Effect of light and ontogenetic stage on sink strength in bean leaves

Light (about 3,000 foot-candles) neither increased nor decreased the sink strength of young, rapidly expanding leaves of Phaseolus vulgaris L. cv. Black Valentine, as measured by the comparative rates of import of ¹⁴C-labeled photosynthates by sink leaves in the light versus dark in short term experiments. Although irradiated sink leaves accumulated more ¹⁴C activity, the difference was fully accounted for by photosynthetic reabsorption of respiratory CO₂ derived from substrates translocated to the sink leaves.

Maximum sink strength was attained when the sink leaf reached 7 to 8 cm² in area (9 to 10% of its fully expanded size). Thereafter sink strength declined rapidly and asymptotically to a near zero value at about 45% final area. During this period, however, the rapid decline in translocation was offset by a rapid rise in the photosynthetic rate of the sink leaf, maintaining a near constant relative rate of dry weight increase until the sink leaf had expanded to about 17% of its final area. Although the increasing photosynthetic capacity was associated with a decreasing import capacity, suggesting that the rate of translocation to the sink leaf was controlled by the developing capacity of the sink leaf for photosynthesis, it was not possible to vary the total (true) translocation rate to the sink leaf by varying the photosynthetic rate of the sink leaf in short term light-dark experiments. Despite a high ratio of source to sink in these experiments, no evidence accrued that translocation into young bean leaves was ever sink-limited.

     

Comparative Phloem mobility of nickel in nonsenescent plants

⁶³Ni was applied to nonsenescent source leaves and found to be transported to sink tissues in pea (Pisum sativum L.) and geranium plants (Pelargonium zonale L.). The comparative mobilities (percent tracer transported out of source leaf ÷% ⁸⁶Rb transported) for ⁶³Ni in peas was 2.12 and in geranium 0.25. The value for the phloem mobile ⁸⁶Rb was 1.00. By contrast, the comparative mobility of ⁴⁵Ca, which is relatively immobile in the phloem, was low (0.05 in peas, 0.00 in geranium). Interruption of the phloem pathway between source and sink leaves by steam girdling almost completely inhibited ⁶³Ni accumulation in the sink leaves of both species. We conclude that Ni is transported from nonsenescent source leaves to sink tissues via the phloem of leguminous and nonleguminous plants.     

Effect of altered sink: source ratio on photosynthetic metabolism of source leaves

When seven crop species were grown under identical environmental conditions, decreased sink:source ratio led to a decreased photosynthetic rate within 1 to 3 days in Cucumis sativus L., Gossypium hirsutum L., and Raphanus sativus L., but not in Capsicum annuum L., Solanum melongena L., Phaseolus vulgaris L., or Ricinus communis L. The decrease was not associated with stomatal closure. In cotton and cucumber, sink removal led to an increase in starch and sugar content, in glucose 6-phosphate and fructose 6-phosphate pools, and in the proportion of ¹⁴C detected in sugar phosphates and UDPglucose following ¹⁴CO₂ supply. When mannose was supplied to leaf discs to sequester cytoplasmic inorganic phosphate, promotion of starch synthesis, and inhibition of CO₂ fixation, were observed in control discs, but not in discs from treated plants. Phosphate buffer reduced starch synthesis in the latter, but not the former discs. The findings suggest that sink removal led to a decreased ratio inorganic phosphate:phosphorylated compounds. In beans ¹⁴C in sugar phosphates increased following sink removal, but without sucrose accumulation, suggesting tighter feedback control of sugar level. Starch accumulated to higher levels than in the other plants, but CO₂ fixation rate was constant for several days.     

Phenotyping using semi-automated BIOLOG and conventional PCR for identification of Bacillus isolated from biofilm of sink drainage pipes

