Cloning of Watermelon ClSHMT8 and Its Functional Analysis under Cold Stress and Salt Stress

doi:10.7668/hbnxb.20196251

Abstract

Abstract:

Serine hydroxy methyl transferase(SHMT) is a conserved pyridoxal phosphate-dependent enzyme that catalyzes the reversible conversion between serine and glycine. It is also an essential enzyme for nucleotide biosynthesis, folate, and amino acid metabolism, playing a critical role in plant development and enhancing the ability of crops to resist stress.The watermelon inbred line TN07011 was used as the material.ClSHM78 was cloned by RT-PCR technology and subjected to bioinformatics analysis.An overexpression vector was constructed and transformed into Arabidopsis thaliana. The T₃ transformed lines were subjected to low-temperature and salt stress treatments,by determining phenotypic and physiological indices, the function of ClSHMT8 in abiotic stress was explored.The results showed that ClSHMT8 was 1 749 bp in full length, encoding 582 amino acids. NCBI-CDD conserved domain prediction indicated that the protein contained a typical SHMT domain.Phylogenetic tree analysis showed that this protein shares a close genetic relationship with its homologous proteins from cucumber and bitter melon.Heterologous expression of ClSHMT8 in Arabidopsis thaliana yielded three T₃ lines, which exhibited significantly larger rosette leaf area, earlier flowering period, and larger seed size compared to the wild type. Under low-temperature and salt stress, the T₃ plants showed significantly higher contents of soluble protein(SP) and chlorophyll(Chl), as well as activities of antioxidant enzymes such as peroxidase(POD), superoxide dismutase(SOD), and catalase(CAT), while the content of malondialdehyde(MDA) was significantly lower than that of wild type. Taken together, ClSHMT8 not only positively regulates plant growth and development, but also enhances plant tolerance to abiotic stresses.These results demonstrate that ClSHMT8 plays an important role in the response of Arabidopsis thaliana to abiotic stress environments.

Key words: Watermelon, ClSHMT8, Plant overexpression vector, Abiotic stress, Functional analysis

CLC Number:

JIN Yuxuan, SU Chaijing, ZHANG Man, ZHOU Xinming, LI Xinru, CHEN Kunling, ZHANG Weihua, WU Ying. Cloning of Watermelon ClSHMT8 and Its Functional Analysis under Cold Stress and Salt Stress[J]. Acta Agriculturae Boreali-Sinica, 2026, 41(3): 25-31. doi: 10.7668/hbnxb.20196251.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.20196251

http://www.hbnxb.net/EN/Y2026/V41/I3/25

Figures/Tables 6

Fig.1 RT-PCR amplification of ClSHMT8 gene(A)and double enzyme digestion verification of overexpression vector(B)

Fig.2 Bioinformatics prediction of ClSHMT8 A.The hydrophilicity and hydrophobicity;B.Phosphorylation site;C.Signal peptide;D.Transmembrane structure; E.Conserved domain;F.Protein secondary structure;G.Protein tertiary structure.

Fig.3 Phylogenetic tree analysis of SHMT proteins in different species

Fig.4 Seed particle size statistics of Arabidopsis thaliana Different letters indicate significant differences at the 0.05 level.The same as Fig.5-6.

Fig.5 Statistical analysis of flowering and rosette leaves of T₃ Arabidopsis and wild type

Fig.6 Determination of physiological indices in T₃Arabidopsis thaliana and wild-type under abiotic stress

