GUS基因转入烟草细胞通过科间细胞共培养在胡萝卜再生植株中的表达

用Ａｇｒｏｂａｃｔｅｒｉｕｍｔｕｍｅｆａｃｉｅｎｓ菌株ＬＢＡ４４０４（含有质粒ｐＢＩ１２１和ｐＡＬ４４０４）介导转化普通烟草（Ｎｉｃｏｔｉａｎａｔａｂａｃｕｍ）的悬浮细胞，获得卡那霉素抗性（ＫｍＲ）材料，其再生植株中表现ＧＵＳ活性。以这一继代半年以上的转基因烟草细胞系为材料，与胡萝卜（Ｄａｕｃｕｓｃａｒｏｔａ）细胞进行科间细胞共培养和低渗活化等处理，在含２００ｍｇ·ｌ－１卡那霉素（Ｋｍ）的培养基上筛选，得到具有ＫｍＲ的胡萝卜田胞系（ｃｅＲ），其再生的胡萝卜植株中也有一部分表现为ＫｍＲ，并在部分植株中检测到ＧＵＳ活性。这一结果表明Ｔ—ＤＮＡ可以通过科间细胞的共培养而从转基因烟草细胞中通过胞间通道转移到胡萝卜细胞中，并在胡萝卜细胞中表达。从这一结果来看，细胞共培养和有关处理的技术（ＡＣＨＴ）可能会成为植物基因转移的新技术。同时，发现共培养可以改进原来不亲和细胞之间的亲和性，这为植物细胞的识别和亲和性研究、亲和性的改善提供了方法和技术。

CUS Gene Can Be Transferred from Transgenic Tobacco Callus Cell into Carrot Callus Cell via Interfamilial Plant Cell Co Culture and Expressed in the Carrot Regeneration Plants

Tobacco(Nicotiana tabacum) suspension cells were transformed with Agrobacterium tumefaciens strin LBA 4404 containing pBI 121(Jeferson,1987)and pAL 4404, and kanamycin ─resistant(KmR) calli were produced on solid selective medium containing 250～300mg/L of kanamycin(Km) and 500～600/L of carbenicillin(Cb). A stable transgenic tobacco cell line was obtained after six months'subculture of the KmR tobacco calli on the selective medium containing Km and Cb. β─Glucuronidase(GUS) activities were detected histochemically in transgenit tobacco plants regenerated and in regenerating calli from the transgenic tobacco cell line. Carrot (Daucus carota) callus cells were co─cultured interfamilially with the stable transgenic tobacco cell line, treated with activated k+ hypotonic solution, and selected on the solid medium containing 200mg/L of Km,and KmR carrot cell lines were obtained. Some of the carrot plants regenerated from the KmR carrot cell lines were kanamycin resistant, and some exhibited GUS activity.These results indicate that the GUS reporter gene can be transferred interfamilially through intercellular channel (cytomictic channel) from transgenic tobacco cells to carrot cells via interfamilial plant cell co─culture and expressed in the carrot cells, and the method of cell co ─ culture in combination with activated hypotonic treatement developed here might be a useful approach to transfer or introduce genes from a species of line to another. In addition, it was found that the incompatibility between cells from different parents could be improved via cell co─culture, so the method presented here may be used to study plant cell recognition,plant cell compatibility, and improvement in incompatibility.