The presence of Bacillus in natural biofilms which develop in sink drainage pipes is not widely studied. Therefore, the main aim of this study was to isolate and identify Bacillus spp. using the BIOLOG GEN III system as a phenotypic fingerprint and polymerase chain reaction (PCR). A total of 61 biofilms samples were collected from sink drainage pipes in a kitchen and bathroom of different households in Helwan area and both laboratory and hospital collected from National Research Centre (NRC). Bacillus was isolated from the biofilms using HiCrome Bacillus Agar followed by isolates identification by both BIOLOG to the species level and PCR using genus specific primers to the genera level. Bacillus was detected in all tested biofilm samples (61 samples). The highest counts were observed in hospital sink drainage pipes (10⁵?CFU/10?cm²) while; the lowest counts were observed in both bathroom and laboratory sink drainage pipes (10²?CFU/10 cm^?2). In total, 61% Bacillus isolates were identified by BIOLOG while, 67% isolates were confirmed by PCR. The diversity of Bacillus among species level using BIOLOG were 34% B. cereus, 23% B. subtilis ss subtilis, 17% B. thuringiensis, 16% B. licheniformis and 13% B. amyloliquefaciens. It can be concluded that; PCR is more sensitive than BIOLOG for identification of Bacillus. However, BIOLOG can identify Bacillus at species level and test 94 carbon and chemical sources on a microplate in one shot. Thus, the combination between phenotyping by BIOLOG and molecular approaches such as PCR for identification of bacterial isolates is recommended.     

Phloem Unloading in Developing Leaves of Sugar Beet : I. Evidence for Pathway through the Symplast

Physiological and transport data are presented in support of a symplastic pathway of phloem unloading in importing leaves of Beta vulgaris L. (`Klein E multigerm'). The sulfhydryl reagent p-chloromercuribenzene sulfonic acid (PCMBS) at concentration of 10 millimolar inhibited uptake of exogenous [¹⁴C]sucrose by sink leaf tissue over sucrose concentrations of 0.1 to 5.0 millimolar. Inhibited uptake was 24% of controls. The same PCMBS treatment did not affect import of ¹⁴C-label into sink leaves during steady state labeling of a source leaf with ¹⁴CO₂. Lack of inhibition of import implies that sucrose did not pass through the free space during unloading. A passively transported xenobiotic sugar, l-[¹⁴C]glucose, imported by a sink leaf through the phloem, was evenly distributed throughout the leaf as seen by whole-leaf autoradiography. In contrast, l-[¹⁴C]glucose supplied to the apoplast through the cut petiole or into a vein of a sink leaf collected mainly in the vicinity of the major veins with little entering the mesophyll. These patterns are best explained by transport through the symplast from phloem to mesophyll.     

Reserve carbohydrate in maize stem : [C]glucose and [C]sucrose uptake characteristics

Maize (Zea mays L.) stem is thought to function alternately as a net importing and net exporting organ during ontogeny, depending on whole plant photosynthetic source and sink status. The [¹⁴C]sucrose and [¹⁴C]glucose uptake capacity of stem tissues was investigated to increase our understanding of the transport factors which may influence sink status.     

Photoperiodic and genetic control of carbon partitioning in peas and its relationship to apical senescence

Apical senescence but not flower initiation is delayed by short days (SD) compared to long days (LD) in pea plants (Pisum sativum L.) of genotype E Sn Hr. We recently reported that delay of senescence correlated with slower reproductive development, suggesting that fruits are weaker sinks for assimilates under delayed senescence conditions. Thus, we have examined assimilate partitioning in peas to determine if genotype and photoperiod regulate relative sink strength. Assimilate diversion by developing fruit has been implicated in senescence induction. A greater percentage of leaf-exported ¹⁴C was transported to fruits and a smaller percentage to the apical bud of G2 peas (genotype E Sn Hr) in LD than in SD. Relatively more of the ¹⁴C delivered to the apical bud of G2 peas was transported to flower buds than to young leaves in LD as compared to SD. There was no striking photoperiodic difference in carbon partitioning in genetic lines without the Sn Hr allele combination. The Sn Hr allele combination and photoperiod may regulate the relative strength of reproductive and vegetative sinks. Photoperiodic differences in sink strength early in reproduction suggest that these genes regulate sink strength by affecting the physiology of the whole plant. High vegetative sink strength in SD may maintain assimilate supply to the apical bud, delaying senescence.     

Response of Bean (Vicia faba L.) Plants to Low Sink Demand by Measuring the Gas Exchange Rates and Chlorophyll a Fluorescence Kinetics

Background

The decline of photosynthesis in plants under low sink demand is well known. Previous studies focused on the relationship between stomatal conductance (g _s) and net photosynthetic rate (P _n). These studies investigated the effect of changes in Photosystem II (PSII) function on the P _n decline under low sink demand. However, little is known about its effects on different limiting steps of electron transport chain in PSII under this condition.

