红松5个家系未成熟种子的胚性愈伤组织诱导研究

陈士刚 秦彩云 王聪慧 才巨锋 陶晶

摘 要：红松是重要的用材树种和坚果经济林树种。以组织培养的方法，选用三岔子红松种子园5个家系的未成熟种子作为外植体进行胚性愈伤组织诱导研究。结果表明，不同家系未成熟种子在添加萘乙酸（NAA）2 mg/L + 6-苄氨基嘌呤（6-BA ）1 mg/L的培养基上胚性愈伤组织诱导率不同，最佳诱导率为5.68 %。红松愈伤组织诱导的最适蔗糖浓度为30 g/L。选取长势良好的胚性愈伤组织进行超低温保存，在恢复培养10 d后可观察到不同细胞系均有明显新生愈伤组织，15 d后愈伤组织大量增殖。本研究为三岔子红松种子园红松优良种质资源的规模化繁殖和保存奠定了基础。

关键词：红松;红松种子园;胚性愈伤组织;超低温保存;激素组合

中图分类号：S791    文献标识码：A   文章编号：1006-8023（2021）01-0013-05

Abstract：Korean pine is an important timber tree species and nut economic forest tree species. The method of tissue culture was used， the immature seeds of five families from Sanchazi Korean pine seed orchard were selected as explants by tissue culture to study the embryogenic callus induction and cryopreservation. The results showed that immature seeds of different families had different induction rates of embryogenic callus on the medium supplemented with naphthaleneacetic acid （NAA） 2 mg/L + 6-benzylaminopurine （6-BA） 1 mg/L， and the best induction percentage was 5.68 %. The optimal sucrose concentration for the callus induction of Korean pine was 30 g/L. Embryogenic callus with good growth was selected for cryopreservation. After 10 days of recovery culture， it was observed that there were obvious new callus in different cell lines. After 15 days， a large number of callus proliferated. This study laid a foundation for large-scale propagation and conservation of fine germplasm resources of Pinus koraiensis in Sanchazi seed orchard.

Keywords：Pinus koraiensis; Korean pine seed orchard; embryonic callus; cryopreservation; hormone combination

0 引言

植物体细胞胚发生是指体细胞在特定条件下，未经性细胞融合而通过与合子胚发生类似的途径发育成新个体的形态发育过程[1-4]。体细胞胚发生具有以最低成本实现迅速繁殖大量高价值苗木的潜能[5-7]。结合传统的繁殖技术和分子细胞生物技术，体胚发生为促进商业树种的基因改善提供了一个很有价值的工具[8-9]。

红松（Pinus koraiensis Sieb. et Zucc.）为松科松属树种，树高可达40 m。红松又名朝鲜松、果松、海松，是第三纪孑遗树种，同时也是东北温带地带性顶极群落——阔叶红松林的建群种[10-14]。通过体细胞胚胎发生途径使红松优良基因型大规模繁殖，并可实现优良种质资源超低温保存，从而为优良种质资源选育、大规模繁殖以及长期保存提供依据。……