TECHNIQUE AND METHOD

Establishment of Regeneration System of Chrysanthemum vestitum with Three Floret Forms

  • Xiaoyun Wu ,
  • Minling Liao ,
  • Xueru Li ,
  • Zichun Shu ,
  • Jiatong Xin ,
  • Bohan Zhang ,
  • Silan Dai
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  • National Engineering Research Center for Floriculture, Beijing Key Laboratory of Ornamental Plants Germplasm Innovation & Molecular Breeding, Beijing Laboratory of Urban and Rural Ecological Environment, School of Landscape Architecture, Beijing Forestry University, Beijing 100083, China

Received date: 2023-06-25

  Accepted date: 2023-09-19

  Online published: 2023-10-10

Abstract

Chrysanthemum vestitum, a closely related hexaploid wild species of cultivated C. × morifolium has the same and typical flat, tubular and mixed floret forms in its natural population as cultivated chrysanthemum and is an ideal material for studying the petal variation in chrysanthemum. The development progress of this ray floret is affected by key differential genes such as auxin and floral organ development, but the lack of stable and efficient regeneration systems for different floret forms has restricted the study of petal type-related genes in C. vestitum. In this paper, the authors used three floret forms of C. vestitum collected from Funiu Mountain in Henan Province to establish regeneration systems using leaves and transverse thin cell layers (tTCLs) as explants. The results showed that the optimal media for callus induction and adventitious bud differentiation were MS+1 mg∙L-1 NAA+2 mg∙L-1 6-BA with flat floret leaves as explants, and the callus induction rate was 100% at 20 days after inoculation. The differentiation rate of adventitious buds was 100%. The optimal rooting medium was 1/2MS+0.2 mg∙L-1 NAA, and the rooting rate of regenerated plants was 100%. The optimal leaf regeneration system for the flat floret strain was also suitable for determining both the tubular and mixed floret strains. The differentiation rates of adventitious buds were 83.46% and 91.67%, respectively, and the rooting rates were 100%. The flowering plants were observed after transplanting, and it was found that the flower types of the regenerated plants with three different floret forms obtained by the leaf regeneration system were stable, which provided a technical method for the subsequent analysis of the morphological variation mechanism of ray florets by using different floret forms of C. vestitum.

Cite this article

Xiaoyun Wu , Minling Liao , Xueru Li , Zichun Shu , Jiatong Xin , Bohan Zhang , Silan Dai . Establishment of Regeneration System of Chrysanthemum vestitum with Three Floret Forms[J]. Chinese Bulletin of Botany, 2024 , 59(2) : 245 -256 . DOI: 10.11983/CBB23084

