银杉愈伤组织诱导植株再生的影响因素

doi:10.11707/j.1001-7488.LYKX20230141

摘要/Abstract

摘要：

目的: 探索银杉成熟胚愈伤组织诱导植株再生的影响因素，为阐明影响诱导银杉愈伤组织、不定芽等的机制提供帮助。方法: 以广西金秀和广西桂林2个种源的银杉成熟胚为试验材料，利用1/2 MS、DCR、P6培养基，分别添加0.5、1.0、1.5 mg·L^?1的6-BA，0.2、0.4、0.6 mg·L^?1的NAA和0.5、1.0、1.5 mg·L^?1的IBA，诱导银杉愈伤组织、不定芽和不定根，得到银杉再生植株；分析银杉种源、培养基、植物生长调节剂等对银杉愈伤组织、不定芽、不定根诱导率和增殖壮芽率的影响。结果: 广西桂林种源银杉愈伤组织、不定根诱导率和增殖壮芽率最大，分别为66.48%，21.43%和55.00%。广西金秀种源银杉不定芽诱导率最大，为73.33%。适合银杉愈伤组织诱导植株再生的培养基和植物生长调节剂对于愈伤组织为DCR+0.5 mg·L^?16-BA，对于不定芽为DCR+1.5 mg·L^?16-BA，对于不定根为1/2 MS+0.2 mg·L^?1NAA+0.5 mg·L^?1IBA，对于增殖壮芽为DCR+1.0 mg·L^?16-BA+0.4 mg·L^?1NAA。结论: 广西桂林种源银杉整体诱导率和增殖率相比广西金秀种源高。DCR培养基对银杉愈伤组织和不定芽诱导具有显著促进效果，1/2 MS培养基对不定根诱导具有显著促进效果。6-BA既可诱导银杉愈伤组织和又可诱导不定芽，NAA和IBA组合可诱导出银杉不定根，6-BA和NAA组合可对银杉不定芽进行增殖壮芽。

关键词: 银杉, 濒危物种, 成熟胚, 愈伤组织, 不定芽诱导, 植株再生

Abstract:

Objective: Cathaya argyrophylla is a rare and endangered plant unique to China, however it is difficult to breed. In this study, the factors influencing the induction callus of plant regeneration from mature embryo of C. argyrophylla were studied, so as to provide assistance in further elucidating the mechanisms that influence the induction of callus and adventitious buds in C. argyrophylla. Method: The mature embryos of C. argyrophylla from two provenances in Jinxiu and Guilin, Guangxi, were used as experimental materials. The media of 1/2 MS, DCR, and P6, with adding 0.5, 1.0, and 1.5 mg·L^?16-BA, 0.2, 0.4, and 0.6 mg·L^?1NAA, and 0.5, 1.0, 1.5 mg·L^?1IBA, respectively, were used to induce callus tissue, adventitious buds, and roots of C. argyrophylla, in order to obtain regenerated C. argyrophylla plants. The effects of two provenances, culture medium, plant growth regulators, etc. on the induction rate of callus, adventitious buds, adventitious roots, and proliferation rate of breeding and strengthening buds of C. argyrophylla were analyzed. Result: The callus induction rate, adventitious root induction rate, and breeding and strengthening buds rate of C. argyrophylla from Guilin provenance were significantly higher, with values of 66.48%, 21.43%, and 55.00%, respectively. The induction rate of adventitious buds in the C. argyrophylla from Jinxiu provenance was significantly higher, at 73.33%. The suitable culture medium and plant growth regulators for inducing plant regeneration from C. argyrophylla callus were: DCR+0.5 mg·L^?16-BA for callus induction, DCR+1.5 mg·L^?16-BA for adventitious bud induction, 1/2 MS+0.2 mg·L^?1NAA+0.5 mg·L^?1IBA for adventitious root induction, and DCR+1.0 mg·L^?16-BA+0.4 mg·L^?1NAA for bud proliferation and strengthening. Conclusion: The overall induction and proliferation rates of C. argyrophylla from Guilin, Guangxi are higher than those from Jinxiu, Guangxi. DCR medium has a significant promoting effect on the induction of callus and adventitious buds in C. argyrophylla, while 1/2 MS medium has a significant promoting effect on the induction of adventitious roots of C. argyrophylla. The 6-BA can induce both callus tissue and adventitious buds of C. argyrophylla. The combination of NAA and IBA can induce adventitious roots of C. argyrophylla. The combination of 6-BA and NAA can proliferate and strengthen adventitious buds of C. argyrophylla.

