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首页> 外文期刊>Breeding science >Development and bioassay of transgenic Chinese cabbage expressing potato proteinase inhibitor II gene.
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Development and bioassay of transgenic Chinese cabbage expressing potato proteinase inhibitor II gene.

机译：表达马铃薯蛋白酶抑制剂II基因的转基因大白菜的开发和生物测定。

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摘要

Lepidopteran larvae are the most injurious pests of Chinese cabbage production. We attempted the development of transgenic Chinese cabbage expressing the potato proteinase inhibitor II gene (pinII) and bioassayed the pest-repelling ability of these transgenic plants. Cotyledons with petioles from aseptic seedlings were used as explants for Agrobacterium-mediated in vitro transformation. Agrobacterium tumefaciens C58 contained the binary vector pBBBasta-pinII-bar comprising pinII and bar genes. Plants showing vigorous PPT resistance were obtained by a series concentration selection for PPT resistance and subsequent regeneration of leaf explants dissected from the putative chimera. Transgenic plants were confirmed by PCR and genomic Southern blotting, which showed that the bar and pinII genes were integrated into the plant genome. Double haploid homozygous transgenic plants were obtained by microspore culture. The pinII expression was detected using quantitative real time polymerase chain reaction (qRT-PCR) and detection of PINII protein content in the transgenic homozygous lines. Insect-feeding trials using the larvae of cabbage worm (Pieris rapae) and the larvae of the diamondback moth (Plutella xylostella) showed higher larval mortality, stunted larval development, and lower pupal weights, pupation rates, and eclosion rates in most of the transgenic lines in comparison with the corresponding values in the non-transformed wild-type line.
机译：鳞翅目幼虫是大白菜生产中最有害的害虫。我们尝试开发表达马铃薯蛋白酶抑制剂II基因（pinII）的转基因大白菜，并生物测定了这些转基因植物的害虫排斥能力。子叶和来自无菌幼苗的叶柄被用作农杆菌介导的体外转化的外植体。根癌农杆菌C58含有包含pinII和bar基因的二元载体pBBBasta-pinII-bar。通过对PPT抗性进行一系列的浓度选择以及随后从推定的嵌合体解剖的叶外植体的再生，获得显示出强烈的PPT抗性的植物。通过PCR和基因组Southern印迹证实了转基因植物，这表明bar和pinII基因已整合到植物基因组中。通过小孢子培养获得双单倍体纯合的转基因植物。使用定量实时聚合酶链反应（qRT-PCR）检测pinII表达，并检测转基因纯合株系中PINII蛋白含量。使用白菜蠕虫幼虫和小菜蛾幼虫（小菜蛾）的昆虫喂养试验表明，在大多数转基因动物中，幼虫死亡率更高，发育迟缓的幼虫发育，and重量，化脓率和出苗率较低。与未转换的野生型行中的相应值进行比较。

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