Reducing the content of nornicotine in tobacco via targeted mutation breeding
Reducing the content of nornicotine in tobacco via targeted mutation breeding
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DOI：
10.1007/s11032-007-9138-2
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发表时间：
2008-04-01
影响因子：
3.1
通讯作者：
de Borne, Francois Dorlhac
中科院分区：
文献类型：
--
作者：
Julio, Emilie;Laporte, Frederic;de Borne, Francois Dorlhac
种植烟草产生次生生物碱，参与亚硝胺的形成，对健康有影响。最近发现了尼古丁和去甲尼古丁生物合成途径中的靶基因，现在可以通过生物技术来控制它们。我们在这里证明，突变育种可以作为转基因(GM)植物的替代品，用于生产不含去甲基尼古丁的烟草。通过对1,311个M2烟草乙基甲烷磺酸(EMS)家系突变株的筛选，鉴定出NtabCYP82E4基因(烟碱N-脱甲基酶)的10个等位基因。生物碱分析表明，携带NtabCYP82E4无义或错义等位基因的纯合子M2植株的去甲烟碱含量很低或接近于零。与烟草优良品种回交产生的Bc1植株表型与亲本难以区分。烟草育种者在过去几十年中的这一主要目标可以在不到1.5年的时间内实现，包括创建高度诱变的烟草突变品种，并使用大多数育种公司和作物物种都能获得的简单和通用的检测技术(毛细管电泳-单链构象多态，CE-SSCP)检测突变的等位基因。
Cultivated tobacco produces secondary alkaloids involved in the formation of nitrosamines with health concerns. The recent identification of target genes in nicotine and nornicotine biosynthetic pathways now allows biotechnological approaches for their control. We demonstrate here that mutation breeding can be used as an alternative to genetically modified (GM) plants for generating nornicotine-free tobacco. Ten alleles of the NtabCYP82E4 gene (nicotine N-demethylase) were identified by screening 1,311 M2 families of tobacco ethylmethane sulphonate (EMS) mutants. Alkaloid analysis indicated that the nornicotine contents of homozygous M2 plants carrying nonsense or missense alleles of NtabCYP82E4 were very low or near-null. Backcrossing with tobacco elite varieties yielded BC1 plants phenotypically undistinguishable from parental lines. This major objective of tobacco breeders in the last few decades could be reached in a period of less than 1.5 years, including the creation of highly mutagenised tobacco mutant collections and the detection of mutated alleles using a simple and versatile detection technology (capillary electrophoresis-single strand conformation polymorphism, CE-SSCP) accessible to most breeding companies and crop species.