J.

Agronomy & Crop Science 191, 8187 (2005) 2005 Blackwell Verlag, Berlin ISSN 0931-2250

Department of Biochemistry and Chemistry, Punjab Agricultural University, Ludhiana, India

Seed Priming Increases Crop Yield Possibly by Modulating Enzymes of Sucrose Metabolism in Chickpea
S. Kaur, A. K. Gupta, and N. Kaur
Authors address: Dr S. Kaur, Dr A. K. Gupta and Dr N. Kaur (corresponding author; e-mail: nkaur@redimail.com), Department of Biochemistry and Chemistry, Punjab Agricultural University, Ludhiana 141004, India With 1 gure and 5 tables Received October 20, 2003; accepted January 27, 2004

Abstract
The number of seeds and seed yield per plant were higher in chickpea crops raised from water and mannitol (4 %) primed seeds in comparison with the control non-primed crops. In primed plants, an enhanced acid invertase activity in the apical part of the main stem and the part immediately below it at 100 and 130 days after sowing (DAS) might result in an increased availability of hexoses to these plant parts. In addition, decreased acid invertase activity at the point of initiation of branches and in the internodes of stem observed in primed plants indicated restricted hydrolysis of sucrose during its transport through the stem, resulting in its more supply to the actively growing sinks. The activities of sucrose-cleaving enzymes, i.e. invertase and sucrose synthase (SS) in podwall of primed plants were higher at 110 DAS. At 140 DAS, a stage of rapid seed lling, increased activities of SS and sucrose phosphate synthase (SPS) were observed in seeds of primed plants. Increased SPS activity in seeds of primed crop could meet the increased assimilate requirements of the developing seeds. Higher activity of SS in seeds of primed crop may facilitate seed lling. These data suggest that enzymes of sucrose metabolism play an important role in increasing the yield of chickpea crops raised from primed seeds.

Key words: Cicer arietinum invertase priming sucrose phosphate synthase sucrose synthase

Introduction
Crop productivity is generally regarded to be dependent on photosynthesis and sink activity. Photosynthesis determines the strength of assimilate supply from the leaves and the sink activity depends upon the utilization of this assimilate by the developing seeds. Increased photosynthesis by increasing level of atmospheric CO2 (Hardman and Brun 1971) or irradiance (Schou et al. 1978) increased pod and seed numbers while decreased photosynthesis by shading (Egli and Zhen-Wen

1991, Andrade and Ferreiro 1996) or defoliation (Board and Tan 1995) reduced pod and seed numbers. However, depodding, to increase assimilate supply to the remaining seeds, usually increased seed size and weight per seed in grain legumes but did not always change individual seed growth rate (Egli et al. 1985, Munier-Jolain et al. 1998). In soyabean, seed growth rate was generally sink limited if photosynthesis increased during seed lling but source limited if photosynthesis was reduced. It was suggested that in cotyledon sucrose concentration during later stages of seed growth might be near a critical level resulting in sink limitations of seed growth rate if assimilate availability was increased (Egli and Bruening 2001). Defoliation and water stress have been reported to cause a rapid reduction in assimilate availability resulting in 5075 % decrease in levels of apoplastic sucrose in soyabean (Giord and Thorne 1985, Westgate and Grant 1989). Increased total and reducing sugar content in grains of water stress tolerant wheat genotypes was observed during grain development (Nayyar and Walia 2004). We had reported that water decit and salt stress adversely aect the seedling growth of chickpea and this eect could be counteracted by adding GA3 or kinetin in the growth medium (Kaur et al. 1998a,b). It was also observed that osmo- and hydropriming of chickpea seeds with mannitol and water alleviated the adverse eects of water decit and salt stress on seedling growth (Kaur et al. 2002a, 2003). The benecial eect of phytohormones and priming on seedling growth in chickpea were reported to be due to modulation of enzymes of carbohydrate metabolism (Kaur et al. 2000, 2002a). Studies with primed chickpea seeds were

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conducted in the eld and it was observed that priming increased plant biomass, numbers of branches, owers, pods and seeds per plant leading to higher seed yield (Kaur et al. 2002b). Seed priming has been reported to increase the yield of chickpea, maize, rice and wheat under semiarid conditions (Harris et al. 1999, 2001, Musa et al. 1999, 2001). The benecial eects of priming reported in these crops were: faster emergence of crop, better drought tolerance, early owering and higher grain yield. Although seed priming improved the physiological parameters on yield but there is paucity of information regarding the biochemical basis responsible for benecial eects of priming during crop development. In this paper the results of enzymatic studies on sucrose metabolism as inuenced by seed priming have been reported in dierent vegetative parts of plants, pod wall and seeds of chickpea.

