ZHOU Xueqing,CHEN Xueda,WANG Yongxia,et al.Seed Morphology and Asymbiotic Germination of Vanda bicolor Griff.[J].Northern Horticulture,2026,(8):73-81.[doi:10.11937/bfyy.20254053]
双色万代兰种子形态及无菌萌发研究
- Title:
- Seed Morphology and Asymbiotic Germination of Vanda bicolor Griff.
- 文章编号:
- 1001-0009(2026)08-0073-09
- Keywords:
- Vanda bicolor; seed morphology; TTC; asymbiotic germination
- 分类号:
- S682.31
- 文献标志码:
- A
- 摘要:
- 以-25 ℃下冷藏135 d的双色万代兰成熟种子为试材，采用体式显微镜观测、0.5%的TTC进行活力检测及无菌播种正交方法，研究了双色万代兰种子形态特征、活力水平及以MS为基本培养基，设置6-BA、NAA、椰子汁三因素三水平组合，并在光照与暗培养2种条件下培养180 d，以期建立适宜的非共生萌发体系，为濒危双色万代兰的种质保存与人工繁育提供参考依据。结果表明：双色万代兰种子为棕褐色，种长（0.301±0.019）mm，种宽（0.098±0.009）mm，胚长（0.228±0.031）mm，胚宽（0.096±0.013）mm；种胚几乎占满种皮腔；TTC 检测表明种子经长期低温储藏后仍保持60.3%的活力。非共生萌发试验中，最佳萌发培养基组合为1.0 mg·L-1 6-BA+0 mg·L-1 NAA+100 mL·L-1椰子汁；光照显著提高萌发率，而暗培养有利于原球茎膨大与分化。极差分析表明，影响萌发的主导因素依次为6-BA>椰子汁>NAA。综上，6-BA与椰子汁是促进双色万代兰种子萌发的关键因素，光照与暗培养的阶段性配置有利于提高萌发及原球茎的生长。
- Abstract:
- Taking V.bicolor mature seeds refrigerated at -25 ℃ for 135 days as test materials,the seed morphological characteristics and viability were investigated using stereomicroscopy and 0.5% TTC staining，respectively.An orthogonal experiment was conducted using MS as the basic medium to study the effects of different combinations of 6-BA，NAA，and coconut water (CW) under light and dark conditions for 180 days,to establish a suitable asymbiotic germination system,in order to provide a reference for the germplasm conservation and artificial breeding of the endangered Vanda bicolor.The results showed that the seeds of V.bicolor were brownish-black.The seed length and width were (0.301±0.019) mm and (0.098±0.009) mm，respectively，while the embryo length and width were (0.228±0.031) mm and (0.096±0.013) mm，respectively.The embryo occupied almost the entire seed coat cavity.TTC testing indicated that the seeds maintained a viability of 60.3% after long-term low-temperature storage.In the asymbiotic germination experiment，the optimal medium combination was MS+1.0 mg·L-1 6-BA+0 mg·L-1 NAA+100 mL·L-1 CW.Light conditions significantly improved the germination rate，while dark culture was conducive to protocorm expansion and differentiation.Range analysis revealed that the primary factors affecting germination were in the order of 6-BA>CW>NAA.In conclusion，6-BA and CW were the key factors promoting the seed germination of V.bicolor.A phased configuration of light and dark culture was beneficial for improving germination and protocorm growth.
