BIOLOGIA PLANTARUM 54 (1): 145-148, 2010
石家庄雅思考试145
BRIEF COMMUNICATION
Effects of exogenous spermine on sweet sorghum during germination under salinity
compassY .Y . CHAI 1,2, C.D. JIANG 1, L. SHI 1, T.S. SHI 2 and W.B. GU 1*
Institute of Botany, The Chine Academy of Sciences, Beijing, 100093, P .R. China 1
Bioengineering Department, Zhengzhou University, Zhengzhou, Henan, 450001, P .R. China 2
Abstract
Seedlings of Sorghum bicolor (L.) Moench were subjected to 180 mM NaCl with or without 0.25 mM spermine (SPM) for 7 d. NaCl treatment resulted in the inhibition of growth and incread the content of free proline, soluble protein and malondialdehyde (MDA). Additionally, it also enhanced the activity of catala (CAT), peroxida (POX) in both shoots and roots, while decread that of glutathione redu
cta (GR). When exogenous spermine was added to the test solution, the growth of sweet sorghum edlings was improved, and a smaller increa in the free proline and MDA contents was obrved. The addition of spermine also partially incread the activities of POX and GR, but had no effects on soluble protein content or the activity of CAT.
oceansAdditional key words : antioxidant enzymes, oxidative stress, Sorghum bicolor .
⎯⎯⎯⎯
Polyamines (PAs), a low molecular organic polycations, interact with some negatively charged molecules, including DNA, RNA, proteins, phospholipids and pectic polysaccharides (D’Orazi and Bagni 1987, Martin-Tanguy 2001, Groppa and Benavides 2008). Therefore they affect growth, development and nescence, as well as enzyme activities (Galston and Sawhney 1990, Shevyakova et al . 2006, Groppa and Benavides 2007). PAs also affect plant respon to salinity (Tang and Newton 2005). Putrescine (PUT), spermidine (SPD) and spermine (SPM) are the major PAs in higher plants. It was documented that SPM plays a leading role in the development of plant stress tolerance (Kuznetsov et al . 2006). So far, little is known about the effects of SPM in sweet sorghum. The aim of the prent study was to determine whether exogenous SPM has any protectiv
e effect against salt stress in sorghum edlings. Sweet sorghum [Sorghum bicolor (L.) Moench ] cv. M-81E eds were surface sterilized with 3 % (v/v) sodium hypochlorite solution for about 15 min and then were thoroughly rind veral times with double distilled
tpp是什么意思water. Fifty eds were placed in Petri dishes in the dark between two sheets of filter paper at 25 °C. The Petri dishes were supplied with 3 cm 3 of one of the four test solutions: 1) double distilled water (control); 2) 0.25 mM SPM; 3) 180 mM NaCl; 4) 180 mM NaCl + 0.25 mM SPM. Germination counts were taken every 24 h. Further 2 cm 3 of solutions were added to the dishes every 2 d after sowing. The experiments were done in triplicates. The shoots and roots of edlings were parated after 7 d and their length and fresh mass were measured. The dry mass was obtained by placing the samples in an oven at 70 °C for 2 d. Samples of fresh shoots and roots were frozen quickly in liquid nitrogen in order to assay antioxidant enzymes, lipid peroxidation, soluble protein and free proline contents.
The proline content was determined spectrophoto- metrically at 520 nm by following the method described by Bates et al . (1973). The degree of lipid peroxidation was carried out by measuring the malondialdehyde (MDA) content according to the method of Madahava Rao and Sresty (2000). Total soluble protein content was measured
⎯⎯⎯⎯
Received 22 April 2008, accepted 12 September 2008.
Abbreviations : CAT - catala; GR - glutathion reducta; MDA - malondialdehyde; POX - peroxida; ROS - reactive oxygen species; SPD - spermidine; SPM - spermine.