Materials and Methods

Seeds of chickpea cultivar GPF-2 were washed with water, dipped in 0.1 % mercuric chloride for 5 min and then washed thoroughly with sterilized water. The washed seeds were divided into two lots. One was fully immersed in 4 % mannitol for osmopriming and the second in water for hydropriming and then kept in an incubator at 25 1 C for 24 h. The seeds were then washed with distilled water and completely dried on lter papers at room temperature (27 C). The seeds without any treatment were termed as non-primed. The crop was sown in November in three dierent plots under recommended agronomic practices. Each plot had ve rows and there were 6070 plants in each row. The plants in the crop raised from water and mannitol primed seeds were termed as primed plants whereas plants in the crop raised from control, non-primed seeds were termed as non-primed plants. The sucrose metabolizing enzymes were studied in plant parts, podwall and seeds of primed and non-primed plants at dierent stages of crop development. At 100 and 130 days after sowing (DAS), the plants were uprooted from the eld and brought to laboratory in crushed ice in an ice container. The dierent plant parts taken for enzymatic studies were: apical portion (top 12 cm) of the main stem including small leaves, a 2-cm portion immediately below the apical portion; a portion (0.5 cm) of the stem on either side of the branching point including point of initiation of branch and an internodal portion of the stem irrespective of node position (Fig. 1). Acid and alkaline invertases were extracted at 4 C and assayed from these tissues by methods described earlier (Kaur et al. 2002a). Podwall and seeds were taken for enzymatic studies at two stages of development, i.e. at 110 and 140 DAS corresponding to an early stage of seed development and later stages of seed development (but 20 days before maturity).

Fig. 1: Dierent parts of chickpea plant: (A) Apical portion (top 12 cm) of the main stem including small leaves; (B) 2 cm portion immediately below the apical portion; (C) a portion (0.5 cm) of the stem on either side of the branching point including point of initiation of branch; (D) an internodal portion of the stem irrespective of node position

Invertases (acid and alkaline), sucrose synthase (SS) (cleavage) and sucrose phosphate synthase (SPS) were extracted and assayed in podwall and seeds of primed and non-primed plants by the methods described earlier (Chopra et al. 2000, Kaur et al. 2002a). From each row randomly selected plants constituted one sample. Triplicate samples were taken from three dierent rows for extracting and assaying the enzymes in triplicate. The protein content of the enzyme extract was determined (Lowry et al. 1951). All the plants from each eld were taken out at maturity at 160 DAS and number and weight of seeds per plant were noted.

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83

Apical portion (top 12 cm) of the main stem including small leaves

Table 1: Eect of priming of chickpea seeds with water and 4 % mannitol on the performance of crop in the eld Priming treatment Non-primed Water Mannitol (4 %) Number of Seed mass Seed yield seeds per plant (g per seed) per plant (g) 28 2.2 37 1.4 47 3.2 0.138 0.02 3.86 0.94 0.148 0.01 5.47 1.61 0.146 0.01 6.86 0.47

DAS

Values are mean S.D. of 225250 plants from each treatment.

Mannitol 4 %

Non-primed

Priming treatment

Water

100 130 100 130 100 130

1455 796 1718 1863 1579 1499

In the present study, the average seed yields/plant in control, water and mannitol primed plants were 3.86, 5.47 and 6.86 g respectively (Table 1). It resulted in an increase of 41 and 77 % in yield on priming with water and mannitol respectively. The number of seeds increased from 28 (control) to 37 (water primed) and 47 (mannitol primed). The increase in plant biomass, enhanced number of branches, owers, pods and seeds/plant and seed yield has earlier been reported by us in chickpea (Kaur et al. 2002b). Increased yield on priming has also been reported in maize, rice and chickpea in western India (Harris et al. 1999), chickpea in high Barind Tract of Bangladesh (Musa et al. 1999, 2001) and wheat in India, Nepal and Pakistan (Harris et al. 2001). The modication of source activity during owering and pod setting has been reported to result in corresponding changes in pod and seed numbers (Egli and Bruening 2001). To examine the basis of increased plant biomass on priming, the activities of acid invertase which has been known to determine sink strength, were determined in dierent plant parts as marked in Fig. 1 in the primed and non-primed plants at 100 and 130 DAS. These results are based on 1 year experiments. Acid invertase activity was high in the apical portion and portion immediately below it but low in the internodal portion and on either side of the branching points in the primed plants in comparison with controls (Table 2). Alkaline invertase activity was also high in apical portion of the primed plants but these activities were lower than those of acid invertases (Table 3). The growth stages at 100 and 130 DAS correspond to initiation of owering and pod development. Acid invertase has been established an enzyme related to growth of plants (Sturm 1999). Enhanced invertase activities in growing parts of primed plants might result in an increased supply