Key words: Cathaya argyrophylla, endangered species, mature embryo, callus, adventitious bud induction, plant regeneration

中图分类号:

买凯乐,李荣珍,刘宏,覃琨,兰健花,朱昌叁,冯立新,苏杰南,周全连. 银杉愈伤组织诱导植株再生的影响因素[J]. 林业科学, 2024, 60(7): 56-64.

Kaile Mai,Rongzhen Li,Hong Liu,Kun Qin,Jianhua Lan,Changsan Zhu,Lixin Feng,Jienan Su,Quanlian Zhou. Factors Influencing the Callus Induction and Plant Regeneration of Cathaya argyrophylla[J]. Scientia Silvae Sinicae, 2024, 60(7): 56-64.

图/表 4

图1

表1

表2

表3

参考文献 0

	曹基武, 刘春林, 张　斌, 等. 珍稀植物银杉的种子萌发特性. 生态学报, 2010, 30 (15): 4027- 4034.
	Cao J W, Liu C L, Zhang B, et al. Seed germination of endangered Cathaya argyrophylla Chun & Kuang. Acta Ecologica Sinica, 2010, 30 (15): 4027- 4034.
	陈维伦, 郭东红. 银杉(Cathaya argyrophylla)的胚培养, 愈伤组织诱导和愈伤组织上芽的分化. 植物学报, 1981, 23 (3): 249- 250.
	Chen W L, Guo D H. Embryo culture, callus induction and bud formation on callus of Cathaya argyrophylla. Acta Botanica Sinica, 1981, 23 (3): 249- 250.
	董丽芬, 肖　颖, 邵崇斌. 氮、磷、钾元素形态配比及浓度对油松胚培养的影响. 西北林学院学报, 2006, 21 (3): 64- 66.
	Dong L F, Xiao Y, Shao C B. The effect of form and concentration of nitrogen, phosphorus, potassium on inducing embryos of Chinese pine. Journal of Northwest Forestry University, 2006, 21 (3): 64- 66.
	冯育才. 贵州大沙河自然保护区银杉种群结构初步研究. 山地农业生物学报, 2006, 25 (6): 489- 492.
	Feng Y C. A preliminary study on population structure of Cathaya argyrophylla in Dashahe nature reserve of Guizhou. Journal of Mountain Agriculture and Biology, 2006, 25 (6): 489- 492.
	傅立国. 1992. 中国植物红皮书(第一册). 北京: 科学出版社.
	Fu L G. 1992. Red book of Chinese plants (Volume I). Beijing: Science Press. ［in Chinese］
	管中天, 陈　尧. 四川金佛山银杉混交林的初步研究. 植物学报, 1986, 28 (6): 646- 656.
	Guan Z T, Chen Y. A preliminary study on the Cathaya argyrophylla mixed forest in Jinfushan, Sichuan. Acta Botanica Sinica, 1986, 28 (6): 646- 656.
	胡仁传, 谢彦军, 何忠伟, 等. 花坪银杉群落主要木本植物种群生态位特征. 广西师范大学学报(自然科学版), 2014, 32 (1): 149- 155.
	Hu R C, Xie Y J, He Z W, et al. The niche of dominant woody species population in Cathaya argyrophylla forest in Huaping, China. Journal of Guangxi Normal University (Natural Science Edition), 2014, 32 (1): 149- 155.
	黄正福, 李瑞高, 石金华, 等. 银杉种子繁殖试验初报. 广西植物, 1981, 1 (2): 40- 44.
	Huang Z F, Li R G, Shi J H, et al. Preliminary report of reproductive experiment of Cathaya argyrophylla. Guihaia, 1981, 1 (2): 40- 44.
	季孔庶, 王潘潘, 王金铃, 等. 松科树种的离体培养研究进展. 南京林业大学学报(自然科学版), 2015, 39 (1): 142- 148.
	Ji K S, Wang P P, Wang J L, et al. Review on in vitro culture of tree species in Pinaceae. Journal of Nanjing Forestry University (Natural Science Edition), 2015, 39 (1): 142- 148.
	贾彩凤, 李　悦, 张　榕. 华山松成熟胚的器官发生研究初探. 北京林业大学学报, 2006, 28 (4): 100- 105.
	Jia C F, Li Y, Zhang R. Organ culture of mature embryos in Pinus armandii Franch. Journal of Beijing Forestry University, 2006, 28 (4): 100- 105.
	孔冬梅, 万　婷. 2010. 油松成熟胚培养直接器官发生与植株再生(英语). 植物研究, 30(6): 668−673.
	Kong D M, Wan T. 2010. Direct organogenesis and plantlet regeneration from mature zygotic embryos of Chinese pine (Pinus tabulaeformis Carr. ). Bulletin of Botanical Research, 30(6): 668−673. ［in English］
	李科友, 唐德瑞, 李　林, 等. 2008. 离体条件下西黄松成熟合子胚不定芽的诱导及植株再生(英语). 林业科学, 44(5): 38−45, 175.
	Li K Y, Tang D R, Li L, et al. 2008. Adventitious bud induction and plantlet regeneration in vitro from mature zygotic embryos of Ponderosa pine. Scientia Silvae Sinicae, 44(5): 38−45, 175. ［in English］
