作物学报 ›› 2024, Vol. 50 ›› Issue (9): 2408-2414.doi: 10.3724/SP.J.1006.2024.34204
赵美1,2(), 刘一玮1,2, 石翔天1,2, 李亚俐1,2, 申树林1,2, 尹能文1,2, 赵会彦1,2, 傅鹰3,*(
), 曲存民1,2,*(
)
ZHAO Mei1,2(), LIU Yi-Wei1,2, SHI Xiang-Tian1,2, LI Ya-Li1,2, SHEN Shu-Lin1,2, YIN Neng-Wen1,2, ZHAO Hui-Yan1,2, FU Ying3,*(
), QU Cun-Min1,2,*(
)
摘要:
针对甘蓝组织培养过程中再生体系难度大, 培养效率低的问题, 本研究以白花芥蓝下胚轴为外植体, 对离体培养关键影响因素——无菌苗光/暗培养时间、NaClO消毒时间、激素配比对甘蓝种子、不定芽分化及再生的影响进行了比较分析。结果表明, 75%无水乙醇40 s+5% NaClO 15 min消毒后, 将种子暗培养3 d, 再光培养3 d, 无菌苗生长最好; 预培养时培养基最佳激素组合是0.05 mg L-1 NAA (α-萘乙酸)+4 mg L-1 6-BA (6-苄氨基嘌呤); 筛选培养基的最佳激素组合为0.05 mg L-1 NAA+4 mg L-1 6-BA+250 mg L-1 Cb (羧苄青霉素), 愈伤发生率91.1%, 不定芽分化率57.3%; 生根培养基MS+30 g L-1蔗糖+7 g L-1琼脂+0.1 mg L-1 NAA+0.1%活性炭, 不定根产生数量多, 获得植株根系发达、生长势强。本研究建立的甘蓝组织培养优化体系能快速获得再生植株, 有助于加速甘蓝良种的繁育与评价。
[1] | 秦耀国, 杨翠芹, 程明军, 曹必好, 陈国菊, 雷建军. 培养基pH值对芥蓝离体再生的影响. 中国蔬菜, 2010, (6): 47-49. |
Qin Y G, Yang C Q, Cheng M J, Cao B H, Chen G J, Lei J J. Effect of pH value of culture medium on Brassica albograbra Bailey in vitro regeneration. China Veget, 2010, (6): 47-49 (in Chinese with English abstract). | |
[2] | 张智晖, 韩春梅. 芥蓝常见病虫害及其防治技术. 四川农业科技, 2011, (2): 39. |
Zhang Z H, Han C M. Common diseases and insect pests of Chinese Kale and their control techniques. Sichuan Agric Sci Technol, 2011, (2): 39 (in Chinese). | |
[3] | 薛生玲. 芥蓝八氢番茄红素脱氢酶基因BaPDS1和BaPDS2功能差异分析. 四川农业大学硕士学位论文, 四川成都, 2018. |
Xue S L. The Analysis of Functional Difference of BaPDS1 and BaPDS2 in Chinese Kale. MS Thesis of Sichuan Agricultural University, Chengdu, Sichuan, China, 2018 (in Chinese with English abstract). | |
[4] | 王俞丹. 芥蓝CRISPR/Cas9基因编辑体系建立及BocAOP2s的功能初步验证. 华南农业大学硕士学位论文, 广东广州, 2020. |
Wang Y D. Establishment of CRISPR/Cas 9 Gene Editing System and Preliminary Functional Identification of BocAOP2s in Chinese Kale. MS Thesis of South China Agricultural University, Guangzhou, Guangdong, China, 2020 (in Chinese with English abstract). | |
[5] | 何承坤. 芥蓝的分类地位. 见: 中国园艺学会首届青年学术讨论会论文集. 浙江杭州, 1994. pp 155-159. |
He C K. On the atxonomy of kale. In:Proceedings of the First Youth Symposium of Chinese Horticultural Society. Hangzhou, Zhejiang, 1994. pp 155-159 (in Chinese). | |
[6] | 秦耀国, 杨翠芹, 曹必好, 陈国菊, 雷建军. 芥蓝遗传育种与生物技术研究进展. 中国农学通报, 2009, 25(18): 296-299. |
Qin Y G, Yang C Q, Cao B H, Chen G J, Lei J J. Advances in genetic breeding and biotechnology of Chinese Kale. Chin J Agron, 2009, 25(18): 296-299 (in Chinese with English abstract). | |
[7] | 陈汉才, 吴增祥, 林悦欣, 沈卓, 黎庭耀, 杨易, 周轩, 张艳. 广东菜心、芥蓝研究现状与展望. 广东农业科学, 2021, 48(9): 62-71. |
Chen H C, Wu Z X, Lin Y X, Shen Z, Li T Y, Yang Y, Zhou X, Zhang Y. Research status and prospect of flowering Chinese cabbage and Chinese kale in Guangdong. Guangdong Agric Sci, 2021, 48(9): 62-71 (in Chinese with English abstract). | |
