J Integr Plant Biol. ›› 2001, Vol. 43 ›› Issue (10): 1055-1057.

• Research Articles • Previous Articles     Next Articles

Microinjection of Genes into in Vitro Ovaries of Maize and Identification of Transformed Plants

SHEN Shi-Hua, ZHANG Xiu-Jun, GUO Yi-Ming and JING Yu-Xiang   

Abstract:

Transformation of gene into maize ( Zea mays L.) ovaries by microinjection has been successfully achieved. However, this method was laborious and was carried out in the field which led to serious contamination of the ovaries, low ratio of transformation and was significantly influenced by lower atmospheric temperature causing seed immaturity. The method of in vitro injection was used in which the maize ears were removed from the plant after they were pollinated for 24 h, then, the ear coats were stripped off to expose the ovaries in which several μL of transformed gene solution were injected in clean bench area using glass microneedle. After all the ovaries were injected, the ear was cut into several segments, which then were put into a glass bottle containing culture medium and incubated in a growth chamber with fluorescent lamps. Seeds could be directly obtained from the segments in three to four weeks after culture. This method overcomes the shortcomings of the in vivo seed culture and increases the transformation efficiency. Transformed plants after the seed germination were screened by antibiotic selection and identified at molecular levels, indicating that transgenic plants with positive antibiotic resistance, and confirmed by PCR, Southern blotting were obtained.

玉米基因转化的离体子房注射及其转基因植株的鉴定
沈世华 张秀君 郭奕明 荆玉祥*

(中国科学院植物研究所,北京100093)


摘要:用微量注射法对玉米( Zea mays L.)进行基因转化已获得成功, 但这种方法是在田间进行的,污染较大, 操作时劳动强度大,转化率低,而且受气温影响,气温下降则种子不能成熟.采用离体注射培养法,取下授粉24 h后的玉米雌穗,剥去苞叶,在超净工作台进行微量注射,然后切成小块放在培养瓶中,在光照培养箱内培养,3~4周可直接获得种子.这种方法克服了非离体培养的缺点,提高了转化效率.种子萌发后进行植株抗性筛选和分子检测, 得到了具抗性的、PCR 和Southern杂交结果均为阳性的转基因植株.

关键词: 玉米;基因转化;离体子房注射;基因鉴定

Key words: Zea mays , gene transformation, in vitro injection of maize ovaries, gene identification

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