	李乔明, 张耀尹, 冯育才, 等. 银杉播种苗木不同年份生长量研究. 种子, 2019, 38 (1): 90- 93.
	Li Q M, Zhang Y Y, Feng Y C, et al. Growth research on Cathaya argyrophylla in different growth years. Seed, 2019, 38 (1): 90- 93.
	刘彩霞, 焦如珍, 董玉红, 等. 模拟氮沉降对杉木林土壤氮循环相关微生物的影响. 林业科学, 2015, 51 (4): 96- 102.
	Liu C X, Jiao R Z, Dong Y H, et al. Response of the N-cycling associated soil microorganism to simulated N deposition in a plantation of Cunninghamia lanceolata. Scientia Silvae Sinicae, 2015, 51 (4): 96- 102.
	陆菱妹, 姚　琪, 黄有英, 等. 几种木本植物愈伤组织的诱导. 广西植物, 1980, (2): 41- 44.
	Lu L M, Yao Q, Huang Y Y, et al. Callus induction of several woody plants. Guihaia, 1980, (2): 41- 44.
	罗仲春, 陈湘健. 湿地松嫁接银杉成功. 植物杂志, 1981, (1): 28.
	Luo Z C, Chen X J. Successful grafting of Pinus elliottii with Cathaya argyrophylla. Journal of Plants, 1981, (1): 28.
	买凯乐, 李荣珍, 朱昌叁, 等. 2023. 一种银杉的组培方法和繁殖方法. 广西壮族自治区: CN115039698B, 2023-05-12.
	Mai K L, Li R Z, Zhu C S, et al. 2023. Tissue culture method and propagation method of Cathaya argrophylla. Chinese invention patent. The Guangxi Zhuang Autonomous Region: CN115039698B, 2023-05-12 ［in Chinese］
	买凯乐, 邵砾群, 李荣珍, 等. 油松成熟胚培养不定芽状态对其生根状况的影响. 林业科学, 2014, 50 (5): 147- 153.
	Mai K L, Shao L Q, Li R Z, et al. Effect of the status of adventitious buds in vitro cultured from mature embryo of Pinus tabulaeformis on its rooting. Scientia Silvae Sinicae, 2014, 50 (5): 147- 153.
	买凯乐, 朱昌叁, 冯立新, 等. 珍稀植物银杉濒危因素及种群扩大研究进展. 广西林业科学, 2022, 51 (6): 872- 876.
	Mai K L, Zhu C S, Feng L X, et al. Research progress on endangered factors and population expansion of rare plant Cathaya argyrophylla. Guangxi Forestry Science, 2022, 51 (6): 872- 876.
	普晓兰, 林　萍, 李乡旺. 五针白皮松离体胚培养初步研究. 西南林学院学报, 1997, 17 (1): 18- 21.
	Pu X L, Lin P, Li X W. A study on the excised embryo culture of Pinus sfuamata. Journal of Southwest Forestry College, 1997, 17 (1): 18- 21.
	阙国宁, 房建军, 葛万川, 等. 火炬松、湿地松、晚松组培繁殖研究. 林业科学研究, 1997, 10 (3): 227- 232.
	Que G N, Fang J J, Ge W C, et al. Studies on the tissue culture of Pinus taeda, P. elliottii and P. serotina. Forest Research, 1997, 10 (3): 227- 232.
	舒春节. 2016. 湿地松不同家系苗期特性及组织培养研究. 南昌: 江西农业大学.
	Shu C J. 2016. Studies on tissue culture and seedling characteristics of different families in Pinus elliottii. Nanchang: Jiangxi Agricultural University. ［in Chinese］
	苏乐怡, 赵万义, 张记军, 等. 湖南八面山银杉群落特征及其残遗性和保守性分析. 植物资源与环境学报, 2016, 25 (4): 76- 86. doi: 10.3969/j.issn.1674-7895.2016.04.10
	Su L Y, Zhao W Y, Zhang J J, et al. Analyses on community characteristics and its relict and conservation of Cathaya argyrophylla at Bamianshan in Hu’nan Province. Journal of Plant Resources and Environment, 2016, 25 (4): 76- 86. doi: 10.3969/j.issn.1674-7895.2016.04.10
	孙　敏, 汤绍虎, 陈安和, 等. 银杉成熟胚离体培养及愈伤组织诱导的研究. 西南林业大学学报, 1994, 14 (3): 155- 158.
	Sun M, Tang S H, Chen A H, et al. Embryo of Cathaya argyrophylla in vitro and inducing of callus. Journal of Southwest Forestry College, 1994, 14 (3): 155- 158.
	孙　雪. 2006. 银杉再生体系、拟菌根诱导系及秦岭冷杉菌根诱导系的建立. 哈尔滨: 东北林业大学.
	Sun X. 2006. Establishment of in vitro plantlet regeneration, mycorrhiza-like stuctures of Cathaya argrophylla and mycorrhization of Abies chensiensis. Harbin: Northeast Forestry University. ［in Chinese］