[8] | Sheng X, Yu H, Wang J, Shen Y, Gu H. Establishment of a stable, effective and universal genetic transformation technique in the diverse species of Brassica oleracea. Front Plant Sci, 2022, 13: 1021669. |
[9] | 申静静, 廖玉才, 李和平. 甘蓝型油菜叶片组织培养体系的优化. 湖北农业科学, 2014, 53: 447-450. |
Shen J J, Liao Y C, Li H P. The optimization of regeneration system of Brassica napus L. leaf. Hubei Agric Sci, 2014, 53: 447-450 (in Chinese with English abstract). | |
[10] | 黄科, 余小林, 吴秋云, 沈迎春, 向珣, 曹家树. 芥蓝植株再生体系的优化. 细胞生物学杂志, 2004, 26: 313-316. |
Huang K, Yu X L, Wu Q Y, Shen Y C, Xiang X, Cao J S. Optimization of the regeneration system for Brassica albograbra Bailey plants. J Cell Biol, 2004, 26: 313-316 (in Chinese with English abstract). | |
[11] | 胡志群, 江远彩, 陈淳. 芥蓝子叶离体培养再生植株的研究. 华南农业大学学报, 2006, 27(4): 113-114. |
Hu Z Q, Jiang Y C, Chen C. Plant regeneration in vitro from cotyledon of Brassica albograbra Bailey. J South China Agric Univ, 2006, 27(4): 113-114 (in Chinese with English abstract). | |
[12] | Horeau N, Arora R, Bhojwani S S. A comparative study of in vitro shoot regeneration from cotyledon and root explants of four varieties of Brassica oleracea L. Curr Sci, 1988, 57: 1349-1351. |
[13] | 李光远, 王宜娜, 王凤华. 不同激素对结球甘蓝下胚轴离体培养的影响. 安徽农学通报, 2007, 13(15): 59-60. |
Li G Y, Wang Y N, Wang F H. Influence of hormones on in vitro culture of under hypocotyls of Brassica oleracea L. Anhui Agric Sci Bull, 2007, 13(15): 59-60 (in Chinese with English abstract). | |
[14] | 周林, 曾国平, 章崇玲, 邓智泉. 芥蓝的组织培养研究. 长江蔬菜, 2006, (8): 45-46. |
Zhou L, Zeng G P, Zhang C L, Deng Z Q. Study on tissue culture of Brassica albograbra Bailey. J Changjiang Veget, 2006, (8): 45-46 (in Chinese with English abstract). | |
[15] | 唐君, 张蜀宁, 侯喜林, 王淑敏, 孙成振. 芥蓝高频离体再生体系的建立. 南京农业大学学报, 2011, 34(2): 29-32. |
Tang J, Zhang S N, Hou X L, Wang S M, Sun C Z. Establishment of high frequency in vitro regeneration system of Brassica albograbra Bailey. J Nanjing Agric Univ, 2011, 34(2): 29-32 (in Chinese with English abstract). | |
[16] | 许端祥, 方淑桂, 陈文辉. 甘蓝雄性不育系组培快繁技术研究. 福建热作科技, 2007, 32(1): 15-16. |
Xu D X, Fang S G, Chen W H. Study on tissue culture and rapid propagation of male sterile line of Brassica oleracea L. Fujian Sci Technol Trop Crops, 2007, 32(1): 15-16 (in Chinese with English abstract). | |
[17] | 黄文华, 庄东红, 郑汉藩, 邵舜梦. 芥蓝的组织培养和快速繁殖. 植物生理学通讯, 1999, 35(2): 130. |
Huang W H, Zhuang D H, Zheng H F, Shao S M. Tissue culture and rapid propagation of Brassica albograbra Bailey. Plant Physiol Commun, 1999, 35(2): 130 (in Chinese with English abstract). | |
