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MARIAN J. MCKENZIE RONAN K. Y. CHEN MATTHEW J. ASHWORTH DAVID A. BRUMMELL 《Plant, cell & environment》2013,36(1):176-185
Cold‐induced sweetening (CIS) is a serious post‐harvest problem for potato tubers, which need to be stored cold to prevent sprouting and pathogenesis in order to maintain supply throughout the year. During storage at cold temperatures (below 10 °C), many cultivars accumulate free reducing sugars derived from a breakdown of starch to sucrose that is ultimately cleaved by acid invertase to produce glucose and fructose. When affected tubers are processed by frying or roasting, these reducing sugars react with free asparagine by the Maillard reaction, resulting in unacceptably dark‐coloured and bitter‐tasting product and generating the probable carcinogen acrylamide as a by‐product. We have previously identified a vacuolar invertase inhibitor (INH2) whose expression correlates both with low acid invertase activity and with resistance to CIS. Here we show that, during cold storage, overexpression of the INH2 vacuolar invertase inhibitor gene in CIS‐susceptible potato tubers reduced acid invertase activity, the accumulation of reducing sugars and the generation of acrylamide in subsequent fry tests. Conversely, suppression of vacuolar invertase inhibitor expression in a CIS‐resistant line increased susceptibility to CIS. The results show that post‐translational regulation of acid invertase by the vacuolar invertase inhibitor is an important component of resistance to CIS. 相似文献
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Soluble acid invertase determines the hexose-to-sucrose ratio in cold-stored potato tubers 总被引:21,自引:0,他引:21
Cold storage of potato (Solanum tuberosum L.) tubers is known to cause accumulation of reducing sugars. Hexose accumulation has been shown to be cultivar-dependent and proposed to be the result of sucrose hydrolysis via invertase. To study whether hexose accumulation is indeed related to the amount of invertase activities, two different approaches were used: (i) neutral and acidic invertase activities as well as soluble sugars were measured in cold-stored tubers of 24 potato cultivars differing in the cold-induced accumulation of reducing sugars and (ii) antisense potato plants with reduced soluble acid invertase activities were created and the soluble sugar accumulation in cold-stored tubers was studied. The cold-induced hexose accumulation in tubers from the different potato cultivars varied strongly (up to eightfold). Large differences were also detected with respect to soluble acid (50-fold) and neutral (5-fold) invertase activities among the different cultivars. Although there was almost no correlation between the total amount of invertase activity and the accumulation of reducing sugars there was a striking correlation between the hexose/sucrose ratio and the extractable soluble invertase activitiy. To exclude the possibility that other cultivar-specific features could account for the obtained results, the antisense approach was used to decrease the amount of soluble acid invertase activity in a uniform genetic background. To this end the cDNA of a cold-inducible soluble acid invertase (EMBL nucleicacid database accession no. X70368) was cloned from the cultivar Desirée, and transgenic potato plants were created expressing this cDNA in the antisense orientation under control of the constitutive 35S cauliflower mosaic virus promotor. Analysis of the harvested and cold-stored tubers showed that inhibition of the soluble acid invertase activity leads to a decreased hexose and an increased sucrose content compared with controls. As was already found for the different potato cultivars the hexose/sucrose ratio decreased with decreasing invertase activities but the total amount of soluble sugars did not significantly change. From these data we conclude that invertases do not control the total amount of soluble sugars in coldstored potato tubers but are involved in the regulation of the ratio of hexose to sucrose.The authors are grateful to Heike Deppner and Christiane Prüßner for tuber harvest and technical assistance during the further analysis. We thank Andrea Knospe for taking care of tissue culture, Birgit Schäfer for patient photographic work, Hellmuth Fromme and the greenhouse personnel for attending plant growth and development and Astrid Basner for elucidating the sequence of clone INV-19. The work was supported by the Bundesministerium für Forschung und Technologie (BMFT). 相似文献
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Key message
High soil temperature during bulking and maturation of potatoes alters postharvest carbohydrate metabolism to attenuate genotypic resistance to cold-induced sweetening and accelerate loss of process quality.Abstract
