text:
|Publication number
|Publication type
|Publication date
|May 26, 1964
|Filing date
|Jan 19, 1962
|Priority date
|Jan 19, 1962
|Publication number
|Criscuolo James M
|Original Assignee
|Criscuolo James M
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (3), Referenced by (12), Classifications (4)
|External Links: USPTO, USPTO Assignment, Espacenet
y 26, 1964 J. M. CRISCUOLO 3,134,119
SEWER CLEANING DEVICE Filed Jan. 19, 1962 Iii! INVENTOR. JAMES M. CRIS CUOLO BY aoamymflpwzm ATTORNEYS United States Patent 3,134,119 3EWER CLEANING DEVEQCE James M. Criscuolo, 1938 Wallace Ave, Bronx, NY. Filed Jan. 19, 1962, Ser. No. 167,392 4- Ciaims. (Cl. 1il4.3)
This application is related to my coapending applications Serial No. 9,846, filed February 119, 1960*, now U.S. Patent 3,048,870, which latter application is a continuation-in-part of my application Serial No. 617,395, filed October 22, 1956, now abandoned.
My invention relates to cleaning sewers and drain pipes and, more particularly, provides an easily assembled and readily transportable sewer cleaner equipped to withstand the stresses, especially angular stresses, imposed on such devices by obstructions in sewers and drains.
The sewer cleaning machines commonly in use today are made up primarily of a cleaning instrument, usually a rotating cutting tool strong enough to cut through tree roots and similar obstructions found in sewer pipe, a rotating drive device, and "a connecting link for connecting the drive device to the tool. Such connecting links are most commonly braided steel cables or tightly wound steel wire coils, or a combination of both, attached at one end to the cutting tool and at the other end to the drive device, which usually is an electric motor. Various arrangements are commonly used for attaching the drive device to the cable link, such as belts, pulleys, gears, and chains, which permit a portion of the link to be retained on a drum or reel while an extended portion, attached to the cutting tool, is rotated.
The forward progress of the cutting tool along the internal length of the sewer or drain is usually made by passing the relatively stiff cable or wire coil hand over hand into the conduit to be cleaned to drive the cutting tool forward so long as no obstructions interfere or so long as there is cable or coil left on the reel or in the cable drum. As a result, the operator is rarely out of close contact with his sewer cleaning equipment and must almost continuously hazard the dangers inherent in being close to or in actually handling a continuously moving steel cable.
The greatest hazard to the operator and his equipment, and one which often befalls even the most experienced operator arises when his rotating cutting tool strikes an obstruction in the sewer line which does not give way and which causes the cutting blade to stop rotating, despite the fact that torque is still applied to the cable. This sort of stoppage causes the cable to twist, kink, and perhaps break. Consequently, a potentially destructive strain is put on an expensive piece of equipment, no matter how quickly power may be shut off. More important, however, is the danger to the operator of having the twisting cable torn from his hands and, just as easily, of being struck by the lashing cable or its broken ends. It is an object of my invention substantially to eliminate this danger without the necessity of completely cutting off the power supply.
Another disadvantage common to sewer cleaning devices is their bulk and weight. The cable is particularly cumbersome, since it is not uncommon to employ it in 100 lengths weighing from about 60* to 80 pounds which are normally stored on reels or drum cable containers having about a 2 radius and which are most often themselves of metal. It is an object of my invention to make the transportation of such cable and its containers as safe and as convenient as possible by substantially eliminating the need for lifting them.
It is also an object of my invention to make the assembly of the equipment, particularly the connection between the cable and the drive power source for rotating the cutting tool, as simple as possible by eliminating the need for belts, pulleys, gears or chains and excessive manipulations by the operator.
These and other objects are accomplished by my sewer cleaner which has two larger components. These are a rotatable drum cable container mounted upon a horizontal axis which serves not only to store and transport the cable or wire to be used, but also to impart to the cable the rotational force ultimately to be communicated to the cutting tool, and a support means upon which the drum and the power source for rotating the drum can be firmly secured. The two components are so constructed as to afford the operator as much ease as possible in transporting the device. For a more complete understanding of the practical application of the features and principles of my invention, reference is made to the appended drawings in which:
FIGURES l and 2 are front elevations of an embodiment of the sewer cleaning system of my invention; and
FIGURE 3 is an enlarged side view of the embodiment of my invention shown in FIGURES 1 and 2.
In the embodiment of my invention depicted in FIG- URES 1, 2 and 3, a freely rotatable curved cable conduit 1112 is rotatably mounted on axle 104 by means of bushing 106. The conduit 102 is a hollow, bent pipe which curves from the bushing 16% into the drum '1 and outwardly along a radius of the drum. A cable 3 with a cutting tool 16 fastened to its outer end is led out of the drum, through the conduit, toward the obstructed pipe .to be cleaned. Drum 1 is also rotatably mounted on axle 104 as by a bushing, not shown, bolted to sidewall 17. As can be seen in FIGURE 3, the axle 134 extends beyond the side-wall 1'7 of drum 1. A support member 108 is mounted for rotation about this portion of axle 104. Finally, at the end of axle 104- remote from the conduit W2, a motor mounting member 116' is also mounted for rotation about axle 164. The conduit 102, the support member 1118 and the motor mounting member 110, along with the axle 1114, are held together by a pair of washers 112 and a pair of bolts 114- threadedly received in each end of axle 1124.
As can be seen from the drawings, particularly FIG- URES 1 and 2, support member 1118 is of a generally triangular shape and has two legs 116 and 118 attached thereto and extending radially outward from the axis of axle 1M. Legs 116 and 113 are provided with feet 120 and 122, respectively, disposed parallel to the axis of axle 104. The legs 116 and 11 8 are of such length that they extend sufiiciently beyond the periphery ofdr'um 1 and tire 2 so that when the sewer cleaner is assembled it can stand on feet 129 and 122 and a suificient clearance between tire 2 and the surface upon which the feet 120 and 122 are standing is provided to permit rotation of the drum 1.
Motor mounting member 111 is provided with three cylindrical holes 124, 126, and 128 located at the end of motor mounting member 111) remote from axle 104.
set screw 134 tightened to hold it in place.
arrangemenhihandle 136 functions as a third support Holes 124 and 126 are disposed such that their axes extend radially from the axis of axle 1'34. Hole 128, however, is disposed so that its axis intersects the axes of both hole 126 and axle 104. Each of these cylindrical holes 124, 126 and 128 is provided with a wing head set screw 131i), 132and 134, respectively.
FIGURES 1 and '2 show a handle 136 with one end thereof inserted in cylindrical hole 124. The other end of handle 136 is provided with a crossbar 138.
An electric motor 146 having an output shaft 142 is mounted on plate 144 having extension 146 for supporting output shaft 142. Plate 144 is in turn mounted on cylindrical post 148 adapted to be received slidably in cylindrical hole 126.
Referring now to FIGURES 1 and 2, it can be seen that a coil spring 150 is connected at one end thereof to. support member 1118 and at the other end thereof to motor mounting member 110. It will also be seen that when coil spring 150 is in its extended position as shown in FIGURE 1 it will bias support member 1% to rotate toward motor mounting member 110 until such time as coil spring 150 is in its fully retracted position as shown in FIGURE 2. As can also be seen, a small tip 152 extends from the end of motor mounting member 110 remote from motor 146 and overlies a portion of support member 1198. Tip152 is provided with another wing head set screw 154 which is adapted to be threadedly received by tip 152 and bear against support member 108.
FIGURE 1 shows this'particular embodiment of my invention supported on feet 120 and 122 of legs 116 and In order to transport this embodiment 118, respectively. of my invention, it is merely necessary for the operator to loosen wing head set screw 154- so that it does not bear against support member 193 and then pull on handle 136 by means of cross bar 133. As soon as the weight of the machine has been removed from feet 124 and 122 and the machine is resting on tire 2, the coil spring 150 V will cause support member 108 to rotate into the position shown in FIGURE 2. Wing head set screw 154 can then be tightened so as to secure support member 185 in this position when transporting the machine. The
. machine can then be transported by rolling it along tire head set screw 13h can belloosened and handle 136 removedfrom the cylindrical hole 124. The end of handle 136 removed from the cylindrical hole 124 can now be placed into cylindrical hole 128 and wing head leg as illustrated in FIGURE 3.