Acknowledgements : The authors are also very thankful to two anonymous reviewers and Doc. Jeremy for their constructive comments to the early versions of the manuscript. This rearch was financially supported by National Science and Technology Supporting
Projects (2006BAD07A04 ), National Natural Science Foundation (30871455,30770223), the Major Project of Knowledge Innovation Program, Chine Academy of Sciences (KSCX1-YW-09-04) and Kaidi Sustainable Bio-energy Investment Company. * Corresponding author; fax: (+86) 10 82592431, e-mail: guweibin@ibcas.ac
Y .Y . CHAI et al.
146
using the procedure developed by Bradford (1976) using bovine rum albumin as a standard. Catala (CAT) activity was performed by the method of Chance and Maehly (1955) and determined by monitoring the reduction of H 2O 2 at 240 nm and 25 °C. Peroxida (POX) activity was bad upon the method described by Chance and Maehly (1955) and measured by the H 2O 2-dependent oxidation of guaiacol at 470 nm. Glutathione reducta (GR) activity was determined by the procedure described by Halliwell and Foyer (1978) and measured by monitoring the reduction of NADPH at 340 nm.
The application of SPM caud no obvious effect on germination under the salinity stress (data not shown). There were no distinct improvement in the elongation of shoots and roots with the application of SPM compared with the control. However, the FM and DM of shoots were slightly reduced by SPM, while there was a small increment in the FM and DM of the roots. When 180 mM NaCl was added to the germination solution, the length, FM and DM of both shoots and roots were significantly lower when compared with the control. The treatment of SPM + NaCl caud an increment of the length of both shoots and roots (43 and 11 %, respectively) together with a large increa in the FM and root DM comparing with the M-81E edlings expod to NaCl alone, but had no distinct effect on the DM of the shoots (Table 1). The prent study provided strong evidence
that SPM improved the growth of sweet sorghum under salinity. The findings were in agreement with a previous report which found that spermine was partially responsible for restoration of growth in edlings treated with 150 mM NaCl, and was more effective than the putrescine (Benavides et al . 1997).
The total soluble protein of M-81E edlings showed no distinct difference between the control and edlings treated with SPM in either the shoots or the roots (Fig. 1A ). When NaCl was applied, the protein content was substantially higher than that found in the control group. There was no difference between the group treated with SPM + NaCl and NaCl alone. In addition, shoots had higher total soluble protein content than roots.
It is well known that proline is a nitrogen source available for the recovery from stress and restoration of growth (Trotel et al . 1996). In addition, proline also acts as an osmolyte, which reduce the osmotic potential of the cell and the uptake of toxic ions (Woodward and Bennett 2005).
The content of free proline incread significantly in both in shoots and roots of sweet sorghum edlings treated with 180 mM NaCl when comparing with the control groups. However, the less increa in the proline content after SPM + NaCl treatment in both shoots and roots might be due to
alleviation of the NaCl-stress (Fig. 1B ). This was consistent with the results of Jiménez-Bremont et al . (2006), who found less accumulation of the proline in bean tissues under saline condition after application of poly- amines. In addition, the content of free proline was greater in the shoots than in roots, which is also in accordance with the result of Jiménez-Bremont et al . (2006).
Tang and Newton (2005) reported that polyamine plays a vital role in reducing the oxidative stress in Virginia pine. Additionally, Hsu and Kao (2007) showed that spermine protected rice against Cd toxicity by preventing the increa of MDA and enhancing the activities of antioxidant enzymes. Similarly, we obrved the increa in MDA content in edlings treated by NaCl alone, but much less in tho treated by SPM + NaCl (Fig. 1D ). Moreover, Roberts et al . (1985) demonstrated that spermine had the beneficial effects of reducing membrane fluidity and protecting rigidity. This fact strengthens the possibility that the appropriate concentrations of added spermine could induce a stabilization of the plasmalemma (Hsu and Kao 2007).
