Brassica oleracea cv. Pride of India is one of the most promising vegetable cultivars commercially grown as cash crop in Himachal Pradesh, India. However, its overall production is severely hampered by diamondback moth (Plutella xylostella), a notorious pest. To avoid yield losses caused by this pest, cryIAa gene was introduced into cabbage cv. Pride of India using Agrobacterium-mediated transformation method. In an attempt to maximize the transformation frequency, critical determinant factors such as explant type, pre-incubation and co-cultivation period, and acetosyringone effect were successfully optimized. The highest transformation frequency (4.67% and 14.50%) in cotyledon and hypocotyl explant was achieved with a pre-incubation period of 72 h and co-cultivation period of 48 h. Furthermore, transformation frequency was enhanced in cotyledon (18.66%) and hypocotyl (32.00%) explants, when selective regeneration medium was fortified with 100 µM acetosyringone, respectively. The transgene (cryIAa) integration and copy number were confirmed using PCR and Southern blotting. Reverse transcriptase PCR and quantitative real-time PCR analyses were performed that proved transcriptional expression of cryIAa gene in PCR-positive transgenic events. Transgenic cabbage-fed diamondback moth larvae showed significantly higher mortality, thereby proving transgene effectiveness against insect pest control.

中文翻译：

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卷心菜中cry1Aa基因的表达赋予了对小菜蛾（Plutella xylostella）的抗性

甘蓝cv. 印度的骄傲是在印度喜马偕尔邦作为经济作物商业种植的最有前途的蔬菜品种之一。然而，它的整体生产受到小菜蛾 ( Plutella xylostella ) 的严重阻碍，这是一种臭名昭著的害虫。为了避免这种害虫造成的产量损失，将 cryIAa基因引入卷心菜 cv. 印度使用农杆菌的骄傲介导的转化方法。为了最大限度地提高转化频率，成功优化了外植体类型、预孵育和共培养时间以及乙酰丁香酮效应等关键决定因素。子叶和下胚轴外植体的最高转化频率（4.67%和14.50%）是在72 h的预培养期和48 h的共培养期实现的。Furthermore, transformation frequency was enhanced in cotyledon (18.66%) and hypocotyl (32.00%) explants, when selective regeneration medium was fortified with 100 µM acetosyringone, respectively. 转基因（cryIAa) 整合和拷贝数使用 PCR 和 Southern 印迹进行确认。进行了逆转录酶 PCR 和定量实时 PCR 分析，证明了cryIAa基因在 PCR 阳性转基因事件中的转录表达。转基因卷心菜喂养的小菜蛾幼虫表现出显着更高的死亡率，从而证明了转基因对害虫控制的有效性。