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

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
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DOI:
10.1111/j.1467-7652.2008.00375.x
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发表时间:
2009-01-01
影响因子:
13.8
通讯作者:
Yoshida, Kaoru T.
Yoshida, Kaoru T.
中科院分区:
工程技术1区
文献类型:
--
作者:
Kuwano, Mio;Mimura, Tetsuro;Yoshida, Kaoru T.

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植酸是植物种子中磷的主要储存形式,单胃动物难以消化。植酸在动物饲料中的降解是克服环境和营养问题所必需的。1d-肌醇-3-磷酸[Ins(3)P-1]合酶(EC 5.5.1.4)催化肌醇生物合成的第一步并指导种子中植酸的生物合成。水稻Ins(3)P-1合成酶基因(RINO 1)在发育中的种子胚和糊粉层中高度表达,其中植酸在糊粉层中合成和储存。在水稻种子中,18-kDa油质蛋白(Ole 18)以种子特异性方式表达,其转录物仅限于胚和糊粉层。因此,为了有效地抑制植酸的生物合成,反义RINO 1 cDNA在Ole 18启动子的控制下表达,在转基因水稻植物的种子中指导与RINO 1相同的空间模式。所产生的转基因水稻植株表现出强烈的低植酸(lpa)表型,其中种子植酸减少了68%,同时游离有效磷增加。未观察到对种子重量、发芽或植物生长的负面影响。稳定转基因植株的有效磷水平超过了目前可利用的水稻lpa突变体。
Phytic acid acts as the major storage form of phosphorus in plant seeds and is poorly digested by monogastric animals. The degradation of phytic acid in animal diets is necessary to overcome both environmental and nutritional issues. The enzyme 1d-myo-inositol 3-phosphate [Ins(3)P-1] synthase (EC 5.5.1.4) catalyses the first step of myo-inositol biosynthesis and directs phytic acid biosynthesis in seeds. The rice Ins(3)P-1 synthase gene (RINO1) is highly expressed in developing seed embryos and in the aleurone layer, where phytic acid is synthesized and stored. In rice seeds, 18-kDa oleosin (Ole18) is expressed in a seed-specific manner, and its transcripts are restricted to the embryo and the aleurone layer. Therefore, to effectively suppress phytic acid biosynthesis, antisense RINO1 cDNA was expressed under the control of the Ole18 promoter, directing the same spatial pattern in seeds as RINO1 in transgenic rice plants. The generated transgenic rice plants showed strong 'low phytic acid' (lpa) phenotypes, in which seed phytic acid was reduced by 68% and free available phosphate was concomitantly increased. No negative effects on seed weight, germination or plant growth were observed. The available phosphate levels of the stable transgenic plants surpassed those of currently available rice lpa mutants.