Molecular mechanism of the recognition of the style to pollen with discriminate S-genotype in gametophytic self-incompatibility of apple

苹果配子体自交不亲和性S基因型花粉花柱识别的分子机制

基本信息

批准号：
15580088
负责人：
OKUNO Toshikatsu
金额：
$ 2.43万
依托单位：
Hirosaki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

To elucidate a molecular mechanism of gametophytic self-incompatibility of apple the interactive proteins to the S-RNase were screened from the pollen with distinctive S-genotype. The used varieties of apples is Stalking-Delicious (SD)(S-genotype : Sc, Se), Golden-Delicious (GD)(Sa, Sb).○Se-and Sc-RNase as S-RNase from SD were purified by our previously described method.○From germinated pollen of the varieties, SD and GD, crude extracts of soluble proteins and total RNA fractions were prepared.○The specific activities of the purified RNases were compared with other RNases (RNase A and T2) by using various RNA as substrate. The S-RNases showed very low values in specific activity than that of RNase A.○An affinity column of which Se-RNase bound to Hi-Trap (Pharmacia) was prepared. The crude pollen extracts were treated with the column and two absorbed proteins showed molecular mass of 48 and 66 KD on SDS-PAGE were obtained These proteins were confirmed to be a substance which interacts s … More pecifically with Se S-RNase by the immunodetection using Se-RNase antibody. Both substances, however, did not change the activity of the S-RNase to the tube elongation of pollen with the same S-genotype or alternative one.○The proteins, 48 and 66 KD of Mr, were also obtained by immune precipitation method.○The interaction of Se-RNase with soluble pollen proteins was examined by surface plasm on resonance method using Biacore 2000 (Biacore) with a Se-RNase binding sensor tip. For all fractions separated by DE chromatography of pollen crude extracts the protein interactions were measured. Only two fractions containing 48 or 66 KD of Mr were showed large Ka values and slow depression of Kd.In this studies the presence of interactive proteins to S-RNase from pollen extracts were confirmed by affinity column, immune precipitation, and surface plasmon resonance methods. These substances, however, did not inhibit the RNase activity of S-RNase in vitro an assay system, and therefore, additional substances derived from pollen may need to recognize the difference between self and cross pollen. Less

为了阐明苹果配子体自交不亲和的分子机制，从具有独特S基因型的花粉中筛选了与S-RNase相互作用的蛋白，所用的苹果品种是Stalking-Delicious（SD）（S基因型：Sc， Se)、金冠 (GD)(Sa、Sb)。○Se-和 Sc-RNase 作为来自 SD 的 S-RNase，通过我们之前描述的方法纯化。○来自发芽的种子制备了品种花粉、SD 和 GD、可溶性蛋白粗提物和总 RNA 组分。○以各种 RNA 为底物，将纯化的 RNase 与其他 RNase（RNase A 和 T2）的比活性进行比较。与RNase A相比，RNases的比活性值非常低。○制备了Se-RNase与Hi-Trap（Pharmacia）结合的亲和柱，用该柱处理粗花粉提取物并进行处理。获得了两种被吸收的蛋白质，其在 SDS-PAGE 上显示分子量为 48 和 66 KD。通过使用 Se-RNase 抗体的免疫检测，确认这些蛋白质是与 Se S-RNase 特异性相互作用的物质。然而，并没有改变具有相同S基因型或替代基因型的花粉管伸长的S-RNase活性。○Mr的48和66 KD蛋白也通过免疫沉淀法。○使用带有 Se-RNase 结合传感器尖端的 Biacore 2000 (Biacore) 通过表面等离子体共振法检查 Se-RNase 与可溶性花粉蛋白的相互作用。测量了蛋白质相互作用。仅含有 48 或 66 KD Mr 的两个级分显示出较大的 Ka 值和缓慢的 Kd 降低。在这项研究中，花粉中存在与 S-RNase 相互作用的蛋白质。通过亲和柱、免疫沉淀和表面等离振子共振方法证实了提取物，但是，这些物质在体外测定系统中并不抑制S-RNase的RNase活性，因此，可能需要从花粉中提取的其他物质来识别。自花粉和杂交花粉之间的差异较小。

项目成果

期刊论文数量（30）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Biosynthesis of Lambertellols Based on the High Specific Incorporation of the 13-C labeled Acetate and Their Biological Properties

基于13-C标记乙酸盐高特异性掺入的兰伯特醇生物合成及其生物学性质

DOI：
发表时间：
2004
期刊：
J.Amer.Chem.Soc. 126(6)
影响因子：
0
作者：
Ken-ichi Nihei;Kimiko Hashimoto;Kazuo Miyairi;Toshikatsu Okuno;T.Murakami et al.
通讯作者：
T.Murakami et al.

S-RNase from Self-incompatible and - compatible Cultivars : purification, Cloning Enzymic Properties, and Pollen tube Growth

来自自交不亲和和亲和品种的 S-RNase：纯化、克隆酶特性和花粉管生长

DOI：
发表时间：
2002
期刊：
Biosci.Biotechnol.Biochem. 66(6)
影响因子：
0
作者：
Takanori Murakami;Yoshifumi Morikawa;Masaru Hashimoto;Toshikatsu Okuno;Yukio Harada;J.Fujita et al.;N.katoh et al.
通讯作者：
N.katoh et al.

Synthesis of thiosaccharides employing the Pummerer rearrangement of tetrahydrothiopyran oxides

DOI：
10.1016/j.tet.2004.06.006
发表时间：
2004-08
期刊：
Tetrahedron
影响因子：
2.1
作者：
Junji Fujita;H. Matsuda;Kazunori Yamamoto;Yasuharu Morii;M. Hashimoto;T. Okuno;K. Hashimoto
通讯作者：
Junji Fujita;H. Matsuda;Kazunori Yamamoto;Yasuharu Morii;M. Hashimoto;T. Okuno;K. Hashimoto

Lamberteroll C a labile and novel Biosynthetic congener of lambertellols A and B.

Lamberteroll C 是兰伯特醇 A 和 B 的不稳定且新颖的生物合成同系物。

DOI：
发表时间：
2005
期刊：
Bioorg Med.Chem.Lett. 15
影响因子：
0
作者：
Takanori Murakami;Akane Sasaki;Ed Fukushi;Jun Kawabata;Masaru Hashimoto;Toshikatsu Okuno
通讯作者：
Toshikatsu Okuno

Enantioselective Synthesis of Four Isomers of 3-Hydoroxy-4-Methyltertradecanoic Acid, the Constituent of antifungal Cyclodepsipeptides W493 A and B

抗真菌环缩肽 W493 A 和 B 的组成部分 3-羟基-4-甲基十四烷酸的四种异构体的对映选择性合成

DOI：
发表时间：
2005
期刊：
Biosci.Biotechnol.Biochem. 69
影响因子：
0
作者：
Nabeta;K.;katoh.K;Nabeta;K.;Takanori Murakami et al.;Takanori Murakami et al.;Ken-ichi Nihei et al.
通讯作者：
Ken-ichi Nihei et al.