Free Standard AU & NZ Shipping For All Book Orders Over $80!
Register      Login
Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
Shopping Cart: ( empty )
You are here: Home > Journals > FP > FP23210 > Fulltext
RESEARCH ARTICLE
Contents Vol 51(3)

Overexpression of the alfalfa (Medicago sativa) gene, MsKMS1, negatively regulates seed germination in transgenic tobacco (Nicotiana tabacum)

Yiqi Wan A # , Yuman Cao A # , Zhiqiang Zhang A B , Bo Han A C , Maojin Lu A , Zijie Zhuo A , Xinyi Gao A , Peizhi Yang https://orcid.org/0000-0002-5937-837X A * and Yafang Wang https://orcid.org/0000-0002-8104-0375 A *
+ Author Affiliations
- Author Affiliations

A College of Grassland Agriculture, Northwest A&F University, Yangling 712100, China.

B College of Grassland, Resources and Environment, Inner Mongolia Agricultural University, Hohhot 010011, China.

C College of Animal Science and Technology, Yunnan Agriculture University, Kunming 650201, China.

* Correspondence to: yafang.wang@nwsuaf.edu.cn, Yangpeizhi@126.com
# These authors contributed equally to this paper

Handling Editor: Joseph Dubrovsky

Functional Plant Biology 51, FP23210 https://doi.org/10.1071/FP23210
Submitted: 10 September 2023  Accepted: 14 February 2024  Published: 12 March 2024

© 2024 The Author(s) (or their employer(s)). Published by CSIRO Publishing

Abstract

Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-associated proteins are a class of transmembrane proteins involved in intracellular trafficking pathways. However, the functions of many SNARE domain-containing proteins remain unclear. We have previously identified a SNARE-associated gene in alfalfa (Medicago sativa) KILLING ME SLOWLY1 (MsKMS1), which is involved in various abiotic stresses. In this study, we investigated the function of MsKMS1 in the seed germination of transgenic tobacco (Nicotiana tabacum). Phylogenetic analysis showed that MsKMS1 was homologous to the SNARE-associated or MAPR component-related proteins of other plants. Germination assays revealed that MsKMS1 negatively regulated seed germination under normal, D-mannitol and abscisic acid-induced stress conditions, yet MsKMS1-overexpression could confer enhanced heat tolerance in transgenic tobacco. The suppressive effect on germination in MsKMS1-overexpression lines was associated with higher abscisic acid and salicylic acid contents in seeds. This was accompanied by the upregulation of abscisic acid biosynthetic genes (ZEP and NCED) and the downregulation of gibberellin biosynthetic genes (GA20ox2 and GA20ox3). Taken together, these results suggested that MsKMS1 negatively regulated seed germination by increasing abscisic acid and salicylic acid contents through the expression of genes related to abscisic acid and gibberellin biosynthesis. In addition, MsKMS1 could improve heat tolerance during the germination of transgenic tobacco seeds.

Keywords: abiotic stress, abscisic acid, germination, gibberellin, Medicago sativa, phytohormone, salicylic acid, SNARE.

References

Abdul-Baki AA, Anderson JD (1973) Vigor determination in soybean seed by multiple criteria. Crop Science 13, 630-633.
| Crossref | Google Scholar |

An J, Liu Y, Han J, He C, Chen M, Zhu X, Hu W, Song W, Hu J, Guan Y (2022) Transcriptional multiomics reveals the mechanism of seed deterioration in Nicotiana tabacum L. and Oryza sativa L. Journal of Advanced Research 42, 163-176.
| Crossref | Google Scholar | PubMed |

Bao Y-M, Wang J-F, Huang J, Zhang H-S (2008) Cloning and characterization of three genes encoding Qb-SNARE proteins in rice. Molecular Genetics and Genomics 279, 291-301.
| Crossref | Google Scholar | PubMed |

Bao Y-M, Sun S-J, Li M, Li L, Cao W-L, Luo J, Tang H-J, Huang J, Wang Z-F, Wang J-F, Zhang H-S (2012) Overexpression of the Qc-SNARE gene OsSYP71 enhances tolerance to oxidative stress and resistance to rice blast in rice (Oryza sativa L.). Gene 504, 238-244.
| Crossref | Google Scholar | PubMed |

Besserer A, Burnotte E, Bienert GP, Chevalier AS, Errachid A, Grefen C, Blatt MR, Chaumont F (2012) Selective regulation of maize plasma membrane aquaporin trafficking and activity by the SNARE SYP121. The Plant Cell 24, 3463-3481.
| Crossref | Google Scholar | PubMed |