Furthermore, this embodiment of my invention pro- .vides a means'for adjusting the friction drive between output shaft 142 and tire 2. This is accomplished by loosening wing head set screw1132 and raising or lower ing post 148 within cylindrical hole 126. When post 148 is in the desired position, thereby placing output shaft 142 in thedesired position relative to tire 2, wing head set screw 132 can be tightened to hold post 143 in 'place. This'arrangement can also be employed to re move output shaft 142 from contact with tire 2 when in such drum 1 will not begin to rotate immediately, since the tire 2 and the output shaft 142 are so contacted as to permit slippage. However, as the operator begins by hand to feed the cable 3 to which a cutting tool 16 has been a'i'fixed into the sewer or drain pipe, the friction between the output shaft 142 and the tire 2 will be put 1 to use in therotation of the drum 1 and the application of torque to the cable 3. The rotation of conduit 102, on
the other hand, depends entirely on the positioning of the cable remaining in drum 1 Whenever the further feeding of cable into the sewer is stopped or prevented, the clutch-like action between a the 2 and output shaft 142 comes into play and the transporting the machine in the manner described above.
Advantageously, the handle 136 and cross bar 138 are equipped with brake means for contact with tire 2.
' Although the embodiment of'the invention shown in FIGURES 1, 2 and 3 show. the support means movably jointed at the drum axle, the support means can be movably jointed at any point less than a drum radius distant from the axis of rotation of the drum to permit rotation drum 1 stops its rotation. This, in turn, prevents the application of any torque to cable 3 and, as a consequence, the cable is not liable to twist, kink, or even, break to the detriment of the equipment or the operator. ther, the motor need not be turned off. The same is true when the cable is being withdrawn from the sewer pipe.
I claim:
1. A sewer pipe cleaning device comprising a drum for receiving flexible cables and wires annularly coiled therein, a curved cable conduit one end of which is rotatably mounted at one end of said drum adjacent the axial line thereof with the other end of said conduit disposed within said drum and extending outwardly from said axial line, an elongated support means having a pair of spacedapart feet at one end and a handle at the other end of said support means, said drum beingrnounted upon said support means intermediate the ends thereof for rotation about an axis perpendicular thereto, drive means movably mounted on said support means intermediate the handle and the mounted drum, said'drive means being capable of peripheral contact with the said drum for rotating said drum, said support means being movably jointed at a point less than a 'drurn radius distant from the axis of rotation of the drum to permit rotation in a plane perpendicular to the axis of rotation of the drum and to permit support of the device by the drum, and means for preventing rotation of said support means about the joint.
2. A device according to claim 1 which further comprises a tire positioned about said drum and in which said drive means includes a rotatable output shaft peripherally contacting said tire.
3. A device according to claim 1 in which the support means comprises a support member from which two legs depend and on which legs feet aremounted, the support'membe'r being in the same general plane'as the legs, an axle perpendicular to said plane, said support member being joined at one end to said legs at a common point and at the other end enclosing said axle, a motor mounting member having ahandle at one end and at the other end enclosing saidaxle, and means associated with saidsupport member and motor mounting member to hold them rigidly aligned.
4. A sewer pipe cleaning device comprising a drum for receiving flexible cables and wires annularly coiled therein, a cable annularly coiled in said drum, a tire positioned about said drum, a curved cable conduit one end of which is rotatably mounted atone end of said 'drum adjacent the axial line thereof with the other end of said conduit disposed within said drum and extending Furgeneral plane as said legs, an axle perpendicular to said plane, said support member being joined at one end to said legs at a common point and at the other end enclosing said axle, a motor mounting member having a handle at one end and at the other end enclosing said axle, means associated with said support member and motor mounting member to hold them rigidly aligned, spring means associated with said support member and motor mounting member biasing said support member and motor mounting member to rotate about said axle relative to one another, said drum rotatably mounted on said axle, and drive means movably mounted on said motor mounting member including a rotatable output shaft peripherally contacting said tire, the spring means rotating said support member and motor mounting member when not rigidly aligned to permit support and movement of 5 the sewer pipe cleaning device on the drum tire.
References Cited in the file of this patent UNITED STATES PATENTS 2,468,490 Di Joseph Apr. 26, 1949 10 3,007,186 Olsson Nov. 7, 1961 3,048,870 Criscuolo Aug. 14, 1962
|Cited Patent
|Filing date
|Publication date
|Mar 15, 1945
|Apr 26, 1949
|Di Joseph John
|Pipe cleaning power cable feeder
|Feb 10, 1955
|Nov 7, 1961
|H D Conkey & Company
|Sewer cleaning machines
|Feb 19, 1960
|Aug 14, 1962
|Criscuolo James M
|Sewer cleaning device
|Citing Patent
|Filing date
|Publication date
|Feb 20, 1990
|Jul 16, 1991
|Emerson Electric Co.
|Drain cleaning machine
|Jun 13, 2000
|Oct 29, 2002
|Emerson Electric Co.
|Cart and stand for supporting and transporting metal working apparatus
|Nov 1, 2011
|Emerson Electric Co.
|Drain cleaning apparatus with electronic cable counter
|May 15, 2012
|Emerson Electric Co.
|Drain cleaning apparatus with electronic cable monitoring system
|Apr 16, 2012
|Apr 9, 2013
|Emerson Electric Co.
|Drain cleaning apparatus with electronic cable monitoring system
|Dec 21, 2006
|Jun 26, 2008
|Emerson Electric Co.
|Cable feeding device with indicator
|May 21, 2009
|Nov 26, 2009
|Emerson Electric Co.
|Drain cleaning apparatus with electronic cable monitoring system
|Feb 11, 2010
|Emerson Electric Co.
|Drain cleaning apparatus with electronic cable counter
|Dec 12, 2007
|Nov 19, 2009
|Emerson Electric Co.
|Kabelzuf�hrvorrichtung mit Indikator
|Dec 14, 1990
|Aug 28, 1991
|Emerson Electric Co.
|Drain cleaning machine
|Nov 20, 2009
|May 1, 2013
|Emerson Electric Co.
|Graphical representation of enclosed inspection area
|Nov 20, 2009
|May 1, 2013
|Emerson Electric Co.
|Graphical representation of enclosed inspection area
|U.S. Classification
|International Classification
|Cooperative Classification
|European Classification


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Berries of grape (Vitis vinifera) contain three major tissue types (skin, pulp and seed) all of which contribute to the aroma, color, and flavor characters of wine. The pericarp, which is composed of the exocarp (skin) and mesocarp (pulp), not only functions to protect and feed the developing seed, but also to assist in the dispersal of the mature seed by avian and mammalian vectors. The skin provides volatile and nonvolatile aroma and color compounds, the pulp contributes organic acids and sugars, and the seeds provide condensed tannins, all of which are important to the formation of organoleptic characteristics of wine. In order to understand the transcriptional network responsible for controlling tissue-specific mRNA expression patterns, mRNA expression profiling was conducted on each tissue of mature berries of V. vinifera Cabernet Sauvignon using the Affymetrix GeneChip® Vitis oligonucleotide microarray ver. 1.0. In order to monitor the influence of water-deficit stress on tissue-specific expression patterns, mRNA expression profiles were also compared from mature berries harvested from vines subjected to well-watered or water-deficit conditions.