A portion (0.5 cm) on either side of branching point including point of initiation of branch

A (2 cm) portion immediately below the apical portion

205 (154 12) 144 (30 5) 209 (174 20) 515* (92 20) 336 (169 15) 72* (86 8)

2593 822 3876 2434 3343 1929

399 (358 84) 24 (87+8) 87* (598 82) 741** (256 84) 264** (448 102) 129* (217 60)

882 1051 675 477 466 648

62 (175 6) 112 (152 24) 16* (154 15) 34* (82 3) 78* (66 9) 9* (93 22)

198 298 130 253 122 276

19 (78 9) 78 (62 16) 38** (46 11) 25 (56 3) 69 (48 2) 13 (57 11)

Results and Discussion

DAS, days after sowing. Values with parentheses are nanomoles of sucrose hydrolysed min)1 mg)1 protein. Values without parentheses are nanomoles of sucrose hydrolysed min)1 g)1 FW. Values are mean S.D. of three replicates. Dierences signicant in comparison with non-primed at *P < 0.01 or **P < 0.05 (Students t-test).

Table 2: Eect of priming of chickpea seeds with water and 4 % mannitol on acid invertase activity in dierent parts of the chickpea plant at dierent DAS

An internodal portion of the stem irrespective of node position

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of hexoses to them resulting in increased source of energy and growth of the plant. However, acid invertase activity at the point of initiation of branches and in the internodes of the stem was lower in primed plants, indicating that sucrose escapes hydrolysis during its transport from the stem to the actively growing sinks. These results suggest that it is the higher activity of invertases in the growing meristematic tissue which is responsible for increasing the plant growth and biomass in primed plants. Sucrose being the main carbon link between source and sinks, sucrose metabolism could be the regulatory step of both source and sink activities (Daie 1996, Foyer and Galtier 1996). The activities of invertases (acid and alkaline), SS (cleavage) and SPS were examined in podwall and seeds of primed and non-primed plants at early and late stages of seed development, i.e. at 110 and 140 DAS respectively (Tables 4 and 5). At 110 DAS, the stage at which the development of podwall is dominant, the activities of sucrose cleaving enzymes, i.e. SS as well as acid and alkaline invertases were higher in podwall of primed plants (Tables 4 and 5). The activity of alkaline invertase was found to be lesser in comparison with acid invertase. It has been reported that during early phase of owering, starch is accumulated in the podwall which is utilized to provide carbon to the developing seed (Chopra et al. 2000). High activities of invertases and SS in podwall of primed plants could result in more availability of hexoses as well as sugar nucleotides for starch synthesis in podwall that can be utilized for seed lling. However, in podwall, at 140 DAS, the activity of invertase although signicantly lesser than that observed at 110 DAS was still higher in primed plants (Table 4). These results indicate that increased sink strength of podwall because of higher invertase and SS activities at both stages of pod development may lead to better seed lling. The high SS activity during active sink development has been positively correlated with an increase in the grain weight of wheat under water-decit conditions (Yang et al. 2001). There was not much dierence in the invertase activity in seeds of primed and nonprimed plants at 110 DAS. However, at 140 DAS, the late stage of seed development, the specic activity of invertase in seeds of primed plants was much higher in comparison with that in seeds of non-primed plants (Table 4). The SS activity was more in seeds of primed plants than that of non-primed plants at 140 DAS (Table 5). On

DAS, days after sowing. Values with parentheses are nanomoles of sucrose hydrolysed min)1 mg)1 protein. Values without parentheses are nanomoles of sucrose hydrolysed min)1 g)1 FW. Values are mean S.D. of three replicates. Dierences signicant in comparison with non-primed at *P < 0.01 (Students t-test).