	王伏雄, 陈祖铿. 银杉的胚胎发育. 植物学报, 1974, 16 (1): 64- 72.
	Wang F X, Chen Z K. The embryogeny of Cathaya (Pinaceae). Acta Botanica Sinica, 1974, 16 (1): 64- 72.
	王永波, 唐德瑞, 马　佩. 2010. 培养基和生长调节剂对油松离体胚不定芽诱导的影响. 北方园艺, 219(12): 149-151.
	Wang Y B, Tang D R, Ma P. 2010. The influence of medias and growth regulators on explants to induce adventitious buds of Pinus tabulaeformis. Northern Horticulture, 219(12): 149-151. ［in Chinese］
	谢宗强, 陈伟烈, 胡　东, 等. 濒危植物银杉的结实特性及动物对果实的危害性. 植物生态学报, 1998, 22 (4): 319- 326.
	Xie Z Q, Chen W L, Hu D, et al. The fruiting characteristics of an endangered plant, Cathaya argyrophylla and the impact of animals on fruits. Acta Phytoecologica Sinica, 1998, 22 (4): 319- 326.
	谢宗强, 陈伟烈, 路　鹏, 等. 1999a. 濒危植物银杉的种群统计与年龄结构. 生态学报, 19(4): 523-528.
	Xie Z Q, Chen W L, Lu P, et al. 1999a. The demography and age structure of the endangered plant population of Cathaya argyrophylla, Acta Ecologica Sinica, 19(4): 523-528. ［in Chinese］
	谢宗强, 陈伟烈. 中国特有植物银杉的濒危原因及保护对策. 植物生态学报, 1999b, 23 (1): 1- 7. doi: 10.3321/j.issn:1005-264X.1999.01.001
	Xie Z Q, Chen W L. The endangering causes and preserving strategies for Cathaya Argyrophylla, a plant endemic to China. Acta Phytoecologica Sinica, 1999b, 23 (1): 1- 7. doi: 10.3321/j.issn:1005-264X.1999.01.001
	徐　筱. 不同种源柔枝松组织培养苗移栽后的成活率及生长情况比较. 中国城市林业, 2014, 12 (3): 25- 26, 36. doi: 10.3969/j.issn.1672-4925.2014.03.008
	Xu Y. Comparative study of survival rate and growth of Pinus flexilis tissue culture seedlings after transplanted in different provenances. Journal of Chinese Urban Forestry, 2014, 12 (3): 25- 26, 36. doi: 10.3969/j.issn.1672-4925.2014.03.008
	张　蕊, 王　艺, 金国庆, 等. 2015. 模拟氮沉降对低磷胁迫下3个种源木荷幼苗生长及叶片氮磷含量的影响. 林业科学, 51(4): 36−43.
	Zhang R, Wang Y, Jin G Q, et al. 2015. Effects of simulated N deposition on growth and leaf N and P content in seedlings of three provenances of Schima superba under phosphorous deficit stress. Scientia Silvae Sinicae, 51(4): 36−43. ［in Chinese］
	张耀尹, 李乔明, 冯育才, 等. 2017. 贵州大沙河自然保护区天然银杉优良母树选择初探. 贵州林业科技, 45(1): 34−38.
	Zhang Y Y, Li Q M, Feng Y C, et al. 2017. First exploration in selection of Cathaya argyrophylla natural elite tree in Dashahe nature reserve of Guizhou. Guizhou Forestry Science and Technology, 45(1): 34−38. ［in Chinese］
	张祖荣, 唐建中. 重庆金佛山野生银杉人工繁殖技术初步研究. 四川林业科技, 2004, 25 (3): 56- 59. doi: 10.3969/j.issn.1003-5508.2004.03.013
	Zhang Z R, Tang J Z. A preliminary study of artificial breeding technique in wild Cathaya argrophylla in Jinfu mountain Chongqing. Journal of Sichuan Forestry Science and Technology, 2004, 25 (3): 56- 59. doi: 10.3969/j.issn.1003-5508.2004.03.013
	郑涵予, 金河延, 冯艺川, 等. 2020. 氮和磷对膜荚黄芪不定根芒柄花苷积累的影响. 北方园艺, 454(7): 137−141.
	Zheng H Y, Jin H Y, Feng Y C, et al. 2020. Effects of nitrogen and phosphorus on the accumulation of arbutin in adventitious roots of Astragalus membranaceus, Northern Horticulture, 454(7): 137−141. ［in Chinese］
	郑均宝, 潘冬梅, 陈正华. 油松离体胚子芽的组织分化和无根试管苗的形成. 河北林学院学报, 1994, 9 (2): 97- 101.
	Zheng J B, Pan D M, Chen Z H. In vitro culture of cotyledon of Pinus tabulaeformis embryo and formation of rootless plantlets. Journal of Hebei Forestry College, 1994, 9 (2): 97- 101.
	郑万均. 1985. 中国树木志(第二卷). 北京: 中国林业出版社.
	Zheng W J. 1985. Dendrography of China (Volume II). Beijing: China Forestry Publishing House. ［in Chinese］