References

[1] Annadana S, Rademaker W, Ramanna M, Udayakumar M, de Jong J (2000). Response of stem explants to screening and explant source as a basis for methodical advancing of regeneration protocols for chrysanthemum. Plant Cell Tissue Organ Cult 62, 47-55.
[2] Aswath CR, Mo SY, Kim SH, Kim DH (2004). IbMADS 4 regulates the vegetative shoot development in transgenic chrysanthemum (Dendrathema grandiflora (Ramat.) Kitamura). Plant Sci 166, 847-854.
[3] Chakrabarty D, Mandal AKA, Datta SK (1999). Management of chimera through direct shoot regeneration from florets of chrysanthemum (Chrysanthemum morifolium Ramat.). J Hortic Sci Biotechnol 74, 293-296.
[4] Chen H, Mo LW, Wang ZF, Wang YC, Chen YZ, Zhang W, Song XQ, Meng XY (2023). Establishment of tissue culture and plant regeneration system of ‘Zhenfen’ Chrysanthemum morifolium. Mol Plant Breed 21, 1633-1640. (in Chinese)
  陈好, 莫丽文, 王泽峰, 王亚沉, 陈英转, 张雯, 宋希强, 孟新亚 (2023). 地被菊花‘珍粉’组织培养与植株再生体系的建立. 分子植物育种 21, 1633-1640.
[5] Chen H, Wang YF, Chen SM, Liu SY, Teng NJ, Lan W, Chen FD (2009). Study on tissue culture of five wild species closely related to Dendranthema × grandiflorum. J Nanjing Agric Univ 32, 30-35. (in Chinese)
  晨卉, 王艳芳, 陈素梅, 刘思余, 滕年军, 兰伟, 陈发棣 (2009). 五种菊花近缘植物组织培养研究. 南京农业大学学报 32, 30-35.
[6] Chen PY (2010). Study on Regeneration of Chrysanthemum koraiense Nakai. Master’s thesis. Dalian: Liaoning Normal University. pp. 17-28. (in Chinese)
  陈鹏彦 (2010). 朝鲜野菊再生系统建立的研究. 硕士论文. 大连: 辽宁师范大学. pp. 17-28.
[7] Dai SL, Chen JY (1997). A cladistic study on some Dendranthema spp. in China. Wuhan Bot Res J 15, 27-34. (in Chinese)
  戴思兰, 陈俊愉 (1997). 中国菊属一些种的分支分类学研究. 武汉植物学研究 15, 27-34.
[8] Dai SL, Chen JY, Li WB (1998). Application of RAPD analysis in the study on the origin of Chinese cultivated chrysanthemum. Acta Bot Sin 40, 1053-1059. (in Chinese)
  戴思兰, 陈俊愉, 李文彬 (1998). 菊花起源的RAPD分析. 植物学报 40, 1053-1059.
[9] Dai SL, Wang WK, Huang JP (2002). Advances of researches on phylogeny of Dendranthema and origin of chrysanthemum. J Beijing For Univ 24(5), 230-234. (in Chinese)
  戴思兰, 王文奎, 黄家平 (2002). 菊属系统学及菊花起源的研究进展. 北京林业大学学报 24(5), 230-234.
[10] Dai SL, Zhong Y, Zhang XY (1995). Study on numerical taxonomy of some Chinese species of Dendranthema (DC) Des Moul. J Beijing For Univ 17(4), 9-15. (in Chinese)
  戴思兰, 钟扬, 张晓艳 (1995). 中国菊属植物部分种的数量分类研究. 北京林业大学学报 17(4), 9-15.
[11] Ding XX (2018). Study progress of tissue culture regeneration system of chrysanthemum. J Liaoning For Sci Tech (4), 64-65, 68. (in Chinese)
  丁晓霞 (2018). 菊花组培再生体系研究进展. 辽宁林业科技 (4), 64-65, 68.
[12] Editorial Committee of the Floral of China, Chinese Academy of Sciences(1983). Floral of China, Vol. 76. Beijing: Science Press. pp. 29-42. (in Chinese)
  中国科学院中国植物志编辑委员会(1983). 中国植物志, 第76卷. 北京: 科学出版社. pp. 29-42.
[13] Fan GX (2017). Transcriptome Analysis of Ray Floret Variation in Chrysanthemum vestitum. Master’s thesis. Beijing: Beijing Forestry University. pp. 20-21. (in Chinese)
