ความ เกี่ยวข้องของ antioxidant enzymes และ lipid ... - CRDC

Agricultural Sci. J. 41(3/1)(Suppl.): 553-556 (2010) ว. วิทย์. กษ. 40(3/1)(พิเศษ): 553-556 (2553)
ความเกี ่ยวข้องของ antioxidant enzymes และ lipid peroxidation กับความแข็งแรงของเมล็ดพันธุ ์ข้าวโพด
พันธุ ์สุวรรณ 5 ที ่ hydropriming
Involvement of Antioxidant Enzymes and Lipid Peroxidation on Seed Vigour of Maize Seeds cv. Suwan 5
Through Hydropriming
ภภัสสร วัฒนกุลภาคิน 1 คิน เลย์ คู1 กนก รัตนะกนกชัย 1 ภาณุมาศ ฤทธิชัย 2 และ ทรงศิลป์ พจน์ชนะชัย 3 *
Wattanakulpakin, P. 1 , Kyu, K.L. 1 , Ratanakhanokchai, K. 1 , Ritthichai, P. 2 and Photchanachai, S. 3 *
Abstract
Maize seeds (Zea mays L.) ‘Suwan 5’ from Nation Corn and Sorghum Research Center (NCSRC) was
hydroprimed for 6-18 h at 25C. Hydropriming did not effect to seed germination compared to the control seeds.
Hydropriming for 6 and 12 h improved the seed vigour as indicated by a higher germination percentage after
seed ageing at 42C and 100% RH for 96 h. However, the longer hydropriming times by 18 h significantly
reduced seed vigour compared to the seeds primed for 6 and 12 h. While, the declined malondialdehyde (MDA)
content in seeds primed for 18 h was significantly different to the other treatments. Hydropriming for 12 and 18 h
decreased the superoxide dismutase (SOD) activity compared to the control seeds. Ascobate (APX) activity was
independent by the different priming treatments. The experiment suggests that the seed vigour of maize primed
seeds do not involve with lipid peroxidation but declining seed vigour may be related to a reduction of SOD
activity.
Keywords: seed vigour, priming, antioxidant enzymes, lipid peroxidation
บทคัดย่อ
เมล็ดพันธุ ์ข้าวโพดพันธุ ์สุวรรณ 5 จากสถานีวิจัยข้าวโพดและข้าวฟ่ างแห่งชาติ ที่ hydropriming เป็ นเวลา 6-18 ชม. ที่
อุณหภูมิ 25C พบว่า hydropriming ไม่มีผลต่อความงอกของเมล็ดพันธุ ์เมื่อเปรียบเทียบกับชุดควบคุม แต่พบว่าเมล็ดพันธุ ์ที่
hydropriming เป็ นเวลา 6 และ 12 ชม. มีความแข็งแรงสูงขึ ้นเมื่อทดสอบโดยการเร่งอายุที่ 42C ความชื ้นสัมพัทธ์ 100% เป็ น
เวลา 96 ชม. แต่ hydropriming เป็ นเวลา 18 ชม. ท าให้ความแข็งแรงของเมล็ดพันธุ ์ลดลงอย่างมีนัยส าคัญกับเมล็ดที่
hydropriming เป็ นเวลา 6 และ 12 ชม. และพบว่าปริมาณ malondialdehyde (MDA) ลดลงอย่างมีนัยส าคัญทางสถิติเมื่อ
เปรียบเทียบกับชุดการทดลองอื่นๆ ในขณะที่กิจกรรมของ superoxide dismutase (SOD) ของเมล็ดข้าวโพดที่ hydropriming
เป็ นเวลา 12 และ 18 ชม. ลดลงเมื่อเปรียบเทียบกับชุดควบคุม แต่ hydropriming ไม่มีผลต่อกิจกรรมของ ascobate
peroxidase (APX) จากผลการทดลองพบว่าความแข็งแรงของเมล็ดพันธุ ์ข้าวโพดที่ hydropriming ไม่มีความเกี่ยวข้องกับ lipid
peroxidation แต่การลดลงของความแข็งแรงอาจมีความสัมพันธ์กับการลดลงของกิจกรรม SOD
ค าส าคัญ: ความแข็งแรงของเมล็ดพันธุ ์, priming, antioxidant enzymes, lipid peroxidation
1
สายวิชาเทคโนโลยีชีวเคมี คณะทรัพยากรชีวภาพและเทคโนโลยี ม.เทคโนโลยีพระจอมเกล้าธนบุรี 49 ม.8 ถ.เทียนทะเล 25 แขวงท่าข้าม เขตบางขุนเทียน กทม.10150
1
Division of Biochemical Technology, School of Bioresources and Technology, King Mongkut’s University of Technology Thonburi, 49 Moo. 8,Tientalay 25 Rd.,
Thakham, Bangkhuntien, Bangkok 10150, Thailand.
