利用卡那霉素间接鉴定法进行大规模的棉花转基因育种技术

:( 1 )将脱脂棉撕成小条沾取 0 .0 5%的卡那霉素溶液 ,粘附于棉花植株倒 2新生叶上。 5d后所有沾有卡那霉素溶液的感虫棉株的叶片均出现明显的黄色斑块 ,而转 Bt基因抗虫棉的叶片仍为正常绿色 ,无任何症状。 ( 2 )转 Bt基因抗虫棉种子接种于加有 1 0 0 0 mg· l- 1卡那霉素的 MS培养基中。四天后 ,抗虫棉子叶均表现为正常绿色 ,而常规品种子叶全为黄色 ,表明卡那霉素间接鉴定法是鉴定转 Bt基因抗虫棉的简便而又准确的方法

Selection Technique for Transgenic Bt Cotton Using Kanamycin as an Indirect Identification Marker

A simple, cheap, and efficient breeding technique for transgenic Bt cotton using kanamycin as an indirect identification marker both in the field and in the laboratory is presented. For field selection, small strips of absorbent cotton were soaked with 0.05% kanamycin solution, and placed on second neonate expanded leaves. Five days later, all of the leaves of the susceptible plants showed apparent yellow patches whereas the leaves of the Bt resistant plants remained green without any symptom.Kanamycin can also be used in the laboratory to test whether or not a plant is resistant. Seed of transgenic Bt cotton were cultured on MS medium containing 1000 mg·L-1 kanamycin solution for four days.The cotyledons of resistant varieties remained normal green color, whilst those of the commercial varieties all turned yellow. This technique is especially useful for the purity test of transgenic Bt hybrid before the seed is sold.A homozygous transgenic line for target genes could be developed quickly using both field and laboratory tests.