The effects of soil temperature during tuber development on physiological processes affecting retention of postharvest quality in low-temperature sweetening (LTS) resistant and susceptible potato cultivars were investigated. ‘Premier Russet’ (LTS resistant), AO02183-2 (LTS resistant) and ‘Ranger Russet’ (LTS susceptible) tubers were grown at 16 (ambient), 23 and 29 °C during bulking (111–164 DAP) and maturation (151–180 DAP). Bulking at 29 °C virtually eliminated yield despite vigorous vine growth. Tuber specific gravity decreased as soil temperature increased during bulking, but was not affected by temperature during maturation. Bulking at 23 °C and maturation at 29 °C induced higher reducing sugar levels in the proximal (basal) ends of tubers, resulting in non-uniform fry color at harvest, and abolished the LTS-resistant phenotype of ‘Premier Russet’ tubers. AO02183-2 tubers were more tolerant of heat for retention of LTS resistance. Higher bulking and maturation temperatures also accelerated LTS and loss of process quality of ‘Ranger Russet’ tubers, consistent with increased invertase and lower invertase inhibitor activities. During LTS, tuber respiration fell rapidly to a minimum as temperature decreased from 9 to 4 °C, followed by an increase to a maximum as tubers acclimated to 4 °C; respiration then declined over the remaining storage period. The magnitude of this cold-induced acclimation response correlated directly with the extent of buildup in sugars over the 24-day LTS period and thus reflected the effects of in-season heat stress on propensity of tubers to sweeten and lose process quality at 4 °C. While morphologically indistinguishable from control tubers, tubers grown at elevated temperature had different basal metabolic (respiration) rates at harvest and during cold acclimation, reduced dormancy during storage, greater increases in sucrose and reducing sugars and associated loss of process quality during LTS, and reduced ability to improve process quality through reconditioning. Breeding for retention of postharvest quality and LTS resistance should consider strategies for incorporating more robust tolerance to in-season heat stress. 相似文献6.
Exposure to low but nonfreezing temperatures induces the breakdown of starch and the accumulation of sucrose, glucose and fructose in potato tubers, a complex phenomenon known as low-temperature sweetening (LTS). A kinetic model for the degradation of starch to sucrose, fructose, glucose, hexose phosphates and carbon dioxide in 2 degrees C-stored mature Solanum tuberosum cv. Norchip (LTS-sensitive) and Solanum tuberosum seedlling ND860-2 (LTS-tolerant) tubers is presented in this work. Analysis of sugar accumulation data in tubers grown in 1993 and 1994 showed no significant differences in the rates of conversion of starch to hexose phosphates and hexose phosphates to sucrose for both cultivars (P > 0.05). The rate constant corresponding to invertase activity was 2.3 day(-1) for Norchip tubers and 1.1 day(-1) for ND860-2 tubers grown in 1993 (P < or = 0.05); however, no significant differences were observed in invertase activity for 1994-grown tubers (P > 0.05). The accumulation of the reducing sugars fructose and glucose was found to be dependent on the relative difference in rate constants corresponding to invertase activity and glycolytic/respiratory capacity. This difference was 3-4 fold greater for Norchip in 1993, and 4-6 fold greater for Norchip in 1994, than for ND860-2 (P < or = 0.05). Results from the analysis also suggest that the amount of available starch for degradation was greater in Norchip tubers than ND860-2 tubers (P < or = 0.05). Our analysis suggests that tubers with decreased invertase activity coupled to increased glycolytic/respiratory capacity should be more tolerant to low-temperature stress. 相似文献
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Datir SS Latimer JM Thomson SJ Ridgway HJ Conner AJ Jacobs JM 《Molecular genetics and genomics : MGG》2012,287(6):451-460
In planta the enzymatic activity of apoplastic and vacuolar invertases is controlled by inhibitory proteins. Although these invertase inhibitors (apoplastic and vacuolar forms) have been implicated as contributing to resistance to cold-induced sweetening (CIS) in tubers of potato (Solanum tuberosum L.), there is a lack of information on the structure and allelic diversity of the apoplastic invertase inhibitor genes. We have PCR-isolated and sequenced the alleles of the apoplastic invertase inhibitor gene (Stinh1) from three tetraploid potato genotypes: 1021/1 (a genotype with very high tolerance to CIS), 'Karaka' and 'Summer Delight' (two cultivars that are highly susceptible to CIS). In total, five alleles were identified in these genotypes, of which four (Stinh1-c, Stinh1-d, Stinh1-e, Stinh1-f) were novel. An analysis of allele diversity was conducted by incorporating previously published sequences of apoplastic invertase inhibitors from potato. Eight alleles were assessed for sequence polymorphism in the two exons and the single hypervariable intron. Contrary to the hypervariable intron, only 65 single nucleotide polymorphisms were observed in the exons, of which 42 confer amino acid substitutions. Phylogenetic analysis of amino acid sequences indicates that the alleles of the invertase inhibitor are highly conserved amongst members of the Solanaceae family. 相似文献