An internodal portion of the stem irrespective of node position A portion (0.5 cm) on either side of branching point including point of initiation of branch A (2 cm) portion immediately below the apical portion Apical portion (top 12 cm) of the main stem including small leaves DAS

Table 3: Eect of priming of chickpea seeds with water and 4 % mannitol on alkaline invertase activity in dierent parts of plants at dierent DAS

Mannitol 4 %

Non-primed

Priming treatment

Water

100 130 100 130 100 130

745 394 1039 647 1130 720

15 (82 19.7) 97 (16 4.7) 119* (105 9.2) 53* (32 3.1) 283 (121 4.9) 125* (42 2.4)

413 340 488 154 454 179

41 (57 07) 30 (36 2.0) 135 (74 18.3) 13* (16 1.7) 37 (30 2.1) 73* (18 5.0)

251 190 300 102 320 102

23 (50 6.2) 9 (28 2.8) 66 (66 14.2) 12 (17 2.4) 52 (45 3.7) 3 (15 4.2)

228 148 130 146 298 103

70 43 39 21 18 12

(87 (30 (46 (33 (75 (23

15.6) 8.7) 11.5) 2.2) 7.1) 4.0)

Table 4: Eect of priming of chickpea seeds with water and 4 % mannitol on acid and alkaline invertase activity in seeds and podwall of plants at dierent DAS Acid invertase Water primed 371 387 1538 458 84 (35 13.3) 18 (50 6.2) 61* (175 5.6) 13* (60 19.7) 236 419 1727 438 27* (22 6.2) 17* (50 12.5) 455* (212 5.4) 77* (47 12.6) 161 114 139 89 52 18 24 17 (10 2.2) (7.5 1.8) (10 0.9) (8 0.5) 124 89 135 94 52 (11 3.8) 15 (10 2.9) 6.3 (15 0.6) 11 (11 2.1) 160 146 262 229 Mannitol primed Non-primed Water primed Alkaline invertase Mannitol primed 22 (14 0.4) 32 (18 0.8) 13* (35 11.8) 44* (25 8.8)

Tissue

DAS

Non-primed

Seed Priming and Crop Yield

Seeds

110 140 Podwall 110 140

480 305 955 186

43 (31 0.9) 65 (20 6.0) 132 (71 3.8) 26 (18 5.0)

DAS, days after sowing. Values with parentheses are nanomoles of sucrose hydrolysed min)1 mg)1 protein. Values without parentheses are nanomoles of sucrose hydrolysed min)1 g)1 FW. Values are mean S.D. of three replicates. Dierences signicant in comparison with non-primed at *P < 0.01 (Students t-test).

Table 5: Eect of priming of chickpea seeds with water and 4 % mannitol on sucrose phosphate synthase and sucrose synthase activities in seeds and podwalls of plants at dierent DAS SPS Water primed Mannitol primed Non-primed 559 57 (3.9 1.4) 800 23 (8.7 0.7) 85 3.4 (1.1 0.06) 264 95 (2.0 0.4) SS (Cleavage) Water primed 379 25* (1.8 0.5) 1310 341** (11.7 2.1) 264 12* (4.3 0.7) 181 8 (2.1 0.3) Mannitol primed

Tissue

DAS

Non-primed

Seeds

110

1227 313 (8.9 4.5) 807 90** (3.7 0.9)

140

407 4.6 (4.4 0.2) 727 32* (6.7 0.8)

Podwall

110

760 39 (10.4 0.4) 380 179* (5.7 2.0)

140

804 156 (5.8 0.2) 423 82* (5.2 0.9)

604 14* (2.8 0.5) 874 32* (7.6 1.5) 329 75* (5.5 1.9) 218 15* (2.5 0.1)

404 36* (2.9 0.8) 1165 39* (10.2 2.0) 222 12* (3.7 0.7) 185 13 (2.1 0.1)

DAS, days after sowing; SPS, sucrose phosphate synthase; SS, sucrose synthase. Values without parentheses are nanomoles of sucrose synthesized min)1 mg)1 protein for SPS; and nanomoles of sucrose hydrolysed min)1 mg)1 protein for SS (cleavage). Values with parentheses are nanomoles of sucrose synthesized min)1 mg)1 FW for SPS; and nanomoles of sucrose hydrolysed min)1 mg)1 FW for SS (cleavage). Values are mean S.D. of three replicates. Dierences signicant in comparison with non-primed at *P < 0.01 or **P < 0.05 (Students t-test).
85

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Kaur et al.

