Altitudinal differentiation in the leaf wax-mediated flowering bud protection against frost in a perennial Arabidopsis

Oecologia (2021)Cite this article

Abstract

An altitudinal gradient of leaf water repellency is often observed between and within species. In a previous study of Arabidopsis halleri, cauline leaves (stem leaves that wrap flowering buds) showed higher water repellency in exposed semi-alpine plants than in understory low-elevation plants. Here, we examined altitudinal variations in the cuticular wax content of the leaf surface and experimentally evaluated the role of high water repellency of cauline leaves. Leaf cuticular wax was analysed using comprehensive two-dimensional gas chromatography (GC)-mass spectrometry and a GC-flame ionisation detector. Young flowering buds wrapped by cauline leaves were exposed to freezing temperatures with or without water, and frost damage to the flowering buds was compared between plants from semi-alpine and low-elevation habitats. Higher amounts of C29, C31, and C33 alkanes were observed in the cauline leaves of semi-alpine plants than in those of low-elevation plants. In the freezing experiment, water application increased damage to the flowering buds of low-elevation plants, and the extent of damage to the flowering buds was lower in semi-alpine plants than in low-elevation plants when water was applied to the plant surface. Genetic variations in the amounts of alkanes on the leaf surface depending on the altitude occurred specifically in cauline leaves. Our results indicate that the water repellency of cauline leaves presumably minimises frost damage to flowering buds at high altitudes.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

US$ 39.95

Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Availability of data and material

All data generated or analysed during this study are included in this published article [and in Online Resource 1].

References

  1. Aryal B, Neuner G (2010) Leaf wettability decreases along an extreme altitudinal gradient. Oecologia 126:1–9. https://doi.org/10.1007/s00442-009-1437-3

    Article  Google Scholar 

  2. Aryal B, Neuner G (2012) Leaf wettability in bilberry Vaccinium myrtillus L. as affected by altitude and openness of the growing site. Phyton 52:245–262

    Google Scholar 

  3. Aryal B, Shinohara W, Honjo MN, Kudoh H (2018) Genetic differentiation in cauline-leaf-specific wettability of a rosette-forming perennial Arabidopsis from two contrasting montane habitats. Ann Bot 121:1351–1360. https://doi.org/10.1093/aob/mcy033

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. Bernard A, Joubès J (2013) Arabidopsis cuticular waxes: Advances in synthesis, export and regulation. Prog Lipid Res 52:110–129. https://doi.org/10.1016/j.plipres.2012.10.002

    CAS  Article  PubMed  Google Scholar 

  5. Bhushan B, Jung YC (2008) Wetting, adhesion and friction of superhydrophobic and hydrophilic leaves and fabricated micro/nanopatterned surfaces. J Phys Condens. https://doi.org/10.1088/0953-8984/20/22/225010

    Article  Google Scholar 

  6. Brewer CA, Nuñez CI (2007) Patterns of leaf wettability along an extreme moisture gradient in western Patagonia, Argentina. Int J Plant Sci 168:555–562. https://doi.org/10.1086/513468

    Article  Google Scholar 

  7. Brewer CA, Smith K (1997) Patterns of leaf surface wetness for montane and subalpine plants. Plant Cell Environ 20:1–11. https://doi.org/10.1046/j.1365-3040.1997.d01-15.x

    Article  Google Scholar 

  8. Brewer CA, Smith WK, Vogelmann TC (1991) Functional interaction between leaf trichomes, leaf wettability and the optical properties of water droplets. Plant Cell Environ 14:955–962. https://doi.org/10.1111/j.1365-3040.1991.tb00965.x

    Article  Google Scholar 

  9. Cary JW, Lindow SE (1986) The effect of leaf surface water variables on ice nucleating Pseudomonas syringae in beans. HortScience 21:1417–1418

    Google Scholar 

  10. Clausen J, Keck DD, Hiesey HM (1940) Experimental studies on the nature of species. I. Effect of varied environments on western North American plants, Carnegie Institute of Washington, Washington DC

