Establishment of high-efficiency regeneration system from in vitro leaves of ‘Jingzao 39’

Gao Yi; Bo Wenhao; Li Yingyue; Cao Ming; Pang Xiaoming

doi:10.12171/j.1000-1522.20210370

Journal of Beijing Forestry University > 2023 > 45(2): 68-77. > DOI: 10.12171/j.1000-1522.20210370

Gao Yi, Bo Wenhao, Li Yingyue, Cao Ming, Pang Xiaoming. Establishment of high-efficiency regeneration system from in vitro leaves of ‘Jingzao 39’[J]. Journal of Beijing Forestry University, 2023, 45(2): 68-77. DOI: 10.12171/j.1000-1522.20210370

Citation:

PDF (1526 KB)

Establishment of high-efficiency regeneration system from in vitro leaves of ‘Jingzao 39’

1.
School of Biological Sciences and Biotechnology, National Engineering Research Center of Tree Breeding and Ecological Restoration, Beijing Forestry University, Beijing 100083, China
2.
National Foundation for Improved Cultivars of Chinese Jujube, Cangzhou 061000, Hebei, China

More Information

Received Date: September 21, 2021
Revised Date: October 31, 2021
Accepted Date: November 13, 2022
Available Online: November 15, 2022
Published Date: February 24, 2023

Graphical Abstract

Abstract

Abstract

Objective ‘Jingzao 39’ is a new high-quality jujube variety. There is no effective regeneration system in vitro at present, which affects the cultivation of self-rooted seedlings and the purity of seedlings. In order to solve the problems of low genetic transformation efficiency of jujube, different influencing factors of jujube leaf regeneration were explored in this study, in order to establish an efficient leaf regeneration system.

Method Using the leaves of tissue cultured seedlings of ‘Jingzao 39’ as experimental materials, the effects of different inoculation methods, types of basic culture medium, types and concentration ratio of plant growth regulators on the direct induction of adventitious bud regeneration from detached leaves were studied.

Result The results showed that the regeneration rate of the inoculation method with the leaves being placed upward was much higher than that of back up, which was the best placement method for the regeneration of adventitious buds from leaves of ‘Jingzao 39’. The effect of basic culture medium on leaf regeneration of ‘Jingzao 39’ was greater than that of plant growth regulator, and the optimal basic medium and plant growth regulator were 1/2 MS and cytokinin 6-BA, respectively. The optimal regeneration system for leaves of ‘Jingzao 39’ tissue cultured seedlings established in this study was 1/2 MS medium + 1.0 mg/L thiabenduron + 0.5 mg/L 6-BA + 0.2 mg/L NAA + 30 g/L maltose + 6 g/L agar, an average of 4.67 adventitious buds could be regenerated for each leaf. Then, the optimal propagation system of ‘Jingzao 39’ tissue culture seedlings was initially explored and established: MS + 20 g/L maltose + 6 g/L agar + 3.00 mg/L 6-BA + 0.10 mg/L NAA, with the branching rate up to 88%, and the average branch number of 2.76; the most suitable rooting medium was: 1/2MS + 20 g/L sucrose + 8 g/L agar + 1.50 mg/L IBA + 0.30 mg/L NAA, with rooting rate of 90.91%, and the average root length up to 4.08 cm.

Conclusion The leaf regeneration efficiency of ‘Jingzao 39’ is greatly influenced by external factors, especially the basic medium type and inoculation method, and each factor does not play an independent role. This study establishes a complete cultivation system for the functional verification of interested gene with the leaves of ‘Jingzao 39’ as transgenic materials, and lays a foundation for accelerating the jujube research process.
- ‘Jingzao 39’ leaf,
- direct regeneration,
- adventitious bud,
- optimal medium

FullText(HTML)

References (28)