Methodology/Principal Finding

Two-month-old bean plants were processed by removing pods and flowers (low sink demand). On the 1^st day after low sink demand treatment, a decline of P _n was accompanied by a decrease in g _s and internal-to-ambient CO₂ concentration ratio (C _i/C _a). From the 3^rd to 9^th day, P _n and g _s declined continuously while C _i/C _a ratio remained stable in the treatment. Moreover, these values were lower than that of control. W_k (a parameter reflecting the damage to oxygen evolving complex of the donor side of PSII) values in the treatment were significantly higher than their corresponding control values. However, RC_QA (a parameter reflecting the number of active RCs per excited cross-section of PSII) values in the treatment were significantly lower than control from the 5^th day. From the 11^th to 21^st day, P _n and g _s of the treatment continued to decline and were lower than control. This was accompanied by a decrease of RC_QA, and an increase of W_k. Furthermore, the quantum yield parameters φ _Po, φ _Eo and ψ _Eo in the treatment were lower than in control; however, C _i/C _a values in the treatment gradually increased and were significantly higher than control on the 21^st day.

Conclusions

Stomatal limitation during the early stage, whereas a combination of stomatal and non-stomatal limitation during the middle stage might be responsible for the reduction of P _n under low sink demand. Non-stomatal limitation during the late stages after the removal of the sink of roots and pods may also cause P _n reduction. The non-stomatal limitation was associated with the inhibition of PSII electron transport chain. Our data suggests that the donor side of PSII was the most sensitive to low sink demand followed by the reaction center of PSII. The acceptor side of PSII may be the least sensitive.     

Incorporation of C-photosynthate into major chemical fractions of source and sink leaves of cottonwood

The incorporation and distribution of photosynthetically fixed ¹⁴CO₂ was followed for 48 hours in a recently matured source leaf (LPI 7) and in young expanding source and sink leaves (LPI 4) of cottonwood (Populus deltoides Bartr.). The major chemical constituents of leaf laminae and petioles were separated by sequential solvent extractions and enzyme hydrolyses. Two hours after labeling, about 80% of the ¹⁴C was found in water-alcohol-soluble constituents in the mature source lamina as compared to about 45% in those of the young expanding leaf. In both mature and expanding source leaves the water-alcohol-soluble constituents decreased while the CHCl₃-soluble and -insoluble compounds increased with time. After 48 hours, 7 and 37% of the total ¹⁴C was recovered from structural carbohydrates and from protein + CHCl₃-soluble fractions, respectively, in the mature source leaf; and 4 and 65%, respectively, in the young source leaf. When the distribution of ¹⁴C among major chemical fractions was calculated on per cent dpm/mg basis, the data showed that a young sink leaf incorporated over twice as much ¹⁴C into structural carbohydrates as a young source leaf (11% versus 4%). However, when calculated on an absolute dpm/mg basis, activity in this fraction of the young source leaf exceeded that in the sink leaf by a ratio of about 11:1 (9528 versus 845 dpm/mg). Thus, most of the material for synthesis of structural carbohydrates was derived from in situ photosynthate.     

Evidence for thiol-induced enhanced in situ translocation of 14C-sucrose from source to sink in Brassica juncea

The translocation of ¹⁴C-sucrose to the different parts of the mustard (Brassica juncea) crop has been evaluated in the context of understanding the source to sink relationship in the thiol-induced enhanced crop yield. The foliar application of thiols like TU, TGA and DTT to the plant gave maximum sucrose phosphate synthase activity, which was found to have direct correlation with the movement of sucrose. The distribution pattern of ¹⁴C-sucrose follows the path from internode and node to pod via leaf. The translocation of ¹⁴C-sucrose was found to be a light dependent process. Among the nucleotides ATP and GTP, only ATP was able to promote the translocation and GTP was ineffective. In this unique in situ tracer experiment using ¹⁴C-sucrose, we could establish that thiols are able to enhance the translocation of sucrose from source to sink.     

Evidence for thiol-induced enhanced in situ translocation of 14C-sucrose from source to sink in Brassica juncea

The translocation of ¹⁴C-sucrose to the different parts of the mustard (Brassica juncea) crop has been evaluated in the context of understanding the source to sink relationship in the thiol-induced enhanced crop yield. The foliar application of thiols like TU, TGA and DTT to the plant gave maximum sucrose phosphate synthase activity, which was found to have direct correlation with the movement of sucrose. The distribution pattern of ¹⁴C-sucrose follows the path from internode and node to pod via leaf. The translocation of ¹⁴C-sucrose was found to be a light dependent process. Among the nucleotides ATP and GTP, only ATP was able to promote the translocation and GTP was ineffective. In this unique in situ tracer experiment using ¹⁴C-sucrose, we could establish that thiols are able to enhance the translocation of sucrose from source to sink.     