    Google Scholar 

  11. Feakins SJ, Bentley LP, Salinas N, Shenkin A, Blonder B, Goldsmith GR, Ponton C, Arvin LJ, WuMS PT, West AJ, Martin RE, Enquist BJ, Asner GP, Malhi Y (2016b) Plant leaf wax biomarkers capture gradients in hydrogen isotopes of precipitation from the Andes and Amazon. Geochim Cosmochim Acta 182:155–172. https://doi.org/10.1016/j.gca.2016.03.018

    CAS  Article  Google Scholar 

  12. Feakins SJ, Peters T, Wu MS, Shenkin A, Salinas N, Girardin CAJ, Bentley LP, Blonder B, Enquist BJ, Martin RE, Asner GP, Malhi Y (2016a) Production of leaf wax n-alkanes across a tropical forest elevation transect. Org Geochem 100:89–100. https://doi.org/10.1016/j.orggeochem.2016.07.004

    CAS  Article  Google Scholar 

  13. Fernández V, Guzmán-Delgado P, Graça J, Santos S, Gil L (2016) Cuticle structure in relation to chemical composition: re-assessing the prevailing model. Front Plant Sci 7:427. https://doi.org/10.3389/fpls.2016.00427

    Article  PubMed  PubMed Central  Google Scholar 

  14. Fogg GE (1947) Quantitative studies on the wetting of leaves by water. Proc Royal Soc B 134:503–522. https://doi.org/10.1098/rspb.1947.0028

    CAS  Article  Google Scholar 

  15. Fuller M, Hamed F, Wisniewski ME, Glenn DM (2003) Protection of plants from frost using hydrophobic particle film and acrylic polymer. Ann Appl Biol 143:93–97. https://doi.org/10.1111/j.1744-7348.2003.00093.x

    CAS  Article  Google Scholar 

  16. George FM, Becwar MR, Burke MJ (1982) Freezing avoidance by deep undercooling of tissue water in winter-hardy plants. Cryobiology 19:628–639. https://doi.org/10.1016/0011-2240(82)90192-4

    CAS  Article  PubMed  Google Scholar 

  17. Goldsmith GR, Bentley LP, Shenkin A, Salinas N, Blonder B, Martin RE, Castro-Ccossco R, Chambi-Porroa P, Diaz S, Enquist BJ, Asner GP, Malhi Y (2017) Variation in leaf wettability traits along a tropical montane elevation gradient. New Phytol 214:989–1001. https://doi.org/10.1111/nph.14121

    Article  PubMed  Google Scholar 

  18. Gonzalo-Turpin H, Hazard L (2009) Local adaptation occurs along altitudinal gradient despite the existence of gene flow in the alpine plant species. J Ecol 97:742–751. https://doi.org/10.1111/j.1365-2745.2009.01509.x

    Article  Google Scholar 

  19. Guo N, Gao J, He Y, Guo Y (2016) Compositae plants differed in leaf cuticular waxes between high and low altitudes. Chem Biodivers 13:710–718. https://doi.org/10.1002/cbdv.201500208

    CAS  Article  PubMed  Google Scholar 

  20. Gusta LV, Wisniewski ME (2013) Understanding plant cold hardiness: an opinion. Physiol Plant 147:4–14. https://doi.org/10.1111/j.1399-3054.2012.01611.x

    CAS  Article  PubMed  Google Scholar 

  21. Hacker J, Neuner G (2008) Ice propagation in dehardened alpine plant species studied by infrared differential thermal analysis (IDTA). Arct Antarct Alp Res 40:660–670. https://doi.org/10.1657/1523-0430(07-077)[HACKER]2.0.CO;2

    Article  Google Scholar 

  22. Hegebarth D, Buschhaus C, Wu M, Bird D, Jetter R (2016) The composition of surface wax on trichomes of Arabidopsis thaliana differs from wax on other epidermal cells. Plant J 88:762–774. https://doi.org/10.1111/tpj.13294

    CAS  Article  PubMed  Google Scholar 

  23. Holder CD (2011) The relationship between leaf water repellency and leaf traits in three distinct biogeographical regions. Plant Ecol 212:1913–1926. https://doi.org/10.1007/s11258-011-9963-6

    Article  Google Scholar 

  24. Holloway PJ (1969) The effects of superficial wax on leaf wettability. Ann Appl Biol 63:145–153. https://doi.org/10.1111/j.1744-7348.1969.tb05475.x