2
ภาควิชาเทคโนโลยีการเกษตร คณะวิทยาศาสตร์และเทคโนโลยี ม.ธรรมศาสตร์ ศูนย์รังสิต 99 ถ.พหลโยธิน อ.คลองหลวง จ.ปทุมธานี 12121
2
Department of Agricultural Technology, Faculty of Science and Technolog, Thammasat University Rangsit Center, 99 Paholyothin Rd., Klongluang, Patumtani
12121, Thailand.
3
หลักสูตรเทคโนโลยีหลังการเก็บเกี ่ยว คณะทรัพยากรชีวภาพและเทคโนโลยี ม.เทคโนโลยีพระจอมเกล้าธนบุรี 49 ม.8 ถ.เทียนทะเล 25 แขวงท่าข้าม เขตบางขุน
เทียน กทม.10150
3
Postharvest Programme, School of Bioresources and Technology, King Mongkut’s University of Technology Thonburi, 49 Moo. 8, Tientalay 25 Rd., Thakham,
Bangkhuntien, Bangkok 10150, Thailand.

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Introduction
Priming has been widely used to improve seed germination and vigour in many crop species. The advantages
of priming, improved seed vigour, are reported to involve with the repair and build up of nucleic acids, the
increased synthesis of proteins as well as the repair of membranes (McDonald, 1999). Priming also enhances the
activities of scavenging enzymes and reduces lipid peroxidation, indicating by MDA content, result to the increase
in seed germination and vigour in purple coneflower (Chiu et al., 2006) and bitter gourd seeds (Wang et al.,
2003). However, improper priming processes, due to the unfavorable imbibitions and re-drying steps, generate
reactive oxygen species (ROS) such as superoxide radical (O 2 - ), hydrogen peroxide (H 2 O 2 ) and hydroxyl radical
(Leprince et al., 1994). ROS can initiate injurious degradation causing lipid peroxidation, membrane deterioration,
protein and nucleic acids degradation leading to reduce seed vigour and cell death (McDonald, 1999).The poorer
longevity of primed sh-2 sweet corn seeds than unprimed seeds was found (Chang and Sung, 1998). The effect of
priming on seed vigour and biochemical mechanisms is depended on seed species and priming methods
(McDonald, 1999). The aim of this research was to study the involvement of antioxidant enzymes and lipid
peroxidation on seed vigour of maize seeds cv. Suwan 5 through the variations of hydropriming times.
Material and Methods
Maize seeds (Zea mays L.) ‘Suwan 5’, harvested in December, 2008, was purchased from Nation Corn and
Sorghum Research Center (NCSRC). The seeds were hydroprimed by placing them on metal wire sieve container
laying on distilled water surface for 6-18 h at 25C. Thereafter, the imbibed seeds were immediately dried at 40C
for 24 h to reduce the moisture content down to initial moisture content. The unprimed seeds were used as a
control sample. Moisture content (%), germination (%), vigour test (accelerated ageing test at 42C and 100%RH
for 96 h) of primed and unprimed seeds was investigated followed by ISTA rule (2007). The lipid peroxidation was
explained in term of malondialdehyde (MDA) content in the seeds (Tamagnone et al., 1998). Superoxide
dismutase (SOD) (Beauchamp and Fridovish, 1971) and ascobate peroxidase (APX) (Nakano and Asada, 1987)
were analyzed. The experiment was arranged as a completely randomized design (CRD) with four replications.
Data were subjected to analysis of variance (ANOVA) and significant differences among means were determined
by Duncan’s New Multiple Range Test at P

Germination(%) and
Germination after AA-test (%)
MDA content (mol/g DW)
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activity was independent by the different hydropriming times (Table 1). The experiment suggests that the reduced
seed vigour by hydropriming for 18 h may relate to the decline in SOD activity but these two enzymes did not
involve with the increased seed vigour.
The reduction of SOD activity may lead to accumulate MDA content in the seeds priming for longer period. The
comprehensive understanding of the relationship between MDA content and seed vigour by longer hydropriming
was investigated. Germination of primed seeds (24 and 30 h) decreased significantly compared to control
samples, nevertheless, the MDA content remained unchanged (Fig. 1). Thus, the reduction of vigour of primed
maize seed is not related to lipid peroxidation. Unlike our results, the decrease of the germination rate as well as
seed vigour correlated to an increase of the MDA content found in sunflower (Corbineau et al., 2002) and cotton
seeds (Goel et al., 2003). Non-enzymatic antioxidants or protein degradation may contribute to the reduction of
the maize primed seed vigour.
Table 1 Moisture content (%MC), germination (%G), germination after accelerated ageing (%AA), MDA content
and the activities of SOD and APX in maize seeds ‘Suwan 5’ hydroprimed for 6-18 h at 25C.
Priming MC G AA MDA SOD APX
times (h) (%) (%) (%) mol/g DW unit/kernel unit/mg protein
0
6
11.2
11.5
96ab
99a
84b
90a
6.68a
6.27a
5.099ab
5.388a
13.01
13.11
12 11.5 98a 92a 6.05a 4.427b 9.29
18 11.4 92b 80b 3.29b 3.130c 11.29
The different mean in column separated by small letters indicate statistical significance according to the Duncan’s New Multiple
Range Test (P < 0.05)
.