References

[1]	Liu M J, Cheng C Y. A taxonomy study on the genus Ziziphus[J]. Acta Horticulturae, 1995, 390: 161−165.
[2]	Liu M J. Chinese jujube: botany and horticulture[J]. Horticultural Review, 2006, 32: 229−298.
[3]	何业华, 熊细满, 谢碧霞, 等. 枣树组织培养愈伤组织诱导的研究[J]. 中南林学院学报, 1997, 17(1): 13−18. He Y H, Xiong X M, Xie B X, et al. Inducement of calluses in the tissue culture of jujube trees[J]. Journal of Central South Forestry College, 1997, 17(1): 13−18.
[4]	周瑞金, 刘孟军. 枣离体叶片高效再生植株的研究[J]. 园艺学报, 2006, 33(3): 625−628. doi: 10.3321/j.issn:0513-353X.2006.03.034 Zhou R J, Liu M J. Establishment of high-efficient in vitro leaf regeneration system in Chinese jujube[J]. Acta Horticulturae Sinica, 2006, 33(3): 625−628. doi: 10.3321/j.issn:0513-353X.2006.03.034
[5]	李云, 王宇, 田砚亭, 等. 赞皇大枣叶片再生植株的初步研究[J]. 核农学报, 2003, 17(3): 187−190. doi: 10.3969/j.issn.1000-8551.2003.03.006 Li Y, Wang Y, Tian Y T, et al. A preliminary study on the plant regeneration from leaves of Zanhuang jujube[J]. Acta Agriculturae Nu-cleatae Sinica, 2003, 17(3): 187−190. doi: 10.3969/j.issn.1000-8551.2003.03.006
[6]	胡影, 崔建国, 施满, 等. 枣叶片愈伤组织的诱导和器官分化[J]. 甘肃林业科技, 1993, 18(4): 18−19. Hu Y, Cui J G, Shi M, et al. Induction and organ differentiation of calluses in jujube leaves[J]. Gansu Forestry Science and Technology, 1993, 18(4): 18−19.
[7]	张春雨. 黑果腺肋花楸离体叶片再生体系建立与原生质体分离培养研究[D]. 长春: 吉林农业大学, 2005. Zhang C Y. Study on the establishment of ex vivo leaf regeneration system and protoplast isolation and culture of black fruit glandular rib rowan[D]. Changchun: Jilin Agricultural University, 2005.
[8]	李晓青, 张晓申. 灰枣试管苗移栽技术研究[J]. 河南农业科学, 2007, 36(8): 97−98. doi: 10.3969/j.issn.1004-3268.2007.08.032 Li X Q, Zhang X S. Research on transplanting technology of gray jujube test tube seedlings[J]. Journal of Henan Agricultural Sciences, 2007, 36(8): 97−98. doi: 10.3969/j.issn.1004-3268.2007.08.032
[9]	陈宗礼, 延志莲, 齐龙. 枣叶片离体培养再生植株[J]. 植物生理学通讯, 1996, 32(1): 27−28. Chen Z L, Yan Z L, Qi L. Plantlet regeneration from leaves cultures of Ziziphus jujuba[J]. Plant Physiology Communications, 1996, 32(1): 27−28.
[10]	孙清荣, 孙洪雁, 郑红军, 等. 酸枣叶片不定植株的诱导[J]. 果树科学, 2000(1): 48−51. Sun Q R, Sun H Y, Zheng H J, et al. Study on the plant regeneration from leaves of sour jujube (Zizyphus spinosus Hu.)[J]. Journal of Fruit Science, 2000(1): 48−51.
[11]	王玖瑞, 刘孟军, 代丽. 枣树组织培养研究进展[J]. 果树学报, 2002(5): 336−339. doi: 10.3969/j.issn.1009-9980.2002.05.011 Wang J R, Liu M J, Dai L. Advances in tissue culture of Chinese jujube[J]. Journal of Fruit Science, 2002(5): 336−339. doi: 10.3969/j.issn.1009-9980.2002.05.011
[12]	陈宗礼, 延志莲, 薛皓, 等. 沾化冬枣叶片培养和植株再生[J]. 植物生理学通讯, 2002, 38(6): 584. Chen Z L, Yan Z L, Xue H, et al. In vitro culture of leaves and plantlet regeneration of Ziziphus jujuba var. zhanhua-dongzao[J]. Plant Physiology Communications, 2002, 38(6): 584.
[13]	胡桂兵, 陈大成, 郑启发, 等. 抗生素对台湾青枣茎段和愈伤组织生长的影响[J]. 华南农业大学学报, 2001, 22(2): 21−23. doi: 10.3969/j.issn.1001-411X.2001.02.007 Hu G B, Chen D C, Zheng Q F, et al. Effect of antibiotics on growth of stem segments and callus of ber (Ziziphus mauritiana Lam.)[J]. Journal of South China Agricultural University, 2001, 22(2): 21−23. doi: 10.3969/j.issn.1001-411X.2001.02.007
[14]	何振艳, 王玉国, 石武良, 等. 山西特有品种梨枣叶片的组织培养及植株再生[J]. 植物生理学通讯, 2002, 38(5): 457. He Z Y, Wang Y G, Shi W L, et al. Tissue culture and plant regeneration of leaves of Lizao endemic variety in Shanxi[J]. Plant Physiology Communications, 2002, 38(5): 457.
[15]	尹美强, 王玉国, 温银元, 等. 梨枣叶片和茎段再生体系的建立[J]. 西北植物学报, 2005, 25(10): 1954−1959. doi: 10.3321/j.issn:1000-4025.2005.10.005 Yin M Q, Wang Y G, Wen Y Y, et al. The construction of the regeneration system for the Chinese date cultivar Lizao to regenerate from its leaves and stems[J]. Acta Botanica Boreali-Occidentalia Sinica, 2005, 25(10): 1954−1959. doi: 10.3321/j.issn:1000-4025.2005.10.005