Overall, berry tissues were found to express approximately 76% of genes represented on the Vitis microarray. Approximately 60% of these genes exhibited significant differential expression in one or more of the three major tissue types with more than 28% of genes showing pronounced (2-fold or greater) differences in mRNA expression. The largest difference in tissue-specific expression was observed between the seed and pulp/skin. Exocarp tissue, which is involved in pathogen defense and pigment production, showed higher mRNA abundance relative to other berry tissues for genes involved with flavonoid biosynthesis, pathogen resistance, and cell wall modification. Mesocarp tissue, which is considered a nutritive tissue, exhibited a higher mRNA abundance of genes involved in cell wall function and transport processes. Seeds, which supply essential resources for embryo development, showed higher mRNA abundance of genes encoding phenylpropanoid biosynthetic enzymes, seed storage proteins, and late embryogenesis abundant proteins. Water-deficit stress affected the mRNA abundance of 13% of the genes with differential expression patterns occurring mainly in the pulp and skin. In pulp and seed tissues transcript abundance in most functional categories declined in water-deficit stressed vines relative to well-watered vines with transcripts for storage proteins and novel (no-hit) functional assignments being over represented. In the skin of berries from water-deficit stressed vines, however, transcripts from several functional categories including general phenypropanoid and ethylene metabolism, pathogenesis-related responses, energy, and interaction with the environment were significantly over-represented.
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Individual berries in a grape (Vitis vinifera L.) cluster enter the ripening phase at different times leading to an asynchronous cluster in terms of ripening. The factors causing this variable ripening initiation among berries are not known. Because the influence via hormonal communication of the seed on fruit set and growth is well known across fruit species, differences in berry seed content and resultant quantitative or qualitative differences in the hormone signals to the pericarp likely influence the relative timing of ripening initiation among berries of the cluster.
At the time of the initiation of cluster ripening (véraison), underripe green berries have higher seed content compared to the riper berries and there is a negative correlation between the seed weight-to-berry weight ratio (SB) and the sugar level in berries of a cluster. Auxin levels in seeds relative to the pericarp tissues are two to 12 times higher at pre-ripening stages. The pericarp of berries with high-SB had higher auxin and lower abscisic acid (ABA) levels compared to those with low-SB from two weeks before véraison. In the prevéraison cluster, the expression of auxin-response factor genes was significantly higher in the pericarp of high-SB berries and remained higher until véraison compared to low-SB berries. The expression level of auxin-biosynthetic genes in the pericarp was the same between both berry groups based upon similar expression activity of YUC genes that are rate-limiting factors in auxin biosynthesis. On the other hand, in low-SB berries, the expression of ABA-biosynthetic and ABA-inducible NCED and MYB genes was higher even two weeks before véraison.
Differences in the relative seed content among berries plays a major role in the timing of ripening initiation. Towards the end of berry maturation phase, low and high levels of auxin are observed in the pericarp of low- and high-SB berries, respectively. This results in higher auxin-signaling activity that lasts longer in the pericarp of high-SB berries. In contrast, in low-SB berries, concomitant with an earlier decrease of auxin level, the features of ripening initiation, such as increases in ABA and sugar accumulation begin earlier.
Electronic supplementary material
The online version of this article (doi:10.1186/s12870-015-0440-6) contains supplementary material, which is available to authorized users.
Seed; Auxin; Fruit ripening; Vitis vinifera; Asynchronous ripening
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Berry skin development in Norton grape: Distinct patterns of transcriptional regulation and flavonoid biosynthesis
Ali, Mohammad B
Howard, Susanne
Chen, Shangwu
Wang, Yechun
Yu, Oliver
Kovacs, Laszlo G
Qiu, Wenping
BMC Plant Biology
The complex and dynamic changes during grape berry development have been studied in Vitis vinifera, but little is known about these processes in other Vitis species. The grape variety 'Norton', with a major portion of its genome derived from Vitis aestivalis, maintains high levels of malic acid and phenolic acids in the ripening berries in comparison with V. vinifera varieties such as Cabernet Sauvignon. Furthermore, Norton berries develop a remarkably high level of resistance to most fungal pathogens while Cabernet Sauvignon berries remain susceptible to those pathogens. The distinct characteristics of Norton and Cabernet Sauvignon merit a comprehensive analysis of transcriptional regulation and metabolite pathways.
A microarray study was conducted on transcriptome changes of Norton berry skin during the period of 37 to 127 days after bloom, which represents berry developmental phases from herbaceous growth to full ripeness. Samples of six berry developmental stages were collected. Analysis of the microarray data revealed that a total of 3,352 probe sets exhibited significant differences at transcript levels, with two-fold changes between at least two developmental stages. Expression profiles of defense-related genes showed a dynamic modulation of nucleotide-binding site-leucine-rich repeat (NBS-LRR) resistance genes and pathogenesis-related (PR) genes during berry development. Transcript levels of PR-1 in Norton berry skin clearly increased during the ripening phase. As in other grapevines, genes of the phenylpropanoid pathway were up-regulated in Norton as the berry developed. The most noticeable was the steady increase of transcript levels of stilbene synthase genes. Transcriptional patterns of six MYB transcription factors and eleven structural genes of the flavonoid pathway and profiles of anthocyanins and proanthocyanidins (PAs) during berry skin development were analyzed comparatively in Norton and Cabernet Sauvignon. Transcriptional patterns of MYB5A and MYB5B were similar during berry development between the two varieties, but those of MYBPA1 and MYBPA2 were strikingly different, demonstrating that the general flavonoid pathways are regulated under different MYB factors. The data showed that there were higher transcript levels of the genes encoding flavonoid-3'-O-hydroxylase (F3'H), flavonoid-3',5'-hydroxylase (F3'5'H), leucoanthocyanidin dioxygenase (LDOX), UDP-glucose:flavonoid 3'-O-glucosyltransferase (UFGT), anthocyanidin reductase (ANR), leucoanthocyanidin reductase (LAR) 1 and LAR2 in berry skin of Norton than in those of Cabernet Sauvignon. It was also found that the total amount of anthocyanins was markedly higher in Norton than in Cabernet Sauvignon berry skin at harvest, and five anthocyanin derivatives and three PA compounds exhibited distinctive accumulation patterns in Norton berry skin.
This study provides an overview of the transcriptome changes and the flavonoid profiles in the berry skin of Norton, an important North American wine grape, during berry development. The steady increase of transcripts of PR-1 and stilbene synthase genes likely contributes to the developmentally regulated resistance during ripening of Norton berries. More studies are required to address the precise role of each stilbene synthase gene in berry development and disease resistance. Transcriptional regulation of MYBA1, MYBA2, MYB5A and MYBPA1 as well as expression levels of their putative targets F3'H, F3'5'H, LDOX, UFGT, ANR, LAR1, and LAR2 are highly correlated with the characteristic anthocyanin and PA profiles in Norton berry skin. These results reveal a unique pattern of the regulation of transcription and biosynthesis pathways underlying the viticultural and enological characteristics of Norton grape, and yield new insights into the understanding of the flavonoid pathway in non-vinifera grape varieties.
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Vv-AMP1, a ripening induced peptide from Vitis vinifera shows strong antifungal activity
de Beer, Abré
Vivier, Melané A
BMC Plant Biology
Latest research shows that small antimicrobial peptides play a role in the innate defense system of plants. These peptides typically contribute to preformed defense by developing protective barriers around germinating seeds or between different tissue layers within plant organs. The encoding genes could also be upregulated by abiotic and biotic stimuli during active defense processes. The peptides display a broad spectrum of antimicrobial activities. Their potent anti-pathogenic characteristics have ensured that they are promising targets in the medical and agricultural biotechnology sectors.
A berry specific cDNA sequence designated Vv-AMP1, Vitis vinifera antimicrobial peptide 1, was isolated from Vitis vinifera. Vv-AMP1 encodes for a 77 amino acid peptide that shows sequence homology to the family of plant defensins. Vv-AMP1 is expressed in a tissue specific, developmentally regulated manner, being only expressed in berry tissue at the onset of berry ripening and onwards. Treatment of leaf and berry tissue with biotic or abiotic factors did not lead to increased expression of Vv-AMP1 under the conditions tested. The predicted signal peptide of Vv-AMP1, fused to the green fluorescent protein (GFP), showed that the signal peptide allowed accumulation of its product in the apoplast. Vv-AMP1 peptide, produced in Escherichia coli, had a molecular mass of 5.495 kDa as determined by mass spectrometry. Recombinant Vv-AMP1 was extremely heat-stable and showed strong antifungal activity against a broad spectrum of plant pathogenic fungi, with very high levels of activity against the wilting disease causing pathogens Fusarium oxysporum and Verticillium dahliae. The Vv-AMP1 peptide did not induce morphological changes on the treated fungal hyphae, but instead strongly inhibited hyphal elongation. A propidium iodide uptake assay suggested that the inhibitory activity of Vv-AMP1 might be associated with altering the membrane permeability of the fungal membranes.
