AGROBACTERIUM-MEDIATED TRANSFORMATION OF APPLE (MALUS X DOMESTICA BORKH) - AN ASSESSMENT OF FACTORS AFFECTING GENE-TRANSFER EFFICIENCY DURING EARLY TRANSFORMATION STEPS

被引:111
作者
DEBONDT, A [1 ]
EGGERMONT, K [1 ]
DRUART, P [1 ]
DEVIL, M [1 ]
GODERIS, I [1 ]
VANDERLEYDEN, J [1 ]
BROEKAERT, WE [1 ]
机构
[1] CTR RECH AGRON ETAT,CULTURES FRUITIERES & MARAICHERES STN,B-5800 GEMBLOUX,BELGIUM
关键词
D O I
10.1007/BF00234517
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
The factors influencing transfer of an intron - containing beta-glucuronidase gene to apple leaf explants were studied during early steps of an Agrobacterium tumefaciens-mediated transformation procedure, The gene transfer process was evaluated by counting the number of beta-glucuronidase expressing leaf zones immediately after cocultivation, as well as by counting the number of beta-glucuronidase expressing calli developing on the explants after 6 weeks of postcultivation in the presence of 50 mg/l kanamycin. Of three different tested disarmed A. tumefaciens strains, EHA101(pEHA101) was the most effective for apple transformation. Cocultivation of leaf explants with A. tumefaciens on a medium with a high cytokinin level was more conducive to gene transfer than cocultivation on media with high auxin concentrations, Precultivation of leaf explants, prior to cocultivation, slightly increased the number of beta-glucuronidase expressing zones measured immediately after cocultivation, but it drastically decreased the number of transformed calli appearing on the explants 6 weeks after infection. Other factors examined were: Agrobacterium cell density during infection, bacterial growth phase, nature of the carbon source, explant age, and explant genotype.
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页码:587 / 593
页数:7
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