Abstract
The alleviation effects of exogenous putrescine treatment on the ultrastructure of and calcium ion flow rate in lettuce (Lactuca sativa L.) leaf cells under drought stress were studied. Lettuce seedlings were treated with foliar sprays of 0.1 mM putrescine for 8 days, after which drought stress was simulated by using 10% polyethylene glycol 6000. The morphological characteristics of the seedlings and the calcium ion flow rate across stomatal guard cells were subsequently determined, and the leaf cell ultrastructure was observed via transmission electron microscopy. Under drought stress, the morphological characteristics of the seedlings decreased, and calcium ion influx was predominant in the guard cells. In addition, compared to that under control conditions, the stomatal density under drought stress conditions increased significantly, the open/closed stoma ratio was lower, and the degree of stomatal opening was smaller. Exogenous putrescine sprays effectively reduced the stomatal density, increased the degree of stomatal opening, and increased the proportion of open stomata. In addition, the chloroplasts became round in shape, the thylakoid structure became blurry in appearance, the number of starch grains decreased, many osmium granules were produced, and plasmolysis occurred in the mesophyll cells. However, the chloroplasts were elongated, the thylakoid structure was clear, the starch grains were abundant, few osmium granules were produced, and plasmolysis did not occur. The above results show that, by altering the leaf cell ultrastructure as well as the flow rate and direction of calcium ions in guard cells, exogenous putrescine effectively improves the drought tolerance of lettuce.
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References
Aasamaa K, Sõber A, Rahi M (2001) Leaf anatomical characteristics associated with shoot hydraulic conductance, stomatal conductance and stomatal sensitivity to changes of leaf water status in temperate deciduous trees. Funct Plant Biol 28:765–774. https://doi.org/10.1071/PP00157
Aksakal O, Algur OF, Aksakal FI, Aysin F (2017) Exogenous 5-aminolevulinic acid alleviates the detrimental effects of UV-B stress on lettuce (Lactuca sativa L.) seedlings. Acta Physiol Plant 39:55. https://doi.org/10.1007/s11738-017-2347-3
Assmann SM, Simoncini L, Schroeder JI (1985) Blue light activates electrogenic ion pumping in guard cell protoplasts of Vicia faba. Nature 318:285–287. https://doi.org/10.1038/318285a0
Baier M, Kandlbinder A, Golldack D, Dietz KJ (2005) Oxidative stress and ozone: perception, signalling and response. Plant Cell Environ 28:1012–1020. https://doi.org/10.1111/j.1365-3040.2005.01326.x
Barrs HD, Weatherley PE (1962) A re-examination of the relative turgidity technique for estimating water deficits in leaves. Aust J Bio Sci 15:413–428. https://doi.org/10.1071/BI9620413
Bauerle TL, Centinari M, Bauerle WL (2011) Shifts in xylem vessel diameter and embolisms in grafted apple trees of differing rootstock growth potential in response to drought. Planta 234:1045–1054. https://doi.org/10.1007/s00425-011-1460-6
Borges R, Miguel EC, Dias JMR, da Cunha M, Bressan-Smith RE, de Oliveira JG, de Souza FGA (2004) Ultrastructural, physiological and biochemical analyses of chlorate toxicity on rice seedlings. Plant Sci 166:1057–1062. https://doi.org/10.1016/j.plantsci.2003.12.023
Boyer J (1976) Photosynthesis at low water potentials. Philos Trans R Soc Lon B 273:501–512. https://doi.org/10.1098/rstb.1976.0027
Chartzoulakis K, Patakas A, Kofidis G, Bosabalidis A, Nastou A (2002) Water stress affects leaf anatomy, gas exchange, water relations and growth of two avocado cultivars. Sci Hortic 95:39–50. https://doi.org/10.1016/S0304-4238(02)00016-X
Chen JH, Li RH, Guo PG, Xian YS, Tian CE, Miu SY (2011) Impact of drought stress on the ultrastructure of leaf cells in three barley genotypes differing in level of drought tolerance. Chin Bull Bot 46:28–36. https://doi.org/10.3724/SP.J.1259.2011.00028
Cornic G (2000) Drought stress inhibits photosynthesis by decreasing stomatal aperture – not by affecting ATP synthesis. Trends Plant Sci 5:187–188. https://doi.org/10.1016/S1360-1385(00)01625-3
Das A, Mukhopadhyay M, Sarkar B, Saha D, Mondal TK (2015) Influence of drought stress on cellular ultrastructure and antioxidant system in tea cultivars with different drought sensitivities. J Environ Biol 36:875–882