comparing activities of SS and acid invertase in podwall and seeds, activity of acid invertase was high in podwall whereas activity of SS was more in developing seeds. These data showed the importance of SS in seed lling. Higher activity of SS at 140 DAS in seeds of primed crop may facilitate seed lling. Acid invertase was reported to be the dominant sucrolytic enzyme during early pod elongation phases, whereas SS became dominant during seed dry matter accumulation in snap bean pods (Sung et al. 1989). On priming, an increase in SS activity of seeds could result in increased seed growth and seed dry weight in comparison with seeds of non-primed plants. It is suggested that priming regulates the sink strength of developing seeds by increasing the activities of SS and invertase. A major factor establishing the sink strength is the utilization of imported assimilates in the sink tissues. As sucrose is the main transported form of assimilates in plants, the immediate metabolism of imported sucrose is regarded as key regulatory step in sink strength establishment (Weber et al. 1995, Zrenner et al. 1995). At 110 DAS, the activity of SPS in primed seeds was observed to be low in comparison with nonprimed plants. During owering and pod set, the pod and seed number responds to changes in photosynthesis (Hardman and Brun 1971, Schou et al. 1978, Egli and Zhen-Wen 1991, Board and Tan 1995). Increase in number of pods and seeds has been reported to be due to increased photosynthate supply to the plant and hence higher sucrose concentration (Egli and Bruening 2001). Similar changes could have occurred in plants on priming because priming also resulted in increased number of pods and seeds. The higher concentration of sucrose (not determined), in the seeds at this stage, because of increased supply of assimilates could have limited the activity of SPS. However, at 140 DAS, a stage of rapid seed development, the activity of SPS was higher in the primed seeds. The higher requirement of assimilate at this stage might have been met by more ecient conversion of hexoses into sucrose by increased SPS activity and the higher sink strength by the increased SS activity. The results of this study indicate that increased chickpea yield on priming might be due to modulation of enzymes of sucrose metabolism in actively growing plant parts, the podwall and the seeds during development. In addition, priming of seeds could make the plant more resistant to various abiotic and biotic stresses resulting possibly in better growth and yield (Conrath et al. 2002).

Acknowledgements
The senior author is thankful to the Council of Scientic and Industrial Research (New Delhi, India) for the award of Senior Research Associateship under Scientists pool scheme.

References
Andrade, F. H., and M. A. Ferreiro, 1996: Reproductive growth of maize, sunower and soybean at dierent source levels during grain lling. Field Crops Res. 48, 155165. Board, J. E., and Q. Tan, 1995: Assimilatory capacity eects on soybean yield components and pod number. Crop Sci. 35, 846851. Chopra, J., N. Kaur, and A. K Gupta, 2000: Ontogenic changes in enzymes of carbon metabolism in relation to carbohydrate status in developing mungbean reproductive structures. Phytochemistry 53, 549553. Conrath, U., C. M. J. Pieterse, and B. Mauch-Mani, 2002: Priming in plant-pathogen interactions. Trends Plant Sci. 7, 210216. Daie, J., 1996: Metabolic adjustments, assimilate partitioning and alterations in source-sink relations in drought stressed plants. In: E. Zamski, and A. A. Schaer, eds. Photoassimilate Distribution in Plants and Crops, Source-Sink Relationships, pp. 407420. Marcel Dekker Inc., New York, NY. Egli, D. B., and W. P. Bruening, 2001: Source-sink relationships, seed sucrose levels and seed growth rates in soybean. Ann. Bot. 88, 235242. Egli, D. B., and Y. Zhen-Wen, 1991: Crop growth rate and seed number per unit area in soybean. Crop Sci. 31, 439442. Egli, D. B., R. D. Guy, L.W. Meckel, and J. E. Leggett, 1985: The eect of source sink alterations on soybean seed growth. Ann. Bot. 55, 395402. Foyer, C. H., and N. Galtier, 1996: Source-sink interaction and communication in leaves. In: E. Zamski, and A. A. Schaer, eds. Photoassimilate Distribution in Plants and Crops, Source-Sink Relationships, pp. 311340. Marcel Dekker Inc., New York, NY. Giord, R. M., and J. H. Thorne, 1985: Sucrose concentration at the apoplastic interface between seed coat and cotyledons of developing soybean seeds. Plant Physiol. 77, 863868. Hardman, L. L., and W. A. Brun, 1971: Eects of atmospheric carbon dioxide enrichment at dierent development stages on growth and yield components of soybeans. Crop Sci. 11, 886888. Harris, D., A. Joshi, P. A. Khan, P. Gothkar, and P. S. Sodhi, 1999: On farm seed priming in semi-arid agriculture: development and evaluation in maize, rice and chickpea in India using participatory methods. Exp. Agric. 35, 1529. Harris, D., B. S. Raghuwenshi, J. S. Gangwar, S. C. Singh, K. B. Joshi, A. Rashid, and P. A. Hollington,

