百合鳞茎发育过程中淀粉合成相关酶基因的克隆及表达分析

该研究通过同源克隆技术克隆腺苷二磷酸葡萄糖焦磷酸化酶(AGPase)、颗粒结合淀粉合酶(GBSS)和可溶性淀粉合酶(SSS) 3类百合淀粉合成关键酶基因,分析这三类淀粉合成关键酶基因的表达变化,测定百合鳞茎膨大发育中淀粉含量变化。结果表明:(1) AGPase具有GlgC家族蛋白PLN02241蛋白结构特征及cl11394家族蛋白ADP_Glucose_PP与NTP_transferase结构域,获登录号KP751443; GBSS与SSS具有cl10013家族蛋白Glyco_transf_5,GT1_Glycogen_synthase_DULL1_like结构域,获登录号分别为KP751444、KP751445。(2)百合鳞茎形成与膨大发育过程中,淀粉含量呈现递增趋势,鳞茎盘开始分化茎杆时其淀粉含量最高,达到44.52%。鳞茎与叶片部位的三个淀粉合成相关酶基因表达量均逐渐增加;在鳞茎膨大后茎杆分化阶段,三个淀粉合成相关酶基因表达量达到最高,AGPase、GBSS、SSS在鳞片中的表达量分别为10.79,6.92和5.12,叶片中的表达量分别为6.79,5.22和4.41,鳞片中的表达量大幅度高于叶片;淀粉合成相关酶基因的表达量变化与淀粉含量、鳞茎的膨大发育成正相关。这为鳞茎的繁殖生产提供了可通过调节淀粉合成关键酶基因表达促进百合鳞茎膨大发育的思路。     

多效唑对不同品质类型花生产量、品质及相关酶活性的影响

选用高蛋白品种KB008（KB008）、高脂肪品种花17(H17)和高油酸/亚油酸（O/L）品种农大818（818）,在大田栽培条件下,研究了盛花后期叶面喷施多效唑(PBZ)对不同品质类型花生产量、品质及相关碳、氮代谢酶活性的影响.结果表明：喷施PBZ显著增加了3种品质类型花生荚果产量,原因是增加了单株结果数,降低了千克果数而提高了双仁果率.喷施PBZ不同程度地提高了3种类型花生籽仁脂肪和可溶性糖含量,降低了蛋白质含量,显著增加了高脂肪品种H17的O/L值.PBZ使高O/L值品种818的脂肪含量增加显著,同时其蛋白质含量显著降低,而对其他两品种的蛋白质和脂肪含量影响较小.喷施PBZ均降低了3种类型花生结荚期叶片硝酸还原酶（NR）活性及结荚期和饱果期叶片谷氨酰胺合成酶和谷氨酸脱氢酶活性,818的3种酶活性降低幅度最大,KB008和H17的酶活性降幅较小;喷施PBZ均降低了3种类型花生结荚期和饱果期叶片谷草转氨酶和谷丙转氨酶活性.说明氮代谢酶活性的降低是喷施PBZ降低3种类型花生籽仁蛋白质含量的主要原因.喷施PBZ均提高了3品种结荚期和饱果期叶片蔗糖合成酶和磷酸蔗糖合成酶活性,其中显著提高了818的2种酶活性,而对KB008和H17的活性提高不显著;喷施PBZ提高了3品种结荚期和饱果期的磷酸烯醇式丙酮酸羧化酶和1,5-二磷酸核酮糖羧化酶活性,其中对818在结荚期的活性提高最显著,对H17活性提高较小.碳代谢酶活性的增强是喷施PBZ提高花生籽仁脂肪含量的生理基础.     

Effect of Putrescine and Paclobutrazol on Growth, Physiochemical Parameters, and Nutrient Acquisition of Salt-sensitive Citrus Rootstock Karna khatta (Citrus karna Raf.) under NaCl Stress

