紫花苜蓿NFF2高效组织培养再生体系的建立

Establishment of efficient tissue Culture and generation system for Alfalfa NFF2

生物技术是紫花苜蓿等牧草产量和品质遗传改良的重要途径之一。高效、稳定的再生体系是苜蓿基因工程技术利用的关键,而紫花苜蓿的再生受基因型和环境条件等影响很大,苜蓿不能稳定的高频再生始终是苜蓿基因工程跨越发展的瓶颈。本试验以易于被农杆菌转染的紫花苜蓿品种NFF2无菌苗的叶柄为外植体,研究了不同激素种类和配比等不同培养条件对其愈伤组织诱导的影响,以及不同分化培养基对愈伤组织分化的影响,以期建立一套紫花苜蓿NFF2的高频、稳定的再生体系。结果表明:以叶柄为外植体,最佳愈伤组织诱导培养基为改良SH(SH2K)+1.0mg/L 2,4-D+0.2mg/L KT;最佳愈伤组织继代培养基为BOi2Y;最佳愈伤组织分化培养基为MSO;成苗培养基可以用1/2MS。

Modern biotechnology,especially genetic engineering,is one of the effective methods for alfalfa to improve its forage yield and quality,while a prerequisite for using this technology is the efficient solutions to tissue culture and regeneration system of Alfalfa.The bottleneck of alfalfa genetic engineering attributes to lack of stalbe and hige frequency regeneration,because the tissue culture and regeneration are highly affected by genotype of alfalfa and environmental conditions.A solution for rapid and highly efficient tissue regeneration of alfalfa NFF2was established by studying the influence of hormone concentration,and different combinations of medium on alfalfa callus induction and differentiation in the experiment.The result showed that the best medium for callus induction was the modified SH(SH2K)with 1.0 mg/L 2,4-D+0.2mg/L KT,and the best subculture medium for callus induction was BOi2Y;additionally,the best differentiation medium for callus induction was MSO,while the best medium for plant development of alfalfa was 1/2 MS.