Scientia Agricultura Sinica ›› 2025, Vol. 58 ›› Issue (8): 1617-1626.doi: 10.3864/j.issn.0578-1752.2025.08.012

• HORTICULTURE • Previous Articles     Next Articles

Cloning of CmASMT and Its Role in Thermotolerance of Chrysanthemum

MENG Hui(), LUO BingYu, LU ZhengYu, WANG Peng, KANG DongRu, ZHENG ChengShu, WANG WenLi()   

  1. College of Horticulture Science and Engineering, Shandong Agricultural University/Chrysanthemum Research Center of China, Japan and Korea in Shandong Province, Tai’an 271018, Shandong
  • Received:2024-10-14 Accepted:2024-12-27 Online:2025-04-16 Published:2025-04-21
  • Contact: WANG WenLi

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Abstract:

【Background】 Chrysanthemum is one of the ten Chinese traditional flowers and four most important cut flowers in the world, originating from China and widely cultivated throughout the world. As chrysanthemum thrives in mild and cool climates, the summer high-temperature weather leads to continuous high temperature in protected cultivation environments, which seriously affects the yield and quality of chrysanthemum. 【Objective】Exogenous application of melatonin helps to regulate plant response to various abiotic stresses, including high-temperature stress. N-acetyl-5-hydroxytryptamine methyltransferase (ASMT) is the rate-limiting enzyme for melatonin biosynthesis. This study investigated the function of CmASMT, examined its effect on chrysanthemum growth under high-temperature stress, and explored how endogenous melatonin synthesis influences thermotolerance. This paper provides a theoretical basis for the molecular mechanism of melatonin-regulated thermotolerance in plants and the molecular breeding of chrysanthemum. 【Method】The CmASMT involved in melatonin biosynthesis was cloned from Chrysanthemum morifolium Jinba and analyzed for bioinformatics, subcellular localization, and spatiotemporal expression properties. CmASMT gene-silenced plants were generated by using virus-induced gene silencing (VIGS) technology to investigate the effects of CmASMT on heat tolerance of chrysanthemum through photosynthesis, the stability of the membrane system, and antioxidant system. 【Result】The ORF of CmASMT is 1 059 bp in length, encodes 352 amino acids, and belongs to the O-methyltransferases family. During the vegetative growth stage, CmASMT is expressed in roots, stems, leaves and buds of chrysanthemum, with the highest levels observe in the roots. Expression of CmASMT was induced by high temperature, low temperature, waterlogging, salt and drought stress, and CmASMT was most responsive to high-temperature stress. The CmASMT protein localized to the cell membranes, cytoplasm and nucleus. The content of endogenous melatonin in CmASMT-silenced plants of chrysanthemum was significantly reduced. Under high-temperature stress, CmASMT-silenced plants exhibited inhibition of photosynthesis, reduction of membrane system stability, aggravation of oxidative stress, weakening of antioxidant enzyme activity, and an increase of denatured proteins. CmASMT may enhance the photosynthetic efficiency of chrysanthemum by regulating the synthesis of endogenous melatonin, while improving the activity of antioxidant enzymes, scavenging excess reactive oxygen species, alleviating membrane structure damage and degradation of photosynthetic pigments, thereby improving chrysanthemum thermotolerance. 【Conclusion】CmASMT plays an important role in responding to high-temperature stress in chrysanthemum by regulating endogenous melatonin synthesis.

Key words: chrysanthemum, high temperature stress, melatonin, CmASMT, virus induced gene silencing

Fig. 1

Expression pattern analysis and subcellular localization of CmASMT A: Expression levels of CmASMT in different tissue organs of Chrysanthemum; B: Expression levels of CmASMT in chrysanthemum leaves under different stresses; C: Subcellular localization of CmASMT, Bar=20 μm. CK: Control check; HS: Heat stress; LS: Low temperature stress; WS: Waterlogging stress; SS: Salt stress; DS: Drought stress. Different letters indicate significant differences (P<0.05). The same as below"

Fig. 2

Validation of CmASMT gene silencing efficiency A: Expression levels of CmASMT gene in untreated, control and experimental groups; B: Changes in melatonin content of chrysanthemum by CmASMT silencing. WT: Wild type; TRV: Control group; 1-10: TRV-CmASMT silencing plant"

Fig. 3

Effects of CmASMT silencing on morphology and photosynthesis in chrysanthemum under heat stress A: External form of chrysanthemum; B: Net photosynthetic rate; C: Maximum photochemical efficiency of PSⅡ; D: Chlorophyll content"

Fig. 4

Effects of CmASMT silencing on cell membrane permeability and lipid peroxidation in chrysanthemum under heat stress"

Fig. 5

Effects of CmASMT silencing on ROS content in chrysanthemum under heat stress"

Fig. 6

Effects of CmASMT silencing on antioxidant enzyme activities and related gene expression profiles in chrysanthemum under heat stress A: SOD activity; B: APX activity; C: POD activity; D: CAT activity; E: GR activity; F: The expression level of CmSOD; G: The expression level of CmCAT; H: The expression level of CmPOD"

Fig. 7

Effects of CmASMT silencing on protein content in chrysanthemum under heat stress"

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