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将含有抗溴苯腈基因bxn和雄性不育基因的重组载体pTA29-Barnase/bxn导入农杆菌(Agrobacterium tumefaciens)后转化烟草(Nicotiana tabacum L.),得到33个转基因植株。在27℃/23℃培养的16株中,有7株表现部分不育,另9株全部可育。而在20℃/15℃培养的17株中,12株表现不育,5株表现部分不育。部分不育的植株上同时开放可育花朵和不育花朵,不过其中的不育花朵中的花粉萌发活力很低。将在20℃/15℃条件下表现不育的12个不育株从20℃/15℃温室移至27℃/23℃温室后30d左右,其中9株表现程度不同的育性恢复现象:5株表现部分可育,另4株表现完全可育;但仍有3株表现雄性不育。雄性不育花朵的花丝变短,花药皱瘪,不散粉。细胞学观察证明,转基因植株的花药绒毡层降解提早于四分体时期,至单核小孢子时期降解殆尽。
Thirty-three transgenic plants were obtained by transformation of tobacco (Nicotiana tabacum L.) with the recombinant vector pTA29-Barnase / bxn containing the anti-bromoxynil gene bxn and the male sterility gene introduced into Agrobacterium tumefaciens. Among the 16 strains cultured at 27 ℃ / 23 ℃, 7 showed partial infertility and the other 9 were fertile. Of the 17 strains cultured at 20 ℃ / 15 ℃, 12 showed infertility and 5 showed partial infertility. Some sterile plants also have fertile and sterile flowers open at the same time, but pollen germination activity in sterile flowers is very low. Twelve sterile lines, sterile at 20 ℃ / 15 ℃, were moved from the greenhouse at 20 ℃ / 15 ℃ to 30d at 27 ℃ / 23 ℃. Nine of them showed fertility restoration with different degrees: Strain was partially fertile and the other 4 plants were completely fertile; however, 3 were still male sterile. Male sterile silk flowers become shorter, anther wrinkled, not loose powder. Cytological observation proved that the anther tapetum of transgenic plants degraded earlier than the tetrad stage and degraded to the mononuclear microspore stage.