References 23

[1]	靳丹丹, 张卫华, 杨立芳, 刘颖, 李天杰, 刘海学, 吴颖. 西瓜转录因子基因ClERF5的克隆及表达分析[J]. 分子植物育种, 2025, 23(1):53-59.doi:10.13271/j.mpb.023.000053.
	Jin D D, Zhang W H, Yang L F, Liu Y, Li T J, Liu H X, Wu Y. Cloning and expression analysis of watermelon transcription factor gene ClERF5[J]. Molecular Plant Breeding, 2025, 23(1):53-59.
[2]	Anees M, Gao L, Umer M J, Yuan P L, Zhu H J, Lu X Q, He N, Gong C S, Kaseb M O, Zhao S J, Liu W G. Identification of key gene networks associated with cell wall components leading to flesh firmness in watermelon[J]. Frontiers in Plant Science, 2021, 12:630243.doi:10.3389/FPLS.2021.630243. URL
[3]	李琼, 李丽丽, 侯娟, 罗忍忍, 王瑞丹, 胡建斌, 黄松. 瓜类作物响应低温胁迫机制的研究进展[J]. 园艺学报, 2022, 49(6):1382-1394.doi:10.16420/j.issn.0513-353x.2021-0575.
	Li Q, Li L L, Hou J, Luo R R, Wang R D, Hu J B, Huang S. Advances on mechanism of cucurbit crops in response to low-temperature stress[J]. Acta Horticulturae Sinica, 2022, 49(6):1382-1394.
[4]	Sheikh S, Noh J, Seong M H, Jung G T, Kim J M. Consequences of chilling stress on watermelon [Citrullus lanatus(Thunb.) Matsum.and Nakai]germplasm lines at seedling stage[J].Horticulture,Environment,and Biotechnology, 2015, 56(1):79-88.doi:10.1007/s13580-015-0174-2.
[5]	Nogués I, Sekula B, Angelaccio S, Grzechowiak M, Tramonti A, Contestabile R, Ruszkowski M. Arabidopsis thaliana serine hydroxymethyltransferases:functions,structures,and perspectives[J]. Plant Physiology and Biochemistry, 2022, 187:37-49.doi:10.1016/j.plaphy.2022.07.025. URL
[6]	侯学文, 李英杰, 钟琪, 彭新湘. 光呼吸代谢途径及其调控的研究进展[J]. 植物生理学报, 2019, 55(3):255-264.doi:10.13592/j.cnki.ppj.2018.0551.
	Hou X W, Li Y J, Zhong Q, Peng X X. Recent progress of photorespiration pathway and its regulation[J]. Plant Physiology Communications, 2019, 55(3):255-264.
[7]	赵玉琪, 周志林, 唐君, 姚改芳, 胡康棣, 张华. 甘薯、番茄和拟南芥中丝氨酸羟甲基转移酶基因家族功能研究[J]. 合肥工业大学学报(自然科学版), 2022, 45(12):1705-1714.doi:10.3969/j.issn.1003-5060.2022.12.018.
	Zhao Y Q, Zhou Z L, Tang J, Yao G F, Hu K D, Zhang H. Study on the function of serine hydroxymethyltransferase gene family in sweet potato,tomato and Arabidopsis[J].Journal of Hefei University of Technology(Natural Science), 2022, 45(12):1705-1714.
[8]	Gao R, Luo Y Y, Pan X J, Wang C L, Liao W B. Genome-wide identification of SHMT family genes in cucumber(Cucumis sativus L.) and functional analyses of CsSHMTs in response to hormones and abiotic stresses[J].3 Biotech, 2022, 12(11):305.doi:10.1007/s13205-022-03378-x.
[9]	Lakhssassi N, Patil G, Piya S, Zhou Z, Baharlouei A, Kassem M A, Lightfoot D A, Hewezi T, Barakat A, Nguyen H T, Meksem K. Genome reorganization of the GmSHMT gene family in soybean showed a lack of functional redundancy in resistance to soybean cyst nematode[J]. Scientific Reports, 2019, 9(1):1506.doi:10.1038/s41598-018-37815-w. pmid: 30728404
[10]	Liu H, Li N, Zhao Y, Kang G Z, Zhao Y H, Xu H W. Serine hydroxymethyltransferase(SHMT)gene family in wheat(Triticum aestivum L.):identification,evolution,and expression analysis[J]. Agronomy, 2022, 12(6):1346.doi:10.3390/agronomy12061346. URL
[11]	Mishra P, Jain A, Takabe T, Tanaka Y, Negi M, Singh N, Jain N, Mishra V, Maniraj R, Krishnamurthy S L, Sreevathsa R, Singh N K, Rai V. Heterologous expression of serine hydroxymethyltransferase-3 from rice confers tolerance to salinity stress in E.coli and Arabidopsis[J].Frontiers in Plant Science, 2019, 10:217.doi:10.3389/fpls.2019.00217.