Calvo-Garrido J, Carilla-Latorre S, Escalante R (2008) Vacuole membrane protein 1, autophagy and much more. Autophagy 4, 835-837.
| Crossref | Google Scholar | PubMed |

Chen YA, Scheller RH (2001) SNARE-mediated membrane fusion. Nature Reviews Molecular Cell Biology 2, 98-106.
| Crossref | Google Scholar | PubMed |

Clark HF, Gurney AL, Abaya E, Baker K, Baldwin D, Brush J, Chen J, Chow B, Chui C, Crowley C, Currell B, Deuel B, Dowd P, Eaton D, Foster J, Grimaldi C, Gu Q, Hass PE, Heldens S, Huang A, Kim HS, Klimowski L, Jin Y, Johnson S, Lee J, Lewis L, Liao D, Mark M, Robbie E, Sanchez C, Schoenfeld J, Seshagiri S, Simmons L, Singh J, Smith V, Stinson J, Vagts A, Vandlen R, Watanabe C, Wieand D, Woods K, Xie M-H, Yansura D, Yi S, Yu G, Yuan J, Zhang M, Zhang Z, Goddard A, Wood WI, Godowski P (2003) The secreted protein discovery initiative (SPDI), a large-scale effort to identify novel human secreted and transmembrane proteins: a bioinformatics assessment. Genome Research 13, 2265-2270.
| Crossref | Google Scholar | PubMed |

Collins NC, Thordal-Christensen H, Lipka V, Bau S, Kombrink E, Qiu J-L, Huckelhoven R, Stein M, Freialdenhoven A, Somerville SC, Schulze-Lefert P (2003) SNARE-protein-mediated disease resistance at the plant cell wall. Nature 425, 973-977.
| Crossref | Google Scholar | PubMed |

Deng Y, Yuan F, Feng Z, Ding T, Song J, Wang B (2014) Comparative study on seed germination characteristics of two species of Australia saltbush under salt stress. Acta Ecologica Sinica 34, 337-341.
| Crossref | Google Scholar |

Ebine K, Okatani Y, Uemura T, Goh T, Shoda K, Niihama M, Morita MT, Spitzer C, Otegui MS, Nakano A, Ueda T (2008) A SNARE complex unique to seed plants is required for protein storage vacuole biogenesis and seed development of Arabidopsis thaliana. The Plant Cell 20, 3006-3021.
| Crossref | Google Scholar | PubMed |

Ehlers MRW, Riordan JF (1991) Membrane proteins with soluble counterparts: role of proteolysis in the release of transmembrane proteins. Biochemistry 30, 10065-10074.
| Crossref | Google Scholar | PubMed |

Eisenach C, Chen Z-H, Grefen C, Blatt MR (2012) The trafficking protein SYP121 of Arabidopsis connects programmed stomatal closure and K+ channel activity with vegetative growth. The Plant Journal 69, 241-251.
| Crossref | Google Scholar | PubMed |

Frey A, Audran C, Marin E, Sotta B, Marion-Poll A (1999) Engineering seed dormancy by the modification of zeaxanthin epoxidase gene expression. Plant Molecular Biology 39, 1267-1274.
| Crossref | Google Scholar | PubMed |

Funakoshi K, Suzuki H, Takeuchi S (2006) Lipid bilayer formation by contacting monolayers in a microfluidic device for membrane protein analysis. Analytical Chemistry 78, 8169-8174.
| Crossref | Google Scholar | PubMed |

Guan L, Scandalios JG (1995) Developmentally related responses of maize catalase genes to salicylic acid. Proceedings of the National Academy of Sciences of the United States of America 92, 5930-5934.
| Crossref | Google Scholar |

Hakoyama T, Oi R, Hazuma K, Suga E, Adachi Y, Kobayashi M, Akai R, Sato S, Fukai E, Tabata S, Shibata S, Wu G-J, Hase Y, Tanaka A, Kawaguchi M, Kouchi H, Umehara Y, Suganuma N (2012) The SNARE protein SYP71 expressed in vascular tissues is involved in symbiotic nitrogen fixation in Lotus japonicus nodules. Plant Physiology 160, 897-905.
| Crossref | Google Scholar | PubMed |