  樊光迅 (2017). 毛华菊舌状花形态变异的转录组分析. 硕士论文. 北京: 北京林业大学. pp. 20-21.
[14] Fan GX, Qi S, Wang WK, Dai SL (2016). Mathematical analysis of morphological traits of Chrysanthemum vestitum. Advances in Ornamental Horticulture of China 2016. Changsha: Ornamental Horticulture Professional Committee, Chinese Society for Horticultural Science. pp. 134-141. (in Chinese)
  樊光迅, 亓帅, 王文奎, 戴思兰 (2016). 毛华菊形态性状变异的数学分析. 中国观赏园艺研究进展2016. 长沙: 中国园艺学会观赏园艺专业委员会. pp. 134-141.
[15] Fu JX, Zhao C, Wang Y, Dai SL (2012). Establishing an in vitro regeneration system from hypocotyls of Chrysanthemum lavandulifolium. J Beijing For Univ 34(3), 91-96. (in Chinese)
  付建新, 张超, 王翊, 戴思兰 (2012). 甘菊下胚轴离体再生体系的建立. 北京林业大学学报 34(3), 91-96.
[16] Gao YK, Zhao B, Ding GX, Zhang QX (2001). Shoot regeneration from stem and leaf explant of Dendrathema grandiflorum. J Beijing For Univ 23, 32-33. (in Chinese)
  高亦珂, 赵勃, 丁国勋, 张启翔 (2001). 菊花茎叶外植体再生体系的研究. 北京林业大学学报 23, 32-33.
[17] Han KT, Wang J, Dai SL (2009). Adventitious shoot regeneration from internode transverse thin cell layers of cut spray chrysanthemum. J Beijing For Univ 31, 102-107. (in Chinese)
  韩科厅, 王娟, 戴思兰 (2009). 多头切花菊横切薄层不定芽再生的研究. 北京林业大学学报 31, 102-107.
[18] Hill GP (1968). Shoot formation in tissue cultures of Chrysanthemum ‘Bronze Pride’. Physiol Plant 21, 386-389.
[19] Kaul V, Miller RM, Hutchinson JF, Richards D (1990). Shoot regeneration from stem and leaf explants of Dendranthema grandiflora Tzvelev (syn.Chrysanthemum morifolium Ramat.). Plant Cell Tissue Organ Cult 21, 21-30.
[20] Ledger SE, Deroles SC, Given NK (1991). Regeneration and Agrobacterium-mediated transformation of chrysanthemum. Plant Cell Rep 10, 195-199.
[21] Li YJ, Li Y, Huang H, Dai SL (2018). Establishment of an efficient regeneration system for the cut flower Chrysanthemum ‘Fenguiren’. Advances in Ornamental Horticulture of China 2018. Harbin: Ornamental Horticulture Professional Committee, Chinese Society for Horticultural Science. pp. 427-434. (in Chinese)
  李亚军, 李悦, 黄河, 戴思兰 (2018). 切花菊‘粉贵人’高效再生体系的建立. 中国观赏园艺研究进展2018. 哈尔滨: 中国园艺学会观赏园艺专业委员会. pp. 427-434.
[22] Liao ML, Pu Y, Wu XY, Ma CF, Wang WK, Dai SL (2023). Establishment of regeneration system of Chrysanthemum indicum in Pingtan with various ligulate floret forms. Chin Bull Bot 58, 449-460. (in Chinese)
  廖敏凌, 蒲娅, 武晓云, 马朝峰, 王文奎, 戴思兰 (2023). 平潭野菊混合瓣型株系再生体系的建立. 植物学报 58, 449-460.
[23] Liu QQ, Dai SL (2007). Application of cytology methods on researches on chrysanthemum origin and Dendranthema spp. evolution. Nonwood For Res 25(3), 86-89. (in Chinese)
  刘倩倩, 戴思兰 (2007). 细胞学在探讨菊花起源与菊属植物进化中的应用. 经济林研究 25(3), 86-89.
[24] Liu YT, Huang H, Ye F, Dai SL (2018). The construction of regeneration system for hypocotyl of Senecio cruentus. Advances in Ornamental Horticulture of China 2018. Harbin: Ornamental Horticulture Professional Committee, Chinese Society for Horticultural Science. pp. 493-501. (in Chinese)