学韩语视频Reactive oxygen species (ROS) are responsible for salinity-induced damage to macromolecules and cellular structures (Özdemir et al . 2004). Therefore, the roles of antioxidative enzymes, including CAT, POX and GR assume great importance. As it was shown in our study, NaCl resulted in an increa in the activity of CAT (Fig. 1C ) and POX (Fig. 1E ) but a decrea in GR activity (Fig. 1F ).
The results are in accordance with tho of Mandhania et al . (2006) who reported the enhancement in the POX and CAT activities in wheat under salinity. Similarly, POX and CAT activities were incread in potato edlings (Rahnama and Ebrahimzadeh 2005) and barley edlings (Fedina et al. 2009). The lowering of GR activity in sweet sorghum edlings under salt stress was consistent with the result of Özdemir et al . (2004). When exogenous SPM was added to the test solution, the activities of POX and GR incread, but no effect on the CAT activity was obrved. Whereas, the salicylic acid treatments significantly inhibited CAT activity and incread POX activity under salinity (Muthu et al . 2009).
Table1. The effects of SPM on growth of sweet sorghum edlings under 180 mM NaCl stress. Values are means ± SE bad on 15 replicates for length and four replicates for fresh and dry mass.
Parameter Control SPM NaCl NaCl + SPM Length [cm ] shoot 7.86 ± 0.04 a 7.86 ± 0.04 a 2.11 ± 0.02 c 3.02 ± 0.01 b
root 12.35 ± 0.10 a 11.94 ± 0.05 b 4.59 ± 0.10 d 5.09 ± 0.02 c FM [mg ] shoot 80.09 ± 0.45 a 74.46 ± 0.31 b 21.13 ± 0.29 d 24.71 ± 0.29 c
root 4.94 ± 0.08 a 7.29 ± 0.18 b 1.36 ± 0.40 d 7.74 ± 0.27 c DM [mg ] shoot 7.48 ± 0.12 a 6.71 ± 0.08 b 2.84 ± 0.23 c 5.06 ± 0.62 c
root
3.06 ± 0.34 a
3.32 ± 0.13 a
routinely1.22 ± 0.02 c
2.82 ± 0.11 b
国际音标发音表下载EFFECT OF SPERMINE ON SALT-STRESSED SORGHUM
147
Fig.1. The effects of exogenous SPM on sweet sorghum edlings protein content (A ), proline content (B ), CAT activity (C ), MDA content (D ) POX activity (E ) and GR activity (F ) under salinity stress.
The incread activity of the POX and GR under the SPM treatment indicated that SPM alleviated oxidative stress. Exogenous SPM might reduce NaCl-induced oxidative damage by restraining the Na + uptake from the medium. In conclusion, exogenous SPM partially protected sweet sorghum during the germination stage under salinity stress.
References
Bates, L.S., Waldren, R.P., Teare, I.D.: Rapid determination of
free proline for water-stress studies. - Plant Soil 39: 205-207, 1973.
Benavides, M.P., Aizencang, G ., Tomaro, M.L.: Polyamine in
Helianthus annuus L. during germination under salt stress. - J. Plant Growth Regul. 16: 205-211, 1997.
Bradford, M.M.: A rapid and nsitive method for the
quantification of microgram quantities of protein utilizing the principle of protein-dye binding. - Anal. Biochem. 72: 248-254, 1976.
Chance, B., Maehly, A.C.: Assay of catalas and peroxidas. -
In: Colowick, N.O., Kaplan, S.P. (ed.): Methods Enzymology. Pp. 764-775. Academic Press, New York 1955.
D’Orazi, D., Bagni, N.: In vitro interactions between polyamines
afloat
and pectic substances. - Biochem. biophys. Res. Commun. 148: 1259-1263, 1987.
Fedina. I.S., Nedeva, D., Çiçek, N.: Pre-treatment with H 2O 2
induces salt tolerance in barley edlings. - Biol. Plant. 53: 321-324, 2009.
