Plant Cell Culture Technology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400 085, India.
Biol Trace Elem Res. 2011 Dec;144(1-3):1219-28. doi: 10.1007/s12011-011-9092-z. Epub 2011 May 31.
Pineapple (Ananas comosus L. Merr., cv. "Queen") leaf bases were transformed with Agrobacterium tumefaciens strain EHA 105 harboring the pSF and pEFESF plasmids with soybean ferritin cDNA. Four to eight percent of the co-cultivated leaf bases produced multiple shoots 6 weeks after transfer to Murashige and Skoog's medium supplemented with α-naphthalene acetic acid 1.8 mg/l, indole-3-butyric acid 2.0 mg/l, kinetin 2.0 mg/l, cefotaxime 400 mg/l, and kanamycin 50 mg/l. Putatively transformed shoots (1-2 cm) were selected and multiplied on medium of the same composition and elongated shoots (5 cm) were rooted on liquid rooting medium supplemented with cefotaxime 400 mg/l and kanamycin 100 mg/l. The rooted plants were analyzed through PCR, genomic Southern analysis, and reverse transcription PCR. The results clearly confirmed the integration and expression of soybean ferritin gene in the transformed plants. Atomic absorption spectroscopic analysis carried out with six independently transformed lines of pSF and pEFE-SF revealed a maximum of 5.03-fold increase in iron and 2.44-fold increase in zinc accumulation in the leaves of pSF-transformed plants. In pEFE-SF-transformed plants, a 3.65-fold increase in iron and 2.05-fold increase in zinc levels was observed. Few of the transgenic plants were hardened in the greenhouse and are being grown to maturity to determine the enhanced iron and zinc accumulation in the fruits. To the best of our knowledge this is the first report on the transformation of pineapple with soybean ferritin for enhanced accumulation of iron and zinc content in the transgenic plants.
菠萝（Ananas comosus L. Merr.，cv. “Queen”）叶片基部被携带大豆铁蛋白 cDNA 的根癌农杆菌菌株 EHA105 转化。在含有 1.8mg/lα-萘乙酸、2.0mg/l 吲哚丁酸、2.0mg/l 激动素、400mg/l 头孢噻肟和 50mg/l 卡那霉素的 Murashige 和 Skoog 培养基上，4%至 8%的共培养叶片基部在转移后 6 周产生多个芽。将疑似转化的芽（1-2cm）选择并在相同组成的培养基上增殖，将伸长的芽（5cm）在含有 400mg/l 头孢噻肟和 100mg/l 卡那霉素的液体生根培养基上生根。通过 PCR、基因组 Southern 分析和反转录 PCR 对生根植物进行分析。结果清楚地证实了大豆铁蛋白基因在转化植物中的整合和表达。对 pSF 和 pEFE-SF 的六个独立转化系进行原子吸收光谱分析表明，pSF 转化植物叶片中铁的积累增加了 5.03 倍，锌的积累增加了 2.44 倍。在 pEFE-SF 转化植物中，铁的积累增加了 3.65 倍，锌的积累增加了 2.05 倍。一些转基因植物在温室中进行了硬化，并正在生长成熟，以确定果实中铁和锌的积累增加。据我们所知，这是首次报道利用大豆铁蛋白转化菠萝，以提高转基因植物中铁和锌的含量。