A berry specific cDNA clone, Vv-AMP1, was isolated and characterized and shown to encode a plant defensin. Recombinant Vv-AMP1 displayed non-morphogenic antifungal activity against a broad spectrum of fungi, probably altering the membrane permeability of the fungal pathogens. The expression of this peptide is highly regulated in Vitis vinifera, hinting at an important defense role during berry-ripening.
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Genomic and transcriptomic analysis of the AP2/ERF superfamily in Vitis vinifera
Licausi, Francesco
Giorgi, Federico M
Zenoni, Sara
Osti, Fabio
Pezzotti, Mario
Perata, Pierdomenico
BMC Genomics
The AP2/ERF protein family contains transcription factors that play a crucial role in plant growth and development and in response to biotic and abiotic stress conditions in plants. Grapevine (Vitis vinifera) is the only woody crop whose genome has been fully sequenced. So far, no detailed expression profile of AP2/ERF-like genes is available for grapevine.
An exhaustive search for AP2/ERF genes was carried out on the Vitis vinifera genome and their expression profile was analyzed by Real-Time quantitative PCR (qRT-PCR) in different vegetative and reproductive tissues and under two different ripening stages.
One hundred and forty nine sequences, containing at least one ERF domain, were identified. Specific clusters within the AP2 and ERF families showed conserved expression patterns reminiscent of other species and grapevine specific trends related to berry ripening. Moreover, putative targets of group IX ERFs were identified by co-expression and protein similarity comparisons.
The grapevine genome contains an amount of AP2/ERF genes comparable to that of other dicot species analyzed so far. We observed an increase in the size of specific groups within the ERF family, probably due to recent duplication events. Expression analyses in different aerial tissues display common features previously described in other plant systems and introduce possible new roles for members of some ERF groups during fruit ripening. The presented analysis of AP2/ERF genes in grapevine provides the bases for studying the molecular regulation of berry development and the ripening process.
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Day and night heat stress trigger different transcriptomic responses in green and ripening grapevine (vitis vinifera) fruit
Rienth, Markus
Torregrosa, Laurent
Luchaire, Nathalie
Chatbanyong, Ratthaphon
Lecourieux, David
Kelly, Mary T
Romieu, Charles
BMC Plant Biology
Global climate change will noticeably affect plant vegetative and reproductive development. The recent increase in temperatures has already impacted yields and composition of berries in many grapevine-growing regions. Physiological processes underlying temperature response and tolerance of the grapevine fruit have not been extensively investigated. To date, all studies investigating the molecular regulation of fleshly fruit response to abiotic stress were only conducted during the day, overlooking possible critical night-specific variations. The present study explores the night and day transcriptomic response of grapevine fruit to heat stress at several developmental stages. Short heat stresses (2 h) were applied at day and night to vines bearing clusters sequentially ordered according to the developmental stages along their vertical axes. The recently proposed microvine model (DRCF-Dwarf Rapid Cycling and Continuous Flowering) was grown in climatic chambers in order to circumvent common constraints and biases inevitable in field experiments with perennial macrovines. Post-véraison berry heterogeneity within clusters was avoided by constituting homogenous batches following organic acids and sugars measurements of individual berries. A whole genome transcriptomic approach was subsequently conducted using NimbleGen 090818 Vitis 12X (30 K) microarrays.
Present work reveals significant differences in heat stress responsive pathways according to day or night treatment, in particular regarding genes associated with acidity and phenylpropanoid metabolism. Precise distinction of ripening stages led to stage-specific detection of malic acid and anthocyanin-related transcripts modulated by heat stress. Important changes in cell wall modification related processes as well as indications for heat-induced delay of ripening and sugar accumulation were observed at véraison, an effect that was reversed at later stages.
This first day - night study on heat stress adaption of the grapevine berry shows that the transcriptome of fleshy fruits is differentially affected by abiotic stress at night. The present results emphasize the necessity of including different developmental stages and especially several daytime points in transcriptomic studies.
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Interactions between ethylene and auxin are crucial to the control of grape (Vitis vinifera L.) berry ripening
Böttcher, Christine
Burbidge, Crista A
Boss, Paul K
Davies, Christopher
BMC Plant Biology
Fruit development is controlled by plant hormones, but the role of hormone interactions during fruit ripening is poorly understood. Interactions between ethylene and the auxin indole-3-acetic acid (IAA) are likely to be crucial during the ripening process, since both hormones have been shown to be implicated in the control of ripening in a range of different fruit species.
Grapevine (Vitis vinifera L.) homologues of the TRYPTOPHAN AMINOTRANSFERASE RELATED (TAR) and YUCCA families, functioning in the only characterized pathway of auxin biosynthesis, were identified and the expression of several TAR genes was shown to be induced by the pre-ripening application of the ethylene-releasing compound Ethrel. The induction of TAR expression was accompanied by increased IAA and IAA-Asp concentrations, indicative of an upregulation of auxin biosynthesis and conjugation. Exposure of ex planta, pre-ripening berries to the ethylene biosynthesis inhibitor aminoethoxyvinylglycine resulted in decreased IAA and IAA-Asp concentrations. The delayed initiation of ripening observed in Ethrel-treated berries might therefore represent an indirect ethylene effect mediated by increased auxin concentrations. During berry development, the expression of three TAR genes and one YUCCA gene was upregulated at the time of ripening initiation and/or during ripening. This increase in auxin biosynthesis gene expression was preceded by high expression levels of the ethylene biosynthesis genes 1-aminocyclopropane-1-carboxylate synthase and 1-aminocyclopropane-1-carboxylate oxidase.
In grape berries, members of both gene families involved in the two-step pathway of auxin biosynthesis are expressed, suggesting that IAA is produced through the combined action of TAR and YUCCA proteins in developing berries. The induction of TAR expression by Ethrel applications and the developmental expression patterns of auxin and ethylene biosynthesis genes indicate that elevated concentrations of ethylene prior to the initiation of ripening might lead to an increased production of IAA, suggesting a complex involvement of this auxin and its conjugates in grape berry ripening.
Aminoethoxyvinylglycine; Auxin; Biosynthesis; Ethrel; Ethylene; Interaction; Vitis vinifera; Ripening
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Colour variation in red grapevines (Vitis vinifera L.): genomic organisation, expression of flavonoid 3'-hydroxylase, flavonoid 3',5'-hydroxylase genes and related metabolite profiling of red cyanidin-/blue delphinidin-based anthocyanins in berry skin
Castellarin, Simone D
Di Gaspero, Gabriele
Marconi, Raffaella
Nonis, Alberto
Peterlunger, Enrico
Paillard, Sophie
Adam-Blondon, Anne-Francoise
Testolin, Raffaele
BMC Genomics
Structural genes of the phenyl-propanoid pathway which encode flavonoid 3'- and 3',5'-hydroxylases (F3'H and F3'5'H) have long been invoked to explain the biosynthesis of cyanidin- and delphinidin-based anthocyanin pigments in the so-called red cultivars of grapevine. The relative proportion of the two types of anthocyanins is largely under genetic control and determines the colour variation among red/purple/blue berry grape varieties and their corresponding wines.
