On the production, fusion, and function of pollen protoplasts

花粉原生质体的产生、融合和功能

• 批准号: 05640748
• 负责人: ITO Michio
• 金额: $ 1.28万
• 依托单位: Shizuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05640748/
• 关键词: Cell fusion; Gene introduction; Pollen; protoplasts; 花粉プロトプラスト; 受粉; 花粉管

Enzymatic isolation of protoplasts from mature pollen in various species of higher plants, their culture and fusion and their functional expression after the regeneration of cell wall were studied.1.Isolation of protoplasts from pollen in Angiosperms. The protoplasts were obtained from 8 dicotyledon species within more than 100 species tried, whereas 27 within 31 species in monocotyleadons.2.In Lilium longiflorum, pollen protoplasts were cultured for 2 days and placed on the stigma. Pollen tubes elongated into their pistil through ovary, and resulted in the seed formation.3.In fusion of lily protoplasts, the regeneration of cell wall, elongation of pollen tubes and the division of generative cells showed normal behavior. The number of of pollen tubes was usally one, and vegetative nuclei preceeded in the tubes. In protoplast fusion between defferent species, usually one pollen tube elongated from the fusion body.4.In fusion between pollen and mesohyll protoplasts, the function as pollen was expressed after the regeneration of cell wall.5.A b-glucuronidase gene was successfully expressed in pollen of Lilium longiflorum using a pneumatic particle gun.

本文研究了高等植物成熟花粉原生质体的酶法分离、培养、融合及细胞壁再生后的功能表达。1.被子植物花粉原生质体的分离。在100余种植物中，双子叶植物有8种获得了原生质体，单子叶植物有31种有27种获得了原生质体。在百合原生质体融合过程中，细胞壁再生、花粉管伸长和生殖细胞分裂均表现正常。花粉管数目多为1个，营养核早于花粉管。在不同种间原生质体融合中，融合体中通常只长出一根花粉管; 4.在花粉与叶肉原生质体融合中，细胞壁再生后具有花粉功能; 5.利用气动粒子枪在麝香百合花粉中成功表达了β-葡萄糖醛酸酶基因。

项目成果

Nisihara, M., Ito,M., Tanaka, I., et al.: "Expression of the b-glucunonidase gene in pollen of lily, Nicotiana rustica,and peony by particle bombardment" Plant Physiology. 102. 357-361 (1993)

Nisihara, M.、Ito,M.、Tanaka, I. 等人：“通过粒子轰击在百合、烟草和牡丹花粉中表达 β-葡萄糖苷酶基因”植物生理学。

伊藤道夫: "花粉プロトプラストの育種的意義" 植物遺伝. 4. 137-145 (1994)

伊藤道雄：“花粉原生质体的育种意义”《植物遗传学》4. 137-145 (1994)。

Ito, M., Koizumi, M., Nishihara, M.: "Germination of pollen protoplasts on stigma in Lilium longiflorum" Plant Cell. 27. 89-94 (1994)

Ito, M.、Koizumi, M.、Nishihara, M.：“长花百合柱头上花粉原生质体的萌发”植物细胞。

M.Nishihara, M.Ito, I.Tanaka, M.Kyo, K.Ono and H.Morikawa: "Expression of the b-glucuronidase gene in pollen of lily, tabacco, Nicotiana restica, and peony by particle bombardment." Plant Physiology. 102. 357-361 (1993)

M.Nishihara、M.Ito、I.Tanaka、M.Kyo、K.Ono 和 H.Morikawa：“通过粒子轰击在百合、烟草、烟草和牡丹花粉中表达 β-葡萄糖醛酸酶基因。”

Nisihara,M.Ito,M.et al.: "Expresionof the b-glucuronidase gene in pollen of lily,tobacco,Nicotiana rustica,and peony by particle bombardment" Plant Physiology. 102. 357-361 (1993)

Nisihara，M.Ito，M.et al.：“通过粒子轰击在百合、烟草、烟草和牡丹花粉中表达 β-葡萄糖醛酸酶基因”植物生理学。