藜麦愈伤组织诱导和分化条件初探

Preliminary study on callus induction and redifferentiation of Chenopodium quinoa

[目的]探索藜麦愈伤组织诱导和分化条件,为建立其遗传转化体系提供参考。[方法]以下胚轴为外植体进行根癌农杆菌侵染,在诱导阶段分别比较品种、光照、培养基碳源和外源基因对藜麦愈伤组织诱导的影响;在分化阶段分别比较培养基中KNO_(3)和6-BA浓度对愈伤组织分化的影响。[结果]从18个藜麦品种中筛选出9个较易诱导出愈伤组织的品种;光照较黑暗有利于诱导愈伤组织;以(20 g麦芽糖+10 g葡萄糖)为碳源时,愈伤组织形态最好;愈伤组织过表达SERK基因时,生长较快,体积较大。愈伤组织在含3 800 mg·L^(-1) KNO_(3)、不含6-BA或1.00 mg·L^(-1) 6-BA的再生培养基中形态不佳,难以分化。[结论]品种、光照、碳源、外源基因均影响藜麦愈伤组织诱导,KNO_(3)、6-BA浓度过高不利于其愈伤组织分化。

[Purpose]To explore the conditions of callus induction and redifferentiation,and to provide guiding information for the establishment of stable transformation system in quinoa(Chenopodium quinoa).[Method]The hypocotyl explants of different cultivars of quinoa were infected with Agrobacterium tumefaciens,and then the callus were induced under different lighting conditions,carbon sources and exogenous genes.The concentrations of KNO 3 and 6-BA were also explored for callus redifferentiation.[Result]A total of 9 quinoa cultivars which were easier to be induced into callus were screened out from 18 cultivars.The optimal carbon source was comprised of 20 g maltose and 10 g glucose,and lighting was more conductive to callus induction than darkness.Callus developed faster when the SERK gene was overexpressed,and grew bigger than the others.In regeneration medium comprising 3800 mg·L^(-1) KNO 3 and 1 mg·L^(-1)6-BA or without 6-BA,callus was poorly shaped and failed to redifferentiate.[Conclusion]Cultivar,light,carbon source and exogenous gene had different effects on callus induction in quinoa,and high concentration of KNO 3 and 6-BA impaired callus redifferentiation.