Eight F<sub>1</sub>-hybrid cultivars of broccoli were studied.We obtained cell division,celled colonies and p-calli in 5 cultivars,roots and shoots regeneration in one cultivar.The leavesof propagated plan...Eight F<sub>1</sub>-hybrid cultivars of broccoli were studied.We obtained cell division,celled colonies and p-calli in 5 cultivars,roots and shoots regeneration in one cultivar.The leavesof propagated plantlets in vitro were cut into 1—2mm pieces,isolated with an enzyme solutioncontaining 2% cellulase and 1%macerase on a rotary shaker（50 rpm,21℃,3h,2500 lux light）,and purified with a 0.5M sucrose solution.The purified protoplasts were placed on a drop of 1%agarose.2—3 ml liquid medium was added around the agarose drops,and all of the cultures wereincubated at 25℃ under light（4000 lux）for 16 hours.3—5 days after isolation the cell divisionwas found.About 7 days after incubation 4 multicellular colonies were formed.After 3—5 wksome p-calli were developed.When the p-calli were 2—3 mm in diameter it was transferred to asolidified medium.Once they were developed to 1 cm in diameter they were transferred on a re-generation medium.About 5 months after incubation some roots and shoots grown from the calliwere展开更多
Using three varieties of Brassica campestris, Hauarad (708), Maoshan-3 (714) and Youbai (715),as the maternal plants and one variety of Brassica oleracea Jingfeng-1 (6012) as paternalplants, crosses were made to produ...Using three varieties of Brassica campestris, Hauarad (708), Maoshan-3 (714) and Youbai (715),as the maternal plants and one variety of Brassica oleracea Jingfeng-1 (6012) as paternalplants, crosses were made to produce interspecific hybrids through ovary culture techniques.The ovaries from the cross between B. campestrisB.oleracea (7086012 and 7146012) werecultured and ovary culture was more effective in terms of obtained seeds when ovaries werecultured in vitro at 9 d after pollination (DAP). While for the cross of 7156012, it wasbetter when ovaries in vitro cultured at 12 DAP. Among three cross combinations, the cross of7146012 showed the best response and 43 seeds per ovary were obtained. Among the mediastudied, the ovaries from the cross of 7086012 cultured on MS media supplemented with 3.0 mgL-1 BA0.1 mg L-1 NAA showed better response, and its rate of seeds per ovary reached 44.0%.While the ovaries from the other two crosses (7146012 and 7156012) showed the best responsewhen cultured on B5 media supplemented with 3.0 mg L-1 BA + 0.2 mg L-1 NAA, and the rates of seedsper ovary reached 72.0 and 60.0%, respectively. All seeds obtained from the three crosscombinations were cultured on the MS media supplemented with 1.0 mg L-1 BA + 0.05 mg L-1 NAA,and the seeds from the cross of 7156012 showed the best germination response and thepercentage of germinations reached 66.7%. The regenerated plantlets were obtained from theseseedlings after cultured on the MS media supplemented with 0.05 mg L-1 NAA. Cytological studyshowed that these regenerated plants were all true hybrids of B.campestrisB.oleracea.展开更多
皱叶甘蓝(Brassica oleracea L. var. subauda)“SA61”(SV)的叶及下胚轴分离的原生质体在 MS_1(修改的MS)培养基上细胞壁再生和分裂启动较快。叶原生质体在 DPD_1(修改的 DPD)培养基上获得了最高的分裂率和植板率;下胚轴原生质体在MS_...皱叶甘蓝(Brassica oleracea L. var. subauda)“SA61”(SV)的叶及下胚轴分离的原生质体在 MS_1(修改的MS)培养基上细胞壁再生和分裂启动较快。叶原生质体在 DPD_1(修改的 DPD)培养基上获得了最高的分裂率和植板率;下胚轴原生质体在MS_1上获得最佳的培养效果。叶原生质体培养3—4天后见到一次分裂;下胚轴原生质体在48小时左右即可发生一次分裂。原生质体培养 20—30天后形成肉眼可见的微愈伤颗粒,40天左右即可达1mm大小。在7种不同培养基上增殖微愈伤组织,MB_2、MB_3表现了优良的效果。在MS_2培养基上的芽分化效果最为理想。在不加任何激素的MS培养基上诱导生根,2周后得到再生植株。展开更多