    Article  Google Scholar 

  25. Honjo MN, Kudoh H (2019) Arabidopsis halleri: a perennial model system for studying population differentiation and local adaptation. AoBP 11:plz076. https://doi.org/10.1093/aobpla/plz076

    Article  Google Scholar 

  26. Ikeda H, Setoguchi H, Morinaga S (2010) Genomic structure of lowland and highland ecotypes of Arabidopsis halleri subsp. gemmifera (Brassicaceae) on Mt Ibuki. APG 61:21–26. https://doi.org/10.18942/apg.KJ00006537151

    Article  Google Scholar 

  27. Klamerus-Iwan A, Błońska E (2018) Canopy storage capacity and wettability of leaves and needles: the effect of water temperature changes. J Hydrol 559:534–540. https://doi.org/10.1016/j.jhydrol.2018.02.032

    Article  Google Scholar 

  28. Koch K, Bhushan B, Barthlott W (2008) Diversity of structure, morphology and wetting of plant surfaces. Soft Matter 4:1943–1963. https://doi.org/10.1039/B804854A

    CAS  Article  Google Scholar 

  29. Kondo S, Hori K, Sasaki-Sekimoto Y, Kobayashi A, Kato T, Yuno-Ohta N, Nobusawa T, Ohtaka K, Shimojima M, Ohta H (2016) Primitive extracellular lipid components on the surface of the charophytic alga Klebsormidium flaccidum and their possible biosynthetic pathways as deduced from the genome sequence. Front Plant Sci 7:952. https://doi.org/10.3389/fpls.2016.00952

    Article  PubMed  PubMed Central  Google Scholar 

  30. Körner C (2003) Alpine plant life: Functional Plant Ecology of High Mountain Ecosystems, 2nd edn. Springer, Berlin

    Google Scholar 

  31. Krauss P, Markstadter C, Riederer M (1997) Attenuation of UV radiation by plant cuticles from woody species. Plant Cell Environ 20:1079–1085. https://doi.org/10.1111/j.1365-3040.1997.tb00684.x

    Article  Google Scholar 

  32. Kubota S, Iwasaki T, Hanada K, Nagano AJ, Fujiyama A, Toyoda A, Sugano S, Suzuki Y, Hikosaka K, Ito M, Morinaga S (2015) A genome scan for genes underlying microgeographic-scale local adaptation in a wild Arabidopsis species. PLoS Genet 11:e1005361. https://doi.org/10.1371/journal.pgen.1005361

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  33. Kuprian E, Munkler C, Resnyak A, Zimmermann S, Tuong TD, Gierlinger N, Müller T, Livingston DP, Neuner G (2017) Complex bud architecture and cell-specific chemical patterns enable supercooling of Picea abies bud primordia. Plant Cell Environ 40:3101–3112. https://doi.org/10.1111/pce.13078

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  34. Ladinig U, Hacker J, Neuner G, Wagner J (2013) How endangered is sexual reproduction of high-mountain plants by summer frosts? Frost resistance, frequency of frost events and risk assessment. Oecologia 171:743–760. https://doi.org/10.1007/s00442-012-2581-8

    Article  PubMed  PubMed Central  Google Scholar 

  35. Larcher W, Kainmüller C, Wagner J (2010) Survival types of high mountain plants under extreme temperatures. Flora 205:3–18. https://doi.org/10.1016/j.flora.2008.12.005

    Article  Google Scholar 

  36. Lee SB, Suh MC (2015) Advances in the understanding of cuticular waxes in Arabidopsis thaliana and crop species. Plant Cell Rep 34:557–572. https://doi.org/10.1007/s00299-015-1772-2

    CAS  Article  PubMed  Google Scholar 

  37. Li Y, Hou X, Li X, Zhao X, Wu Z, Xiao Y, Guo Y (2019) Will the climate of plant origins influence the chemical profiles of cuticular waxes on leaves of Leymus chinensis in a common garden experiment? Ecol and Evol 10:543–556. https://doi.org/10.1002/ece3.5930