[16]	陈宗礼, 张向前, 齐向英, 等. 枣树叶片愈伤组织培养与再生植株的研究[J]. 福建林业科技, 2007, 34(3): 23−27. doi: 10.3969/j.issn.1002-7351.2007.03.006 Chen Z L, Zhang X Q, Qi X Y, et al. Callus culture from leaves and plant regeneration of jujube trees[J]. Journal of Fujian Forestry Science and Technology, 2007, 34(3): 23−27. doi: 10.3969/j.issn.1002-7351.2007.03.006
[17]	王国平, 李晓梅, 陈秋芳. 枣不定芽再生体系的研究[J]. 中国农学通报, 2006, 22(4): 281−283. doi: 10.3969/j.issn.1000-6850.2006.04.074 Wang G P, Li X M, Chen Q F. Study on jujube adventitious bud regeneration system[J]. Chinese Agricultural Science Bulletin, 2006, 22(4): 281−283. doi: 10.3969/j.issn.1000-6850.2006.04.074
[18]	黄健, 马锋旺, 樊军锋, 等. 枣树离体叶片不定芽再生体系建立的研究[J]. 西北植物学报, 2006, 26(5): 942−948. doi: 10.3321/j.issn:1000-4025.2006.05.012 Huang J, Ma F W, Fan J F, et al. In vitro plant regeneration with adventitious buds of Zizyphus jujuba leaves[J]. Acta Botanica Boreali-Occidentalia Sinica, 2006, 26(5): 942−948. doi: 10.3321/j.issn:1000-4025.2006.05.012
[19]	Ma C H, Ye X, Chen Y H. Anatomical observations of adventitious bud regenetation from leaf explants of Ziziphus jujuba Mill. ‘Huizao’[J]. Horticulture Environment and Biotechnology, 2012, 53(4): 316−319. doi: 10.1007/s13580-012-0081-8
[20]	潘青华, 白金, 王保强. 枣鲜食优质新品种‘京枣39’[J]. 中国果树, 2003(4): 3−4. doi: 10.3969/j.issn.1000-8047.2003.04.002 Pan Q H, Bai J, Wang B Q. New high-quality jujube variety ‘Jingzao 39’[J]. China Fruits, 2003(4): 3−4. doi: 10.3969/j.issn.1000-8047.2003.04.002
[21]	程佑发, 安黎哲, 浦铜良, 等. 枣愈伤组织诱导和再生植株[J]. 西北植物学报, 2000, 20(3): 364−369. doi: 10.3321/j.issn:1000-4025.2000.03.008 Cheng Y F, An L Z, Pu T L, et al. Induction of callus and plant regeneration of Zizyphus jujuba Mill.[J]. Acta Botanica Boreali-Occidentalia Sinica, 2000, 20(3): 364−369. doi: 10.3321/j.issn:1000-4025.2000.03.008
[22]	王慧瑜, 马锋旺, 张晓申, 等. 鸡心枣试管苗叶片再生植株的研究[J]. 中国南方果树, 2007, 36(4): 48−50. doi: 10.3969/j.issn.1007-1431.2007.04.024 Wang H Y, Ma F W, Zhang X S, et al. Study on leaf regeneration plants of Jixinzao test tube seedlings[J]. South China Fruits, 2007, 36(4): 48−50. doi: 10.3969/j.issn.1007-1431.2007.04.024
[23]	肖蓉, 王国平, 李春燕, 等. 辣椒枣试管苗叶片再生植株研究[J]. 山西农业科学, 2012, 40(3): 193−196, 216. doi: 10.3969/j.issn.1002-2481.2012.03.02 Xiao R, Wang G P, Li C Y, et al. Research of leaf regeneration plant in Ziziphus jujuba Mill. Lajiaozao test tube seedlings[J]. Journal of Shanxi Agricultural Sciences, 2012, 40(3): 193−196, 216. doi: 10.3969/j.issn.1002-2481.2012.03.02
[24]	周瑞金. 枣(Ziziphus jujuba Mill.)离体叶片高效再生体系的建立[D]. 保定: 河北农业大学, 2004. Zhou R J. Establishment of an efficient regeneration system for ex vivo leaves of Ziziphus jujuba Mill.[D]. Baoding: Agricultural University of Hebei, 2004.
[25]	宁强, 刘晓光, 刘孟军. 冬枣叶片愈伤组织诱导研究[J]. 河北林果研究, 2008, 23(4): 348−352. doi: 10.3969/j.issn.1007-4961.2008.04.002 Ning Q, Liu X G, Liu M J. Callus inducement in tissue culture of jujube[J]. Hebei Forest Fruit Research, 2008, 23(4): 348−352. doi: 10.3969/j.issn.1007-4961.2008.04.002
[26]	徐明, 代培红, 罗淑萍, 等. 哈密大枣叶片不定芽再生体系研究[J]. 新疆农业大学学报, 2014, 37(3): 185−190. doi: 10.3969/j.issn.1007-8614.2014.03.003 Xu M, Dai P H, Luo S P, et al. Study on adventitious bud regeneration system of Hami-Dazao leaves[J]. Journal of Xinjiang Agricultural University, 2014, 37(3): 185−190. doi: 10.3969/j.issn.1007-8614.2014.03.003
[27]	李静, 张景臣, 郭伟, 等. 毛白杨叶片愈伤组织的诱导、芽的分化及不定根的再生[J]. 山东林业科技, 2007, 168(1): 52−53. doi: 10.3969/j.issn.1002-2724.2007.01.020 Li J, Zhang J C, Guo W, et al. Callus induction, bud differentiation and adventitious root regeneration from leaves of Populus tomentosa[J]. Shandong Forestry Science and Technology, 2007, 168(1): 52−53. doi: 10.3969/j.issn.1002-2724.2007.01.020
[28]	刘晓光, 单梅华, 苏帅, 等. 山杏组培苗生根培养基优化及移栽[J]. 分子植物育种, 2020, 18(6): 1999−2005. doi: 10.13271/j.mpb.018.001999 Liu X G, Shan M H, Su S, et al. Transplanting and optimization of rooting medium for apricot tissue cultured seedlings[J]. Molecular Plant Breeding, 2020, 18(6): 1999−2005. doi: 10.13271/j.mpb.018.001999