  刘钰婷, 黄河, 叶繁, 戴思兰 (2018). 瓜叶菊下胚轴再生体系的建立. 中国观赏园艺研究进展2018. 哈尔滨: 中国园艺学会观赏园艺专业委员会. pp. 493-501.
[25] Luo H, Wen XH, Zhou YY, Dai SL (2020). Establishment of in vitro regeneration system of Helenium aromaticum. Chin Bull Bot 55, 318-328. (in Chinese)
  罗虹, 温小蕙, 周圆圆, 戴思兰 (2020). 芳香堆心菊离体再生体系的建立. 植物学报 55, 318-328.
[26] Luo YL, Huang H, Lian L, Dai SL (2017). The construction of regeneration system for the ray floret of Senecio cruentus. Advances in Ornamental Horticulture of China 2017. Chengdu: Ornamental Horticulture Professional Committee, Chinese Society for Horticultural Science. pp. 252-259. (in Chinese)
  罗怡柳, 黄河, 连璐, 戴思兰 (2017). 瓜叶菊舌状花再生体系的建立. 中国观赏园艺研究进展2017. 成都: 中国园艺学会观赏园艺专业委员会. pp. 252-259.
[27] Ma YP, Chen MM, Wei JX, Zhao L, Liu PL, Dai SL, Wen J (2016). Origin of Chrysanthemum cultivars—evidence from nuclear low-copy LFY gene sequences. Biochem Syst Ecol 65, 129-136.
[28] Ma YP, Zhao L, Zhang WJ, Zhang YH, Xing X, Duan XX, Hu J, Harris A, Liu PL, Dai SL, Wen J (2020). Origins of cultivars of Chrysanthemum—evidence from the chloroplast genome and nuclear LFY gene. J Syst Evol 58, 925-944.
[29] Mao HY, Zhou Y, Liu D, Jia HM (2015). Establishment of receptor and genetic transformation system of ground- cover chrysanthemum ‘China Red’. J Shenyang Agric Univ 46, 672-677. (in Chinese)
  毛洪玉, 周杨, 刘迪, 贾红梅 (2015). 地被菊‘中国红’再生及遗传转化体系的建立. 沈阳农业大学学报 46, 672-677.
[30] Pu Y (2023). Molecular Mechanism of Auxin Regulation on Morphological Difference of Ray Florets in Chrysanthemum vestitum. Doctoral dissertation. Beijing: Beijing Forestry University. pp. 17-23. (in Chinese)
  蒲娅 (2023). 生长素调控毛华菊舌状花形态差异的分子机制. 博士论文. 北京: 北京林业大学. pp. 17-23.
[31] Pu Y, Huang H, Wen XH, Lu CF, Zhang BH, Gu XQ, Qi S, Fan GX, Wang WK, Dai SL (2020). Comprehensive transcriptomic analysis provides new insights into the mechanism of ray floret morphogenesis in chrysanthemum. BMC Genomics 21, 728.
[32] Pu Y, Liao ML, Li JZ, Tian YK, Wang ZM, Song X, Dai SL (2023). Floral development stage-specific transcriptomic analysis reveals the formation mechanism of different shapes of ray florets in chrysanthemum. Genes 14, 766.
[33] Ren JS, Gu XQ, Pu Y, Huang H (2021). Establishment of regeneration system for the ray floret of Chrysanthemum indicum var. huludoensis. Mod Hortic 44(24), 3-4, 7. (in Chinese)
  任江珊, 顾雪琪, 蒲娅, 黄河 (2021). 葫芦岛野菊舌状花再生体系建立. 现代园艺 44(24), 3-4, 7.
[34] Renou JP, Brochard P, Jalouzot R (1993). Recovery of transgenic chrysanthemum (Dendranthema grandiflora Tzvelev) after hygromycin resistance selection. Plant Sci 89, 185-197.
[35] Rout GR, Das P (1997). Recent trends in the biotechnology of Chrysanthemum: a critical review. Sci Hortic 69, 239-257.
