摘要
青稞是青藏高原的主要农作物,研究其原生质体制备的最优体系具有重要意义。以甘露醇浓度、酶解时间、离心速度和不同苗龄为主要影响因素,采用正交试验法,对青稞原生质体制备进行优化。结果表明:最优条件为以培养7 d的青稞幼叶为材料,3%纤维素酶+1%果胶酶+0.5mol/L甘露醇+0.1%MES+1mmol/LCaCl_(2)2H_(2)O+0.1%BSA构成酶解液,25℃,60 r/min,黑暗酶解5 h,900 r/min离心5 min收集细胞,原生质体产量可达3.2×10^(6)个/g,活力为85.42%。
Since Hordeum vulgare L.var.nudum,is the main crop in Qinghai-Tibet Plateau,it is important to study the optimal system of its protoplast preparation.Taking the concentration of mannitol,enzymolysis time,centrifugal speed and leaves of different seedling ages as main influencing factors,the preparation of Hordeum vulgare L var.nudum,protoplast is optimized by orthogonal test.The results show that the optimal condition is to use the seedling leaves cultured for 7 days as materials and the enzymatic hydrolysate composed of 3%cellulase+1%macerozyme+0.5 mol/L mannitol+0.1%MES+1 mmol/L CaCl_(2)2H_(2)0+0.1%BSA.The cells are collected by centrifugation at 900 r/min for 5 mins after 5 h of enzymatic hydrolysis at 25℃,60 r/min in dark.Then the protoplast yield reaches 3.2×10^(6)/g with the viability of about 85.42%.
作者
杨成兰
武雄雄
祁存英
李欣音
张倩倩
段瑞君
YANG Chenglan;WU Xiongxiong;QI Cunying;LI Xinyin;ZHANG Qianqian;DUAN Ruijun(College of Eco-environmental Engineering,Qinghai University,Xining 810016,China;College of Agriculture and Animal Husbandry,Qinghai University,Xining 810016,China)
出处
《青海大学学报》
2021年第4期26-30,共5页
Journal of Qinghai University
基金
国家自然科学基金项目(31560052)
青海省科学技术厅应用基础研究项目(2019-ZJ-7049)。
关键词
青稞
原生质体
制备
正交设计
Hordeum vulgare L.var.nudum.
protoplast
preparation
orthogonal design
作者简介
杨成兰(1996-),女,青海互助人,青海大学在读硕士研究生;通信作者:段瑞君,E-mail:ruijunduan@163.com。