[18] | 黄俊轩, 李双跃, 李建科, 刘艳军, 杨静慧. 花椰菜叶片离体再生技术的研究. 天津农学院学报, 2006, 13(4): 21-23. |
Huang J X, Li S Y, Li J K, Liu Y J, Yang J H. Regeneration by leaf of Brassicao leracea var. Botrytis. J Tianjin Agric Univ, 2006, 13(4): 21-23 (in Chinese with English abstract). | |
[19] | Wong K W, Loh C S. In vitro regeneration of plantlets in Brassica alboglabra. Plant Cell Tissue Organ Cult, 1987, 10: 143-148. |
[20] | Pua E C, Trần T, Chua N H. High frequency plant regeneration from stem explants of Brassica alboglabra Bailey in vitro. Plant Cell Tissue Organ Cult, 1989, 17: 143-152. |
[21] | 陈琼娥, 钟凤林, 林碧英. 芥蓝组织培养研究. 亚热带植物科学, 2011, 40(1): 34-37. |
Chen Q E, Zhong F L, Lin B Y. Study on tissue culture of Brassica albograbra Bailey. Subtrop Plant Sci, 2011, 40(1): 34-37 (in Chinese with English abstract). | |
[22] | 赵军良, 逯保德, 梁爱华, 赵美华. 大白菜原生质体培养再生体系的优化. 西北植物学报, 2005, 25: 546-551. |
Zhao J L, Lu B D, Liang A H, Zhao M H. Optimization of the system protoplast culture and regeneration for Chinese cabbage. Acta Bot Borealli-Occident Sin, 2005, 25: 546-551 (in Chinese with English abstract). | |
[23] |
赵军良, 逯宝德, 赵美华. 大白菜无菌苗子叶组织培养再生植株. 华北农学报, 2004, 19(2): 17-19.
doi: 10.3321/j.issn:1000-7091.2004.02.005 |
Zhao J L, Lu B D, Zhao M H. Plant regeneration from cotyledons of Chinese cabbage cultured in vitro. Acta Agric Boreali Sin, 2004, 19(2): 17-19 (in Chinese with English abstract). | |
[24] | 廖志强, 邬贤梦, 孙娟, 官春云. 甘蓝型油菜下胚轴一步不定芽再生培养及遗传转化应用. 分子植物育种, 2015, 13: 793-799. |
Liao Z Q, Wu X M, Sun J, Guan C Y. One-step shoot regeneration culture established from hypocotyl of Brassica napus L. and applied to transformation. Mol Plant Breed, 2015, 13: 793-799 (in Chinese with English abstract). | |
[25] | 李楠, 张恩慧, 许忠民, 陈丽潇, 姜娇, 曹丽红. 甘蓝根系再生植株技术研究. 西北农林科技大学学报(自然科学版), 2019, 47(2): 103-111. |
Li N, Zhang E H, Xu Z M, Chen L X, Jiang J, Cao L H. Plant Regeneration technology of Brassica oleracea var. capitata root. J Northwest A&F Univ (Nat Sci Edn), 2019, 47(2): 103-111 (in Chinese with English abstract). | |
[26] | Luciani G F, Mary A K, Pellegrini C, Curvetto N R. Effects of explants and growth regulators in garlic callus formation and plant regeneration. Plant Cell Tissue Organ Cult, 2006, 87: 139-143. |
[27] | Pretto F R, Santarém R E. Callus formation and plant regeneration from Hypericum perforatum leaves. Plant Cell Tissue Organ Cult, 2000, 62: 107-113. |
[28] | Ahmad S, Spoor W. Effects of NAA and BAP on callus culture and plant regeneration in curly kale (Brassica oleraces L.). Pak J Biol Sci, 1999, 2: 109-112. |
[29] | Kou Y P, Ma G H, Teixeira da Silva J A, Liu N. Callus induction and shoot organogenesis from anther cultures of Curcuma attenuata Wall. Plant Cell Tissue Organ Cult, 2013, 112: 1-7. |
