VASCULAR BUNDLE-SPECIFIC LOCALIZATION OF CYTOSOLIC GLUTAMINE-SYNTHETASE IN RICE LEAVES
VASCULAR BUNDLE-SPECIFIC LOCALIZATION OF CYTOSOLIC GLUTAMINE-SYNTHETASE IN RICE LEAVES
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DOI：
10.1104/pp.99.4.1481
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发表时间：
1992-08-01
期刊：
影响因子：
7.4
通讯作者：
OJIMA, K
中科院分区：
文献类型：
--
作者：
KAMACHI, K;YAMAYA, T;OJIMA, K
水稻（Oryza sativa L.）胞质谷氨酰胺合成酶（GS 1; EC 6.3.1.2）、叶绿体GS（GS 2）和铁氧还蛋白依赖性谷氨酸合酶（Fd-GOGAT; EC 1.4.7.1）的组织定位使用具有特异性抗体的组织印迹免疫印迹方法研究叶片。将来自中部和基部区域的成熟和衰老叶片的横截面用于组织打印。抗GS1抗体，提出了对合成的17个残基的肽对应于推导的N-末端氨基酸序列的水稻GS1，交叉反应特异性与天然GS1蛋白质，但不与GS 2转移到硝酸纤维素膜上后。组织印迹免疫印迹表明，GS1蛋白位于大，小维管束在所有地区的叶片准备从任何阶段的成熟。GS 2和Fd-GOGAT蛋白主要定位于叶肉细胞中。GS1的膜上的颜色的强度在两个叶龄之间是相似的，而GS 2和Fd-GOGAT在衰老过程中下降。GS 1的组织特异性定位表明，这种GS异构体在谷氨酰胺的合成中是重要的，谷氨酰胺是水稻植株衰老叶片输出氮的主要形式。
Tissue localizations of cytosolic glutamine synthetase (GS1; EC 6.3.1.2), chloroplastic GS (GS2), and ferredoxin-dependent glutamate synthase (Fd-GOGAT; EC 1.4.7.1) in rice (Oryza sativa L.) leaf blades were investigated using a tissue-print immunoblot method with specific antibodies. The cross-sections of mature and senescent leaf blades from middle and basal regions were used for tissue printing. The anti-GS1 antibody, raised against a synthetic 17-residue peptide corresponding to the deduced N-terminal amino acid sequence of rice GS1, cross-reacted specifically with native GS1 protein, but not with GS2 after transfer onto a nitrocellulose membrane. Tissue-print immunoblots showed that the GS1 protein was located in large and small vascular bundles in all regions of the leaf blade prepared from either stage of maturity. On the other hand, GS2 and Fd-GOGAT proteins were mainly located in mesophyll cells. The intensity of the developed color on the membrane for GS1 was similar between the two leaf ages, whereas that for GS2 and Fd-GOGAT decreased during senescence. The tissue-specific localization of GS1 suggests that this GS isoform is important in the synthesis of glutamine, which is a major form of nitrogen exported from the senescing leaf in rice plants.