伯克氏植物的高效再生、愈伤组织和细胞悬浮培养的建立以及再生植株的随机扩增多态性DNA分析

Highly efficient regeneration, establishment of callus and cell suspension cultures and RAPD analysis of regenerants of Burkill.

作者信息

Kshirsagar Parthraj R, Chavan Jaykumar J, Umdale Suraj D, Nimbalkar Mansingraj S, Dixit Ghansham B, Gaikwad Nikhil B

机构信息

Department of Botany, Shivaji University, Kolhapur 416 004, India.

Department of Botany, Yashavantrao Chavan Institute of Science, Satara 415 001, India.

出版信息

Biotechnol Rep (Amst). 2015 Mar 11;6:79-84. doi: 10.1016/j.btre.2015.03.003. eCollection 2015 Jun.

DOI:10.1016/j.btre.2015.03.003

PMID:28626699

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5466263/

Abstract

Highly efficient regeneration system has been developed for Burkill, an important herb used as substitute for . Shoot tips explants were cultured on MS medium with various phytohormones for multiple shoot production. The best shoot production frequency (100%) and maximum shoots (10.4 ± 0.8) were obtained on MS media containing TDZ (3.0 mg l) in combination with IBA (0.3 mg l). Maximum callus induction (95 ± 4.8%) and callus growth (1.7 ± 0.4 gm) was achieved on MS medium with 2, 4-D (3.0 mg l). Cell suspension cultures were established and studied for their growth kinetics. Shoots were rooted best (22.1 ± 2.5) in 1/2 MS medium with IAA (3.0 mg l). The genetic uniformity of the micropropagated clones was assessed using RAPD markers. Out of 405 bands, 400 (98.76%) were monomorphic and rest 5 (1.24%) were polymorphic. High multiplication frequency and low risk of genetic instability ensures the efficacy of this protocol.

摘要

已为Burkill开发了高效的再生系统，Burkill是一种重要的草药替代品。将茎尖外植体接种在添加了各种植物激素的MS培养基上以诱导产生丛生芽。在含有噻苯隆（TDZ，3.0毫克/升）和吲哚丁酸（IBA，0.3毫克/升）的MS培养基上，获得了最佳的丛生芽诱导频率（100%）和最多的丛生芽数（10.4±0.8个）。在含有2,4-二氯苯氧乙酸（2,4-D，3.0毫克/升）的MS培养基上，愈伤组织诱导率最高（95±4.8%），愈伤组织生长量最大（1.7±0.4克）。建立了细胞悬浮培养体系并研究了其生长动力学。在添加吲哚乙酸（IAA，3.0毫克/升）的1/2MS培养基上，生根效果最佳（22.1±2.5条根）。使用随机扩增多态性DNA（RAPD）标记评估微繁殖克隆的遗传一致性。在405条带中，400条（98.76%）为单态性，其余5条（1.24%）为多态性。高增殖频率和低遗传不稳定性风险确保了该方案的有效性。