Gene fragments of VvF3'H and VvF3'5'H, that were isolated from Vitis vinifera 'Cabernet Sauvignon' using degenerate primers designed on plant homologous genes, translated into 313 and 239 amino acid protein fragments, respectively, with up to 76% and 82% identity to plant CYP75 cytochrome P450 monooxygenases. Putative function was assigned on the basis of sequence homology, expression profiling and its correlation with metabolite accumulation at ten different ripening stages. At the onset of colour transition, transcriptional induction of VvF3'H and VvF3'5'H was temporally coordinated with the beginning of anthocyanin biosynthesis, the expression being 2-fold and 50-fold higher, respectively, in red berries versus green berries. The peak of VvF3'5'H expression was observed two weeks later concomitantly with the increase of the ratio of delphinidin-/cyanidin-derivatives. The analysis of structural genomics revealed that two copies of VvF3'H are physically linked on linkage group no. 17 and several copies of VvF3'5'H are tightly clustered and embedded into a segmental duplication on linkage group no. 6, unveiling a high complexity when compared to other plant flavonoid hydroxylase genes known so far, mostly in ornamentals.
We have shown that genes encoding flavonoid 3'- and 3',5'-hydroxylases are expressed in any tissues of the grape plant that accumulate flavonoids and, particularly, in skin of ripening red berries that synthesise mostly anthocyanins. The correlation between transcript profiles and the kinetics of accumulation of red/cyanidin- and blue/delphinidin-based anthocyanins indicated that VvF3'H and VvF3'5'H expression is consistent with the chromatic evolution of ripening bunches. Local physical maps constructed around the VvF3'H and VvF3'5'H loci should help facilitate the identification of the regulatory elements of each isoform and the future manipulation of grapevine and wine colour through agronomical, environmental and biotechnological tools.
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Plant Flavonoids—Biosynthesis, Transport and Involvement in Stress Responses
Petrussa, Elisa
Braidot, Enrico
Zancani, Marco
Peresson, Carlo
Bertolini, Alberto
Patui, Sonia
Vianello, Angelo
International Journal of Molecular Sciences
This paper aims at analysing the synthesis of flavonoids, their import and export in plant cell compartments, as well as their involvement in the response to stress, with particular reference to grapevine (Vitis vinifera L.). A multidrug and toxic compound extrusion (MATE) as well as ABC transporters have been demonstrated in the tonoplast of grape berry, where they perform a flavonoid transport. The involvement of a glutathione S-transferase (GST) gene has also been inferred. Recently, a putative flavonoid carrier, similar to mammalian bilitranslocase (BTL), has been identified in both grape berry skin and pulp. In skin the pattern of BTL expression increases from véraison to harvest, while in the pulp its expression reaches the maximum at the early ripening stage. Moreover, the presence of BTL in vascular bundles suggests its participation in long distance transport of flavonoids. In addition, the presence of a vesicular trafficking in plants responsible for flavonoid transport is discussed. Finally, the involvement of flavonoids in the response to stress is described.
flavonoid transport; bilitranslocase; ABC transporters; secondary metabolites; biotic and abiotic stress
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Transcriptional analysis of late ripening stages of grapevine berry
Guillaumie, Sabine
Fouquet, Romain
Kappel, Christian
Camps, Céline
Terrier, Nancy
Moncomble, Dominique
Dunlevy, Jake D
Davies, Christopher
Boss, Paul K
Delrot, Serge
BMC Plant Biology
The composition of grapevine berry at harvest is a major determinant of wine quality. Optimal oenological maturity of berries is characterized by a high sugar/acidity ratio, high anthocyanin content in the skin, and low astringency. However, harvest time is still mostly determined empirically, based on crude biochemical composition and berry tasting. In this context, it is interesting to identify genes that are expressed/repressed specifically at the late stages of ripening and which may be used as indicators of maturity.
Whole bunches and berries sorted by density were collected in vineyard on Chardonnay (white cultivar) grapevines for two consecutive years at three stages of ripening (7-days before harvest (TH-7), harvest (TH), and 10-days after harvest (TH+10)). Microvinification and sensory analysis indicate that the quality of the wines made from the whole bunches collected at TH-7, TH and TH+10 differed, TH providing the highest quality wines.
In parallel, gene expression was studied with Qiagen/Operon microarrays using two types of samples, i.e. whole bunches and berries sorted by density. Only 12 genes were consistently up- or down-regulated in whole bunches and density sorted berries for the two years studied in Chardonnay. 52 genes were differentially expressed between the TH-7 and TH samples. In order to determine whether these genes followed a similar pattern of expression during the late stages of berry ripening in a red cultivar, nine genes were selected for RT-PCR analysis with Cabernet Sauvignon grown under two different temperature regimes affecting the precocity of ripening. The expression profiles and their relationship to ripening were confirmed in Cabernet Sauvignon for seven genes, encoding a carotenoid cleavage dioxygenase, a galactinol synthase, a late embryogenesis abundant protein, a dirigent-like protein, a histidine kinase receptor, a valencene synthase and a putative S-adenosyl-L-methionine:salicylic acid carboxyl methyltransferase.
This set of up- and down-regulated genes characterize the late stages of berry ripening in the two cultivars studied, and are indirectly linked to wine quality. They might be used directly or indirectly to design immunological, biochemical or molecular tools aimed at the determination of optimal ripening in these cultivars.
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Use of diffusion magnetic resonance imaging to correlate the developmental changes in grape berry tissue structure with water diffusion patterns
Dean, Ryan J
Stait-Gardner, Timothy
Clarke, Simon J
Rogiers, Suzy Y
Bobek, Gabriele
Price, William S
Plant Methods
Over the course of grape berry development, the tissues of the berry undergo numerous morphological transformations in response to processes such as water and solute accumulation and cell division, growth and senescence. These transformations are expected to produce changes to the diffusion of water through these tissues detectable using diffusion magnetic resonance imaging (MRI). To assess this non-invasive technique diffusion was examined over the course of grape berry development, and in plant tissues with contrasting oil content.
In this study, the fruit of Vitis vinfera L. cv. Semillon at seven different stages of berry development, from four weeks post-anthesis to over-ripe, were imaged using diffusion tensor and transverse relaxation MRI acquisition protocols. Variations in diffusive motion between these stages of development were then linked to known events in the morphological development of the grape berry. Within the inner mesocarp of the berry, preferential directions of diffusion became increasingly apparent as immature berries increased in size and then declined as berries progressed through the ripening and senescence phases. Transverse relaxation images showed radial striation patterns throughout the sub-tissue, initiating at the septum and vascular systems located at the centre of the berry, and terminating at the boundary between the inner and outer mesocarp. This study confirms that these radial patterns are due to bands of cells of alternating width that extend across the inner mesocarp. Preferential directions of diffusion were also noted in young grape seed nucelli prior to their dehydration. These observations point towards a strong association between patterns of diffusion within grape berries and the underlying tissue structures across berry development. A diffusion tensor image of a post-harvest olive demonstrated that the technique is applicable to tissues with high oil content.
This study demonstrates that diffusion MRI is a powerful and information rich technique for probing the internal microstructure of plant tissues. It was shown that macroscopic diffusion anisotropy patterns correlate with the microstructure of the major pericarp tissues of cv. Semillon grape berries, and that changes in grape berry tissue structure during berry development can be observed.
Development; Diffusion anisotropy; Diffusion tensor imaging; Grape berry; Nucellus; Nuclear magnetic resonance imaging; Olive; Seeds; Striation patterns; Vitis vinifera
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Transcript and metabolite analysis in Trincadeira cultivar reveals novel information regarding the dynamics of grape ripening
Fortes, Ana M
Agudelo-Romero, Patricia
Silva, Marta S
Ali, Kashif
Sousa, Lisete
Maltese, Federica
Choi, Young H
Grimplet, Jerome
Martinez- Zapater, José M
Verpoorte, Robert
Pais, Maria S
BMC Plant Biology
Grapes (Vitis vinifera L.) are economically the most important fruit crop worldwide. However, the complexity of molecular and biochemical events that lead to the onset of ripening of nonclimacteric fruits is not fully understood which is further complicated in grapes due to seasonal and cultivar specific variation. The Portuguese wine variety Trincadeira gives rise to high quality wines but presents extremely irregular berry ripening among seasons probably due to high susceptibility to abiotic and biotic stresses.