[30] | Demmig-Adams B, Adams W W IIl. Photoprotection and other responses of plants to high light stress. Annu Rev Plant Physiol Plant Mol Biol, 1992, 43: 599-626. |
[31] | 王景雪, 孙毅, 崔贵梅, 刘少翔, 王果萍, 尚勇进, 王卉. 在油菜组织培养中激素及基因型对下胚轴分化的影响. 中国油料作物学报, 2000, 22: 12-14. |
Wang J X, Sun Y, Cui G M, Liu S X, Wang G P, Shang Y J, Wang H. Effects of hormones and genotypes on hypocotyl differentiation in tissue culture. Chin J Oil Crops, 2000, 22: 12-14 (in Chinese with English abstract). | |
[32] | 郑爱红, 张芬, 袁巧, 肖楠, 杨莉媛, 甘颖, 杨朝君, 孙勃. 白花芥蓝离体再生植株的研究. 分子植物育种, 2017, 15: 1022-1028. |
Zheng A H, Zhang F, Yuan Q, Xiao N, Yang L Y, Gan Y, Yang Z J, Sun B. The study on regeneration in vitro of Brassica alboglabra Bailey. Mol Plant Breed, 2017, 15: 1022-1028 (in Chinese with English abstract). | |
[33] | 王慧芳. 甘蓝组织培养再生、转化及筛选系统的优化研究. 山西农业大学硕士学位论文, 山西晋中, 2004. |
Wang H F. Optimization of Tissue Culture Regeneration, Transformation and Screening System of Brassica napus L. MS Thesis of Shanxi Agricultural University, Jinzhong, Shanxi, China, 2004 (in Chinese with English abstract). | |
[34] | 李尊文. 芥蓝悬浮细胞培养的初步研究与ABD2的遗传转化. 福建农林大学硕士学位论文, 福建福州, 2022. |
Li Z W. Preliminary Study on Suspension Cell Culture of Chinese Kale and Genetic Transformation of ABD2. MS Thesis of Fujian Agriculture and Forestry University, Fuzhou, Fujian, China, 2022 (in Chinese with English abstract). | |
[35] | 倪德祥. 光在植物组织培养中的调控作用. 自然杂志, 1986, 9(3): 35-40. |
Ni D X. The regulatory role of light in plant tissue culture. Chin J Nat, 1986, 9(3): 35-40 (in Chinese). | |
[36] | 董飞, 陈运起, 刘世琦, 高莉敏, 王传增. 种子消毒方法对大葱无菌苗培养的影响. 中国蔬菜, 2011, (18): 74-76. |
Dong F, Chen Y Q, Liu S Q, Gao L M, Wang C Z. Influence of seed disinfection methods on Aseptic Welsh onion tissue culture. China Veget, 2011, (18): 74-76 (in Chinese with English abstract). | |
[37] | 王欢, 付小蕾, 毛洪玉, 祝朋芳. 根癌农杆菌介导观赏羽衣甘蓝遗传转化体系优化. 沈阳农业大学学报, 2019, 50: 543-549. |
Wang H, Fu X L, Mao H Y, Zhu P F. Agrobacterium tumefaciens-mediated genetic transformation system in Brassica oleracea var. acephala. J Shenyang Agric Univ, 2019, 50: 543-549 (in Chinese with English abstract). | |
[38] | 刘洁. 结球甘蓝离体遗传转化体系的建立及转FOC1基因植株的获得. 甘肃农业大学硕士学位论文, 甘肃兰州, 2016. |
Liu J. Agrobacterium Mediated Transformation and Regeneration of Transgenic Plants with FOC1 Gene in Brassica oleracea. MS Thesis of Gansu Agricultural University, Lanzhou, Gansu, China, 2016 (in Chinese with English abstract). | |
[39] |
Palmer C E. Enhanced shoot regeneration from Brassica campestris by silver nitrate. Plant Cell Rep, 1992, 11: 541-545.