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Kumar D 《Indian journal of biochemistry & biophysics》2011,48(2):123-127
Developing cold sweetening resistant processing varieties is one of the frontal areas of research all over the world. In India, first potato processing variety was released in the year 1998 and till 2005 three varieties have been developed. But, there is no information available regarding sugar accumulation response of Indian varieties to low temperature storage. Therefore, it is imperative to generate basic information on cold sweetening development in Indian processing varieties for the use of potato breeders. Development of cold-induced sweetening and its relation to phenolic content of the tuber was studied in three Indian potato varieties viz., Kufri Chipsona-1, Kufri Chipsona-3 and Kufri Jyoti. The reducing sugars decreased in initial phase of storage, followed by continuous increase to unacceptably higher levels after around two weeks of storage. The increase in reducing sugar contents took place subsequent to increase in sucrose content. The changes in phenol content were not in a fixed trend. The degree or number of folds increase in reducing sugar content was relatively less in Kufri Jyoti which contained highest phenol content among the three varieties investigated. It is suggested that development of processing varieties with higher anti-oxidant content and lower invertase activity may provide better cold-induced sweetening resistance. 相似文献
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Chen S Hajirezaei MR Zanor MI Hornyik C Debast S Lacomme C Fernie AR Sonnewald U Börnke F 《Plant, cell & environment》2008,31(1):165-176
Storage of potato tubers at low temperatures leads to the accumulation of glucose and fructose in a process called 'cold sweetening'. The aim of this work was to investigate the role of sucrose-phosphatase (SPP) in potato tuber carbohydrate metabolism at low temperature (4 degrees C). To this end, RNA interference (RNAi) was used to reduce SPP expression in transgenic potato tubers. Analysis of SPP specific small interfering RNAs (siRNAs), SPP protein accumulation and enzyme activity indicated that SPP silencing in transgenic tubers was stable during the cold treatment. Analysis of soluble carbohydrates showed that in transgenic tubers, cold-induced hexogenesis was inhibited while, despite strongly reduced SPP activity, sucrose levels exceeded wild-type (WT) values four- to fivefold after 34 d of cold treatment. This led to a drastic change in the hexose-to-sucrose ratio from 1.9 in WT tubers to 0.15 to 0.11 in transgenic tubers, while the total amount of soluble sugars was largely unchanged in both genotypes. Sucrose-6(F)-phosphate (Suc6P), the substrate of SPP, accumulated in transgenic tubers in the cold which most likely enables the residual enzyme to operate with maximal catalytic activity in vivo and thus, in the long term, counterbalances reduced SPP activity in the transformants. Northern analysis revealed that cold-induced expression of vacuolar invertase (VI) was blocked in SPP-silenced tubers explaining a reduced sucrose-to-hexose conversion. Suc6P levels were found to negatively correlate with VI expression. A possible role of Suc6P in regulating VI expression is discussed. 相似文献
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In plants, vacuolar invertase (β-fructofuranosidase, EC 3.2.1.26) is known to play as a key modulator for hexose accumulation and cell expansion. In this study, two cDNA clones (2,013 and 1,945 bp, with 99 % sequence identity) encoding vacuolar invertase isoforms were isolated from a commercially important Indian potato cultivar, Kufri Chipsona-1 by RT-PCR. The corresponding predicted proteins consisted of 635 amino acids (designated as KC-VIN1, lacking a few amino acids at N-terminus) and 639 amino acids (designated as KC-VIN2), respectively. They showed 99 % identity, and found to vary at several locations with mostly non-conservative substitutions. Multiple sequence alignment of vacuolar invertase homologs covering four Solanaceae family members revealed some notable distinguishing sequence features (signature-type sequences). A consensus sequence was predicted using 45 vacuolar invertase sequences from 27 taxonomically different plant species, and a phylogenetic tree was generated to know the evolutionary relation between them. Hydrophobic characters were predicted, and compared in different plant species. All these data are presented in a comprehensive manner which were not documented in the earlier reports. As a preliminary study, vacuolar invertase expression patterns in the tubers of some Indian potato cultivars were analyzed by semi-quantitative RT-PCR and extractable enzyme assay. In all the potato cultivars, the overall expression level of invertase was found to be considerably higher after storage at low temperature as compared to the freshly harvested tubers. 相似文献