Ripening of Trincadeira grapes was studied taking into account the transcriptional and metabolic profilings complemented with biochemical data. The mRNA expression profiles of four time points spanning developmental stages from pea size green berries, through véraison and mature berries (EL 32, EL 34, EL 35 and EL 36) and in two seasons (2007 and 2008) were compared using the Affymetrix GrapeGen® genome array containing 23096 probesets corresponding to 18726 unique sequences. Over 50% of these probesets were significantly differentially expressed (1.5 fold) between at least two developmental stages. A common set of modulated transcripts corresponding to 5877 unigenes indicates the activation of common pathways between years despite the irregular development of Trincadeira grapes. These unigenes were assigned to the functional categories of "metabolism", "development", "cellular process", "diverse/miscellanenous functions", "regulation overview", "response to stimulus, stress", "signaling", "transport overview", "xenoprotein, transposable element" and "unknown". Quantitative RT-PCR validated microarrays results being carried out for eight selected genes and five developmental stages (EL 32, EL 34, EL 35, EL 36 and EL 38). Metabolic profiling using 1H NMR spectroscopy associated to two-dimensional techniques showed the importance of metabolites related to oxidative stress response, amino acid and sugar metabolism as well as secondary metabolism. These results were integrated with transcriptional profiling obtained using genome array to provide new information regarding the network of events leading to grape ripening.
Altogether the data obtained provides the most extensive survey obtained so far for gene expression and metabolites accumulated during grape ripening. Moreover, it highlighted information obtained in a poorly known variety exhibiting particular characteristics that may be cultivar specific or dependent upon climatic conditions. Several genes were identified that had not been previously reported in the context of grape ripening namely genes involved in carbohydrate and amino acid metabolisms as well as in growth regulators; metabolism, epigenetic factors and signaling pathways. Some of these genes were annotated as receptors, transcription factors, and kinases and constitute good candidates for functional analysis in order to establish a model for ripening control of a non-climacteric fruit.
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An in vivo Experimental System to Study Sugar Phloem Unloading in Ripening Grape Berries During Water Deficiency Stress
Annals of Botany
An in vivo experimental system—called the 'berry‐cup' technique—was developed to study sugar phloem unloading and the accumulation of sugar in ripening grape berries. The berry‐cup system consists of a single peeled grape berry immersed in a buffer solution in a cup prepared from a polypropylene syringe. A small cross‐incision (2 mm in length) is made on the stylar remnant of a berry during its ripening phase, the skin of the berry then being easily peeled off, exposing the dorsal vascular bundles without damaging either these or the pulp tissue of the berry. The sites of sugar phloem unloading are thus made directly accessible and may be regulated by the buffer solution. In addition, the unloaded photoassimilates are easily transported into the buffer solution in the berry‐cup. With the berry‐cup technique, it takes 60 min to purge the sugar already present in the apoplast, after which the amount of sugar in the buffer solution is a direct measure of the sugar unloading from the grape berry phloem. The optimum times for sampling were 20 or 30 min, depending on the type of experiment. Sugar phloem unloading was significantly inhibited by the inclusion of either 7·5 mm NaF or 2·5 mm PCMB in the buffer solution. This study indicates that sugar phloem unloading in ripening grape berries is via the apoplastic network and that the process requires the input of energy. The system was shown to be an appropriate experimental system with which to study sugar phloem unloading in ripening grape berries, and was applied successfully to the study of berry sugar unloaded from grapevines subjected to water stress. The results showed that water deficiency inhibits sugar unloading in grape berries.
Sugar phloem unloading; 'berry‐cup' technique; ripening grape berry; inhibitors; water deficiency
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Selective defoliation affects plant growth, fruit transcriptional ripening program and flavonoid metabolism in grapevine
Pastore, Chiara
Zenoni, Sara
Fasoli, Marianna
Pezzotti, Mario
Tornielli, Giovanni Battista
Filippetti, Ilaria
BMC Plant Biology
The selective removal of grapevine leaves around berry clusters can improve the quality of ripening fruits by influencing parameters such as the berry sugar and anthocyanin content at harvest. The outcome depends strongly on the timing of defoliation, which influences the source–sink balance and the modified microclimate surrounding the berries. We removed the basal leaves from Vitis vinifera L. cv Sangiovese shoots at the pre-bloom and veraison stages, and investigated responses such as shoot growth, fruit morphology and composition compared to untreated controls. Moreover, we performed a genome-wide expression analysis to explore the impact of these defoliation treatments on berry transcriptome.
We found that pre-bloom defoliation improved berry quality traits such as sugar and anthocyanin content, whereas defoliation at veraison had a detrimental effect, e.g. less anthocyanin and higher incidence of sunburn damage. Genome-wide expression analysis during berry ripening revealed that defoliation at either stage resulted in major transcriptome reprogramming, which slightly delayed the onset of ripening. However, a closer investigation of individual gene expression profiles identified genes that were specifically modulated by defoliation at each stage, reflecting the uncoupling of metabolic processes such as flavonoid biosynthesis, cell wall and stress metabolism, from the general ripening program.
The specific transcriptional modifications we observed following defoliation at different time points allow the identification of the developmental or metabolic processes affected in berries thus deepening the knowledge of the mechanisms by which these agronomical practices impact the final berry ripening traits.
Vitis vinifera; Defoliation; Berry ripening; Transcriptome; Flavonoid; Source-sink balance
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Berry and phenology-related traits in grapevine (Vitis vinifera L.): From Quantitative Trait Loci to underlying genes
Costantini, Laura
Battilana, Juri
Lamaj, Flutura
Fanizza, Girolamo
Grando, Maria Stella
BMC Plant Biology
The timing of grape ripening initiation, length of maturation period, berry size and seed content are target traits in viticulture. The availability of early and late ripening varieties is desirable for staggering harvest along growing season, expanding production towards periods when the fruit gets a higher value in the market and ensuring an optimal plant adaptation to climatic and geographic conditions. Berry size determines grape productivity; seedlessness is especially demanded in the table grape market and is negatively correlated to fruit size. These traits result from complex developmental processes modified by genetic, physiological and environmental factors. In order to elucidate their genetic determinism we carried out a quantitative analysis in a 163 individuals-F1 segregating progeny obtained by crossing two table grape cultivars.
Molecular linkage maps covering most of the genome (2n = 38 for Vitis vinifera) were generated for each parent. Eighteen pairs of homologous groups were integrated into a consensus map spanning over 1426 cM with 341 markers (mainly microsatellite, AFLP and EST-derived markers) and an average map distance between loci of 4.2 cM. Segregating traits were evaluated in three growing seasons by recording flowering, veraison and ripening dates and by measuring berry size, seed number and weight. QTL (Quantitative Trait Loci) analysis was carried out based on single marker and interval mapping methods. QTLs were identified for all but one of the studied traits, a number of them steadily over more than one year. Clusters of QTLs for different characters were detected, suggesting linkage or pleiotropic effects of loci, as well as regions affecting specific traits. The most interesting QTLs were investigated at the gene level through a bioinformatic analysis of the underlying Pinot noir genomic sequence.
Our results revealed novel insights into the genetic control of relevant grapevine features. They provide a basis for performing marker-assisted selection and testing the role of specific genes in trait variation.
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Proteomic Analysis of Grape Berry Cell Cultures Reveals that Developmentally Regulated Ripening Related Processes Can Be Studied Using Cultured Cells
Sharathchandra, Ramaschandra G.
Stander, Charmaine
Jacobson, Dan
Ndimba, Bongani
Vivier, Melané A.
PLoS ONE
This work describes a proteomics profiling method, optimized and applied to berry cell suspensions to evaluate organ-specific cultures as a platform to study grape berry ripening. Variations in berry ripening within a cluster(s) on a vine and in a vineyard are a major impediment towards complete understanding of the functional processes that control ripening, specifically when a characterized and homogenous sample is required. Berry cell suspensions could overcome some of these problems, but their suitability as a model system for berry development and ripening needs to be established first.