Diao Q, Song Y, Shi D, Qi H (2017) Interaction of polyamines, abscisic acid, nitric oxide, and hydrogen peroxide under chilling stress in tomato (Lycopersicon esculentum Mill.) seedlings. Front Plant Sci 8:203. https://doi.org/10.3389/fpls.2017.00203
Feng HY, Wang ZM, Kong FN, Zhang MJ, Zhou SL (2011) Roles of carbohydrate supply and ethylene, polyamines in maize kernel set. J Integr Plant Biol 53:388–398. https://doi.org/10.1111/j.1744-7909.2011.01039.x
Galmes J, Medrano H, Flexas J (2007) Photosynthetic limitations in response to water stress and recovery in Mediterranean plants with different growth forms. New Phytol 175:81–93. https://doi.org/10.1111/j.1469-8137.2007.02087.x
Hawkins BJ, Robbins S (2010) pH affects ammonium, nitrate and proton fluxes in the apical region of conifer and soybean roots. Physiol Plant 138:238–247. https://doi.org/10.1111/j.1399-3054.2009.01317.x
Hawkins BJ, Boukcim H, Plassard C (2008) A comparison of ammonium, nitrate and proton net fluxes along seedling roots of Douglas-fir and lodgepole pine grown and measured with different inorganic nitrogen sources. Plant Cell Environ 31:278–287. https://doi.org/10.1111/j.1365-3040.2007.01760.x
Hoagland DR, Arnon DI (1950) The water-culture method for growingplants without soil. Calif Agric Exp Stn 347:1–32
Jaffe LF (2010) Fast calcium waves. Cell Calcium 48:102–113. https://doi.org/10.1016/j.ceca.2010.08.007
Jiang HM, Yang JC, Zhang JF (2007) Effects of external phosphorus on the cell ultrastructure and the chlorophyll content of maize under cadmium and zinc stress. Environ Pollut 147:750–756. https://doi.org/10.1016/j.envpol.2006.09.006
Knight H (2000) Calcium signaling during abiotic stress in plants. Int Rev Cytol 195:269–324. https://doi.org/10.1016/S0074-7696(08)62707-2
Koenig H, Goldstone A, Lu CY (1983) Polyamines regulate calcium fluxes in a rapid plasma membrane response. Nature 305:530–534. https://doi.org/10.1038/305530a0
Kubis J, Floryszak-Wieczorek J, Arasimowicz-Jelonek M (2014) Polyamines induce adaptive responses in water deficit stressed cucumber roots. J Plant Res 127:151–158. https://doi.org/10.1007/s10265-013-0585-z
Kusano T, Yamaguchi K, Berberich T, Takahashi Y (2007) The polyamine spermine rescues Arabidopsis from salinity and drought stresses. Plant Signal Behav 2:251–252. https://doi.org/10.4161/psb.2.4.3866
Lage-Pinto F, Oliveira JG, Da Cunha M, Souza CMM, Rezende CE, Azevedo RA, Vitória AP (2008) Chlorophyll a fluorescence and ultrastructural changes in chloroplast of water hyacinth as indicators of environmental stress. Environ Exp Bot 64:307–313. https://doi.org/10.1016/j.envexpbot.2008.07.007
Li B, He L, Guo S, Li J, Yang Y, Yan B, Sun J, Li J (2013) Proteomics reveal cucumber Spd-responses under normal condition and salt stress. Plant Physiol Biochem 67:7–14. https://doi.org/10.1016/j.plaphy.2013.02.016
Liu K, Fu H, Bei Q, Luan S (2000) Inward potassium channel in guard cells as a target for polyamine regulation of stomatal movements. Plant Physiol 124:1315–1326. https://doi.org/10.1104/pp.124.3.1315
Lushbaugh WB, Hofbauer AF, Pittman FE, Pittman JC (1978) Surface ultrastructure of entamoeba histolytica; a study by high voltage transmission electron microscopy (HVTEM) and scanning electron microscopy (SEM). Arch Invest Med (Mex) 9:191–202
Molas J (2002) Changes of chloroplast ultrastructure and total chlorophyll concentration in cabbage leaves caused by excess of organic Ni(II) complexes. Environ Exp Bot 47:115–126. https://doi.org/10.1016/S0098-8472(01)00116-2
Możdżeń K, Bojarski B, Rut G, Migdałek G, Repka P, Rzepka A (2015) Effect of drought stress induced by mannitol on physiological parameters of maize (Zea mays L.) seedlings and plants. J Microbiol Biotech Food Sci 4:86–91. https://doi.org/10.15414/jmbfs.2015.4.special2.86-91
Nautiyal S, Badola HK, Pal M, Negi DS (1994) Plant responses to water stress: changes in growth, dry matter production, stomatal frequency and leaf anatomy. Biol Plant 36:91. https://doi.org/10.1007/BF02921275
Pal M, Szalai G, Janda T (2015) Speculation: polyamines are important in abiotic stress signaling. Plant Sci 237:16–23. https://doi.org/10.1016/j.plantsci.2015.05.003
Sadiqov ST, Akbulut M, Ehmedov V (2002) Role of Ca2+ in drought stress signaling in wheat seedlings. Biochemistry (Moscow) 67:491–497. https://doi.org/10.1023/A:1015298309888