    CAS  Article  Google Scholar 

  38. Neinhuis C, Barthlott W (1997) Characterization and distribution of water-repellent, self-cleaning plant surfaces. Ann Bot 79:667–677. https://doi.org/10.1006/anbo.1997.0400

    Article  Google Scholar 

  39. Neuner G, Beikircher B (2010) Critically reduced frost resistance of Picea abies during sprouting could be linked to cytological changes. Protoplasma 243:145–152. https://doi.org/10.1007/s00709-009-0052-9

    CAS  Article  PubMed  Google Scholar 

  40. Neuner G, Erler A, Ladinig U, Hacker J, Wagner J (2013) Frost resistance of reproductive tissues during various stages of development in high mountain plants. Physiol Plant 147:88–100. https://doi.org/10.1111/j.1399-3054.2012.01616.x

    CAS  Article  PubMed  Google Scholar 

  41. Neuner G, Kreische B, Kaplenig D, Monitzer K, Miller R (2019) Deep supercooling enabled by surface impregnation with lipophilic substances explains the survival of overwintering buds at extreme freezing. Plant Cell Environ 42:2065–2074. https://doi.org/10.1111/pce.13545

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  42. Pu Y, Gao J, Guo Y, Liu T, Zhu L, Xu P, Yi B, Wen J, Tu J, Ma C, Fu T, Zou J, Shen J (2013) A novel dominant glossy mutation causes suppression of wax biosynthesis pathway and deficiency of cuticular wax in Brassica napus. BMC Plant Biol 13:215. https://doi.org/10.1186/1471-2229-13-215

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  43. Quamme HA, Su WA, Veto LJ (1995) Anatomical features facilitating supercooling of the flower within the dormant Peach flower bud. J Am Soc Hortic Sci 120:814–822. https://doi.org/10.21273/JASHS.120.5.814

    Article  Google Scholar 

  44. R Development Core Team. (2005). R: A language and environment for statistical computing. Vienna, Austria: R Foundation for Statistical Computing. URL http://www.R-project.org.

  45. Rasband WS (1997–2012) ImageJ. Maryland, U.S.A.: U. S. National Institutes of Health. URL http://rsb.info.nih.gov/ij/.

  46. Rashotte AM, Jenks MA, Nguyen TD, Feldmann KA (1997) Epicuticular wax variation in ecotypes of Arabidopsis thaliana. Phytochem 45:251–255. https://doi.org/10.1016/s0031-9422(96)00792-3

    CAS  Article  Google Scholar 

  47. Rosado BHP, Holder CD (2013) The significance of leaf water repellency in ecohydrological research: a review. Ecohydrology 6:150–161. https://doi.org/10.1002/eco.1340

    Article  Google Scholar 

  48. Sakai A, Larcher W (1987) Frost survival of plants. Springer, New York

    Google Scholar 

  49. Sasaki-Sekimoto Y, Ohta H (2018) Comprehensive analysis of cuticular wax components using GC×GC-MS. SHIMADZU Application Note No. 50 URL https://www.shimadzu.com/an/literature/gcms/jpo218045.html

  50. Smith WK, McClean TM (1989) Adaptive relationship between leaf water repellency, stomatal distribution and gas exchange. Am J Bot 76:465–469. https://doi.org/10.2307/2444617

    Article  Google Scholar 

  51. Song B, Zhang ZQ, Stöcklin J, Yang Y, Niu Y, Chen JG, Sun H (2013) Multifunctional bracts enhance plant fitness during flowering and seed development in Rheum nobile (Polygonaceae), a giant herb endemic to the high Himalayas. Oecologia 172:359–370. https://doi.org/10.1007/s00442-012-2518-2

    Article  PubMed  Google Scholar 

  52. Song B, Stöcklin J, Peng D, Gao Y, Sun H (2015) The bracts of the alpine ‘glasshouse’ plant Rheum alexandrae (Polygonaceae) enhance reproductive fitness of its pollinating seed-consuming mutualist. Bot J Linn Soc 179:349–359. https://doi.org/10.1111/boj.12312

    Article  Google Scholar 

  53. Taschler D, Neuner G (2004) Summer frost resistance and freezing patterns measured in situ in leaves of major alpine plant growth forms in relation to their upper distribution boundary. Plant Cell Environ 27:737–746. https://doi.org/10.1111/j.1365-3040.2004.01176.x