Seed Priming and Crop Yield

87

2001: Participatory evaluation by farmers of on-farm seed priming in wheat in India, Nepal and Pakistan. Exp. Agric. 37, 403415. Kaur, S., A. K. Gupta, and N. Kaur, 1998a: Gibberellic acid and kinetin partially reverse the eect of water stress on germination and seedling growth in chickpea. Plant Growth Regul. 25, 2933. Kaur, S., A. K. Gupta, and N. Kaur, 1998b: Gibberellin A3 reverses the eect of salt stress in chickpea (Cicer arietinum L.) seedlings by enhancing the amylase activity and mobilization of starch in cotyledons. Plant Growth Regul. 26, 8590. Kaur, S., A. K. Gupta, and N. Kaur, 2000: Eect of GA3, kinetin and indole acetic acid on carbohydrate metabolism in chickpea seedlings germinating under water stress. Plant Growth Regul. 30, 6170. Kaur, S., A. K. Gupta, and N. Kaur, 2002a: Eect of osmo- and hydropriming of chickpea seeds on seedling growth and carbohydrate metabolism under water decit stress. Plant Growth Regul. 37, 1722. Kaur, S., A. K. Gupta, and N. Kaur, 2002b: Eect of osmo- and hydropriming of chickpea seeds on the performance of crop in the eld. Int. Chickpea Pigeonpea Newslett. 9, 1517. Kaur, S., A. K. Gupta, and N. Kaur, 2003: Priming of chickpea Seeds with water and mannitol can overcome the eect of salt stress on seedling growth. Int. Chickpea Pigeonpea Newslett. 10, 1820. Lowry, O. H., N. J. Rosebrough, A. L. Farr, and R. J. Randall, 1951: Protein measurement with folin phenol reagent. J. Biol. Chem. 193, 265268. Munier-Jolain, N. G., N. M. Munier-Jolain, R. Roche, B. Ney, and C. Duthion, 1998: Seed growth rate in grain legumes 1: eect of photoassimilate availability on seed growth rate. J. Exp. Bot. 49, 19631969. Musa, A. M., J. Johansen, J. Kumar, and D. Harris, 1999: Response of chickpea to seed priming in the

high Barind Tract of Bangladesh. Int. Chickpea Pigeonpea Newslett. 6, 2022. Musa, A. M., D. Harris, C. Johansen, and J. Kumar, 2001: Short duration chickpea to replace fallow after Aman Rice: the role of on-farm seed priming in the high Barind Tract of Bangladesh. Exp. Agric. 37, 509521. Nayyar, H., and D. P. Walia, 2004: Genotypic variation in wheat in response to water stress and abscisic acidinduced accumulation of osmolytes in developing grains. J. Agron. Crop Sci. 190, 3945. Schou, J. B., D. L. Jeers, and J. G. Streeter, 1978: Eects of reectors, blackboards or shades applied at dierent stages of plant development on yield of soybeans. Crop Sci. 18, 2934. Sturm, A., 1999: Invertases. Primary structures, functions and roles in plant development and sucrose partitioning. Plant Physiol. 121, 17. Sung, S. S., D. Xu, and C. C. Black, 1989: Identication of actively lling sinks. Plant Physiol. 89, 11171121. Weber, H., L. Borisjuk, U. Heim, P. Buchner, and U. Wobus, 1995: Seed coat associated invertases of Fava bean control both unloading and storage functions: cloning of cDNAs and cell type specic expression. Plant Cell 7, 18351846. Westgate, M. E., and D. T. Grant, 1989: Eects of water decits on seed development in soybean. I. Tissue water status. Plant Physiol. 91, 975979. Yang, J., J. Zhang, Z. Wang, Q. Zhu, and L. Liu, 2001: Water decit induced senescence and its relationship to the remobilization of pre-stored carbon in wheat during grain lling. Agron. J. 93, 196206. Zrenner, R., M. Salanoubat, L. Willmitzer, and U. Sonnewald, 1995: Evidence of the crucial role of sucrose synthase for sink strength using transgenic potato plants (Solanum tuberosum L.). Plant J. 7, 97107.