新疆野生黄花苜蓿愈伤诱导及分化的研究

摘要

Xinjiang wild Medicago falcata L. was selected to study callus induction, differentiation and propagation regeneration. The influence factors of wild M. falcata L. tissue culture such as hard seed, explants, hormones (2,4-D and KT) and media were discussed. Results showed that the combination of 4"C low-temperature treatment and sulfuric acid soaking as the better method of breaking the hard seed of M. falcata L. . This treatment resulted in 90% germination rate. Hypocotyl tissue was the most suitable explant for callus induction. The MS medium was more effective than the improved SH medium for the callus induction. The optimal callus induction medium contained MS+2,4-D 0. 5 mg · L-1 +KT 0. 8 mg · L-1 + sucrose 30 g · L-1-|-agar powder 7 g · L-1 with 80% callus induction rate. Light green and light yellow callus were differentiated in culture medium containing MS+2,4-D 0. 2 mg · L-1 +KT 0. 4 mg · L-1 +CH 1 g · L-1 + sucrose 20 g · L-1 +agar 7 g · L-1 with 30% differentiation rate. This technique will lay a foundation for M. falcata L. biotechnology breeding.%以新疆野生黄花苜蓿(Medicago falcata L.)为材料,通过愈伤组织诱导及分化途径获得再生植株；从种子硬实、外植体、激素(2,4-D和KT)、培养基方面探讨野生黄花苜蓿组织培养的影响因素.结果表明:4℃低温处理与浓硫酸浸种结合是破除黄花苜蓿硬实的较优方法,发芽率达90％；下胚轴为诱导愈伤组织的最佳外植体；MS培养基比改良的SH培养基对愈伤组织的诱导更有效,MS诱导培养基为MS+2,4-D 0.5 mg·L-1+KT 0.8 mg·L-1+蔗糖30 g·L-1+琼脂粉7g·L-1,出愈率达到80％；浅绿色和浅黄色的愈伤组织在MS+2,4-D 0.2 mg·L-1+KT 0.4mg·L-1+CH 1 g·L-1+蔗糖20g·L -1+琼脂粉7 g·L -1的培养基上分化较好,分化率为30％.研究结果将为黄花苜蓿生物技术育种奠定一定的基础.