Methodology/Principal Findings
In this study we report on the proteomic evaluation of the cytosolic proteins obtained from synchronized cell suspension cultures that were established from callus lines originating from green, véraison and ripe Vitis vinifera berry explants. The proteins were separated using liquid phase IEF in a Microrotofor cell and SDS PAGE. This method proved superior to gel-based 2DE. Principal component analysis confirmed that biological and technical repeats grouped tightly and importantly, showed that the proteomes of berry cultures originating from the different growth/ripening stages were distinct. A total of twenty six common bands were selected after band matching between different growth stages and twenty two of these bands were positively identified. Thirty two % of the identified proteins are currently annotated as hypothetical. The differential expression profile of the identified proteins, when compared with published literature on grape berry ripening, suggested common trends in terms of relative abundance in the different developmental stages between real berries and cell suspensions.
The advantages of having suspension cultures that accurately mimic specific developmental stages are profound and could significantly contribute to the study of the intricate regulatory and signaling networks responsible for berry development and ripening.
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Reciprocity between abscisic acid and ethylene at the onset of berry ripening and after harvest
Sun, Liang
Zhang, Mei
Ren, Jie
Qi, Jianxun
Zhang, Guojun
Leng , Ping
BMC Plant Biology
The ripening of grape berry is generally regulated by abscisic acid (ABA), and has no relationship with ethylene function. However, functional interaction and synergism between ABA and ethylene during the beginning of grape berry ripening (véraison) has been found recently.
The expressions of VvNCED1 encoding 9-cis-epoxycarotenoid dioxygenase (NCED) and VvGT encoding ABA glucosyltransferase were all increased rapidly at the stage of véraison and reached the highest level at 9th week after full bloom. However, VvCYP1 encoding ABA 8'-hydroxylase and VvβG1 encoding berry β-glucosidase are different, whose expression peak appeared at the 10th week after full bloom and in especial VvβG1 remained at a high level till harvest. The VvACO1 encoding 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase, the VvETR2 (ethylene response 2) and VvCTR1 (constitutive triple response 1) had a transient expression peak at pre-véraison, while the VvEIN4 (ethylene insensitive 4) expression gradually increased from the véraison to one week before harvest stage. The above mentioned changes happened again in the berry after harvest. At one week before véraison, double block treatment with NiCl2 plus 1-methylcyclopropene (1-MCP) not only inhibited the release of ethylene and the expression of related genes but also suppressed the transcription of VvNCED1 and the synthesis of ABA which all might result in inhibiting the fruit ripening onset. Treatment with ABA could relieve the double block and restore fruit ripening course. However, after harvest, double block treatment with NiCl2 plus 1-MCP could not suppress the transcription of VvNCED1 and the accumulation of ABA, and also could not inhibit the start of fruit senescence.
The trace endogenous ethylene induces the transcription of VvNCED1 and then the generation of ABA followed. Both ethylene and ABA are likely to be important and their interplaying may be required to start the process of berry ripening. When the level of ABA reached the peak value, part of it will be stored in the form of ABA-GE. While after harvest, abiotic stresses principally (such as dehydration, harvest shock) could induce the transcription of VvNCED1 and the accumulation of ABA, thus starting the process of fruit senescence.
Related Articles
Proteome changes in the skin of the grape cultivar Barbera among different stages of ripening
Negri, Alfredo S
Prinsi, Bhakti
Rossoni, Mara
Failla, Osvaldo
Scienza, Attilio
Cocucci, Maurizio
Espen, Luca
BMC Genomics
Grape ripening represents the third phase of the double sigmoidal curve of berry development and is characterized by deep changes in the organoleptic characteristics. In this process, the skin plays a central role in the synthesis of many compounds of interest (e.g. anthocyanins and aroma volatiles) and represents a fundamental protective barrier against damage by physical injuries and pathogen attacks. In order to improve the knowledge on the role of this tissue during ripening, changes in the protein expression in the skin of the red cultivar Barbera at five different stages from véraison to full maturation were studied by performing a comparative 2-DE analysis.
The proteomic analysis revealed that 80 spots were differentially expressed throughout berry ripening. Applying a two-way hierarchical clustering analysis to these variations, a clear difference between the first two samplings (up to 14 days after véraison) and the following three (from 28 to 49 days after véraison) emerged, thus suggesting that the most relevant changes in protein expression occurred in the first weeks of ripening. By means of LC-ESI-MS/MS analysis, 69 proteins were characterized. Many of these variations were related to proteins involved in responses to stress (38%), glycolysis and gluconeogenesis (13%), C-compounds and carbohydrate metabolism (13%) and amino acid metabolism (10%).
These results give new insights to the skin proteome evolution during ripening, thus underlining some interesting traits of this tissue. In this view, we observed the ripening-related induction of many enzymes involved in primary metabolism, including those of the last five steps of the glycolytic pathway, which had been described as down-regulated in previous studies performed on whole fruit. Moreover, these data emphasize the relevance of this tissue as a physical barrier exerting an important part in berry protection. In fact, the level of many proteins involved in (a)biotic stress responses remarkably changed through the five stages taken into consideration, thus suggesting that their expression may be developmentally regulated.
Related Articles
Generation of a predicted protein database from EST data and application to iTRAQ analyses in grape (Vitis vinifera cv. Cabernet Sauvignon) berries at ripening initiation
Lücker, Joost
Laszczak, Mario
Smith, Derek
Lund, Steven T
BMC Genomics
iTRAQ is a proteomics technique that uses isobaric tags for relative and absolute quantitation of tryptic peptides. In proteomics experiments, the detection and high confidence annotation of proteins and the significance of corresponding expression differences can depend on the quality and the species specificity of the tryptic peptide map database used for analysis of the data. For species for which finished genome sequence data are not available, identification of proteins relies on similarity to proteins from other species using comprehensive peptide map databases such as the MSDB.
We were interested in characterizing ripening initiation ('veraison') in grape berries at the protein level in order to better define the molecular control of this important process for grape growers and wine makers. We developed a bioinformatic pipeline for processing EST data in order to produce a predicted tryptic peptide database specifically targeted to the wine grape cultivar, Vitis vinifera cv. Cabernet Sauvignon, and lacking truncated N- and C-terminal fragments. By searching iTRAQ MS/MS data generated from berry exocarp and mesocarp samples at ripening initiation, we determined that implementation of the custom database afforded a large improvement in high confidence peptide annotation in comparison to the MSDB. We used iTRAQ MS/MS in conjunction with custom peptide db searches to quantitatively characterize several important pathway components for berry ripening previously described at the transcriptional level and confirmed expression patterns for these at the protein level.
We determined that a predicted peptide database for MS/MS applications can be derived from EST data using advanced clustering and trimming approaches and successfully implemented for quantitative proteome profiling. Quantitative shotgun proteome profiling holds great promise for characterizing biological processes such as fruit ripening initiation and may be further improved by employing preparative techniques and/or analytical equipment that increase peptide detection sensitivity via a shotgun approach.
Related Articles
Acyl substrate preferences of an IAA-amido synthetase account for variations in grape (Vitis vinifera L.) berry ripening caused by different auxinic compounds indicating the importance of auxin conjugation in plant development
Böttcher, Christine
Boss, Paul K.
