黑毛石斛×鼓槌石斛杂交种组培快繁技术研究

田英男 张天泽 秦晓杰 冯垒 胡淞 师凯辉 黄鑫 韩欣妤 朱永平 和凤美

摘  要：本研究以黑毛石斛×鼓槌石斛杂交种蒴果为外植体，通过对种子无菌萌发和原球茎诱导、原球茎增殖及丛生芽诱导、生根壮苗以及移栽驯化研究，建立杂交石斛兰的组培快繁技术体系。结果表明，最适合种子无菌萌发和原球茎诱导的培养基为MS+6-BA 0.5 mg/L+NAA 0.3 mg/L+琼脂6.8 g/L+蔗糖20 g/L+马铃薯50 g/L;原球茎增殖及丛生芽诱导的最佳培养基为1/2 MS+6-BA 1.0 mg/L+NAA 0.2 mg/L+KT 1.0 mg/L+蔗糖25 g/L+琼脂6.8 g/L+碳粉1.0 g/L+马铃薯50 g/L+香蕉50 g/L，继代培养50 d，增殖系数达6.19;最佳生根壮苗培养基为1/2 MS+6-BA 0.2 mg/L+NAA 0.5 mg/L+花宝1号0.5 g/L+蔗糖25 g/L+琼脂6.8 g/L+碳粉1.0 g/L+马铃薯50 g/L+香蕉50 g/L，培养2个月后，平均株高达7.14 cm，平均根数6.17条，平均根长6.46 cm，生根率达100%。体积比为单一松树皮和松树皮∶兰石=1∶1适于移栽，移栽65 d成活率分别达到96.32%和96.71%。

关键词：黑毛石斛;鼓槌石斛;杂交育种;组织培养

Abstract： In order to establish a rapid propagation for Dendrobium williamsonii × D. chrysotoxum， the immature hybrid seeds were selected as the explants to study the suitable medium for aseptic seed germination， protocorm-like bodies （PLBs） induction， proliferation of protocorm and shoot induction， rooting and growth and domestication by transplanting were also screened. MS + 6-BA 0.5 mg/L + NAA 0.3 mg/L + agar 6.8 g/L + sucrose 20 g/L + potato 50 g/L was the best medium for hybrid seeds germination and protocorm induction. 1/2 MS + 6-BA 1.0 mg/L + NAA 0.2 mg/L + KT 1.0 mg/L + sucrose 25 g/L + agar 6.8 g/L+AC 1.0 g/L + potato 50 g/L + banana 50 g/L was suitable for protocorm proliferation and shoot induction，and the proliferation coefficient reached 6.19 times after 50d. The best rooting and growth medium was 1/2 MS + 6-BA 0.2 mg/L + NAA 0.5 mg/L + Hyponex No.1 0.5 g/L + sucrose 25 g/L + agar 6.8 g/L + AC 1.0 g/L + potato 50 g/L + banana 50 g/L， cultivated for 2 months， root system was long and strong， plant height was 7.14 cm， average number of roots was 6.17， average root length was 6.46 cm， rooting rate was 100%. In the pine bark and mixture of pine bark∶orchid stone =1∶1 （V∶V）， after cultured for 65 d， the survival percentage was 96.32% and 96.71%， respectively.

Keywords： Dendrobium williamsonii; Dendrobium chrysotoxum; cross breeding; tissue culture

石斛屬（Dendrobium）是兰科中最大的一个属，其原种将近2 000种，13 600多个杂交种[1-3]。中国有74种，2变种[4]，分布于西南、华南、台湾等热带、亚热带和秦岭以南各地[5]，以附生为主，生存环境的气候一般都是比较湿润、温暖以及半荫的环境[6-7]。石斛属植物对生长环境要求比较苛刻，自然状态下生长较缓慢且繁育力极低[8-9]，加之具有较高观赏性和药用价值，长期被掠夺性采挖，对石斛属自然资源造成严重破坏，种质资源枯竭严重，培育石斛新品种已变得越来越重要。近年来，石斛杂交育种工作已开展，但报道仍较少[10-12]。

兰科植物繁殖主要靠分株繁殖，繁殖系数低、速度慢，不能满足日益增长的市场需求[13]。兰科植物也可……