生物技术通报 ›› 2016, Vol. 32 ›› Issue (6): 103-110.doi: 10.13560/j.cnki.biotech.bull.1985.2016.06.015

• 研究报告 • 上一篇    下一篇

EMS诱变和NaCl胁迫的正交实验及耐盐纤维亚麻种质的筛选

王卫国, 邓倩, 计巧灵, 王伟   

  1. 新疆大学生命科学与技术学院,乌鲁木齐 830046
  • 收稿日期:2015-08-03 出版日期:2016-06-27 发布日期:2016-06-28
  • 作者简介:王卫国,男,助理研究员,研究方向:植物细胞工程;E-mail:2240658411@qq.com
  • 基金资助:
    国家自然科学基金项目(31160295)

Orthogonal Test of EMS Mutation and NaCl Stress and Screening of Salt-tolerant Fiber Flax Germplasm in Vitro

WANG Wei-guo, DENG Qian, JI Qiao-ling, WANG Wei   

  1. College of Life Science and Technology,Xinjiang University,Urumqi 830046
  • Received:2015-08-03 Published:2016-06-27 Online:2016-06-28

摘要: 旨为经愈伤组织诱导再生的耐盐性纤维亚麻植株奠定基础。以3种纤维亚麻无菌苗的下胚轴段为材,经EMS诱变和盐胁迫的正交实验,再经诱芽和愈伤组织培养,研究其理化特性。结果显示,‘范妮’、‘天鑫3号’和‘双亚5号’下胚轴段不定芽培养的最适处理组合均为0.025% EMS处理2 h(或4 h),之后用100或150 mmol/L NaCl处理12 h;总产芽率由高到低分别为‘范妮’、‘天鑫3号’和‘双亚5号’下胚轴段;对3种亚麻不定芽生长影响的顺序为:NaCl浓度>EMS浓度>EMS处理时间;3种亚麻下胚轴段不定芽诱导的适宜培养基为MS + 0.000 2 mg/L TDZ + 0.02 mg/L KT + 0.005 mg/L NAA + 0.3%蔗糖+ 0.1 g/L肌醇+ 3.3 g/L植物凝胶。经0.025% EMS处理4 h + 150 mmol/L NaCl处理12 h后生长的愈伤组织,经过2次交替筛选后,其理化指标表明,与对照组的相比,3种亚麻处理组愈伤组织脯氨酸、可溶性糖、可溶性蛋白含量、SOD和POD活性均显著提高(P<0.05)。经过0.025% EMS处理4 h+150 mmol/L NaCl处理12 h后生长的愈伤组织具有一定的耐盐性。

关键词: 纤维亚麻, EMS诱变, 盐胁迫, 正交实验, 理化特性

Abstract: This work aims to lay the primary foundation for breeding salt-tolerant fiber flax plant induced from the callus. The hypocotyl segments from sterile seedlings of 3 kinds of fiber flax were treated in the orthogonal test with EMS induction and salt stress,and cultured through bud and callus induction. Further,the physicochemical characteristics of the buds and callus were explored. The optimal treatment combination for adventitious buds derived from “Fanny”,“Tianxin 3” and “Shuangya 5” all were 0.025% EMS for 2 h or 4 h,and then 150 mmol/L NaCl for 12 h. Total bud yield from high to low were the hypocotyls segments of “Fanny”,“Tianxin 3” and “Shuangya 5”. The order of factors affecting the emergence and growth of the adventitious buds was concentration of NaCl>concentration of EMS>treating time with EMS. The optimal medium for the induction of adventitious buds from 3 species fiber flaxes was MS+0.000 2 mg/L TDZ+0.02 mg/L KT+0.005 mg/L NAA+0.3% sucrose+0.1 g/L inositol+3.3 g/L phytogel. The callus was treated with 0.025% EMS for 4 h then 150 mmol/L NaCl for 12 h and screened through salt and salt-free medium twice alternately,and the physicochemical properties of which were determined. The results showed that proline content,soluble sugar content,soluble protein,SOD activity and POD activity in the callus of treated group of 3 species fiber flax all increased remarkably(P<0.05)compared with their control group. Conclusively,the callus of treated group with 0.025% EMS for 4 h then 150 mmol/L NaCl for 12 h possessed solid salt tolerance.

Key words: fiber flax, EMS mutation, NaCl stress, orthogonal test, physicochemical property