[36] Song XB, Gao K, Fan GX, Zhao XG, Liu ZL, Dai SL (2018). Quantitative classification of the morphological traits of ray florets in large-flowered chrysanthemum. HortScience 53, 1258-1265.
[37] Song XB, Xu YH, Gao K, Fan GX, Zhang F, Deng CY, Dai SL, Huang H, Xin HG, Li YY (2020). High-density genetic map construction and identification of loci controlling flower-type traits in chrysanthemum (Chrysanthemum × morifolium Ramat.). Hortic Res 7, 108.
[38] Tanaka K, Kanno Y, Kudo S, Suzuki M (2000). Somatic embryogenesis and plant regeneration in chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura). Plant Cell Rep 19, 946-953.
[39] Tang ZH (1963). A study of the classification of Chinese chrysanthemums. Acta Hortic 2, 411-420. (in Chinese)
  汤忠皓 (1963). 中国菊花品种分类的探讨. 园艺学报 2, 411-420.
[40] Teixeira da Silva JA (2003). Thin cell layer technology for induced response and control of rhizogenesis in chrysanthemum. Plant Growth Regul 39, 67-76.
[41] Wang WK, Zhou CL, Dai SL (1999). Flower morphological variation of Dendranthema vestitum. J Beijing For Univ 21(3), 92-95. (in Chinese)
  王文奎, 周春玲, 戴思兰 (1999). 毛华菊花朵形态变异. 北京林业大学学报 21(3), 92-95.
[42] Wang ZB, Mo GX, Luo HL, Zhang DY (2015). Research of tissue culture and regenerated of the different explants of Chrysanthemum indicum L. North Hortic (18), 106-109. (in Chinese)
  王自布, 莫国秀, 罗会兰, 张德英 (2015). 菊花不同外植体组培快繁及其再生体系的研究. 北方园艺 (18), 106-109.
[43] Wu XY, Wen XH, Ma CF, Dai SL (2018). Rapid induction of plant regeneration from leaf of Chrysanthemum lanvandulifolium. Advances in Ornamental Horticulture of China 2018. Harbin: Ornamental Horticulture Professional Committee, Chinese Society for Horticultural Science. pp. 461-468. (in Chinese)
  武晓云, 温小蕙, 马朝峰, 戴思兰 (2018). 快速诱导甘菊叶片再生植株的方法. 中国观赏园艺研究进展2018. 哈尔滨: 中国园艺学会观赏园艺专业委员会. pp. 461-468.
[44] Wu ZP, Gao YK, Fan M, Gao YH (2020). Construction of regeneration and transformation system of Chrysanthemum ‘Jinbudiao’. Mol Plant Breed 18, 150-158. (in Chinese)
  吴志苹, 高亦珂, 范敏, 高耀辉 (2020). 菊花‘金不凋’再生及遗传转化体系的构建. 分子植物育种 18, 150-158.
[45] Xu YX (2005). Analysis on Morphological Diversity and Classification of Chrysanthemum. Master’s thesis. Beijing: Beijing Forestry University. pp. 23-36. (in Chinese)
  许莹修 (2005). 菊花形态性状多样性和品种分类的研究. 硕士论文. 北京: 北京林业大学. pp. 23-36.
[46] Yepes LM, Mittak V, Pang SZ, Gonsalves C, Slightom JL, Gonsalves D (1995). Biolistic transformation of chrysanthemum with the nucleocapsid gene of tomato spotted wilt virus. Plant Cell Rep 14, 694-698.
[47] Zhang SL (1965). Studies on the classification of garden varieties of florist’s chrysanthemum. Acta Hortic 4, 35-46, 61-62.
  张树林 (1965). 菊花品种分类的研究. 园艺学报 4, 35-46, 61-62.
[48] Zhou J (2009). Studies on the Problem of Origin of Chinese Garden Chrysanthemum. Doctoral dissertation. Beijing: Beijing Forestry University. pp. 31-36, 109-113.
  周杰 (2009). 关于中国菊花起源问题的若干实验研究. 博士论文. 北京: 北京林业大学. pp. 31-36, 109-113.
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