Hamaji K, Nagira M, Yoshida K, Ohnishi M, Oda Y, Uemura T, Goh T, Sato MH, Morita MT, Tasaka M, Hasezawa S-I, Nakano A, Hara-Nishimura I, Maeshima M, Fukaki H, Mimura T (2009) Dynamic aspects of ion accumulation by vesicle traffic under salt stress in Arabidopsis. Plant and Cell Physiology 50, 2023-2033.
| Crossref | Google Scholar | PubMed |

Han B, Zhang P, Zhang Z-Q, Wang Y-F, Hu T-M, Yang P-Z (2021) Molecular cloning and characterization of a novel gene MsKMS1 in Medicago sativa. Biologia plantarum 65, 1-9.
| Crossref | Google Scholar |

Hungria M, Kaschuk G (2014) Regulation of N2 fixation and NO3−/NH4+ assimilation in nodulated and N-fertilized Phaseolus vulgaris L. exposed to high temperature stress. Environmental and Experimental Botany 98, 32-39.
| Crossref | Google Scholar |

Hunt L, Holdsworth MJ, Gray JE (2007) Nicotinamidase activity is important for germination. The Plant Journal 51, 341-351.
| Crossref | Google Scholar | PubMed |

Jiang J, Yang BL, Xia T, Yu SM, Wu YN, Jin H, Li JR (2015) Analysis of genetic diversity of salt-tolerant alfalfa germplasms. Genetics and Molecular Research 14, 4438-4447.
| Crossref | Google Scholar | PubMed |

Kato T, Morita MT, Fukaki H, Yamauchi Y, Uehara M, Niihama M, Tasaka M (2002) SGR2, a phospholipase-like protein, and ZIG/SGR4, a SNARE, are involved in the shoot gravitropism of Arabidopsis. The Plant Cell 14, 33-46.
| Crossref | Google Scholar | PubMed |

Kelkar DA, Chattopadhyay A (2007) The gramicidin ion channel: a model membrane protein. Biochimica et Biophysica Acta (BBA)-Biomembranes 1768, 2011-2025.
| Crossref | Google Scholar |

Kim DH, Yamaguchi S, Lim S, Oh E, Park J, Hanada A, Kamiya Y, Choi G (2008) SOMNUS, a CCCH-type zinc finger protein in Arabidopsis, negatively regulates light-dependent seed germination downstream of PIL5. The Plant Cell 20, 1260-1277.
| Crossref | Google Scholar | PubMed |

Koornneef M, Bentsink L, Hilhorst H (2002) Seed dormancy and germination. Current Opinion in Plant Biology 5, 33-36.
| Crossref | Google Scholar | PubMed |

Leshem Y, Melamed-Book N, Cagnac O, Ronen G, Nishri Y, Solomon M, Cohen G, Levine A (2006) Suppression of Arabidopsis vesicle-SNARE expression inhibited fusion of H2O2 containing vesicles with tonoplast and increased salt tolerance. Proceedings of the National Academy of Sciences of the United States of America 103, 18008-18013.
| Crossref | Google Scholar |

Leshem Y, Golani Y, Kaye Y, Levine A (2010) Reduced expression of the v-SNAREs AtVAMP71/AtVAMP7C gene family in Arabidopsis reduces drought tolerance by suppression of abscisic acid-dependent stomatal closure. Journal of Experimental Botany 61, 2615-2622.
| Crossref | Google Scholar | PubMed |

Leyman B, Geelen D, Quintero FJ, Blatt MR (1999) A tobacco syntaxin with a role in hormonal control of guard cell ion channels. Science 283, 537-540.
| Crossref | Google Scholar | PubMed |

Lin W, Sun Y, Qiu X, Huang Q, Kong L, Lu JJ (2021) VMP1, a novel prognostic biomarker, contributes to glioma development by regulating autophagy. Journal of Neuroinflammation 18, 165.
| Crossref | Google Scholar | PubMed |

Lipka V, Kwon C, Panstruga R (2007) SNARE-ware: the role of SNARE-domain proteins in plant biology. Annual Review of Cell and Developmental Biology 23, 147-174.
| Crossref | Google Scholar | PubMed |

Morishita H, Zhao YG, Tamura N, Nishimura T, Kanda Y, Sakamaki Y, Okazaki M, Li D, Mizushima N (2019) A critical role of VMP1 in lipoprotein secretion. eLife 8, e48834.
| Crossref | Google Scholar | PubMed |

Nambara E, Marion-Poll A (2005) Abscisic acid biosynthesis and catabolism. Annual Review of Plant Biology 56, 165-185.
| Crossref | Google Scholar | PubMed |