Assessment of the Soil Organic Carbon Sink in a Project for the Conversion of Farmland to Forestland: A Case Study in Zichang County,Shaanxi, China

The conversion of farmland to forestland not only changes the ecological environment but also enriches the soil with organic matter and affects the global carbon cycle. This paper reviews the influence of land use changes on the soil organic carbon sink to determine whether the Chinese “Grain-for-Green” (conversion of farmland to forestland) project increased the rate of SOC content during its implementation between 1999 and 2010 in the hilly and gully areas of the Loess Plateau in north-central China. The carbon sink was quantified, and the effects of the main species were assessed. The carbon sink increased from 2.26×10⁶ kg in 1999 to 8.32×10⁶ kg in 2010 with the sustainable growth of the converted areas. The black locust (Robinia pseudoacacia L.) and alfalfa (Medicago sativa L.) soil increased SOC content in the top soil (0–100 cm) in the initial 7-yr period, while the sequestration occurred later (>7 yr) in the 100–120 cm layer after the “Grain-for-Green” project was implemented. The carbon sink function measured for the afforested land provides evidence that the Grain-for-Green project has successfully excavated the carbon sink potential of the Shaanxi province and served as an important milestone for establishing an effective organic carbon management program.     

Source/Sink relations of abscising and nonabscising soybean flowers

The partitioning of recently fixed ¹⁴C to setting and abscising flowers within the axillary raceme of `Clark' isoline E1t soybeans (Glycine max L. Merr) was examined as a function of time after anthesis of individual flowers. In such racemes, the first four flowers showed a 17% abscission while the next four flowers showed 47% abscission.

Source/sink relations of flowers I-IV (normally setting) were compared to those of flowers V-VIII (normally abscising) by pulse labeling source leaves with ¹⁴CO₂ and determining the radioactivity of individual flowers after a 4-hour chase period. The relative specific activity (RSA;% disintegrations per minute per% dry weight), sink strength (% disintegrations per minute), and its components, sink size (milligrams dry weight) and sink intensity (% disintegrations per milligram dry weight) were then calculated as a function of days after anthesis.

Sink intensity (i.e. the competitive ability to accumulate photoassimilate per unit mass) was very high prior to anthesis of both setting and abscising flowers. Sink intensity then became very low for the first 3 days following anthesis after which it recovered in normally setting flowers, but failed to recover in normally abscising flowers. It is concluded that soybean reproductive abscission is determined at or very near the day of anthesis.

     

Characterization of the light reaction in promoting the mobilizing ability of rose shoot tips

Mixed fluorescent and incandescent light increased growth and sink strength of the uppermost young shoot of rose plants (Rosa hybrida cv. Marimba) in comparison to pure fluorescent light. This was manifested by increased apical dominance. Monochromatic low-energy red light, given by means of optic fibers for 24 hours to shoot tips that had been previously darkened for 5 days, increased the transport of ¹⁴C-labeled assimilates to the intact tips and the uptake of [¹⁴C]sucrose by detached tips. Far-red had little or no effect, and blue was not effective at all in these reactions. Red light given directly to detached shoot tips, in vitro, increased the uptake of [¹⁴C]sucrose by the isolated tips. Adding far-red to the red greatly promoted the uptake, whereas blue and blue plus far-red were not active. The main character of the light reaction promoting sink activity in the shoot is that it is perceived by the shoot tip itself. It is operated by red light; far-red promotes the red effect but has little or no effect when alone. Light apparently promotes shoot sink activity by increasing the unloading process.     

Decorrelating source and sink determinism of nitrogen remobilization during grain filling in wheat

Background and Aims

Nitrogen (N) remobilization is the major source of N for grain filling in wheat, the other being N uptake after anthesis (N_up); however, variations in remobilization efficiency are not fully understood. It is hard to tell whether the source or the sink effects predominate, because N in the culm at anthesis (N_ant) correlates strongly with both N remobilization (N_rem) and grain number (G_n), respectively the main source and the main sink.