Salinity is a serious problem in arid and semiarid areas and citrus trees are classified as salt-sensitive. Because putrescine (Put) and paclobutrazol (PBZ) are known to act as plant protectants under environmental stresses, we examined the effect of Put and PBZ on the physiochemical parameters of the salt-susceptible citrus rootstock Karna khatta under NaCl stress. PBZ was applied at 0, 250, and 500 mg L⁻¹ as a soil drench 1 week prior to salinization. A computed amount of NaCl salt to develop soil salinity of 3 dS m⁻¹ (3 g NaCl kg⁻¹ soil) and foliar spray of Put at 0 or 50 mg L⁻¹ were applied. The electrical conductivity (EC) of the garden soil (0.35 dS m⁻¹) was used as control. Application of PBZ and/or Put reduced the membrane injury index and increased relative water content, photosynthetic rate, and pigments content under saline conditions compared to what occurred in plants exposed to NaCl in the absence of PBZ or Put. Application of PBZ or Put alone or in combination also improved the activities of SOD and peroxidase and proline content under saline conditions. Application of PBZ and/or Put also increased K⁺ and reduced Na⁺ and Cl⁻ concentrations in leaf tissues. It is proposed that PBZ and/or Put could improve the tolerance of salt-susceptible Karna khatta by regulating absorption and accumulation of ions and improving antioxidant enzyme activities.     

Temperature induced changes in the starch components and biosynthetic enzymes of two rice varieties

The effects of temperature on starch and amylose accumulation, fine structure of amylopectin and activities of some enzymes related to starch synthesis in developing rice endosperms was examined. Two early indica rice varieties were used, differing in amylose concentration (AC, %), namely Jia 935 (low AC) and Jia 353 (high AC). The results showed that the effects of high temperature on AC and amylopectin fine structure were variety-dependent. High temperature caused a reduction in amylose concentration and an increase in the short chain (CL<22) proportion of amylopectin for Jia 935; while opposite was true for Jia 353. High temperature also reduced and increased the activity of granule-bound starch synthase (GBSS) in Jia 935 and in Jia 353, respectively. This suggests that a change in the ratio of amylose/starch due to temperature was attributable to a change in GBSS activity. Moreover, obvious differences between the two rice varieties were detected in the activities of sucrose synthase (SuSy), ADP-glucose pyrophosphorylase (ADPG-Ppase), soluble starch synthase (SSS), starch branching enzyme (SBE), starch de-branching enzyme (SDBE) and starch phosphorylase (SPase) to high temperature. Accumulation rate of amylose was significantly and positively correlated with GBSS for Jia 935, but not for Jia 353. Amylose accumulation was also significantly and positively correlated with the activities of SDBE, SBE, ADPG-Ppase and SuSy for both varieties. The results suggest that the ratio of amylose to starch in rice endosperm is not only related to GBSS, but also affected by the activities of SDBE, SBE, ADPG-Ppase and SuSy.     

The effect of water stress on the activities of key regulatory enzymes of the sucrose to starch pathway in wheat

Developmental changes in the starch and sucrose content of grains andthe activities of enzymes of starch synthesis in wheat were studied under waterstress conditions. Water stress caused a marked reduction in the sucrose andstarch content of the grains. Sucrose synthase (SS) and UDP-glucosepyrophosphorylase (UDP-Gppase), showed higher catalytic activity and moreresistance to water stress compared with amyloplastic enzymes. ADP-glucosepyrophosphorylase (ADP-Gppase) activity was reduced to a low level under bothin situ and osmotic stress conditions in which grainsfailed to accumulate dry matter in vivo. Granule-boundstarch synthase (GBSS) also responded rapidly to in situwater stress treatments as did ADP-Gppase. Reduction in GBSS activity at thetime of growth cessation in situ was less than that ofADP-Gppase and the enzyme did not respond to severe osmotic stress. Solublestarch synthase (SSS) was the enzyme most sensitive to water stress in that itresponded earlier, and to a greater extent, than the other enzymes. However,under severe dehydration conditions, leading to cessation of growth, thedeclinein SSS activity was less than that for ADP-Gppase. SSS showed the lowestin vitro activity followed by GBSS. These results suggestthat SSS is the site of response to water stress by which the rate of graingrowth can be affected, whereas growth cessation is due mainly to theinactivation of ADP-Gppase.     

硫营养对小麦籽粒淀粉合成及相关酶活性的影响

在田间条件下,研究了施硫对小麦籽粒淀粉合成及相关酶活性的影响.结果表明:在0～20 cm土层土壤有效硫含量为5.84 mg/kg的地块上施硫不仅提高了小麦籽粒中蔗糖的含量,而且催化蔗糖降解代谢的蔗糖合成酶(SS)活性提高,利于籽粒蔗糖的降解.施硫显著提高了灌浆期间籽粒可溶性淀粉合成酶(SSS)活性,并使腺苷二磷酸葡萄糖焦磷酸化酶(ADPGPPase)和束缚态淀粉合成酶(GBSS)活性在灌浆中、后期维持在较高水平,对直链和支链淀粉的合成都起促进作用, 使总淀粉积累增加,千粒重提高,产量增加.     