	周　婧, 李卓蓉, 吴江婷. 不同氮形态处理条件下杨树根尖差异表达基因的特征分析. 林业科学研究, 2022, 35 (2): 45- 55.
	Zhou J, Li Z R, Wu J T. Characterization of differentially expressed genes in root tips of poplar under different nitrogen forms. Forest Research, 2022, 35 (2): 45- 55.
	朱丽华, 吴小芹. 湿地松成熟合子胚直接器官发生及植株再生. 林业科学, 2006, 42 (8): 25- 29. doi: 10.3321/j.issn:1001-7488.2006.08.005
	Zhu L H, Wu X Q. Plantlet regeneration via organogenesis from mature embryos of Pinus elliottii. Scientia Silvae Sinicae, 2006, 42 (8): 25- 29. doi: 10.3321/j.issn:1001-7488.2006.08.005
	Aiteken J, Lorgan K J, Thorpe T A. Influence of explant selection on the shoot forming capacity of juvenile tissue of Pinus radiata. Canadian Journal of Forest Research, 1981, 11 (1): 112- 117. doi: 10.1139/x81-015
	David A, David H, Mateille T. In vitro adventitious budding on Pinus pinaster cotyledons and needles. Physiologia Plantarum, 1982, 56 (1): 102- 107. doi: 10.1111/j.1399-3054.1982.tb04906.x
	Fan D Y, Wang X P, Zhang W F, et al. 2020. Does Cathaya argyrophylla, an ancient and threatened Pinaceae species endemic to China, show eco-physiological outliers to its Pinaceae relatives? Conservation Physiology, 8(1): 94.
	Hakman, Fowke L C, Arnold V, et al. The development of somatic embryos in tissue cultures initiated from immature embryos of Picea abies (Norway spruce). Plant Science, 1985, 38 (1): 53- 59. doi: 10.1016/0168-9452(85)90079-2
	Horgan K, Aitken J. Reliable plantlet formation from embryos and seedling shoot tips of radiata pine. Physiologia Plantarum, 1981, 53, 170- 175. doi: 10.1111/j.1399-3054.1981.tb04128.x
	Kaul K. 1987. Plant regeneration from cotyledon-hypocotyl explants of Pinus strobes L. Plant Cell Reports, 6: 5-7.
	Kolevska P B, Jelaska S, et al. Bud and shoot formation in juvenile tissue culture of Pinus nigra. Silvae Genetica, 1983, 32, 115- 119.
	Lara-Chavez A, Flinn B S, Egertsdotter U. Initiation of somatic embryogenesis from immature zygotic embryos of oocarpa pine (Pinus oocarpa Schiede ex Schlectendal). Tree Physiology, 2011, 31 (5): 539- 554. doi: 10.1093/treephys/tpr040
	Liu Y S, Basinger J F. Fossil Cathaya (Pinaceae) pollen from the Canadian High Arctic. International Journal of Plant Sciences, 2000, 161 (5): 829- 847. doi: 10.1086/314296
	Nimsch H, Dorken V M. Morpho-anatomical investigations of cones and pollen in Cathaya argyrophylla Chung & Kuang (Pinaceae, Coniferales) under systematical and evolutional aspects. Feddes Repertorium, 2014, 125 (1/2): 25- 38.
	Qian S H, Yang Y C, Tang C Q, et al. Effective conservation measures are needed for wild Cathaya argyrophylla populations in China: insights from the population structure and regeneration characteristics. Forest Ecology and Management, 2016, 361, 358- 367. doi: 10.1016/j.foreco.2015.11.041
	Ramage C M, Williams R R. Mineral nutrition and plant morphogenesis. In Vitro Cellular & Developmental Biology-Plant, 2002, 38 (2): 116- 124.
	Tremblay L, Tremblay F M. Effects of gelling agents, ammonium nitrate, and light on the development of Picea mariana (Mill) B.S.P. (black spruce) and Picea rubens Sarg. (red spruce) somatic embryos. Plant Science, 1991, 77 (2): 233- 242. doi: 10.1016/0168-9452(91)90092-M