Methods

A pot experiment was thus designed to assess the relative contributions of the source and sink to N_rem regulation. Using two cultivars of winter wheat (Triticum aestivum, ‘Apache’ and ‘Autan’), three pre-anthesis and two post-anthesis N fertilization levels were applied in order to vary the N sources, while ear trimming at anthesis reduced sink size.

Key Results

Unlike results observed at a scale of m², the equation binding N_ant to N_rem exhibited a negative intercept, challenging the concept of nitrogen remobilization efficiency. Before ear trimming, G_n fitted well to N_ant, with a slope dependent on genotype. To obtain a sink variable that was less correlated with N_ant, the difference δG_n was calculated between actual grain number and that which could be predicted from culm N before trimming. A multiple regression then predicted N_rem (r² = 0·95) from N_ant, N_up and δG_n, with fitting unbiased by fertilization treatment, trimming or genotype.

Conclusions

In untrimmed culms, δG_n had a negligible effect, so that N_rem could be fitted to N_ant and N_up only: grain N filling appeared to be determined by sources only (N_ant and N_up), not by sink, and the reduction of N_rem by N_up was quantified. In these ‘normal’ cases, the regulation of N_rem should thus be located within the N sources themselves. In contrast, ear-trimming needs to be considered with caution as it introduced a sink limitation on N_rem; moreover one with an important genotype effect.Key words: Triticum aestivum, winter wheat, source/sink, grain filling, nitrogen uptake, grain number, nitrogen harvest index, nitrogen remobilization efficiency, genotype × environment     

滇西北高原纳帕海湿地湖滨带优势植物生物量及其凋落物分解

选取滇西北高原湿地纳帕海湖滨带优势植物茭草(Zizania caducifolia)、水葱(Scirpus tabernaemontani)和刘氏荸荠(Heleocharis liouana),研究其生物量及其凋落物分解特征,结果表明:水葱、茭草、刘氏荸荠为纳帕海湿地湖滨带单优植物群落,均具有较高的地上生物量,不同植物群落地上生物量不同,其中,茭草地上生物量(853.6±58.2)g·m-2·a-1显著高于水葱(730.7±7.8)g·m-2·a-1与刘氏荸荠(338.9±32.6) g·m-2·a-1的地上生物量.3种植物群落凋落物分解速率不同、并随月平均气温升高均呈增加的趋势,其中,刘氏荸荠分解速率k值最大(0.067±0.0026)、茭草(0.062±0.0072)其次、水葱最小(0.039±0.0062).凋落物经过1年的分解,水葱、茭草和刘氏荸荠凋落物存留率分别为(62.0±8.8)％、(47.5±9.0)％和(44.5±7.9)％.综合3种湖滨带植物地上生物量与凋落物年分解,水葱地上生物量年存留量(453.1±4.9)g·m-2·a-1显著高于茭草(405.4±27.7)g·m-2·a-1和刘氏荸荠(150.9±14.5) g·m-2·a-1.研究进一步表明滇西北高原湿地湖滨带植物具有极高的生物量存留率,成为该类型湿地生态系统碳汇功能的基础,其碳汇过程及其贡献率需要进一步深入研究.     

Effect of the replacement of a native savanna by an African Brachiaria decumbens pasture on the CO2 exchange in the Orinoco lowlands,Venezuela

In the Orinoco lowlands, savannas have been often replaced by pastures composed of the C₄ grass, Brachiaria decumbens Stapf. We addressed following questions: (1) How does the replacement of the native vegetation affect CO₂ exchange on seasonal and annual scales? (2) How do biophysical constraints change when the landscape is transformed? To assess how these changes affect carbon exchange, we determined simultaneously the CO₂ fluxes by eddy covariance, and the soil CO₂ efflux by a chamber-based system in B. decumbens and herbaceous savanna stands. Measurements covered a one-year period from the beginning of the dry season (November 2008) to the end of the wet season (November 2009). During the wet season, the net ecosystem CO₂ exchange reached maximum values of 23 and 10 μmol(CO₂) m^?2 s^?1 in the B. decumbens field and in the herbaceous savanna stand, respectively. The soil CO₂ efflux for both stands followed a temperature variation during the dry and wet seasons, when the soil water content (SWC) increased above 0.087 m³ m^?3 in the latter case. Bursts of CO₂ emissions were evident when the dry soil experienced rehydration. The carbon source/sink dynamics over the two canopies differed markedly. Annual measurements of the net ecosystem production indicated that the B. decumbens field constituted a strong carbon sink of 216 g(C) m^?2 y^?1. By contrast, the herbaceous savanna stand was found to be only a weak sink [36 g(C) m^?2 y^?1]. About 53% of the gross primary production was lost as the ecosystem respiration. Carbon uptake was limited by SWC in the herbaceous savanna stand as evident from the pattern of water-use efficiency (WUE). At the B. decumbens stand, WUE was relatively insensitive to SWC. Although these results were specific to the studied site, the effect of land use changes and the physiological response of the studied stands might be applicable to other savannas.     