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Xiaobiao Zhu Craig Richael Patrick Chamberlain James S. Busse Alvin J. Bussan Jiming Jiang Paul C. Bethke 《PloS one》2014,9(4)
Sugar-end defect is a tuber quality disorder and persistent problem for the French fry processing industry that causes unacceptable darkening of one end of French fries. This defect appears when environmental stress during tuber growth increases post-harvest vacuolar acid invertase activity at one end of the tuber. Reducing sugars produced by invertase form dark-colored Maillard reaction products during frying. Acrylamide is another Maillard reaction product formed from reducing sugars and acrylamide consumption has raised health concerns worldwide. Vacuolar invertase gene (VInv) expression was suppressed in cultivars Russet Burbank and Ranger Russet using RNA interference to determine if this approach could control sugar-end defect formation. Acid invertase activity and reducing sugar content decreased at both ends of tubers. Sugar-end defects and acrylamide in fried potato strips were strongly reduced in multiple transgenic potato lines. Thus vacuolar invertase silencing can minimize a long-standing French fry quality problem while providing consumers with attractive products that reduce health concerns related to dietary acrylamide. 相似文献
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This work investigates the possible causes of cold-induced sweetening in potato by examining the impact of low temperature on carbohydrate metabolism in mature tubers. Metabolism in tuber discs was monitored by determining the redistribution of radiolabel following incubation in [U-(14)C]glucose. Estimates of flux based on the specific activity of hexose phosphates established that while incubation at 4 degrees C resulted in an immediate restriction in pathways of carbohydrate oxidation relative to activity at 25 degrees C, there was no corresponding increase in flux to soluble sugars. In contrast, prior storage at low temperature stimulated flux to sugars at both 4 and 25 degrees C. Comparison of (14)CO(2) release from specifically labeled glucose and gluconate fed to tuber discs at 4 and 25 degrees C indicated that flux through glycolysis was preferentially restricted relative to the oxidative pentose phosphate pathway at low temperature, irrespective of prior storage temperature. However, the degree of randomization of label between positions C1 and C6 in the fructosyl moiety of sucrose following metabolism of [1-(13)C]glucose established that there was no preferential inhibition of the recycling of triose phosphates to hexose phosphates at low temperature. These results indicate that sugar accumulation in tubers during storage in the cold is not a direct consequence of a constraint in carbohydrate oxidation, despite preferential restriction of glycolysis at low temperature. It is concluded that the cold lability of enzymes catalyzing the conversion of fructose 6-phosphate to fructose 1,6-bisphosphate is not a major factor in cold-induced sweetening in plants and that this widely held hypothesis should be abandoned. 相似文献
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Rabot A Henry C Ben Baaziz K Mortreau E Azri W Lothier J Hamama L Boummaza R Leduc N Pelleschi-Travier S Le Gourrierec J Sakr S 《Plant & cell physiology》2012,53(6):1068-1082
Bud burst is a decisive process in plant architecture that requires light in Rosa sp. This light effect was correlated with stimulation of sugar transport and metabolism in favor of bud outgrowth. We investigated whether sugars could act as signaling entities in the light-mediated regulation of vacuolar invertases and bud burst. Full-length cDNAs encoding two vacuolar invertases (RhVI1 and RhVI2) were isolated from buds. Unlike RhVI2, RhVI1 was preferentially expressed in bursting buds, and was up-regulated in buds of beheaded plants exposed to light. To assess the importance of sugars in this process, the expression of RhVI1 and RhVI2 and the total vacuolar invertase activity were further characterized in buds cultured in vitro on 100 mM sucrose or mannitol under light or in darkness for 48 h. Unlike mannitol, sucrose promoted the stimulatory effect of light on both RhVI1 expression and vacuolar invertase activity. This up-regulation of RhVI1 was rapid (after 6 h incubation) and was induced by as little as 10 mM sucrose or fructose. No effect of glucose was found. Interestingly, both 30 mM palatinose (a non-metabolizable sucrose analog) and 5 mM psicose (a non-metabolizable fructose analog) promoted the light-induced expression of RhVI1 and total vacuolar invertase activity. Sucrose, fructose, palatinose and psicose all promoted bursting of in vitro cultured buds under light. These findings indicate that soluble sugars contribute to the light effect on bud burst and vacuolar invertases, and can function as signaling entities. 相似文献