Regulation of Starch and Protein Synthesis in Wheat Grains by Feeding Sucrose and Glutamine to Detached Ears Cultured in vitro

Three experiments of in vitro ear culture were conducted to evaluate how the substrates of C (carbon) and N (nitrogen) supply in liquid medium regulate the grain growth and synthesis of protein and starch in two winter wheat cultivars. Increasing glutamine supply with constant sucrose concentration increased the contents of total protein and protein components of albumin and globulin in grain, and the activity of glutamate pyruvate transaminase (GPT) across most treatments, while markedly reducing the contents of total starch and components of amylose and amylopectin as well as the activities of soluble starch synthase (SSS) and granule bounded starch synthase (GBSS). The opposite patterns were observed in the experiment of increasing sucrose supply at constant glutamine concentration. When simultaneously increasing sucrose and glutamine supply at constant ratio, the contents of total protein, albumin and globulin in grain were slowly enhanced, whereas the contents of total starch, amylose and amylopectin and the activities of SSS, GBSS and GPT increased only to a certain extent and then decreased. Negative correlations were found between the contents of protein or protein components in grains and the relative ratio of sucrose to glutamine concentrations in the culture medium, while positive correlations were seen between the contents of total starch or starch components and the ratio of sucrose to glutamine. These results implied that the composition of protein and starch in wheat grain could be readily manipulated by varying the concentrations of sucrose and glutamine and their ratio in the culture medium.     

氮素水平对冬小麦籽粒灌浆过程中淀粉合成关键酶活性的影响

小麦籽粒灌浆过程中,淀粉合成关键酶腺苷二磷酸葡萄糖焦磷酸化酶(ADPG-PPase)、可溶性淀粉合成酶(SSS)、淀粉分支酶(SBE)和束缚态淀粉合成酶(GBSS)均随着灌浆进程呈单峰曲线变化,峰值出现在花后25d;不同氮肥施用量对灌浆前期酶活性的影响较小,而在花后20d之后影响较大;随着氮肥施用量的增加,4种酶活性均呈增加趋势,但氮肥过量时酶活性下降,表明适当增加施氮量有利于淀粉合成关键酶活性的提高。     

Partial substitution of organic nitrogen with synthetic nitrogen enhances rice yield,grain starch metabolism and related genes expression under the dual cropping system

Improving grain filling in the presernt farming systems is crucial where grain filling is a concern due to the extreme use of chemical fertilizers (CF). A field experiment was conducted at the experimental station of Guangxi University, China in 2019 to test the hypothesis that cattle manure (CM) and poultry manure (PM) combined with CF could improve rice grain filling rate, yield, biochemical and qualitative attributes. A total of six treatments, i.e., no fertilizer (T₁), 100% CF (T₂), 60% CM + 40% CF (T₃), 30% CM + 70% CF (T₄), 60% PM + 40% CF (T₅), and 30% PM + 70% CF (T₆) were used in this study. Results showed that the combined treatment T₆increased starch metabolizing enzymes activity (SMEs), such as ADP-glucose phosphorylase (ADGPase) by 8 and 12%, soluble starch synthase (SSS) by 7 and 10%, granule bound starch synthesis (GBSS) by 7 and 9%, and starch branching enzyme (SBE) by 14 and 21% in the early and late seasons, respectively, compared with T₂. Similarly, higher rice grain yield, grain filling rate, starch, and amylose content were also recorded in combined treatments. In terms of seasons, higher activity of SMEs , grain starch, and amylose content was noted in the late-season compared to the early season. The increment in these traits was mainly attributed to a lower temperature in the late season during the grain filling period. Furthermore, our results suggested that an increment in starch accumulation and grain filling rate were mainly associated with the enhanced sink capacity by regulating key enzyme activities involved in Suc-to-starch conversion. In-addition, RT-qPCR analysis showed higher expression levels of AGPS2b, SSS1, GBSS1, and GBSE11b genes, which resultantly increased the activities of SMEs during the grain filling period under combined treatments. Linear regression analysis revealed that the activity of ADGPase, SSS, GBSS, and SBE were highly positively correlated with starch and amylose accumulation. Thus, we concluded that a combination of 30% N from PM or CM with 70% N from CF is a promising option in terms of improving rice grain yield and quality. Our study provides a sustainable fertilizer management strategy to enhance rice grain yield and quality at the lowest environmental cost.     

