoter instead of any constitutive promoter to avoid any undesirable and detrimental effects on the plant (Kuwano et al 2009;Ali et al 2013).…”

Section: Discussionmentioning

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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Generation of stable ‘low phytic acid’ transgenic rice through antisense repression of the 1d‐myo‐inositol 3‐phosphate synthase gene (RINO1) using the 18‐kDa oleosin promoter

Cited by 104 publications

References 58 publications

InsP6-Sensitive Variants of the Gle1 mRNA Export Factor Rescue Growth and Fertility Defects of the ipk1 Low-Phytic-Acid Mutation in Arabidopsis

InsP6-Sensitive Variants of the Gle1 mRNA Export Factor Rescue Growth and Fertility Defects of the ipk1 Low-Phytic-Acid Mutation in Arabidopsis

Identification of genes necessary for wild-type levels of seed phytic acid in Arabidopsis thaliana using a reverse genetics approach

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

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