100
80
60
40
20
0
G
AA
MDA
6 3
Hydropriming times (h)
8
7
6
5
4
3
2
1
0
Figure 1 %Germination (G),
%germination after accelerated ageing
(AA) and MDA content in maize seeds
‘Suwan 5’ hydroprimed for 6-30 h at
25C. Error bars represent ±S.E.
Summary
Hydropriming for 6 and 12 improved maize seed vigour. The high seed vigour did not associate with MDA
content (lipid peroxidation) and the activity of SOD and APX. Hydropriming for 18 h reduced seed vigour which
may relate to decline in SOD activity. In this study, however, lipid peroxidation did not involve with primed maize
seed vigour.
Acknowledgement
This research was supported by a grant under the program Strategic Scholarships for Frontier Research
Network for the Ph.D. Thai Doctoral degree Program from the Commission on Higher Education, Thailand.

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Literature cited
Beauchamp, C.O. and Fridovich, I., 1971, Superoxide Dismutase: Improved Assays and an Assay Applicable to
Acrylamide Gels, Analytical Biochemistry, 44: 276-287.
Berno Bernal-Lugo, I. and Leopold, A.C., 1995, Seed Stability during Storage: Raffinose Content and Seed
Glassy State, Seed Science Research, 5: 75–80.
Chiu, K.Y., Chen, C.L. and Sung, J.M., 2003, Partial Vacuum Storage Iimproves the Longevity of Primed sh-2
Sweet Corn Seeds, Scientia Horticulturae, 98: 99–111.
Chiu, K.Y., Chuang, S.J. and Sung, J.M., 2006, Both Anti-oxidation and Lipid-carbohydrate Conversion
Enhancements are Involved in Priming-improved Emergence of Echinacea purpurea Seeds that Differ in
Size, Scientia Horticulturae, 108: 220–226.
Chang, S.M. and Sung, J.M., 1998, Deteriorative Changes in Primed Sweet Corn Seeds During Storage, Seed
Science Technology, 26: 613–626.
Corbineau, F., Gay-Mathieu, C., Vinel, D. and Come, D., 2002, Decrease in Sunflower (Helianthus annuus) Seed
Viability Caused by High Temperature as Related to Energy Metabolism, Membrane Damage and Lipid
Composition, Physiologia Plantarum, 116: 489–496.
El-Araby, M.M. and Hegazi, A.Z., 2004, Responses of Tomato Seeds to Hydro- and Osmo-priming, and Possible
Relations of Some Antioxidant Enzymes and Endogenous Polyamine Fractions, Egyptian Journal of Biology,
6: 81-93.
Goel, A., Goe, A.K. and Sheoran, I.S., 2003, Changes in Oxidative Stress Enzymes During Artificial Aging in
Cotton (Gossypium hirsutum L.) Seeds, Journal of Plant Physiology, 160: 1093–1100.
ISTA, 2007, International Rules for Seed Testing Edition, Published by The International Seed Testing
Association, Switzerland.
Leprince, O., Atherton, N.M., Deltour, R. and Hendry, G.A.F., 1994, The Involvement of Respiration in Free
Fadical Processes During Loss of Desiccation Tolerance in Germinating Zea mays L. An Electron
Paramagnetic Resonance Study, Plant Physiology, 104: 1333–1339.
McDonald, M.B., 1999, Seed Deterioration: Physiology, Repair and Assessment, Seed Science Technology, 27,
177–237.
Nakano, Y. and Asada, K., 1987, Purification of Ascorbate Peroxidase in Spinach Chloroplasts: Its Inactivation in
Ascorbatedepleted Medium and Reactivation by Monodehydroascorbate Radical, Plant and Cell
Physiology, 28: 131–140.
Qun, S., Jim-hua, W. and Bao-qi, S., 2007, Advances on Seed Vigour Physiological and Genetic Mechanisms.
Agricultural Sciences in China, 6: 1060-1066.
Tamagnone, L., Merida, A., Stacey, N., Plaskitt, K., Parr, A., Chang, C.F., Lynn, D., Dow, J.M., Roberts, K. and
Martin, C., 1998, Inhibition of Phenolic Acid Metabolism Results in Precocious Cell Death and Altered Cell
Morphology in Leaves of Transgenic Tobacco Plants, Plant Cell, 10: 1801–1816.
Wang, H.Y., Chen, C.L. and Sung, J.M., 2003, Both Warm Water Soaking and Solid Priming Treatments Enhance
Antioxidation of Bitter Gourd Seeds Germinated at Sub-optimal Temperature, Seed Science Technology,
31: 47–56.
Yeh, Y.M., Chiu, K.Y., Chen, C.L. and Sung, J.M., 2005, Partial Vacuum Extends the Longevity of Primed Bitter
Gourd Seeds by Enhancing Their Anti-oxidative Activities During Storage, Scientia Horticulturae, 104: 101–
112.