指标 Index	植物生长调节剂 Growth regulator/(mg·L^?1)	金秀种源 Jinxiu provenance(%)			桂林种源 Guilin provenance(%)
指标 Index	6-BA	1/2 MS	DCR	P6	1/2 MS	DCR	P6
愈伤率 Callus induction rate	0.5	15.56±8.68abc	14.81±14.81abc	59.72±14.50bc	47.22±23.73abc	66.48±11.11c	24.61±4.45abc
	1.0	30.16±8.40abc	8.93±4.49ab	36.62±16.14abc	60.32±24.95bc	20.83±4.17abc	9.78±7.71ab
	1.5	23.61±11.87abc	0.00±0.00a	50.07±17.71abc	58.33±30.05bc	43.33±23.33abc	13.73±2.29ab
不定芽诱导率 The induction rate of adventitious bud	0.5	37.78±6.19abc	42.96±12.92abc	28.24±16.44abc	25.00±14.43ab	7.41±7.41a	39.89±10.63abc
	1.0	30.16±8.40abc	48.21±5.74abc	29.08±15.90abc	25.40±12.99ab	45.83±4.17abc	45.29±10.62abc
	1.5	29.167±18.16abc	73.33±6.67c	32.15±18.91abc	20.83±20.83a	26.67±26.67abc	69.22±1.38bc
干枯率 Shriveling rate	0.5	0.00±0.00a	0.00±0.00a	0.00±0.00a	0.00±0.00a	0.00±0.00a	3.53±1.85a
	1.0	0.00±0.00a	0.00±0.00a	5.80±2.90ab	0.00±0.00a	16.67±8.33b	2.90±2.90a
	1.5	11.11±11.11ab	0.00±0.00a	5.93±1.58ab	8.33±8.33ab	0.00±0.00a	4.62±0.14ab
未发育率Undeveloped rate	0.5	46.67±3.33c	42.22±8.89bc	12.04±6.48ab	27.78±14.70abc	26.11±3.89abc	30.93±12.69abc
	1.0	39.68±3.18bc	42.86±7.14bc	28.50±2.41abc	14.29±14.29ab	16.67±16.67abc	42.03±7.97bc
	1.5	36.11±1.39abc	26.67±6.67abc	11.86±3.16ab	12.50±12.50ab	30.00±10.00abc	7.66±4.16a