[1]	Zhao Hanqing, Wang Qiang, Pang Xiaoming, Bo Wenhao, Li Yingyue. In vitro polyploid induction and optimization of its proliferation and rooting culture system for ‘Jingzao 39’[J]. Journal of Beijing Forestry University, 2024, 46(6): 118-126. DOI: 10.12171/j.1000-1522.20230039
[2]	Yang Weicong, Zhang Chenxing, Zhao Hanqing, Liu Xinru, Pang Xiaoming, Li Yingyue. Optimization of tissue culture system and establishment of high-efficiency adventitious bud regeneration system from leaf explants of tetraploid Ziziphus jujuba var. spinosa[J]. Journal of Beijing Forestry University, 2023, 45(6): 8-18. DOI: 10.12171/j.1000-1522.20220067
[3]	CAO Shan, JIANG Lu-yao, LI Li-hong, YAO Xiao-yun, ZHANG Qiang, HAN Jing-yi, WANG Ying, LI Hui, LU Hai.. Cloning and enzymatic analysis of medium-chain acyl coenzyme A synthetase in Populus trichocarpa.[J]. Journal of Beijing Forestry University, 2016, 38(7): 9-15. DOI: 10.13332/j.1000-1522.20160121
[4]	WANG Si-yao, LI Ming-yang, SHAO Zhan-yuan, YIN Jing. Accumulation of total phenolics, triterpenoids and flavonoids in tufted buds of Tamarix chinensis under different medium and hormone treatment[J]. Journal of Beijing Forestry University, 2014, 36(5): 74-81. DOI: 10.13332/j.cnki.jbfu.2014.05.021
[5]	LI Zhi-jun, JIAO Pei-pei, ZHOU Zheng-li, LI Qian, LI Jian-qiang. Anatomic characteristics of transverse lateral roots and adventitious buds of Populus euphratica.[J]. Journal of Beijing Forestry University, 2011, 33(5): 42-48.
[6]	ZHANG Xiao-xia, DING You-fang, SHI Ling-ling, ZHANG Yun-wei, WANG Fu-hai, YANG Fu-yu, LIU Yu-jun. Tissue culture and rapid propagation of Pueraria lobata.[J]. Journal of Beijing Forestry University, 2010, 32(5): 138-142.
[7]	LI Hui, ZHANG Qian, FU Wen-feng, ZHANG Jin-feng. Adventitious bud formation in vitro from mature zygotic embryos of Pinus tabulaeformis Carr.[J]. Journal of Beijing Forestry University, 2010, 32(5): 111-115.
[8]	ZHANG Qi-chang, LIANG Qi-lan, XIA Xin-li, YIN Wei-lun. In vitro culture and plant regeneration from sprouting buds of Lonicera edulis[J]. Journal of Beijing Forestry University, 2010, 32(4): 126-130.
[9]	TIAN Cui-ting, LÜ Hong-fei, WANG Feng, ZHANG Chun-yu, LIN Jin-xing. Effects of medium components on in vitro pollen germination and pollen tube growth of Picea wilsonii Mast.[J]. Journal of Beijing Forestry University, 2007, 29(1): 47-52. DOI: 10.13332/j.1000-1522.2007.01.009
[10]	GAO Li-ping, BAO Man-zhu. Optimization of Agrobacterium-mediated transformation of Rosa hybrida[J]. Journal of Beijing Forestry University, 2005, 27(4): 60-64.