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Lixiang Cheng Xin Zhang Qingxia Zhao Huijun Li Yuping Wang Dongxia Wang Di Wang Feng Zhang 《Acta Physiologiae Plantarum》2014,36(5):1197-1210
Cold-induced sweetening is one of the major factors limiting the quality of fried potato products. To understand the mechanisms of protein regulation for cold-induced sweetening in potato tubers, a comparative proteomic approach was used to analyse the differentially expressed proteins both during control (25 °C, 30 days) and cold treatment (4 °C, 30 days) using two-dimensional gel electrophoresis. Quantitative image analyses indicated that there were 25 protein spots with their intensities significantly altered more than twofold. Of these proteins, 9 were up-regulated, 13 were down-regulated, 2 were absent, and 1 was induced in the cold-stored tubers. The MALDI-TOF/TOF MS analyses led to the identification of differentially expressed proteins that are involved in several processes and might work cooperatively to maintain metabolic homeostasis in tubers during low-temperature storage. The preponderance of metabolic proteins reflects the inhibition of starch re-synthesis and the accumulation of sugars in carbon fluxes, linking starch–sugar conversion. The respiration-related proteins suggest the transfer of respiratory activity from aerobic respiration to anaerobic respiration in the cold-stored tubers. The proteins associated with defence appear to protect the tuber cells from low-temperature stress. Some heat shock proteins that act as chaperones also displayed a differential expression pattern, suggesting a potentially important role in cold-stored tubers, although their exact contribution remains to be investigated. The proposed hypothetical model might explain the interaction of these differentially expressed proteins that are associated with cold-induced sweetening in tubers. 相似文献
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Ectopic expression of a tobacco invertase inhibitor homolog prevents cold-induced sweetening of potato tubers. 总被引:25,自引:0,他引:25
We have transformed potato with Nt-inhh cDNA, encoding a putative vacuolar homolog of a tobacco cell wall invertase inhibitor, under the control of the CaMV 35S promoter. In transgenic tubers, cold-induced hexose accumulation was reduced by up to 75%, without any effect on potato tuber yield. Processing quality of tubers was greatly improved without changing starch quantity or quality, an important prerequisite for the biotechnological use of Nt-inhh for potato transformation. 相似文献
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Peter Brandt 《Planta》1975,122(1):105-107
Summary Callus cultures derived from tubers of Solanum tuberosum L. cv. Record were transferred from a complete medium to a maintenance medium that lacked sugar and were then incubated at 2° or 25°. Incubation at 2° for 5 days led to an accumulation of sucrose. This accumulation did not occur at the expense of stored reducing sugars and was not found at 25°. Such cultures could be used for the study of cold-induced sweetening in potatoes. 相似文献
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Privat I Foucrier S Prins A Epalle T Eychenne M Kandalaft L Caillet V Lin C Tanksley S Foyer C McCarthy J 《The New phytologist》2008,178(4):781-797
* Coffea arabica (Arabica) and Coffea canephora (Robusta) are the two main cultivated species used for coffee bean production. Arabica genotypes generally produce a higher coffee quality than Robusta genotypes. Understanding the genetic basis for sucrose accumulation during coffee grain maturation is an important goal because sucrose is an important coffee flavor precursor. * Nine new Coffea genes encoding sucrose metabolism enzymes have been identified: sucrose phosphate synthase (CcSPS1, CcSPS2), sucrose phosphate phosphatase (CcSP1), cytoplasmic (CaInv3) and cell wall (CcInv4) invertases and four invertase inhibitors (CcInvI1, 2, 3, 4). * Activities and mRNA abundance of the sucrose metabolism enzymes were compared at different developmental stages in Arabica and Robusta grains, characterized by different sucrose contents in mature grain. * It is concluded that Robusta accumulates less sucrose than Arabica for two reasons: Robusta has higher sucrose synthase and acid invertase activities early in grain development - the expression of CcSS1 and CcInv2 appears to be crucial at this stage and Robusta has a lower SPS activity and low CcSPS1 expression at the final stages of grain development and hence has less capacity for sucrose re-synthesis. Regulation of vacuolar invertase CcInv2 activity by invertase inhibitors CcInvI2 and/or CcInvI3 during Arabica grain development is considered. 相似文献