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华北农学报 1993
Theprotoplastsisolation,cultureandshootsregenerationofbroccoli(Brassicaoleraceavar.italica)*DOI: 10.7668/hbnxb.1993.S2.005, PP. 28-34 Keywords: protoplastsculture,shootsregeneration,broccoli(Brassicaoleraceavar.italica) Abstract: EightF1-hybridcultivarsofbroccoliwerestudied.Weobtainedcelldivision,celledcoloniesandp-calliin5cultivars,rootsandshootsregenerationinonecultivar.Theleavesofpropagatedplantletsinvitrowerecutinto1—2mmpieces,isolatedwithanenzymesolutioncontaining2%cellulaseand1%maceraseonarotaryshaker（50rpm,21℃,3h,2500luxlight）,andpurifiedwitha0.5Msucrosesolution.Thepurifiedprotoplastswereplacedonadropof1%agarose.2—3mlliquidmediumwasaddedaroundtheagarosedrops,andallofthecultureswereincubatedat25℃underlight（4000lux）for16hours.3—5daysafterisolationthecelldivisionwasfound.About7daysafterincubation4multicellularcolonieswereformed.After3—5wksomep-calliweredeveloped.Whenthep-calliwere2—3mmindiameteritwastransferredtoasolidifiedmedium.Oncetheyweredevelopedto1cmindiametertheyweretransferredonare-generationmedium.About5monthsafterincubationsomerootsandshootsgrownfromthecalliwereregen.
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