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ÄÄλÅóÓѰïÎÒ²éÒ»ÏÂÎÄÕÂAn efficient regeneration protocol for Agrobacterium-mediated transformation of melon (Cucumis melo L.)
H.J. Zhang, P. Gao1, X.Z. Wang1 and F.S. Luan
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An efficient selection and regeneration protocol for Agrobacterium-mediated transformation of oriental melon (Cucumis melo L. var. makuwa)
×÷Õß: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
¾í: 110  ÆÚ: 1  Ò³: 133-140
DOI: 10.1007/s11240-012-0137-6
³ö°æÄê: JUL 2012
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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.
¹Ø¼ü´Ê
×÷Õ߹ؼü´Ê:Oriental melon; Transformation; Kanamycin; Geneticin; Selection
KeyWords Plus:COLOCYNTHIS-CITRULLUS L.; PLANT-REGENERATION; SHOOT REGENERATION; GENETIC-TRANSFORMATION; LEAF EXPLANTS; ORGANOGENESIS; RESISTANCE; CULTIVARS; MUSKMELON; GENOTYPE
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ͨѶ×÷ÕßµØÖ·: Hyung, NI (ͨѶ×÷Õß)
        Sangmyung Univ, Dept Plant Sci & Technol, Cheonan 330720, South Korea.
µØÖ·:
        [ 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
µç×ÓÓʼþµØÖ·:nihyung@smu.ac.kr
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SPRINGER, VAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDS
Àà±ð / ·ÖÀà
Ñо¿·½Ïò:Biotechnology & Applied Microbiology; Plant Sciences
Web of Science Àà±ð:Biotechnology & Applied Microbiology; Plant Sciences
ÎÄÏ×ÐÅÏ¢
ÎÄÏ×ÀàÐÍ:Article
ÓïÖÖ:English
Èë²ØºÅ: WOS:000304664300014
ISSN: 0167-6857
ÆäËûÐÅÏ¢
IDS ºÅ: 950QR
Web of Science ºËÐĺϼ¯ÖÐµÄ "ÒýÓõIJο¼ÎÄÏ×": 35
Web of Science ºËÐĺϼ¯ÖÐµÄ "±»ÒýƵ´Î": 3
2Â¥2014-02-15 08:51:41
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zhhuijun

ľ³æ (ÕýʽдÊÖ)

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2Â¥: Originally posted by jssxh at 2014-02-15 08:51:41
An efficient selection and regeneration protocol for Agrobacterium-mediated transformation of oriental melon (Cucumis melo L. var. makuwa)
×÷Õß:Choi, JY (Choi, Jun Young) ; Shin, JS (Shin, Jeong She ...

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3Â¥2014-02-16 11:01:15
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hezhao311

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4Â¥2014-02-17 11:41:17
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