丹参种根和愈伤组织保存方式研究

Preservation Methods of Salvia miltiorrhiza Seed Roots and Callus

摘要

本研究探讨4 ℃低温保存对丹参种根及愈伤组织活力的影响，并分析保存介质、根段大小和保藏时间与种根发芽率的关系，以期为丹参种质资源的中长期保存提供技术依据。试验以一年生丹参不同大小的鲜根根段和初代愈伤组织为材料，将丹参根段按长度与直径分为四组，通过沙培法测定种根发芽率，利用继代培养中愈伤组织的存活率评价其活力。结果表明：4 ℃湿沙保存可有效维持种根质量，显著抑制其提前发芽，保持含水量与活力，发芽率较高；种根长度与直径对低温保存后的成苗率具有显著影响，多数种根低温保存1年内成苗率保持在85%以上，且随着丹参根段长度和直径的增加，丹参出苗率及苗高相应提高；愈伤组织在4 ℃条件下保存9~12个月后仍具备成苗能力，但不同品种间存在保存时间与活力差异。综上所述，4 ℃低温湿沙保存可使丹参较大种根保存1年，为隔年种植降低成本提供可行途径；愈伤组织在低温保存9个月内可维持较好的成苗能力，为减少继代次数、简化保存流程提供依据。

Abstract

This study investigates the effects of low-temperature preservation at 4 ℃ on the viability of Salvia miltiorrhiza seed roots and callus, and analyzes the relationship between preservation medium, root segment size, preservation duration and root germination rate, aiming to provide technical basis for the medium- and long-term preservation of S. miltiorrhiza germplasm resources. Using fresh root segments of different sizes and primary calli from one-year-old S. miltiorrhiza as materials, this experiment divides the root segments into four groups based on length and diameter, determining the seed root germination rate by the sand culture method, and evaluating callus viability through survival rate of calli during subculture. The results show that wet sand preservation at 4 ℃ effectively maintains the seed root quality, significantly inhibits premature germination, preserves water content and viability, and yields a high germination rate. Root segment length and diameter exert a significant effect on seedling establishment rate after low-temperature preservation. Most seed roots maintain a seedling establishment rate above 85% within one year of low-temperature preservation, and with increases in the length and diameter of the S. miltiorrhizaroot segments, both the seedling emergence rate and seedling height correspondingly rise. Calli retain seedling formation capacity after 9~12 months of preservation at 4 ℃, although variations in preservation duration tolerance and viability exist among different varieties. In summary, low-temperature wet sand preservation at 4 ℃ allows large-dimension S. miltiorrhiza seed roots to be stored for up to one year, providing a feasible approach to reduce the cost of biennial planting. Additionally, Calli maintain strong seedling formation capacity within 9 months of low-temperature preservation, offering a basis for reducing subculture frequency and simplifying the preservation protocols.

关键词

丹参 / 低温保存 / 根段 / 愈伤组织

Key words

Salvia miltiorrhiza / low-temperature preservation / root segment / callus

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基金资助

山东省重点研发项目(2024TZXD056)

山东省重点研发项目(2023T2XD085)

国家自然科学基金项目(82373991)