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首页> 外文期刊>Current Science: A Fortnightly Journal of Research >Agrobacterium-me&iated genetictransformation of Nagpur mandarin(Citrus reticulata Blanco)
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Agrobacterium-me&iated genetictransformation of Nagpur mandarin(Citrus reticulata Blanco)

机译：农杆菌介导的那格浦尔柑桔的遗传转化

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摘要

Genetic transformation protocol was developed forNagpur mandarin (Citrus reticulata Blanco), a choicestcitrus variety grown in India and South East Asia.Cotyledon segments from mature seeds were co-culti-vated with Agrobacterium tumefaciens for two daysand cultured on an adventitive embryo induction medium containing MS basal medium supplementedwith 8.87 muM BAP + 2.32 muM kinetin + 5.37 muM NAAwith 60 mug/ml kanamycin and 300 |ag/ml cefotaxime.The putatively transformed adventitive embryos that re-generated on the cotyledon segment were transferredfor complete plantlet development on auxin-rich me-dium having MS basal salts fortified with 9.80 muMIB A and 100 mg 1~(-1)activated charcoal. The transgenicplants were then hardened in glass jars filled withsoilrite and perlite (1:1). GUS activity was detected inputatively transformed plants and maximum positiveGUS assay frequency 34.5 % was noted. Genetic trans-formation was confirmed by PCR analysis and Southernhybridization with alpha-~(32)P-labelled riboprobes of thegus gene.
机译：为印度和东南亚地区生长的柑橘选择种纳格普尔柑（Citrus reticulata Blanco）开发了遗传转化方案，将成熟种子的子叶片段与根癌农杆菌共培养了两天，并在含有MS的不定胚诱导培养基上培养补充8.87μMBAP + 2.32μMkinetin + 5.37μMNAA的基础培养基，其中60杯/毫升卡那霉素和300μg/ ml头孢噻肟。将在子叶节上再生的推定转化的外胚胚转移，以在富含生长素的我上完成完整的植株发育具有9.80 muMIBA A和100​​ mg 1〜（-1）活性炭强化的MS基础盐的钠盐。然后将转基因植物在装满土壤和珍珠岩（1：1）的玻璃罐中硬化。检测到输入转化的植物中的GUS活性，注意到最大阳性GUS分析频率为34.5％。通过PCR分析和α-〜（32）P标记的thegus基因核糖核酸的Southern杂化证实了遗传转化。

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