Galston, A.W., Sawhney, R.K.: Polyamines in plant physiology. -
Plant Physiol. 94: 406-410, 1990.
Groppa, M.D., Benavides, M.P.: Polyamines and abiotic stress:
recent advances. - Amino Acids 34: 35-45, 2008.
Halliwell, B., Foyer, C.H.: Properties and physiological function
四级准考证丢了怎么办
of a glutathione reducta purified from spinach leaves by affinity chromatography. - Planta 139: 9-17, 1978.
Hsu, Y .T., Kao, C.H.: Cadmium-induced oxidative damage in
rice leaves is reduced by polyamines. - Plant Soil 291: 27-37, 2007.
Jiménez-Bremont, J.F., Becerra-Flora, A., Hwenández-Lucero,
E., Rodríguez-Kessler, M., Acosta-Gallegos, J.A., Ramírez- Pimentel, J.G .: Proline accumulation in two bean cultivars
Y .Y . CHAI et al.
148
under salt stress and the effect of polyamines and ornithine. - Biol. Plant. 50: 763-766, 2006.
Kuznetsov, V .V ., Radyukina, N.L., Shevyakova, N.I.:
Polyamines and stress: biological role, metabolism and regulation. - Russ. J. Plant Physiol. 53: 583-
604, 2006.
Madahava Rao, K.V ., Sresty, T.V .S.: Antioxidative parameters in
the edlings of pigeonpea (Cajanus cajan L. Millspaugh) in respon to Zn and Ni stress. - Plant Sci. 157: 113-128, 2000.
Mandhania, S., Madan, S., Sawhney, V .: Antioxidant defen
mechanism under salt stress in wheat edlings. - Biol. Plant. 50: 227-231, 2006.
Martin-Tanguy, J.: Metabolism and function of polyamines in
plants: recent development (new approaches). - Plant Growth Regul. 34: 135-148, 2001.
Muthu, S., Atici, Ö., Nalbantoglu, B.: Effect of salicylic acid and
salinity on apoplastic antioxidant enzymes in two wheat cultivars differeing in salt tolerance. - Biol. Plant. 53: 334-338, 2009.
tyrannosaurus
Özdemir, F., Bor, M., Demiral, T., Türkan, Ì.: Effects of
24-epibrassinolide on ed germination, edling growth, lipid peroxidation, proline content and antioxidative system of rice (Oryza sativa L.) under salinity stress. - Plant Growth Regul. 42: 203-211, 2004.
Rahnama, H., Ebrahimzadeh, H.: The effect of NaCl on
antioxidant enzyme activities in potato edlings. - Biol.
Plant. 49: 93-97, 2005.
Roberts, D.R., Dumbroff, E.B., Thompson, J.E.: Exogenous
polyamines alter membrane fluidity in bean leaves − a basis for potential misinterpretation of their true physiological role. - Planta 167: 395-401, 1985.
Shevyakova, N.I., Shorina, M.V ., Rakitin, V .Y ., Kuznetsov, VI.V .:
Stress-dependent accumulation of spermidine and spermine in the halophyte Membryanthemum crystallinum under salinity conditions. - Russ. J. Plant Physiol. 53: 739-745, 2006.
Tang, W., Newton, R.J.: Polyamines reduce salt-induced
oxidative damage by increasing the activities of antioxidant enzymes and decreasing lipid peroxidation in Virginia pine. - Plant Growth Regul. 46: 31-43, 2005.
Trotel, P., Bouchereau, A., Niogret, M.F., Larher, F.: The fate of
osmo-accumulated proline in leaf discs of rape (Brassica napus L.) incubated in a medium of low osmolarity. - Plant Sci. 118: 31-45, 1996.
Woodward, A.J., Bennett, I.J.: The effect of salt stress and
abscisic acid on proline production, chlorophyll content and growth of in vitro propagated shoots of Eucalyptus camaldulensis . - Plant Cell, Tissue Organ Cult. 82: 189-200, 2005.