参考文献/References:
［1］DEB C R,JAMIR N S,KIKON Z P.Distribution prediction model of a rare orchid species (Vanda bicolor Griff.) using small sample size［J］.American Journal of Plant Sciences,2017(8):1388-1398.［2］邓杰玲,黄昌艳,崔学强,等.中国万代兰属(兰科)一新记录种:双色万代兰［J］.广西植物,2020,40(2):282-284.［3］CHEN X G,WU Y H,LI N Q,et al.What role does the seed coat play during symbiotic seed germination in orchids:An experimental approach with Dendrobium officinale［J］.BMC Plant Biology,2022,22(1):375.［4］陈菁瑛,万学锋,刘保财,等.白及种子非共生培养高效萌发体系的研究［J］.福建农业学报,2018,33(2):131-135.［5］林夏斌,黄华达,黄怀妹,等.白花拟万代兰(Vandopsis undulata)组培苗的增殖、生根与炼苗［J］.江苏农业科学,2016,44(2):56-59.［6］徐佩玲.矮万代兰组培快繁研究初报［J］.热带林业,2011,39(1):25-26.［7］PRASONGSOM S,THAMMASIRI K,PRITCHARD H W.Seed micromorphology and ex vitro germination of Dendrobium orchids［J］.Acta Horticulturae,2017(1167):339-344.［8］张林瀛.大序隔距兰(Cleisostoma paniculatum)传粉生物学及种子萌发特性研究［D］.福州:福建农林大学,2015.［9］李国平,徐自坤.见血清种子形态结构特征［J］.武夷学院学报,2021,40(6):1-4.［10］边子星.濒危附生兰华石斛种子生物学研究［D］.海口:海南大学,2017.［11］CUSTDIO C C,MARKS T R,PRITCHARD W,et al.Improved tetrazolium viability testing in orchid seeds with a thick carapace (Dactylorhiza fuchsii) or dark seed coat (Vanda curvifolia)［J］.Seed Science and Technology,2016,44(1):177-188.［12］张毓,张启翔,赵世伟,等.大花杓兰种子形态特征与生活力测定［J］.北京林业大学学报,2010,32(1):69-73.［13］李宛宣,陈进红,杨旭,等.野生莲瓣兰种子形态特征与生活力测定［J］.云南农业大学学报(自然科学),2019,34(4):671-677.［14］刘海平,陈秀萍,李珺,等.大黄花虾脊兰种子特性及无菌播种［J］.福建农林大学学报(自然科学版),2020,49(1):29-34.［15］何明高,王瑞霞,宋希强,等.束花石斛种子超低温保存的研究［J］.云南植物研究,2010,32(4):334-338.［16］刘靖,胡泽方,程志杰,等.濒危植物曲茎石斛种子无菌萌发研究［J］.种子,2021,40(9):2,48-51,56.［17］JUALANG A G，DEVINA D，HARTINIE M，et al.Asymbiotic seed germination and seedling development of Vanda dearei［J］.Malaysian Applied Biology，2014，43(2)：25-33.［18］MUHARYATI Y,SETIYADI M W,KHATIMAH A,et al.Propagation of the Vanda helvola orchid in vitro［J］.Jurnal Pijar Mipa,2024,19(3):488-492.［19］PEBAM B,KISHOR R,NARMATHA BAI V.In vitro immature embryo germination and propagation of Vanda stangeana Rchb.f.,an orchid endemic to India［J］.Horticulture,Environment,and Biotechnology,2016,57(6):615-624.［20］WU K，YUAN Y，XU L，et al.Effects of basic medium types and plant growth regulators on seed germination and protocorm differentiation of Pleione maculata (Lindl.).(orchidaceae)［J］.Biology Bulletin，2025,52(2)：63.［21］赵海红,贝丽霞,陈祥梅.不同附加物对马铃薯脱毒试管苗快繁的影响［J］.黑龙江八一农垦大学学报,2007,19(2):17-20.［22］张月盈,李枝林,和凤美,等.观赏用微型兰花组织培养最佳配方的优化［J］.北方园艺,2024(13):49-57.［23］陈尔,陈宝玲,杨舒婷,等.带叶兜兰非共生萌发试验［J］.北方园艺,2016(22):100-103.［24］林丹妮,陈之林,段俊,等.云南火焰兰种子无菌萌发和试管育苗［J］.热带亚热带植物学报,2008,16(1):83-88.［25］王瑞霞,何明高,宋松泉.培养基与光照对沼兰种子非共生萌发的影响［J］.植物生态学报,2010,34(4):438-443.［26］李茂金,何荣晓,王鸿荏,等.诱导流苏贝母兰种子无菌萌发的最佳条件［J］.分子植物育种,2025,23(22):7571-7579.［27］初美静.兜兰种质评价及肉饼兜兰无菌播种与组培技术研究［D］.烟台:烟台大学,2019.［28］王亚沉,石镇源,王丹,等.碧玉兰和独占春正反交及种子无菌萌发研究［J］.北方园艺,2013(4):64-67.［29］周学青,陈学达,山晓丹,等.长叶兰和大雪兰杂交种子质量与萌发特性研究［J］.北方园艺,2026（4）：69-75.
备注/Memo
第一作者简介：周学青（1997-），女，硕士研究生，研究方向为西藏兰科植物种质资源与遗传育种。E-mail：1527204311@qq.com.责任作者：邢震(1973-)，男，博士，教授，现主要从事园林植物与观赏园艺等研究工作。E-mail：xztibetan@xza.edu.cn.基金项目：林芝市林业和草原局重点资助项目（LZLCJ603324061）；西藏自治区教育厅资助项目（XK2024-01，XK2024-04，YJSJD2025-6）；西藏农牧学院林学博士点（一期）资助项目（533325001）。收稿日期：2025-11-25