指标Index	植物生长调节剂 Growth regulator/(mg·L^?1)		金秀种源 Jinxiu provenance(%)			桂林种源 Guilin provenance(%)
指标Index	6-BA	NAA	1/2 MS	DCR	P6	1/2 MS	DCR	P6
愈伤率 Callus induction rate	0.5	0.2	100.00±0.00g	31.94±9.11bcd	71.77±9.05ef	100.00±0.00g	25.00±0.00abc	86.67±6.67fg
	1.0	0.4	60.92±8.57ef	57.77±11.28de	33.63±11.25bcd	0.00±0.00a	10.00±0.00ab	25.00±0.00abc
	1.5	0.6	66.67±16.67ef	12.26±6.57ab	48.96±12.20cde	0.00±0.00a	14.76±8.66ab	16.67±16.67ab
不定芽增殖率 Bud proliferation and strengthen rate	0.5	0.2	0.00±0.00a	50.28±0.28g	12.33±2.26abc	0.00±0.00a	35.00±0.00def	0.00±0.00a
	1.0	0.4	6.10±0.51ab	23.72±1.89cd	14.13±1.30bc	20.00±0.00c	55.00±0.00g	0.00±0.00a
	1.5	0.6	33.33±16.67de	46.23±2.07fg	24.85±0.77cd	0.00±0.00a	44.84±2.60efg	48.89±0.56g
干枯率 Shriveling rate	0.5	0.2	0.00±0.00a	17.78±8.94ab	15.90±8.25ab	0.00±0.00a	40.00±0.00bc	13.33±6.67ab
	1.0	0.4	24.03±1.51ab	12.90±7.23ab	8.86±4.44ab	80.00±0.00d	35.00±0.00abc	75.00±0.00d
	1.5	0.6	0.00±0.00a	21.15±3.44ab	21.03±14.30ab	66.67±33.33cd	23.37±11.30ab	34.45±17.22abc
未发育率 Undeveloped rate	0.5	0.2	0.00±0.00a	0.00±0.00a	0.00±0.00a	0.00±0.00a	0.00±0.00a	0.00±0.00a
	1.0	0.4	0.00±0.00a	5.61±5.61ab	43.39±13.76c	0.00±0.00a	0.00±0.00a	0.00±0.00a
	1.5	0.6	0.00±0.00a	20.36±6.51abc	5.16±2.60ab	33.33±33.33bc	17.02±5.82abc	0.00±0.00a