Osakabe Y, Yamaguchi-Shinozaki K, Shinozaki K, Tran L-SP (2013) Sensing the environment: key roles of membrane-localized kinases in plant perception and response to abiotic stress. Journal of Experimental Botany 64, 445-458.
| Crossref | Google Scholar | PubMed |

Park H-Y, Seok H-Y, Park B-K, Kim S-H, Goh C-H, Lee B-H, Lee C-H, Moon Y-H (2008) Overexpression of Arabidopsis ZEP enhances tolerance to osmotic stress. Biochemical and Biophysical Research Communications 375, 80-85.
| Crossref | Google Scholar | PubMed |

Pelham HRB (2001) SNAREs and the specificity of membrane fusion. Trends in Cell Biology 11, 99-101.
| Crossref | Google Scholar | PubMed |

Rajjou L, Belghazi M, Huguet R, Robin C, Moreau A, Job C, Job D (2006) Proteomic investigation of the effect of salicylic acid on Arabidopsis seed germination and establishment of early defense mechanisms. Plant Physiology 141, 910-923.
| Crossref | Google Scholar | PubMed |

Rajjou L, Duval M, Gallardo K, Catusse J, Bally J, Job C, Job D (2012) Seed germination and vigor. Annual Review of Plant Biology 63, 507-533.
| Crossref | Google Scholar | PubMed |

Rawat N, Singla-Pareek SL, Pareek A (2021) Membrane dynamics during individual and combined abiotic stresses in plants and tools to study the same. Physiologia Plantarum 171, 653-676.
| Crossref | Google Scholar | PubMed |

Razem FA, Baron K, Hill RD (2006) Turning on gibberellin and abscisic acid signaling. Current Opinion in Plant Biology 9, 454-459.
| Crossref | Google Scholar | PubMed |

Saier MH, Jr (2000) A functional-phylogenetic classification system for transmembrane solute transporters. Microbiology and Molecular Biology Reviews 64, 354-411.
| Crossref | Google Scholar | PubMed |

Sanderfoot A (2007) Increases in the number of SNARE genes parallels the rise of multicellularity among the green plants. Plant Physiology 144, 6-17.
| Crossref | Google Scholar | PubMed |

Sanz-Sáez Á, Erice G, Aguirreolea J, Irigoyen JJ, Sánchez-Díaz M (2012) Alfalfa yield under elevated CO2 and temperature depends on the Sinorhizobium strain and growth season. Environmental and Experimental Botany 77, 267-273.
| Crossref | Google Scholar |

Sasaki K, Kim M-H, Kanno Y, Seo M, Kamiya Y, Imai R (2015) Arabidopsis COLD SHOCK DOMAIN PROTEIN 2 influences ABA accumulation in seed and negatively regulates germination. Biochemical and Biophysical Research Communications 456, 380-384.
| Crossref | Google Scholar | PubMed |

Schmid C, Bauer S, Muller B, Bartelheimer M (2013) Belowground neighbor perception in Arabidopsis thaliana studied by transcriptome analysis: roots of Hieracium pilosella cause biotic stress. Frontiers in Plant Science 4, 296.
| Crossref | Google Scholar | PubMed |

Shakirova FM, Sakhabutdinova AR, Bezrukova MV, Fatkhutdinova RA, Fatkhutdinova DR (2003) Changes in the hormonal status of wheat seedlings induced by salicylic acid and salinity. Plant Science 164, 317-322.
| Crossref | Google Scholar |

Shi Y, Zhang Y, Yao H, Wu J, Sun H, Gong H (2014) Silicon improves seed germination and alleviates oxidative stress of bud seedlings in tomato under water deficit stress. Plant Physiology and Biochemistry 78, 27-36.
| Crossref | Google Scholar |

Singh D, Yadav NS, Tiwari V, Agarwal PK, Jha B (2016) A SNARE-like superfamily protein SbSLSP from the halophyte Salicornia brachiata confers salt and drought tolerance by maintaining membrane stability, K+/Na+ ratio, and antioxidant machinery. Frontiers in Plant Science 7, 737.
| Crossref | Google Scholar | PubMed |

Sun X, Ji W, Ding X, Bai X, Cai H, Yang S, Qian X, Sun M, Zhu Y (2013) GsVAMP72, a novel Glycine soja R-SNARE protein, is involved in regulating plant salt tolerance and ABA sensitivity. Plant Cell, Tissue and Organ Culture (PCTOC) 113, 199-215.
| Crossref | Google Scholar |