Inhibition of the expression of the gene for granule-bound starch synthase in potato by antisense constructs

Summary Granule-bound starch synthase [GBSS; EC 24.1.21] determines the presence of amylose in reserve starches. Potato plants were transformed to produce antisense RNA from a gene construct containing a full-length granule-bound starch synthase cDNA in reverse orientation, fused between the cauliflower mosaic virus 35S promoter and the nopaline synthase terminator. The construct was integrated into the potato genome by Agrobacterium rhizogenes-mediated transformation. Inhibition of GBSS activity in potato tuber starch was found to vary from 70% to 100%. In those cases where total suppression of GBSS activity was found both GBSS protein and amylose were absent, giving rise to tubers containing amylose-free starch. The variable response of the transformed plants indicates that position effects on the integrated sequences might be important. The results clearly demonstrate that in tubers of potato plants which constitutively synthesize antisense RNA the starch composition is altered.     

Activities of key enzymes for starch synthesis in relation to growth of superior and inferior grains on winter wheat (Triticum aestivum L.) spike

Weight of individual grains is a major yield component in wheat. The non-uniform distribution of single grain weight on a wheat spike is assumed to be closely associated with starch synthesis in grains. The present study was undertaken to determine if the enzymes involved in starch synthesis cause the differences in single grain weight between superior and inferior grains on a wheat spike. Using two high-yield winter wheat (Triticum aestivum L.) varieties differing in grain weight and three nitrogen rates for one variety, the contents of amylose and amylopectin, and activities of enzymes involved in starch synthesis in both superior and inferior grains were investigated during the entire period of grain filling. Superior grains showed generally higher starch accumulation rates and activities of enzymes including SS (sucrose synthase), UDPGPPase (UDP-glucose pyrophosphorylase), ADPGPPase (ADP-glucose pyrophosphorylase), SSS (soluble starch synthase) and GBSS (starch granule bound starch synthase) and subsequently produced much higher single grain weight than inferior grains. Nitrogen increased enzyme activities and starch accumulation rates, and thus improved individual grain weight, especially for inferior grains. The SS, ADPGPPase and SSS were significantly correlated to amylopectin accumulation, while SS, ADPGPPase, SSS and GBSS were significantly correlated to amylose accumulation. This infers that SS, ADPGPPase and starch synthase play key roles in regulating starch accumulation and grain weight in superior and inferior grains on a wheat spike.     

淀粉含量不同的玉米自交系籽粒淀粉积累及其关键酶活性

以2个高淀粉和2个低淀粉玉米自交系为材料,分析了玉米籽粒淀粉的动态积累规律,同时对高低淀粉玉米籽粒灌浆过程中淀粉生物合成关键酶活性的动态变化及其与淀粉积累动态的相关性进行讨论分析。研究结果表明:灌浆过程中4个自交系淀粉含量变化趋势均呈sigmoid型曲线。灌浆过程中ADPG-PPase(腺苷二磷酸葡萄糖焦磷酸化酶)、SSS(可溶性淀粉合成酶)、GBSS(颗粒结合淀粉合成酶)活性均呈单峰曲线变化,峰值都出现在20～30DAP(授粉后天数)。2个高淀粉自交系的Q酶(淀粉分支酶)活性也呈单峰曲线变化,峰值也出现在20DAP,而2个低淀粉自交系的Q酶活性则呈双峰曲线变化,2个峰值分别出现在15～20DAP和30～35DAP。4个自交系籽粒淀粉的积累速率与各自交系ADPG-PPase、SSS和GBSS的活性变化呈极显著正相关。各自交系关键酶活性之间,ADPG-PPase、SSS和GBSS三者间活性变化呈极显著正相关,这3种酶活性变化与Q酶活性变化也呈不同程度的正相关。     