植物生长调节剂Growth regulator/(mg·L^?1)		金秀种源Jinxiu provenance(%)			桂林种源Guilin provenance(%)
IBA	NAA	1/2 MS	DCR	P6	1/2 MS	DCR	P6
0.5	0.2	16.67±8.33b	0.00±0.00a	1.04±1.04a	21.43±3.57b	0.00±0.00a	1.04±1.04a
1.0	0.2	0.00±0.00a	0.00±0.00a	0.00±0.00a	0.00±0.00a	0.00±0.00a	0.00±0.00a
1.5	0.2	0.00±0.00a	0.00±0.00a	4.76±4.76a	0.00±0.00a	0.00±0.00a	4.76±4.76a

[1]	王阳,张付豪,窦敏,许婷,陈昊. 山苍子植株再生体系的优化[J]. 林业科学, 2022, 58(8): 126-135.
[2]	冯莹,林庆良,潘东明. 青钱柳愈伤组织的离体保存[J]. 林业科学, 2020, 56(9): 58-66.
[3]	吴丽芳, 魏晓梅, 陆伟东. 白刺花胚性愈伤组织诱导及体细胞胚发生[J]. 林业科学, 2019, 55(7): 170-177.
[4]	宋跃, 李淑娟, 张含国, 白晓明, 毕显禹, 董实伟, 董昊. 落叶松胚性愈伤组织悬浮培养体系的优化[J]. 林业科学, 2018, 54(7): 146-154.
[5]	许建秀, 吴小芹, 叶建仁, 朱丽华, 吴静. 抗松材线虫病赤松体细胞胚的发育和成熟萌发[J]. 林业科学, 2017, 53(12): 41-49.
[6]	徐进, 祝晨辰, 欧阳磊, 施季森. 柳杉不同优良无性系再生能力的差异[J]. 林业科学, 2016, 52(8): 46-52.
[7]	宋跃, 甄成, 张含国, 李淑娟. 长白落叶松胚性愈伤组织诱导及体细胞胚胎发生[J]. 林业科学, 2016, 52(10): 45-54.
[8]	买凯乐, 邵砾群, 李荣珍, 董丽芬, 兰健花. 油松成熟胚培养不定芽状态对其生根状况的影响[J]. 林业科学, 2014, 50(5): 147-153.
[9]	张建伟, 王军辉, 李青粉, 马建伟. 云杉未成熟合子胚诱导体细胞胚胎发生[J]. 林业科学, 2014, 50(4): 39-46.
[10]	陈金慧;张艳娟;吴亚云;王鹏凯;王光萍;施季森. 植物磺肽素在杂交鹅掌楸体胚发生中的作用[J]. , 2013, 49(2): 33-38.
[11]	武玉珍, 冯睿芝. 褐马鸡圈养种群ISSR标记的遗传多样性[J]. 林业科学, 2013, 49(11): 103-108.
[12]	杨玲;单琳;沈海龙;祁永会. 花曲柳体胚发生和植株再生[J]. 林业科学, 2012, 48(4): 49-55.
[13]	李晓瑞;胡尚连;曹颖;卢学琴;任鹏;吴晓宇;周美娟. 农杆菌介导慈竹4CL基因遗传转化梁山慈竹[J]. 林业科学, 2012, 48(3): 38-44.
[14]	李清清;叶建仁;朱丽华;吴小芹. 黑松未成熟胚的体细胞胚胎发生和植株再生[J]. 林业科学, 2012, 48(12): 39-44.
[15]	习洋;胡瑞阳;王欢;孙鹏;袁存权;李允菲;戴丽;李云. 刺槐未成熟合子胚的体细胞胚胎发生和植株再生[J]. 林业科学, 2012, 48(1): 60-69.