Davies, Christopher
Journal of Experimental Botany
Nine Gretchen Hagen (GH3) genes were identified in grapevine (Vitis vinifera L.) and six of these were predicted on the basis of protein sequence similarity to act as indole-3-acetic acid (IAA)-amido synthetases. The activity of these enzymes is thought to be important in controlling free IAA levels and one auxin-inducible grapevine GH3 protein, GH3-1, has previously been implicated in the berry ripening process. Ex planta assays showed that the expression of only one other GH3 gene, GH3-2, increased following the treatment of grape berries with auxinic compounds. One of these was the naturally occurring IAA and the other two were synthetic, α-naphthalene acetic acid (NAA) and benzothiazole-2-oxyacetic acid (BTOA). The determination of steady-state kinetic parameters for the recombinant GH3-1 and GH3-2 proteins revealed that both enzymes efficiently conjugated aspartic acid (Asp) to IAA and less well to NAA, while BTOA was a poor substrate. GH3-2 gene expression was induced by IAA treatment of pre-ripening berries with an associated increase in levels of IAA-Asp and a decrease in free IAA levels. This indicates that GH3-2 responded to excess auxin to maintain low levels of free IAA. Grape berry ripening was not affected by IAA application prior to veraison (ripening onset) but was considerably delayed by NAA and even more so by BTOA. The differential effects of the three auxinic compounds on berry ripening can therefore be explained by the induction and acyl substrate specificity of GH3-2. These results further indicate an important role for GH3 proteins in controlling auxin-related plant developmental processes.
Auxin; GH3 proteins; ripening; Vitis vinifera
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text: have a john deere 2010 quik trak 23hp kawasaki 169 hours on it. in the mornings it turns over real hard 3rd to 1/4 turns at a time for about 30 seconds and cranks over just fine after that. it will turn over for another 30 seconds to a minutes before it starts to fire and then it spits and sputters for about 10 to 15 second and gas just sprays out, even after it done sputtering and then i shut it down and restart it and all is fine. what wrong carb? chaged oil and it was 50/50 gas to oil in it. i checked oils, valve clearance, spark plugs, tested ignition coils, gas, air filters just started doing it yesterday once it starts it fine. and after each yard it would take a few second to start instead of right away like it has always done. and if i would choke it it would take longer to start and even more gas coming out of muffler.


text:
Relevant offers
Manaia didn't have too many problems against a Huatoki side missing their top two women winning their Soffe Cup interclub tennis match 12-4 in New Plymouth on Saturday.
Huatoki's No 2 woman Lorraine Heerdegen and debutant Hikaru Han at No 3 both had long three setters against higher ranked opponents, before losing to Maree Gopperth and Sarah Kelly respectively.
Sandfords Manaia's top player, Nicole Koch, had no problems against Chris Wilson winning 6-1 6-1, while Manaia's Trudy Vince had a surprise 7-5 6-0 win over the experienced Saori Han.
On the men's side, Manaia's Jordan Fleming and Cameron Koch showed why they are currently ranked one and two in Taranaki, easily dispatching Huatoki's top two men - Andrew Sangster and Chris Popata.
Manaia's Michael Gates at No 3 blunted Alastair Nairn's power, with his consistency, winning 6-3 6-3, while Beauden Fleming at No 4 was easily beaten by Huatoki's Murray Keast. Keast went on to win his doubles with Popata and combined with Hikaru Han to give some respectability to the final match score.
Icon Sports Tariki Pukekura easily accounted for a depleted Central Finance Rotokare side 14-2.
Whizz kid Louise Waite at No 1 was untroubled in her match for Icons Sports Tariki Pukekura, beating Frances Gray 6-0 6-0, while it was business as usual for Kirsten Adam in her 6-1 6-1 win over Beccie Atkins.
Linda Reid had some early problems against hard-hitting Michelle Atkins, finally prevailing 6-4 6-1, while Donna Thorne was taken to the wire by Rotokare's Juliet Farquhar before winning in three sets.
In the men's singles, Pukekura No 1 Callum Old was too consistent for Rotokare's Malcolm Elder, winning 6-2 6-1. Mike Fleming got the better of Kyle Crowley, while Chris Jury at No 3 was too solid for Greg Hareb.
At No 4, Pukekura's Jamie Simpson countered Shane Devlin's big serve to win 6-1 6-3.
Rotokare's wins were a tough three setter in the men's bottom doubles and a top combines win.
Oakura won a close contest 9-7 against Okaiawa at Okaiawa. After being 8-4 up after the singles and doubles, Hareb Deken Oakura could have been excused for thinking they had the match in the bag.
Farmlands Okaiawa siblings Vincent and Jamie Baylis came back from 8-3 down to snatch a thrilling 9-8 victory over Len Adamson and Chris Davies in the No 3 combines to give their team a chance. But the fourth combines went to Oakura's Craig Waite and Miana Walden to seal the win.
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To skip the usual: Yes, I'm black. No, I don't reinforce the stereotype. And yes, I have blue eyes... well, blue/green/silver (they change depending on my mood.)
The fastest way to my heart (or my pants) is through my mind and I'm looking for someone that can keep me intrigued by the things that go through theirs. So seriously, talk to me about the random stuff going through your head. Ask me random questions. Pretend like we've been friends for years. Just for the love of god please don't try to do small talk. Because I REALLY hate small talk.
I'm kind of a difficult person to explain so if you really want to get to know me you'll have to message me and find out.
Poetry, analysis, loyalty, honesty, creating stories, listening, running/hiding/sneaking (we play manhunt a lot), remembering random bits of information, looking out for people, being a smart ass, debating with people, singing, and making people laugh.
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David Lytton-Cobbold, 2nd Baron Cobbold
|This biography of a living person needs additional citations for verification. (February 2011) (Learn how and when to remove this template message)
David Antony Fromanteel Lytton-Cobbold, 2nd Baron Cobbold,[1] DL (born 14 July 1937) is a British hereditary peer and former member of the House of Lords. He was one of the ninety hereditary peers elected to remain in the House of Lords[2] after the passing of the House of Lords Act 1999. A runner-up in the initial election that year, he replaced Myrtle Robertson, 11th Baroness Wharton following her death on 15 October 2000. On 13 October 2014, he became only the second person to resign his membership of the House under the House of Lords Reform Act 2014, and the first elected hereditary peer to do so.[3]
Education and career[edit]
In keeping with family tradition, Cobbold was educated at Eton College and Cambridge University.
David Cobbold was an executive in the International Banking Department of Bank of London and South America (BOLSA) in the late 1960s, under Edward Clifton-Brown. BOLSA was one of the first banks in the Eurodollar market, developed by Sir George Bolton, Chairman of BOLSA. He was increasingly drawn into the management of Knebworth House, for public events,[4] so he left banking and dedicated himself to the house and estate.
Personal life[edit]
Lytton-Cobbold inherited the title Baron Cobbold on 1 November 1987. He married Christine Elizabeth Stucley, on 7 January 1961; they had four children. His heir, and current occupier of the family seat Knebworth House, is Henry Lytton-Cobbold.
He was born David Antony Fromanteel Cobbold, but changed his name by deed poll on 4 December 1960. He is a member of the Lytton family (Earls of Lytton) through his mother.
- ^ Kidd, Charles; Shaw, Christine (2008-07-25). Debrett's peerage & baronetage 2008. Debrett's Limited. p. 299. ISBN 978-1-870520-80-5. Retrieved 20 February 2011.
- ^ Schaefer, Sarah (30 March 1999). "Parliament: House Of Lords: Peers begin two-day debate on `radical, historic' reform". The Independent. Retrieved 20 February 2011.
- ^ http://www.parliament.uk/mps-lords-and-offices/lords/retired-lords/
- ^ Lewis, Dave (2004). Led Zeppelin: The 'Tight But Loose' Files; Celebration 2. Omnibus Press. pp. 65–. ISBN 978-1-84449-056-1. Retrieved 20 February 2011.
|Peerage of the United Kingdom
|Preceded by
Cameron Cobbold
|Baron Cobbold
Heir apparent:
Hon. Henry Lytton-Cobbold
|This biography of a baron in the peerage of the United Kingdom is a stub. You can help Wikipedia by expanding it.


text:
These are all the News & Announcements posted on Monday, 17 September, 2012.
Photo Scavenger Hunt on Facebook
This morning we're holding a photo scavenger hunt on our Facebook page with the chance to win great deals from Jack's Wood Fired Oven! Head on over to our Facebook page to check it out!
Posted By Logan Library at 10:13 AM in Category:Facebook
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