不同类型土壤上种植的小麦籽粒中与淀粉合成相关的酶活性变化

测定池栽条件下灰潮土、水稻土、砂姜黑土上种植的强筋小麦‘郑麦9023’籽粒灌浆过程中腺苷二磷酸葡萄糖焦磷酸化酶（AGPP）、尿苷二磷酸葡萄糖焦磷酸化酶（UGPP）、可溶性淀粉合成酶（SSS）、淀粉粒结合淀粉合成酶（GBSS）、淀粉分支酶（SBE）5个与淀粉合成有关的酶活性变化的结果表明,不同类型土壤上种植的小麦籽粒中AGPP、UGPP、SSS、GBSS、SBE活性均呈单峰曲线变化,花后18d,AGPP、UGPP、SSS和SBE活性达到峰值,而GBSS则在花后24d达到峰值。AGPP、SSS、SBE活性峰值表现为灰潮土〉水稻土〉砂姜黑土,UGPP峰值表现为灰潮土〉砂姜黑土〉水稻土,GBSS峰值则表现为水稻土〉灰潮土〉砂姜黑土。     

不同类型玉米发育籽粒中淀粉合成及相关酶活性比较

以普通玉米、爆裂玉米、甜玉米和糯玉米为试材,分析和比较不同类型的玉米品种之间籽粒发育过程中淀粉合成及相关酶活性的变化。结果表明,淀粉合成速率和蔗糖合成酶(SS)、可溶性淀粉合成酶(SSS)、束缚态淀粉合成酶(GBSS)、淀粉分支酶(SBE)、去分支酶(DBE)活性都呈单峰曲线变化。30～40 DAP,普通玉米的SS活性显著高于其他3种类型;类型间平均和最大SSS活性水平的顺序为普通玉米>糯玉米>爆裂玉米>甜玉米;30～40 DAP,普通玉米GBSS活性最高,糯玉米GBSS活性最低;20～40 DAP,糯玉米SBE活性最高;甜玉米的DBE活性很低,并且在40 DAP完全丧失。淀粉合成速率与SS、SSS、GBSS和SBE活性相关程度比较高,与腺苷二磷酸葡萄糖焦磷酸化酶(AGP酶)和DBE活性相关不显著。推测AGP酶虽然为淀粉合成提供直接前体ADPG,但可能SS活性过低致使其限速作用比AGP酶的还强,AGP酶潜在的限速作用无法表现,SS成为玉米籽粒淀粉合成的限速因子。GBSS对直链淀粉积累起重要的促进作用;SSS和SBE对支链淀粉积累起重要的促进作用。     

云南野生稻籽粒淀粉合成关键酶活性测定

为研究云南3种野生稻直链淀粉合成机制并利用其直链淀粉含量较低的优良品质,以云南3种野生稻和4种当地栽培稻为材料,研究野生稻灌浆期籽粒4种淀粉合成关键酶(包括ADPG焦磷酸化酶、可溶性淀粉合成、颗粒凝结型淀粉合成酶、淀粉分支酶)活性变化。结果表明,野生稻中4种淀粉合成酶的变化趋势与栽培稻相似,但活性有较大差别。颗粒凝结型淀粉合成酶的活性与直链淀粉含量呈正相关,说明在野生稻中同样是由颗粒凝结型淀粉合成酶控制直链淀粉的合成。同时发现在同一灌浆期,同种材料中可溶性淀粉合成酶和淀粉分支酶的活性变化呈相反趋势,推测这两种酶之间可能在淀粉合成过程中存在某种反馈调节机制。     

鲜食糯玉米采后糖代谢相关酶活性变化

鲜食糯玉米采后腺苷二磷酸葡萄糖焦磷酸化酶（ADPGPPase）、尿苷二磷酸葡萄糖焦磷酸化酶（UDPGPPase）、束缚态淀粉合成酶（GBSS）、淀粉脱分支酶（DBE）等活性均呈现单峰曲线变化,第1～2天出现峰值然后略有下降;可溶性淀粉合成酶（SSS）活性呈较大幅度上升趋势,活性远高于采收初期;淀粉分支酶（SBE）在第3天出现极高峰值。与20℃贮藏温度相比,采后0℃低温贮藏可增强UDPGPPase活性,促进蔗糖降解;降低SSS和GBSS活性,延缓淀粉合成进程;抑制SBE活性,增强DBE活性,促进直链淀粉生成。     

A 56-kDa protein is a novel granule-bound starch synthase existing in the pericarps, aleurone layers, and embryos of immature seed in diploid wheat (Triticum monococcum L.)