文摘Eight F<sub>1</sub>-hybrid cultivars of broccoli were studied.We obtained cell division,celled colonies and p-calli in 5 cultivars,roots and shoots regeneration in one cultivar.The leavesof propagated plantlets in vitro were cut into 1—2mm pieces,isolated with an enzyme solutioncontaining 2% cellulase and 1%macerase on a rotary shaker（50 rpm,21℃,3h,2500 lux light）,and purified with a 0.5M sucrose solution.The purified protoplasts were placed on a drop of 1%agarose.2—3 ml liquid medium was added around the agarose drops,and all of the cultures wereincubated at 25℃ under light（4000 lux）for 16 hours.3—5 days after isolation the cell divisionwas found.About 7 days after incubation 4 multicellular colonies were formed.After 3—5 wksome p-calli were developed.When the p-calli were 2—3 mm in diameter it was transferred to asolidified medium.Once they were developed to 1 cm in diameter they were transferred on a re-generation medium.About 5 months after incubation some roots and shoots grown from the calliwere
基金supported by the Science and Technology Department of Zhej iang Province(021102109)Hangzhou Municipality(2003132E32)China-Sweden Cooperation Project(C08).
文摘Using three varieties of Brassica campestris, Hauarad (708), Maoshan-3 (714) and Youbai (715),as the maternal plants and one variety of Brassica oleracea Jingfeng-1 (6012) as paternalplants, crosses were made to produce interspecific hybrids through ovary culture techniques.The ovaries from the cross between B. campestrisB.oleracea (7086012 and 7146012) werecultured and ovary culture was more effective in terms of obtained seeds when ovaries werecultured in vitro at 9 d after pollination (DAP). While for the cross of 7156012, it wasbetter when ovaries in vitro cultured at 12 DAP. Among three cross combinations, the cross of7146012 showed the best response and 43 seeds per ovary were obtained. Among the mediastudied, the ovaries from the cross of 7086012 cultured on MS media supplemented with 3.0 mgL-1 BA0.1 mg L-1 NAA showed better response, and its rate of seeds per ovary reached 44.0%.While the ovaries from the other two crosses (7146012 and 7156012) showed the best responsewhen cultured on B5 media supplemented with 3.0 mg L-1 BA + 0.2 mg L-1 NAA, and the rates of seedsper ovary reached 72.0 and 60.0%, respectively. All seeds obtained from the three crosscombinations were cultured on the MS media supplemented with 1.0 mg L-1 BA + 0.05 mg L-1 NAA,and the seeds from the cross of 7156012 showed the best germination response and thepercentage of germinations reached 66.7%. The regenerated plantlets were obtained from theseseedlings after cultured on the MS media supplemented with 0.05 mg L-1 NAA. Cytological studyshowed that these regenerated plants were all true hybrids of B.campestrisB.oleracea.
文摘皱叶甘蓝(Brassica oleracea L. var. subauda)“SA61”(SV)的叶及下胚轴分离的原生质体在 MS_1(修改的MS)培养基上细胞壁再生和分裂启动较快。叶原生质体在 DPD_1(修改的 DPD)培养基上获得了最高的分裂率和植板率;下胚轴原生质体在MS_1上获得最佳的培养效果。叶原生质体培养3—4天后见到一次分裂;下胚轴原生质体在48小时左右即可发生一次分裂。原生质体培养 20—30天后形成肉眼可见的微愈伤颗粒,40天左右即可达1mm大小。在7种不同培养基上增殖微愈伤组织,MB_2、MB_3表现了优良的效果。在MS_2培养基上的芽分化效果最为理想。在不加任何激素的MS培养基上诱导生根,2周后得到再生植株。
