甘肃农业大学学报
甘肅農業大學學報
감숙농업대학학보
JOURNAL OF GANSU AGRICULTURAL UNIVERSITY
2014年
2期
125-132
,共8页
苜蓿%植株再生%愈伤组织%体细胞胚
苜蓿%植株再生%愈傷組織%體細胞胚
목숙%식주재생%유상조직%체세포배
Medicago sativa%plant regeneration%callus%somatic embryo
以适宜于西北地区生长的‘甘农1号’杂花苜蓿、‘甘农3号’紫花苜蓿,‘新疆和田大叶’紫花苜蓿3个品种的不同部位外植体为供试材料,进行了植株体外再生的研究.结果表明,苜蓿基因型、外植体来源、外源激素种类及浓度、激素处理时间、培养基蔗糖浓度等因素均影响细胞的生长发育和植株再生.‘甘农1号’苜蓿下胚轴愈伤组织诱导率和体细胞胚形成率最高,可达97.2%和70.9%,‘新疆和田大叶’苜蓿下胚轴次之,‘甘农3号’出愈率和体细胞胚形成率均最低；苜蓿愈伤组织诱导以2.0 mg/L 2,4-D单独处理或2.0 mg/L 2,4-D+0.5 mg/L KT组合处理为最佳,在此激素组合处理下,下胚轴愈伤组织诱导率最高可达97.2%,激素处理时间以20 d为宜；体细胞胚的发育则以2 mg/L 6-BA+0.5 mg/L NAA的激素组合处理为宜,在此激素组合处理下,体细胞胚形成率最高可达70.9%.适宜的生根培养基为1/2 MS+1.0 mg/L IBA+1%蔗糖+0.7%琼脂.体细胞胚形成阶段培养基添加25~30 g/L的蔗糖有利于愈伤组织分化.
以適宜于西北地區生長的‘甘農1號’雜花苜蓿、‘甘農3號’紫花苜蓿,‘新疆和田大葉’紫花苜蓿3箇品種的不同部位外植體為供試材料,進行瞭植株體外再生的研究.結果錶明,苜蓿基因型、外植體來源、外源激素種類及濃度、激素處理時間、培養基蔗糖濃度等因素均影響細胞的生長髮育和植株再生.‘甘農1號’苜蓿下胚軸愈傷組織誘導率和體細胞胚形成率最高,可達97.2%和70.9%,‘新疆和田大葉’苜蓿下胚軸次之,‘甘農3號’齣愈率和體細胞胚形成率均最低；苜蓿愈傷組織誘導以2.0 mg/L 2,4-D單獨處理或2.0 mg/L 2,4-D+0.5 mg/L KT組閤處理為最佳,在此激素組閤處理下,下胚軸愈傷組織誘導率最高可達97.2%,激素處理時間以20 d為宜；體細胞胚的髮育則以2 mg/L 6-BA+0.5 mg/L NAA的激素組閤處理為宜,在此激素組閤處理下,體細胞胚形成率最高可達70.9%.適宜的生根培養基為1/2 MS+1.0 mg/L IBA+1%蔗糖+0.7%瓊脂.體細胞胚形成階段培養基添加25~30 g/L的蔗糖有利于愈傷組織分化.
이괄의우서북지구생장적‘감농1호’잡화목숙、‘감농3호’자화목숙,‘신강화전대협’자화목숙3개품충적불동부위외식체위공시재료,진행료식주체외재생적연구.결과표명,목숙기인형、외식체래원、외원격소충류급농도、격소처리시간、배양기자당농도등인소균영향세포적생장발육화식주재생.‘감농1호’목숙하배축유상조직유도솔화체세포배형성솔최고,가체97.2%화70.9%,‘신강화전대협’목숙하배축차지,‘감농3호’출유솔화체세포배형성솔균최저；목숙유상조직유도이2.0 mg/L 2,4-D단독처리혹2.0 mg/L 2,4-D+0.5 mg/L KT조합처리위최가,재차격소조합처리하,하배축유상조직유도솔최고가체97.2%,격소처리시간이20 d위의；체세포배적발육칙이2 mg/L 6-BA+0.5 mg/L NAA적격소조합처리위의,재차격소조합처리하,체세포배형성솔최고가체70.9%.괄의적생근배양기위1/2 MS+1.0 mg/L IBA+1%자당+0.7%경지.체세포배형성계단배양기첨가25~30 g/L적자당유리우유상조직분화.
The study was conducted on plant regeneration in vitro of 3 cultivars of Medicago sativa (Gannong No.1,Gannong No.3,Xinjiang Daye),widely cultivated in the northwest region.The results showed that genotype of alfalfa,explant resource and different exogenous hormone level,hormone induc-tion time and sucrose concentration had obvious effect on cell development and plant regeneration of alfal-fa.The frequency of callus induction and somatic embryogenesis from hypocotyls of ‘Gannong No.1 ’was 97.2% and 70.9% respectively.‘Xinjiang Daye’took the second place.‘Gannong No.3’had lower fre-quency of callus induction and somatic embryogenesis.The former 2 cultivars were optimum for plant re-generation invitro.Hypocotyl was the optimum donor tissues for rapid and higher frequency induction of callus.It was better to induct callus from hypocotyl on solidified MS medium supplemented with 2.0 mg/L 2,4-D or 2.0 mg/L 2,4-D and 0.5 mg/L KT,the highest rate of callus induction reached 97.2%,and to de-velop somatic embryogenesis on solidified MS medium supplemented with 2 mg/L 6-BA and 0.5 mg/L NAA,its highest frequency was 70.9%.It took 20 minutes to finish the callus induction and initiate devel-opment of somatic under hormone treatment mentioned above.The plants rooted in 1/2 MS medium sup-plemented with 1.0 mg/L IBA,1 % sucrose and 0.7 % agar.It is suggested that solidified MS medium supplemented with 25~30 g/L sucrose would be good for somatic embryogenesis.