Cited By

Cited by

Periodical cited type(6)

1.	桂平，吴高殷，黄小琴，李柯萱. 野生空心泡组织培养再生体系的建立. 山地农业生物学报. 2025(01): 65-69 .
2.	马灵，袁轶，杨爽，高艺，陈学勋，焦立军，刘巍巍，薄文浩，庞晓明. ‘京枣39’叶片遗传转化体系的建立. 华中农业大学学报. 2025(01): 43-49 .
3.	赵汗青，王强，庞晓明，薄文浩，李颖岳. ‘京枣39’离体多倍体诱导及其增殖、生根培养体系优化. 北京林业大学学报. 2024(06): 118-126 . 本站查看
4.	冯雪晶，马灵，杨爽，薄文浩，陈学勋，庞晓明. ‘京枣39’愈伤组织遗传转化体系构建. 北京林业大学学报. 2024(10): 74-80 . 本站查看
5.	杨伟聪，张晨星，赵汗青，刘欣茹，庞晓明，李颖岳. 四倍体酸枣组培体系优化及其叶片离体不定芽高效再生体系的建立. 北京林业大学学报. 2023(06): 8-18 . 本站查看
6.	朱高浦. 枣分子育种研究进展. 中南林业科技大学学报. 2023(10): 1-10 .

Other cited types(3)

Get Citation

PDF

XML

Article views (704) PDF downloads (78) Cited by(9)

Turn off MathJax

Article Contents

Abstract

References

Establishment of high-efficiency regeneration system from in vitro leaves of ‘Jingzao 39’