Sutter J-U, Campanoni P, Blatt MR, Paneque M (2006) Setting SNAREs in a different wood. Traffic 7, 627-638.
| Crossref | Google Scholar | PubMed |

Tarte VN, Seok H-Y, Woo D-H, Le DH, Tran HT, Baik J-W, Kang IS, Lee S-Y, Chung T, Moon Y-H (2015) Arabidopsis Qc-SNARE gene AtSFT12 is involved in salt and osmotic stress responses and Na+ accumulation in vacuoles. Plant Cell Reports 34, 1127-1138.
| Crossref | Google Scholar | PubMed |

Thompson AJ, Jackson AC, Symonds RC, Mulholland BJ, Dadswell AR, Blake PS, Burbidge A, Taylor IB (2000) Ectopic expression of a tomato 9-cis-epoxycarotenoid dioxygenase gene causes over-production of abscisic acid. The Plant Journal 23, 363-374.
| Crossref | Google Scholar | PubMed |

Ungar D, Hughson FM (2003) SNARE protein structure and function. Annual Review of Cell and Developmental Biology 19, 493-517.
| Crossref | Google Scholar | PubMed |

Wang P, Hummel E, Osterrieder A, Meyer AJ, Frigerio L, Sparkes I, Hawes C (2011) KMS1 and KMS2, two plant endoplasmic reticulum proteins involved in the early secretory pathway. The Plant Journal 66, 613-628.
| Crossref | Google Scholar | PubMed |

Wang P, Kou D, Le W (2020) Roles of VMP1 in autophagy and ER–membrane contact: potential implications in neurodegenerative disorders. Frontiers in Molecular Neuroscience 13, 42.
| Crossref | Google Scholar | PubMed |

Wang P, Chen X, Wang Y, Jia C, Liu X, Wang Y, Wu H, Cai H, Shen H-M, Le W (2021) Essential role for autophagy protein VMP1 in maintaining neuronal homeostasis and preventing axonal degeneration. Cell Death & Disease 12, 116.
| Crossref | Google Scholar | PubMed |

Waterworth WM, Masnavi G, Bhardwaj RM, Jiang Q, Bray CM, West CE (2010) A plant DNA ligase is an important determinant of seed longevity. The Plant Journal 63, 848-860.
| Crossref | Google Scholar | PubMed |

Wolters A-MA, Uitdewilligen JGAML, Kloosterman BA, Hutten RCB, Visser RGF, van Eck HJ (2010) Identification of alleles of carotenoid pathway genes important for zeaxanthin accumulation in potato tubers. Plant Molecular Biology 73, 659-671.
| Crossref | Google Scholar | PubMed |

Wu L, Guo X, Harivandi MA (1998) Allelopathic effects of phenolic acids detected in buffalograss (Buchloe dactyloides) clippings on growth of annual bluegrass (Poa annua) and buffalograss seedlings. Environmental and Experimental Botany 39, 159-167.
| Crossref | Google Scholar |

Zhang Z, Wang Y, Chang L, Zhang T, An J, Liu Y, Cao Y, Zhao X, Sha X, Hu T, Yang P (2016) MsZEP, a novel zeaxanthin epoxidase gene from alfalfa (Medicago sativa), confers drought and salt tolerance in transgenic tobacco. Plant Cell Reports 35, 439-453.
| Crossref | Google Scholar | PubMed |

Zhang B, Wang H, Zhang Y (2020) SNARE proteins and their role in plant ion channel regulation. Plant Growth Regulation 92, 443-453.
| Crossref | Google Scholar |

Zhang L, Ma J, Liu H, Yi Q, Wang Y, Xing J, Zhang P, Ji S, Li M, Li J, Shen J, Lin J (2021) SNARE proteins VAMP721 and VAMP722 mediate the post-Golgi trafficking required for auxin-mediated development in Arabidopsis. The Plant Journal 108, 426-440.
| Crossref | Google Scholar | PubMed |

Zhou Y, Wang B, Yuan F (2022) The role of transmembrane proteins in plant growth, development, and stress responses. International Journal of Molecular Sciences 23, 13627.
| Crossref | Google Scholar | PubMed |

Zhu ZX, Ye HB, Xuan YH, Yao DN (2014) Overexpression of a SNARE protein AtBS14b alters BR response in Arabidopsis. Botanical Studies 55, 55.
| Crossref | Google Scholar | PubMed |