Alterations in sucrose metabolizing enzyme activities and total phenol

Front. Biol. China 2009, 4(4): 419–423 DOI 10.1007/s11515-009-0037-2

RESEARCH ARTICLE

Alterations in sucrose metabolizing enzyme activities and total phenol content of Curcuma longa L. as affected by different triazole compounds C. Abdul JALEEL1,2, Changxing ZHAO (✉)3, Sedghi MOHAMED4, Hameed Jasim AL-JUBURI2, Helal Ragab MOUSSA5, M. GOMATHINAYAGAM1, R. PANNEERSELVAM1 1 Stress Physiology Lab, Department of Botany, Annamalai University, Tamil Nadu, India 2 DMJM International (Cansult Maunsell/AECOM Ltd.), Consultant of Gardens Sector Projects, Alain Municipality and Eastern Emirates, P.O. Box 1419, Al-Ain, Abu Dhabi, United Arab Emirates 3 College of Agronomy and Plant Protection, Qingdao Agricultural University, Chunyang Road, Chengyang District, Qingdao 266109, China 4 Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Mohaghegh Ardabili, Ardabil, Iran 5 Radioisotope Department, Atomic Energy Authority, Malaeb El-Gamaa St., P.O. 12311, Dokki, Giza, Egypt

© Higher Education Press and Springer-Verlag 2009

Abstract Changes in the sucrose metabolism of Curcuma longa L. plants were studied under treatment with different triazole compounds viz., triadimefon (TDM) and propiconazole (PCZ). Plants were treated with TDM at 15 mg/L and PCZ at 10 mg/L separately by soil drenching on 80, 110, and 140 days after planting (DAP). The plants were harvested randomly on 90, 120, and 150 DAP to determine the effect of both the triazoles on sucrose metabolizing enzymes and phenol content. The sucrose metabolism was studied by analyzing sucrose metabolizing enzymes like sucrose synthase and sucrose phosphate synthase. All the analyses were assayed in leaves and tubers of both control and treated plants. It was found that both of the triazole compounds had profound effects on these parameters. Keywords Curcuma longa, triadimefon, propiconazole, sucrose synthase, sucrose phosphate synthase, total phenol

1

Introduction

Turmeric (Curcuma longa L.), originated from India and found in South American countries, is an important economic crop cultivated for its underground rhizomes which are widely used in drugs, cosmetic industries, condiments, curry stuff and on religious and auspicious occasions (Panneerselvam, 1998; Panneerselvam et al., Received September 11, 2008; accepted December 2, 2008 E-mail: [email protected]

2007). It has been traditionally used in indigenous herbal medicines due to its biological activities (Govindarajan, 1980). Also because of its easy digestibility, turmeric has been used industrially to prepare special food and children’s foods (Kumar et al., 2006). Turmeric has long been known in India and many other countries as important dietary sources in addition to their use in traditional medicine for wound healing, treatment of inflammation and stomach acidity (Jyothi et al., 2003). Triazoles are a group of compounds which have both fungitoxic and plant growth-regulating properties (Jaleel et al., 2006a, 2006b, 2007a, 2007e). In addition, they can also protect plants against various environmental stresses (Jaleel et al., 2007b, 2007f). Triazoles affect the isoprenoid pathway and alter the levels of certain plant hormones by inhibiting gibberellin synthesis, reducing ethylene evolution, and increasing cytokinin levels (Jaleel et al., 2007d, 2007g). Some of the previous work carried out in our lab revealed the morphological and physiological changes associated with triazole-treatment in various plants, including the inhibition of plant growth, decreased internodal elongation, increased chlorophyll levels, enlarged chloroplasts, thickened leaf tissue, increased root to shoot ratio and alkaloid production, and enhancement of carbohydrate metabolism (Jaleel et al., 2007c, 2008a, 2008b). Triadimefon (TDM) and propiconazole (PCZ), which are the triazole group of fungicides and have plant growth regulator (PGR) properties, were reported to inhibit gibberellic acid biosynthesis and increase abscisic acid and cytokinin content (Fletcher et al., 2000). In addition, previous reports have revealed changes in

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photosynthetic characteristics and carbohydrate metabolism in the S. rotundifolius plant under TDM and PCZ applications (Kishorekumar et al., 2007, 2008). Therefore, the current study aims to evaluate the ability of TDM and PCZ to alter the sucrose metabolism and phenol content of Curcuma longa.

2

Materials and methods

Rhizomes of Curcuma longa L. (family: Zingiberaceae) were planted and treated with different concentrations (5, 10, 15, and 20 mg/L) of TDM and PCZ to determine the optimum concentration. Among these concentrations, 15 mg/L of TDM and 10 mg/L PCZ increased the dry weight significantly, and higher concentrations slightly decreased the growth and dry weight. Each plant was treated separately with one liter of aqueous solution containing 15 mg TDM and 10 mg PCZ at vegetative stages such as 80, 110, and 140 days after planting (DAP). The treatments were given by soil drenching. The plants were uprooted on 90, 120, and 150 DAP and separated into shoot and tubers for analyses.

standard curve prepared with different concentrations of gallic acid.