A novel 56-kDa granule-bound starch synthase (GBSS; NDPglucose-starch glucosyltransferase, EC 2.4.1.21) responsible for amylose synthesis was found in the pericarps, aleurone layers and embryos of immature diploid wheat (Triticum monococcum L.). The GBSS and other proteins bound to starch granules of various tissues of immature normal and waxy diploid wheat seeds were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and their activities were examined. In the waxy mutant, the waxy protein (59.5 kDa, GBSSI) was absent, but amylose and GBSS activity were evident in all tissues except the endosperm. Of the proteins bound to starch granules, only the 56-kDa protein was associated with the presence of amylose and GBSS activities in the pericarps, aleurone layers and embryos. Mutations at the waxy locus did not affect the 56-kDa protein in these tissues. Changes in the amount of 56-kDa protein during the course of seed development, and the distribution of the 56-kDa protein in each tissue of immature seeds were quite different from those of the waxy protein. On the other hand, the N-terminal amino acid sequence of the 56-kDa protein had a 40–50% similarity to GBSSI of some other plant species and was antigenically related to the waxy protein. These results strongly suggest that the 56-kDa protein in diploid wheat is a GBSSI class enzyme and, hence, an isoform of the waxy protein. The waxy protein and 56-kDa protein, however, are expressed in different seed tissues and at different stages of seed development. Received: 15 May 1998 / Accepted: 18 June 1998     

Paclobutrazol Improved the Reproductive Growth and the Quality of Seed Oil of Jatropha curcas

Jatropha curcas was treated by soil drench paclobutrazol (PBZ) (0, 2, and 3 g m^?1 of canopy diameter) and foliar spray PBZ (0, 500, 800, and 1,200 ppm). The results showed that PBZ treatments greatly retarded vegetative growth and improved reproductive growth. The lengths of new branches were greatly decreased, whereas the number of fruits per inflorescence, fruit-bearing branches per tree, and total fruit load per tree were increased. Only the 2-g soil drench and the 1,200-ppm foliar PBZ spray significantly increased fruit load. The 2-g soil drench PBZ treatment resulted in a decrease in seed S and Cu contents of J. curcas, whereas Mn and B were greatly or moderately increased. A higher dose (3-g soil drench PBZ) reversed the improvement in reproductive growth and alleviated the negative effects on element contents in seeds compared with the 2-g soil drench PBZ. Finally, soil drench PBZ treatments significantly improved seed oil content and oil quality by reducing the oil acid value, increasing stearic acid and oleic acid contents, and reducing palmitic acid and linoleic acid content. The optimum drench dose was below 2 g m^?1 of canopy diameter. The optimum foliar spray concentration of PBZ was not determined in this study but our results suggest that it is higher than 1,200 ppm.     

Use of polyethylene glycol in isolation and assay of stable, enzymically active starch granules from developing wheat endosperms

A procedure using polyethylene glycol (PEG), molecular weight 1000, was developed for the isolation of starch granules from wheat endosperm. Immature endosperm tissue was cut repeatedly in 300 millimolar PEG 1000 and filtered through Miracloth. Centrifugation separated a pellet from a supernatant with inhibitory activity. The pellet contained several enzyme activities, including soluble and bound components of starch synthase, starch phosphorylase, and sucrose synthase activities. The starch phosphorylase activity was unaffected by several washings with 300 millimolar PEG 1000 but was lost when the granules were washed once without PEG or washed with sucrose, glycerol, or sorbitol (up to 30%, w/v). The fraction of starch synthase, remaining on the granules after a wash without PEG (the `bound' activity) was not affected by the addition of 30% sorbitol to the wash buffer. This fraction became larger with grain development (0.2-0.7).

To obtain high activity, PEG was required not only during isolation of granules but also in the assay of both starch phosphorylase and starch synthase giving optimum activity at 225 to 255 millimolar. PEG reduced the requirement for glycogen as primer with soluble starch synthase. However, the `bound' starch synthase activity was unaffected by PEG. PEG of different size were compared by their effects in the assay of starch granules: with increase in molecular size, the same effect was obtained at ever lower polymer concentration (w/v) down to a limit.

Treatment of granules with Triton X-100 did not affect their starch synthase activity, but it removed the capacity to incorporate label from UDP [¹⁴C]G into non-starch polymers.

It is concluded that PEG, like some other active compounds (ethanol Na₃-citrate, and Ficoll) could mediate enzyme-primer interaction by exclusion.