Saruhan N, Turgut-Terzi R, Kadioglu A (2006) The effects of exogenous polyamines on some biochemical changes during drought stress in Ctenanthe setosa (Rosc.) Eichler. Acta Biol Hung 57:221–229. https://doi.org/10.1556/ABiol.57.2006.2.9
Singh AP, Wi SG, Kang H, Chung GC, Kim YS (2003) Simple and rapid methods for SEM observation and TEM immunolabeling of rubber particles. J Histochem Cytochem 51:1105–1108. https://doi.org/10.1177/002215540305100815
Sirichandra C, Wasilewska A, Vlad F, Valon C, Leung J (2009) The guard cell as a single-cell model towards understanding drought tolerance and abscisic acid action. J Exp Bot 60:1439–1463. https://doi.org/10.1093/jxb/ern340
Smith TA (1970) Putrescine, spermidine and spermine in higher plants. Phytochemistry 9:1479–1486
Tassoni A, van Buuren M, Franceschetti M, Fornalè S, Bagni N (2000) Polyamine content and metabolism in Arabidopsis thaliana and effect of spermidine on plant development. Plant Physiol Biochem 38:383–393. https://doi.org/10.1016/S0981-9428(00)00757-9
Tretyn A (1999) Calcium-dependent signal transduction pathways in plants–phytochrome mechanism of action as an example. Pol J Pharmacol 51:145–151
Vitória AP, Da Cunha M, Azevedo RA (2006) Ultrastructural changes of radish leaf exposed to cadmium. Environ Exp Bot 58:47–52. https://doi.org/10.1016/j.envexpbot.2005.06.014
Wang S, Liang D, Li C, Hao Y, Ma F, Shu H (2012a) Influence of drought stress on the cellular ultrastructure and antioxidant system in leaves of drought-tolerant and drought-sensitive apple rootstocks. Plant Physiol Biochem 51:81–89. https://doi.org/10.1016/j.plaphy.2011.10.014
Wang WH, Yi XQ, Han AD, Liu TW, Chen J, Wu FH, Dong XJ, He JX, Pei ZM et al (2012b) Calcium-sensing receptor regulates stomatal closure through hydrogen peroxide and nitric oxide in response to extracellular calcium in Arabidopsis. J Exp Bot 63:177–190. https://doi.org/10.4161/psb.18882
Wang S, Yang R, Shu C, Zhang X (2016) Screening for cold-resistant tomato under radiation mutagenesis and observation of the submicroscopic structure. Acta Acta Physiol Plant 38:258. https://doi.org/10.1007/s11738-016-2265-9
Xu Z, Zhou G (2008) Responses of leaf stomatal density to water status and its relationship with photosynthesis in a grass. J Exp Bot 59:3317–3325. https://doi.org/10.1093/jxb/ern185
Yamaguchi K, Takahashi Y, Berberich T, Imai A, Takahashi T, Michael AJ, Kusano T (2007) A protective role for the polyamine spermine against drought stress in Arabidopsis. Biochem Biophys Res Commun 352:486–490. https://doi.org/10.1016/j.bbrc.2006.11.041
Yang J, Zhang J, Liu K, Wang Z, Liu L (2007) Involvement of polyamines in the drought resistance of rice. J Exp Bot 58:1545–1555. https://doi.org/10.1093/jxb/erm032
Zeid IM, Shedeed ZA (2006) Response of alfalfa to putrescine treatment under drought stress. Biol Plant 50:635. https://doi.org/10.1007/s10535-006-0099-9
Zeng F, Song C, Guo H, Liu B, Luo W, Gui D, Arndt S, Guo D (2013) Responses of root growth of Alhagi sparsifolia Shap. (Fabaceae) to different simulated groundwater depths in the southern fringe of the Taklimakan Desert, China. J Arid Land 5:220–232. https://doi.org/10.1007/s40333-013-0154-2
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This work was supported by the National Key Research and Development Program of China (2016YFD0201010) and the Beijing Innovation Consortium of Agriculture Research System (BAIC02-2018).
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ZX, YR and WL carried out the experiments, collected and analysed the results. ZX and WL wrote the manuscript. LCJ designed the experiments, and HYY, HJH and FSX helped in the analysis of the results and editing the manuscript. All authors have contributed significantly, and all authors are in agreement with the content of the manuscript.
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Communicated by Jun Gu Lee, Ph.D.
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Zhu, X., Wang, L., Yang, R. et al. Effects of exogenous putrescine on the ultrastructure of and calcium ion flow rate in lettuce leaf epidermal cells under drought stress. Hortic. Environ. Biotechnol. 60, 479–490 (2019). https://doi.org/10.1007/s13580-019-00151-7
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DOI: https://doi.org/10.1007/s13580-019-00151-7