doi: 10.1007/BF00233088 pmid: 24213282 |
[40] | 唐丽. LED光质在植物组织培养和芽苗菜栽培中的调控作用及机理. 南京农业大学硕士学位论文, 江苏南京, 2013. |
Tang L. Regulation and Mechanism of LED Light Quality in Plant Tissue Culture and Sprouts Cultivation. MS Thesis of Nanjing Agricultural University, Nanjing, Jiangsu, China, 2013 (in Chinese with English abstract). | |
[41] | 林小苹, 赖钟雄, 黄浅. 光质对植物离体培养的影响. 亚热带农业研究, 2008, 4(1): 73-80. |
Lin X P, Lai Z X, Huang Q. Effects of light quality on plant tissue culture. Subtrop Agric Res, 2008, 4(1): 73-80 (in Chinese with English abstract). | |
[42] | 尾田羲治. 植物光形态建成. 刘瑞征, 译. 北京: 科学出版社, 1981. pp 1-5. |
Oda X Z. Plant Photomorphogenesis. Liu R Z, trans trans. Beijing: Science Press, 1981. pp 1-5 (in Chinese). | |
[43] | 余叔文, 汤章城. 植物生理与分子生物学. 北京: 科学出版社, 1998. pp 633-654. |
Yu S W, Tang Z C. Plant Physiology and Molecular Biology. Beijing: Science Press, 1998. pp 633-654 (in Chinese). | |
[44] | 陈汝民. 现代植物科学引论. 广州: 广东高等教育出版社, 1995. pp 415-428. |
Chen R M. Introduction to Modern Plant Science. Guangzhou: Guangdong Higher Education Press, 1995. pp 415-428 (in Chinese). | |
[45] | 崔堂兵, 郭勇, 林炜铁. 提高植物细胞培养法生产次级代谢物产量的方法. 植物生理学通讯, 2001, 37: 479-482. |
Cui T B, Guo Y, Lin W T. The ways to enhance the production of secondary metabolites by plant cell culture. Plant Physiol Commun, 2001, 37: 479-482 (in Chinese with English abstract). | |
[46] | Orozco-Mosqueda M D C, Santoyo G, Glick B R. Recent advances in the bacterial phytohormone modulation of plant growth. Plants, 2023, 12: 606. |
[47] | Rademacher W. Plant growth regulators: backgrounds and uses in plant production. J Plant Growth Regul, 2015, 34: 845-872. |
[48] | 王冬梅, 黄学林, 黄上志. 细胞分裂素类物质在植物组织培养中的作用机制. 植物生理学通讯, 1996, 32: 373-377. |
Wang D M, Huang X L, Huang S Z. The action mechanism of cytokinins in plant tissue culture. Plant Physiol Commun, 1996, 32: 373-377 (in Chinese with English abstract). | |
[49] | 王忠. 植物生理学. 北京: 中国农业出版社, 2000. pp 323-330. |
Wang Z. Plant Physiology. Beijing: China Agriculture Press, 2000. pp 323-330 (in Chinese). | |
[50] | Wong W C. In vitro propagation of bananas (Musa spp.): initiation, proliferation and development of shoot-tip cultures on defined media. Plant Cell Tissue Organ Cult, 2004, 6: 159-166. |
[51] |
Gerszberg A. Tissue culture and genetic transformation of cabbage (Brassica oleracea var. capitata): an overview. Planta, 2018, 248: 1037-1048.
doi: 10.1007/s00425-018-2961-3 pmid: 30066219 |
[52] | 曹家树, 余小林, 黄爱军, 徐淑英. 提高白菜离体培养植株再生频率的研究. 园艺学报, 2000, 27: 452-454. |
Cao J S, Yu X L, Huang A J, Xu S Y. Study on improving plant regeneration frequency of Chinese cabbage in vitro. Acta Hortic Sin, 2000, 27: 452-454 (in Chinese with English abstract). | |
[53] | Shin K S, Murthy H N, Heo J W, Hahn E J, Paek K Y. The effect of light quality on the growth and development of in vitro cultured Doritaenopsis plants. Acta Physiol Plant, 2008, 30: 339-343. |
[54] | Ramírez-Mosqueda M A, Iglesias-Andreu L G, Bautista-Aguilar J R. The effect of light quality on growth and development of in vitro plantlet of Stevia rebaudiana Bertoni. Sugar Technol, 2017, 19: 331-336. |
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