Carbon allocation during fruiting in Rubus chamaemorus

Background and Aims

Rubus chamaemorus (cloudberry) is a herbaceous clonal peatland plant that produces an extensive underground rhizome system with distant ramets. Most of these ramets are non-floral. The main objectives of this study were to determine: (a) if plant growth was source limited in cloudberry; (b) if the non-floral ramets translocated carbon (C) to the fruit; and (c) if there was competition between fruit, leaves and rhizomes for C during fruit development.

Methods

Floral and non-floral ramet activities were monitored during the period of flower and fruit development using three approaches: gas exchange measurements, ¹⁴CO₂ labelling and dry mass accumulation in the different organs. Source and sink activity were manipulated by eliminating leaves or flowers or by reducing rhizome length.

Key Results

Photosynthetic rates were lower in floral than in deflowered ramets. Autoradiographs and ¹⁴C labelling data clearly indicated that fruit is a very strong sink for the floral ramet, whereas non-floral ramets translocated C toward the rhizome but not toward floral ramets. Nevertheless, rhizomes received some C from the floral ramet throughout the fruiting period. Ramets with shorter rhizomes produced smaller leaves and smaller fruits, and defoliated ramets produced very small fruits.

Conclusions

Plant growth appears to be source-limited in cloudberry since a reduction in sink strength did not induce a reduction in photosynthetic activity. Non-floral ramets did not participate directly to fruit development. Developing leaves appear to compete with the developing fruit but the intensity of this competition could vary with the specific timing of the two organs. The rhizome appears to act both as a source but also potentially as a sink during fruit development. Further studies are needed to characterize better the complex role played by the rhizome in fruit C nutrition.Key words: Allocation pattern, ¹⁴C labelling, carbon translocation, carbon reserves, cloudberry, defoliation, fruit production, gas exchange, Rubus chamaemorus, source–sink relationship, flowering     

Sink to source translocation in soybean

The possibility that phloem loading may occur in the reproductive sink tissues of soybeans (Glycine max Merr. cv Chippewa 64) was examined. When [¹⁴C]sucrose was applied to seed coat tissues from which the developing embryo had been surgically removed, 0.1% to 0.5% of the radioactivity was translocated to the vegetative plant parts. This sink to source translocation was largely unaffected by destroying a band of phloem with steam treatment on the stem above and below the labeled pod. The same steam treatment, however, completely abolished translocation of [¹⁴C]sucrose between mature leaves and developing fruits. These results indicate that the movement of nutrients from developing seed coats to the vegetative plant parts occur in the xylem and that phloem loading does not occur in this sink tissue.     

Development of a functional assay for Ca2+ release activity of IP3R and expression of an IP3R gene fragment in the baculovirus-insect cell system

Receptor-stimulated phosphoinositide (PI) hydrolysis is an important and ubiquitous mechanism of intracellular signaling. Inositol 1,4,5-trisphosphate (IP₃), generated by phosphoinositide (PI) hydrolysis, binds to and gates an intracellular Ca²⁺ channel, the IP₃ receptor (IP₃R), which is therefore a central component of this signaling cascade. Here we describe the development of a baculovirus (BV)/Sf(S. frugiperda) cell system that can be used to look at IP₃R function. Agonist-evoked changes in intracellular Ca²⁺ levels [Ca²⁺]_i were measured (using Fura2) in Sf cells expressing the gene encoding the muscarinic acetylcholine receptor (vmlAchR). Furthermore, we have constructed a recombinant BV (vlP₃R), with the core of the IP₃R ligand-binding domain from the Drosophila IP₃R, under the polyhedrin promoter. The recombinant protein from such a virus was expected to act as a large ligand sink for IP3, generated by stimulation of vmlAchR. Cells coinfected with recombinant BV carrying the potential dominant-negative vIP₃R construct and vmlAchR have been used to assay the modulation of IP₃R-mediated Ca²⁺ release, by the ligand sink.