    Article  Google Scholar 

  54. Thomas DA, Barber HN (1974) Studies on leaf characteristics of a cline of Eucalyptus urnigera from Mount Wellington, Tasmania. I Water repellency and the freezing of leaves. Aust J Bot 22:501–512. https://doi.org/10.1071/bt9740501

    Article  Google Scholar 

  55. Turesson G (1922) The genotypical response of the plant species to habitat. Hereditas 3:211–350. https://doi.org/10.1111/j.1601-5223.1922.tb02734.x

    Article  Google Scholar 

  56. Wang H, Shi H, Li Y, Wang Y (2014) The effects of leaf roughness, surface free energy and work of adhesion on leaf water drop adhesion. PLoS ONE 9:e107062. https://doi.org/10.1371/journal.pone.0107062

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  57. Wang QW, Nagano S, Ozaki H, Morinaga S, Hidema J, Hikosaka K (2016) Functional differentiation in UV-B-induced DNA damage and growth inhibition between highland and lowland ecotypes of two Arabidopsis species. Environ Exp Bot 131:110–119. https://doi.org/10.1016/j.envexpbot.2016.07.008

    CAS  Article  Google Scholar 

  58. Wisniewski ME, Fuller M, Glenn D, Gusta LV, Duman J, Griffith M (2002) Extrinsic ice nucleation in plants: what are the factors involved and can they be manipulated? In: Li P, Palva E, eds. Plant cold hardiness. Gene regulation and genetic engineering. Kluwer Academic Publishers, New York

  59. Wisniewski ME, Gusta LV, Neuner G (2014) Adaptive mechanisms of freeze avoidance in plants: a brief update. Environ Exp Bot 99:133–140. https://doi.org/10.1016/j.envexpbot.2013.11.011

    CAS  Article  Google Scholar 

  60. Xu X, Fenga J, Lü S, Lohreyc GT, Ana H, Zhoua Y, Jenks MA (2014) Leaf cuticular lipids on the Shandong and Yukon ecotypes of saltwater cress, Eutrema salsugineum, and their response to water deficiency and impact on cuticle permeability. Physiol Plant 151:446–458. https://doi.org/10.1111/ppl.12127

    CAS  Article  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Maasa Hamamura-Yumoto for assistance in field sampling.

Funding

This study was supported by JST CREST JPMJCR15O1 and JSPS KAKENHI (grant number JP19H01001 to HK and grant number JP1072190 to GY).

Author information

Affiliations

  1. Center for Ecological Research, Kyoto University, Hirano 2-509-3, Otsu, 520-2113, Japan

    Genki Yumoto, Biva Aryal & Hiroshi Kudoh

  2. School of Life Science and Technology, Tokyo Institute of Technology, 4259‐B‐65 Nagatsuta, Midori-ku, Yokohama, 226‐8501, Japan

    Yuko Sasaki-Sekimoto & Hiroyuki Ohta

  3. Amrit Campus, Tribhuvan University, Lekhnath Marg, Kathmandu, 44600, Nepal

    Biva Aryal

Authors
  1. Genki Yumoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuko Sasaki-Sekimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Biva Aryal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hiroyuki Ohta
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hiroshi Kudoh
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

GY, BA, and HK conceived the study; GY conducted the field, growth chamber, and freezing experiments; GY, YS, and HO analysed the cuticular wax; GY and HK wrote the paper with input from all authors.

Corresponding author

Correspondence to Hiroshi Kudoh.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Communicated by Louis Stephen Santiago.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file3 (MP4 15739 KB)

Supplementary file4 (MP4 15332 KB)

Supplementary file1 (XLSX 496 KB)

Supplementary file2 (PDF 878 KB)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Yumoto, G., Sasaki-Sekimoto, Y., Aryal, B. et al. Altitudinal differentiation in the leaf wax-mediated flowering bud protection against frost in a perennial Arabidopsis. Oecologia (2021). https://doi.org/10.1007/s00442-021-04870-6

Download citation

Keywords

  • Alkane
  • Cauline leaf
  • Freezing tolerance
  • Gas chromatography
  • Water repellency