3

Results

3.1

Sucrose phosphate synthase

Triazole treatment significantly increased the level of sucrose phosphate synthase activity in the leaves of C. longa up to 150 DAP when compared to the control. In the tuber, the activity of sucrose phosphate synthase was increased to a higher level in the triazole treated plants than in the control. PCZ treatment increased the sucrose phosphate synthase activity than did TDM treatment in the tuber tissue. The tuber tissue showed a higher level of sucrose phosphate synthase activity when compared to the leaves except on 90 DAP. 3.2

Sucrose synthase

Sucrose synthase (UDP-glucose, D-fructose-2-glycosyl transferase (EC: 2.4.1.13) was extracted and estimated using the method of Glavier et al. (1991). The result was expressed in units representing mg of reducing sugars released per min per mg protein. Sucrose phosphate synthase (EC: 3.4.1.14) was extracted and estimated by the method of Huber and Huber (1991).

The sucrose synthase activity of leaves increased with the age of the plant. Triazole treatment significantly increased the sucrose synthase activity in both leaf and tuber. Among the triazoles, PCZ treatment increased the sucrose synthase activity more than that of TDM treated plants. The sucrose synthase activity increased with the age of the plants up to 150 DAP in both control and treated plants, and later it declined in the tuber tissue. The highest sucrose synthase activity was observed in the tuber of PCZ treated plants followed by TDM, and it was 113.26% and 106.53% over the control, respectively, on 150 DAP. Among the organs, tuber tissue showed a higher level of sucrose synthase activity than leaves.

2.2

3.3

2.1

Enzyme extractions

Estimation of total phenols

Total phenol was estimated using the method of Malick and Singh (1980). The total phenol was determined using a

Total phenol in shoot

The total phenol content in shoot increased with the increase in age of both control and triazole treated plants.

Fig. 1 Triazole compounds induced changes in the activity of sucrose phosphate synthase in leaf (B) and tuber (A) of C. longa. Values are the mean of three replicates expressed in specific activity (μg of sucrose min–1 mg–1 protein).

C. Abdul JALEEL et al. Alterations in sucrose metabolizing enzyme activities and total phenol content of Curcuma longa L.

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Fig. 2 Effect of triazole compounds induced changes on the activity of sucrose synthase in leaf (B) and tuber (A) of C. longa (Values are the mean of three replicates expressed in mg of reducing sugar released min–1 mg–1 protein)

Among the triazoles, TDM treatment increased the phenol content to 135.18% over the control and PCZ treatment increased that to 135.71% over the control on 150 DAP in the shoot of C. longa. Triazole treatment also increased the total phenol content in tuber tissue when compared to the control. The phenol content was higher in the shoot than in the tuber. On 150 DAP, PCZ and TDM increased total phenol content to 117.11% and 114.91% over the control, respectively.

4

Discussion

The activity of sucrose synthase increased in the leaves and tubers of the triazole treated plants. Among the plant organs, tubers showed higher levels of sucrose synthase activity when compared to leaves. The major role of sucrose synthase during the active growth phase is splicing the sucrose moiety and providing the basic precursor for

growth and building up the sink structure. Sucrose degradation is the first step for carbon utilization by plant cells (Iragi and Tremblay, 2001). Sucrose synthase may regulate sucrose degradation and thus the rate of dry matter accumulation. The cleavage of sucrose by sucrose synthase in the cytosol provides the substrate for starch synthesis, which is essential to increase the rate of dry matter import as observed in tomato fruit (King et al., 1997). Starch accumulation during early rapid fruit growth parallels with the increase in sucrose synthase activity in tomato (Robinson et al., 1988). Dramatic reduction in the extent of starch accumulation, protein content, and dry weight of transgenic potato tuber deficient in sucrose synthase confirms that this enzyme is essential for effective sucrose metabolism in the developing tuber (Zrenner et al., 1995). The increased activity of invertase and sucrose synthase which breakdown sucrose and provide material for starch synthesis increased to a larger extent in the tubers of triazole treated plants. This increase of sucrolytic enzymes can be well correlated with increased starch

Fig. 3 Triadimefon and propiconazole induced variation in total phenol in leaf (B) and tuber (A) of C. longa (values are the mean of three replicates expressed in mg$g–1 dry weight)

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content and dry weight of tubers of the triazole treated plants. In the leaves, triazole treatment increased the sucrose phosphate synthase activity firstly, and later it declined slightly. In the tubers of C. longa, the sucrose phosphate synthase activity was found increased at all stages. In many plants, the last important regulatory step in sucrose synthesis during photosynthesis is the catalysis by sucrose phosphate synthase (Stitt et al., 1987). The addition of gibberellic acid (GA3) and kinetin to the stressed medium increased both sucrose synthase and sucrose phosphate synthase activities in cotyledons and shoots of chickpea seedlings, while in roots no significant effect was observed (Kaur et al., 2000). The activity of sucrose phosphate synthase in the grain of sorghum increased with ABA in the sucrose medium (Bhatla and Singh, 2002). The increased cytokinin and ABA content induced by the triazoles might have increased the sucrose phosphate synthase activity in C. longa.

5

Conclusions

From the results of this investigation, it is clear that TDM and PCZ treatments increased the activities of sucrose synthase and sucrose phosphate synthase and increased the total phenol contents of C. longa plants. Therefore, it can be concluded that the application of TDM and PCZ could well be used as a potential tool to manipulate sucrose metabolism and phenol content and thereby the rhizome quality in medicinal plants like C. longa. However, the data presented here reflect the importance of a physiological analysis of plant response to fungicide treatments, which should be accompanied by field experiments and evaluation.

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