CAJ | 학술논문

玉米的组织培养比较困难,其再生体系还不完善,建立骨干自交系稳定、高效的遗传转化体系是玉米转基因技术的必要前提。以玉米常用自交系齐319、478、502、黄C和7922不同大小（1.0、1.5和2.0 mm）的幼胚为外植体,探讨不同基因型、胚大小和植物生长调节剂浓度对幼胚再生的影响,从而建立玉米高频再生体系。结果表明：以1.5 mm大小的齐319幼胚为外植体,愈伤诱导率最高,达到了86.2%;1.5 mg/L的24-D有利于胚性愈伤的形成和胚性的保持;两步法分化培养可以提高愈伤组织分化、再生成苗。确定了玉米自交系齐319较适合的培养程序为：将1.5 mm大小的幼胚盾片向上接于诱导培养基（N6＋24-D 1.5 mg/L＋L-脯氨酸0.7 g/L＋蔗糖30 g/L＋琼脂粉8 g/L＋硝酸银5μmol/L）,在25℃下暗培养20 d;挑选胚性愈伤组织在温度25℃、暗培养条件下进行继代培养,继代培养基与诱导培养基相同,每14 d继代1次;将愈伤组织继代42 d后转移至分化培养基1（MS＋肌醇100 mg/L＋蔗糖60 g/L＋gelrite 3 g/L）,继续暗培养10 d左右,待愈伤组织形成象牙白色的块状物且有绿点出现时,将其转移到分化培养基2（MS＋肌醇100 mg/L＋蔗糖30 g/L＋gelrite3 g/L）进行光培养,2～3 d即可分化出苗;待幼苗长到4～5 cm高时,移至生根培养基（1/2 MS＋IBA0.8 mg/L＋蔗糖30 g/L）,14～21 d幼苗长出大量的根系,形成完整的植株。
옥미적조직배양비교곤난,기재생체계환불완선,건립골간자교계은정、고효적유전전화체계시옥미전기인기술적필요전제。이옥미상용자교계제319、478、502、황C화7922불동대소（1.0、1.5화2.0 mm）적유배위외식체,탐토불동기인형、배대소화식물생장조절제농도대유배재생적영향,종이건립옥미고빈재생체계。결과표명：이1.5 mm대소적제319유배위외식체,유상유도솔최고,체도료86.2%;1.5 mg/L적24-D유리우배성유상적형성화배성적보지;량보법분화배양가이제고유상조직분화、재생성묘。학정료옥미자교계제319교괄합적배양정서위：장1.5 mm대소적유배순편향상접우유도배양기（N6＋24-D 1.5 mg/L＋L-포안산0.7 g/L＋자당30 g/L＋경지분8 g/L＋초산은5μmol/L）,재25℃하암배양20 d;도선배성유상조직재온도25℃、암배양조건하진행계대배양,계대배양기여유도배양기상동,매14 d계대1차;장유상조직계대42 d후전이지분화배양기1（MS＋기순100 mg/L＋자당60 g/L＋gelrite 3 g/L）,계속암배양10 d좌우,대유상조직형성상아백색적괴상물차유록점출현시,장기전이도분화배양기2（MS＋기순100 mg/L＋자당30 g/L＋gelrite3 g/L）진행광배양,2～3 d즉가분화출묘;대유묘장도4～5 cm고시,이지생근배양기（1/2 MS＋IBA0.8 mg/L＋자당30 g/L）,14～21 d유묘장출대량적근계,형성완정적식주。
Tissue culture of maize is relatively difficult,and establishing a stable and efficient transformation system is the premise of maize transgene.Using elite inbred lines（Qi 319,478,502,Huang C and 7922）as explants,the effects of different genotypes,embryo sizes and plant growth regulator concentrations on callus induction and regeneration was studied.The results showed that the frequency of callus induction of 1.5 mm Qi 319 embryos was highest,which reached 86.2%;2,4-D 1.5 mg/L solution was favorable to callus induction and maintaining the growth of callus;differentiation in two steps could improve plant regeneration.The proper procedure of Qi 319 tissue culture was determined：putting the immature embryos scutella side up on the induction medium（N6＋2,4-D 1.5 mg/L＋L-proline 0.7 g/L＋sucrose 30 g/L＋agar powder 8 g/L＋AgNO3 5 μmol/L）;incubating at 25 ℃（dark） for 20 d;Selecting the callus to subculture medium（same with induction medium）,once for every 14 d;42 d later,transferring all callus to differentiation medium 1（MS＋ inositol 100 mg/L＋sucrose 60 g/L＋gelrite 3 g/L） and incubating for about 10 d（dark）,till some green spots appeared on the callus;transferring all callus to differentiation medium 2（MS＋inositol 100 mg/L＋sucrose 30 g/L＋gelrite 3 g/L） in light for germination,and somatic embryos would sprout leaves within 2-3 d;when seedlings reach 4-5 cm,transferring them to root medium（1/2 MS＋IBA 0.8 mg/L＋sucrose 30 g/L） and incubating for 14-21 d;when lots of fresh root generated,the seedlings would be ready to transplant.