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【答案】应助回帖
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gruyclewee: 金币+3, 感谢应助，奖励一下。 2014-02-15 10:10:14
zhhuijun: 金币+5, ★★★很有帮助, 谢谢啊 2014-02-16 10:59:46
An efficient selection and regeneration protocol for Agrobacterium-mediated transformation of oriental melon (Cucumis melo L. var. makuwa)
作者:Choi, JY (Choi, Jun Young)[ 1,2 ] ; Shin, JS (Shin, Jeong Sheop)[ 2 ] ; Chung, YS (Chung, Young Soo)[ 3 ] ; Hyung, NI (Hyung, Nam-In)[ 1 ]
PLANT CELL TISSUE AND ORGAN CULTURE
卷: 110 期: 1 页: 133-140
DOI: 10.1007/s11240-012-0137-6
出版年: JUL 2012
查看期刊信息
摘要
An efficient selection and plant regeneration protocol for Agrobacterium-mediated transformation using cotyledon explants of oriental melon (Cucumis melo L. var. makuwa) has been developed. All six oriental melon cultivars evaluated in the study showed a > 90 % shoot regeneration frequency and produced 1.8-3.6 shoots per cotyledon explant when cultured on Murashige and Skoog (MS) medium supplemented with 1.0 mg L-1 benzyladenine and 0.01 mg L-1 indoleacetic acid. Kanamycin (Km) and geneticin (Gt) in the shoot induction medium (SIM) were compared both qualitatively and quantitatively for their efficiency as a selection agent for the selection and regeneration of transgenic plants after Agrobacterium-mediated transformation. Shoot formation was completely inhibited at 50 mg L-1 Km and 10 mg L-1 Gt. Relatively high concentrations of both Gt and Km (> 100 mg L-1 Km and > 25 mg L-1 Gt) were necessary because large numbers of non-transgenic shoots survived during the selection process. The incorporation of a selectable marker (neomycin phosphotransferase II) into the genome of transgenic plants was confirmed using beta-glucuronidase (GUS), PCR and Southern blot analysis. Shoot regeneration frequencies were 41.2 % at 100 mg L-1 Km and 15.2 % at 30 mg L-1 Gt 8 weeks after transformation, whereas the transformation frequencies based on the PCR were 2.9 and 7.1 %, respectively, 16 weeks after transformation. These results demonstrate that a large portion of the regenerated shoots on SIM supplemented with 100 mg L-1 Km consisted of non-transformed or escaped shoots, indicating that 30 mg L-1 Gt is the more suitable for the selection and regeneration of transgenic plants in oriental melon.
关键词
作者关键词:Oriental melon; Transformation; Kanamycin; Geneticin; Selection
KeyWords Plus:COLOCYNTHIS-CITRULLUS L.; PLANT-REGENERATION; SHOOT REGENERATION; GENETIC-TRANSFORMATION; LEAF EXPLANTS; ORGANOGENESIS; RESISTANCE; CULTIVARS; MUSKMELON; GENOTYPE
作者信息
通讯作者地址: Hyung, NI (通讯作者)
Sangmyung Univ, Dept Plant Sci & Technol, Cheonan 330720, South Korea.
地址:
[ 1 ] Sangmyung Univ, Dept Plant Sci & Technol, Cheonan 330720, South Korea
[ 2 ] Korea Univ, Sch Life Sci & Biotechnol, Seoul 136701, South Korea
[ 3 ] Dong A Univ, Dept Genet Engn, Pusan 604714, South Korea
电子邮件地址:nihyung@smu.ac.kr
出版商
SPRINGER, VAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDS
类别 / 分类
研究方向:Biotechnology & Applied Microbiology; Plant Sciences
Web of Science 类别:Biotechnology & Applied Microbiology; Plant Sciences
文献信息
文献类型:Article
语种:English
入藏号: WOS:000304664300014
ISSN: 0167-6857
其他信息
IDS 号: 950QR
Web of Science 核心合集中的 "引用的参考文献": 35
Web of Science 核心合集中的 "被引频次": 3 |
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