[12]	Liu Y P, Mauve C, Lamothe-Sibold M, Guérard F, Glab N, Hodges M, Jossier M. Photorespiratory serine hydroxymethyltransferase 1 activity impacts abiotic stress tolerance and stomatal closure[J]. Plant,Cell & Environment, 2019, 42(9):2567-2583.doi:10.1111/pce.13595. URL
[13]	李冰, 程玉祥. 杨树SHMT基因家族分析及PtrSHMT9突变体创制[J]. 植物研究, 2020, 40(6):906-912.doi:10.7525/j.issn.1673-5102.2020.06.013.
	Li B, Cheng Y X. Analysis of SHMT gene family and production of PtrSHMT9 mutant in Populus trichocarpa[J].Bulletin of Botanical Research, 2020, 40(6):906-912.
[14]	Zhang J, Li M, Bryan A C, Yoo C G, Rottmann W, Winkeler K A, Collins C M, Singan V, Lindquist E A, Jawdy S S, Gunter L E, Engle N L, Yang X H, Barry K, Tschaplinski T J, Schmutz J, Pu Y Q, Ragauskas A J, Tuskan G A, Muchero W, Chen J G. Overexpression of a serine hydroxymethyltransferase increases biomass production and reduces recalcitrance in the bioenergy crop Populus[J].Sustainable Energy & Fuels, 2019, 3(1):195-207.doi:10.1039/C8SE00471D.
[15]	Zhang F, Jin D D, Zhang W H, Liu Y, Liu H X, Pan Q, Wang X Y, Wu Y. Integration of transcriptome and metabolome provides key genes and pathways associated with cold stress in watermelon(Citrullus lanatus)[J]. Plant Molecular Biology Reporter, 2025, 43(2):752-765.doi:10.1007/S11105-024-01508-8.
[16]	粟柴静, 张卫华, 宋嘉欣, 李明轩, 邓满, 池明, 吴颖. 西瓜SHMT基因家族鉴定及表达模式分析[J]. 华北农学报, 2024, 39(6):47-54.doi:10.7668/hbnxb.20195203.
	Su C J, Zhang W H, Song J X, Li M X, Deng M, Chi M, Wu Y. Genome-wide identification and expression analysis of SHMT gene family in watermelon[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(6):47-54. doi: 10.7668/hbnxb.20195203
[17]	蔡庆生. 植物生理学实验[M]. 北京: 中国农业大学出版社,2013:178-185.
	Cai Q S. Plant physiology experiments[M]. Beijing: China Agricultural University Press,2013:178-185.
[18]	Zhao X, Zeng Z D, Cao W J, Khan D, Ikram M, Yang K B, Chen L M, Li K Z. Co-overexpression of AtSHMT1 and AtFDH induces sugar synthesis and enhances the role of original pathways during formaldehyde metabolism in tobacco[J]. Plant Science, 2021, 305:110829.doi:10.1016/J.PLANTSCI.2021.110829. URL
[19]	沈访, 李林, 潘胤池, 张社波, 马激, 徐德林. 白及丝氨酸羟甲基转移酶(SHMT)基因的序列分析[J]. 植物研究, 2018, 38(4):543-550.doi:10.7525/j.issn.1673-5102.2018.04.008.
	Shen F, Li L, Pan Y C, Zhang S B, Ma J, Xu D L. Bioinformatic analysis of serine hydroxymethyltransferase(SHMT)gene in Bletilla striata[J]. Bulletin of Botanical Research, 2018, 38(4):543-550.
[20]	颜梦圆. 水稻调控胚乳发育关键基因SHMT4的图位克隆与功能分析[D]. 北京: 中国农业科学院, 2022.doi:10.27630/d.cnki.gznky.2022.000147.
	Yan M Y. Map-based cloning and functional analysis of SHMT4,a key gene regulating endosperm development in rice[D]. Beijing: Chinese Academy of Agricultural Sciences, 2022.
[21]	高荣. 黄瓜SHMT基因家族分析及CsSHMT3响应盐胁迫的功能研究[D]. 兰州: 甘肃农业大学, 2023.doi:10.27025/d.cnki.ggsnu.2023.000383.
	Gao R. Identification of the SHMT gene family in Cucumis sativus and functional analysis of CsSHMT3 gene in response to salt stress[D]. Lanzhou: Gansu Agricultural University, 2023.
[22]	Fang C X, Zhang P L, Li L L, Yang L K, Mu D, Yan X, Li Z, Lin W X. Serine hydroxymethyltransferase localised in the endoplasmic reticulum plays a role in scavenging H₂O₂ to enhance rice chilling tolerance[J]. BMC Plant Biology, 2020, 20(1):236.doi:10.1186/s12870-020-02446-9.
[23]	张鹏丽. 内质网定位的OsSHMT对Lsi1超表达水稻苗期抗低温的作用机制研究[D]. 福州: 福建农林大学, 2019.doi:10.27018/d.cnki.gfjnu.2019.000778.
	Zhang P L. The possible role of the ER-localized OsSHMT in regulating chilling tolerence of Lsi1-overexpressed rice seedling[D]. Fuzhou: Fujian Agriculture and Forestry University, 2019.

Please choose a citation manager

Content to export