Production of Chimeral Thalli by Protoplast Fusion in Porphyra yezoensis and P.tenera

条斑紫菜和细嫩紫菜原生质体融合产生嵌合菌体

• 批准号: 62560196
• 负责人: ARAKI Toshiyoshi
• 金额: $ 1.28万
• 依托单位: Mie University (1988)Kyushu University (1987)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62560196/
• 关键词: -1,4-Mannanase; -1,3-Xylanase; Porphyranase; Protoplast; Cell fusion; Chimera; Porphyra tenera; キメラ; β-1,3-キシラナーゼ; β-1,4-マンナナーゼ

Cell fusion is the superior technique to improve the quality of plants. In this study was tried to produce chimeral thallus from a red sea algae, Porphyra, by cell fusion. For carrying out cell fusion it is necessary to produce a number of protoplasts. We searched bacteria capable of degrading the cell wall of algae from natural habitats, and isolated 3 strains of bacteria which could produce a lot of cell wall-degrading enzymes ( -1,4-mannanase, -1,3-xylanase, and porphyranase). A green thallus of the spontaneous mutant was also obtained from the wild type of P.tenera. A number of protoplasts from thalli of the wild type of . Yezoensis and the green type mutant of P.tenera by the treatment with -1,4-mannanase, -1,3-xylanase, and porphyranase. These protoplasts were induced to fuse by exposing them to polyethylene glycol (PEG). The PEG-treated protoplasts developed into calli of irregular shapes, and a large nummber of plantlets were formed from the calli. These plantlets grew into sectorially variegated chimeral thalli (chimera-I) composed of brown and green sectors, in addition to brown or green thalli. Microscopical observation revealed that green sector of chimera-I turned into brown mature cells. Some monospores released from chimera-I grew into chimeral thalli composed of brown and green, and others bedame brown or green thalli. A chimeral thallus developed from monospores of chimera-I released carpospores. These carpospores grew into conchocelis and produced conchospores. A 38.4 % of the conchospores grew into chimeral thalli composed of brown and green. Therefore, chimera-I was supposed to be a thallus developed from fusion cell of P.yezoensis and P.tenera.

细胞融合是提高植物品质的先进技术。在这项研究中，尝试通过细胞融合从红海藻类紫菜中产生嵌合叶状体。为了进行细胞融合，需要产生大量原生质体。我们从自然生境中寻找能够降解藻类细胞壁的细菌，分离出3株能够产生大量细胞壁降解酶（-1,4-甘露聚糖酶、-1,3-木聚糖酶、卟啉酶）的细菌。 。自发突变体的绿色菌体也从野生型的柔嫩 P.tenera 中获得。一些来自野生型菌体的原生质体。用-1,4-甘露聚糖酶、-1,3-木聚糖酶和卟啉酶处理的条斑紫菜和细嫩紫菜的绿色型突变体。通过将这些原生质体暴露于聚乙二醇（PEG）来诱导融合。经PEG处理的原生质体发育成不规则形状的愈伤组织，并由愈伤组织形成大量的植株。这些小植株长成扇形杂色的嵌合菌体（嵌合体-I），除了棕色或绿色菌体之外，还由棕色和绿色扇体组成。显微镜观察发现嵌合体-I的绿色部分转变成棕色的成熟细胞。嵌合体-I释放的一些单孢子长成由棕色和绿色组成的嵌合菌体，而另一些则生长为棕色或绿色菌体。嵌合叶状体由嵌合体-I释放的果孢子的单孢子发育而来。这些果孢子长成壳孢子并产生壳孢子。 38.4%的壳孢子长成由棕色和绿色组成的嵌合菌体。因此，chimera-I被认为是由条斑紫菜和柔嫩紫菜融合细胞发育而成的叶状体。

项目成果

Araki Toshiyoshi.: The Japanese Journal of Phycology(SORUI).

荒木俊良：日本藻类学杂志（SORUI）。

Takahiko Aoki: "Purification and Characterization of an Endo- -1,3-Xylanase from Vibrio sp." Nippon Suisan Gakkaishi. 54. 277-281 (1988)

Takahiko Aoki：“弧菌内切--1,3-木聚糖酶的纯化和表征”。