共查询到20条相似文献,搜索用时 31 毫秒
1.
A Protein from Immature Raspberry Fruits which Inhibits Endopolygalacturonases from Botrytis cinerea and other Micro-organisms 总被引:2,自引:0,他引:2
A polygalacturonase-inhibiting protein (PGIP) was purified fromimmature raspberry fruits using ion exchange chromatography.The protein was composed of a single polypeptide chain withMr of 38·5 kDa and a pI residing above pH 10. Kineticstudies suggested that the inhibition was of a non-competitivenature. The PGIP inhibited two endopolygalacturonases (endo-PG)purified from Botrytis cinerea and an endo-PG produced by Aspergillusniger to varying degrees but did not inhibit two exo-PGs purifiedfrom B. cinerea, bacterial endopectate lyases and bacterialendo-PGs. The concentration of PGIP at various stages of flowerand fruit development was determined. The inhibitor was notdetected in the flower, but reached a maximum of 69 units g1in the immature green fruit decreasing to 9 units g1as fruits matured. The N-terminal amino-acid sequence was determined. Key words: Polygalacturonase-inhibiting protein, Rubus idaeus, red raspberry, Botrytis cinerea, pectinases 相似文献
2.
The activities of four fungal polygalacturonases (endo-PGI,endo-PGII, exo-PGI, exo-PGII), detected when Botrytis cinereawas grown on immature fruits of red raspberry (Rubus idaeus),were fractionated into soluble and wall-bound fractions. Westernblots and plate-trapped antigen ELISA showed that endo-PGI andendo-PGII were most abundant in the cell wall-bound fractionsof the host. Immuno-inhibition studies using a polyclonal antiserumagainst polygalacturonase-inhibiting protein (PGIP), purifiedfrom immature raspberry fruits, showed that the low level offungal PG activity detected in fractions containing endo-PGIwas due to the presence of PGIP. When a purified preparationof endo-PGI and endo-PGII from B. cinerea was allowed to reactin vitro with either a crude host cell wall preparation, orone which had previously been treated to remove cell wall-boundproteins, both endo-PG isozymes had a greater binding capacitytowards the former wall preparation. Endo-PGI and endo-PGIIalso had an affinity for fungal cell walls. Exo-PGI and exo-PGIIbound to both fungal and host cell walls. Greater quantitiesof fungal endo-PGs were detected by ELISA in fruits previouslyfrozen and thawed (freeze-thawed) and inoculated,than in fresh inoculated fruit. This result paralleled the extentof fungal growth in these tissues as assessed by chitin assayand suggests that the resistance shown by raspberries is dependenton continual replacement of inhibitory substances or inducedresistance mechanisms. Key words: Polygalacturonase-inhibiting protein, Rubus idaeus, red raspberry, Botrytis cinerea, pectinases 相似文献
3.
Richter A Jacobsen HJ de Kathen A de Lorenzo G Briviba K Hain R Ramsay G Kiesecker H 《Plant cell reports》2006,25(11):1166-1173
The pea (Pisum sativum L.) varieties Baroness (United Kingdome) and Baccara (France) were transformed via Agrobacterium tumefaciens-mediated gene transfer with pGPTV binary vectors containing the bar gene in combination with two different antifungal genes coding for polygalacturonase-inhibiting protein (PGIP) from raspberry (Rubus idaeus L.) driven by a double 35S promoter, or the stilbene synthase (Vst1) from grape (Vitis vinifera L.) driven by its own elicitor-inducible promoter. Transgenic lines were established and transgenes combined via conventional crossing. Resveratrol, produced by Vst1 transgenic plants, was detected using HPLC and the PGIP expression was determined in functional inhibition assays against fungal polygalacturonases. Stable inheritance of the antifungal genes in the transgenic plants was demonstrated. 相似文献
4.
5.
Robert VanBuren Doug Bryant Jill M. Bushakra Kelly J. Vining Patrick P. Edger Erik R. Rowley Henry D. Priest Todd P. Michael Eric Lyons Sergei A. Filichkin Michael Dossett Chad E. Finn Nahla V. Bassil Todd C. Mockler 《The Plant journal : for cell and molecular biology》2016,87(6):535-547
Black raspberry (Rubus occidentalis) is an important specialty fruit crop in the US Pacific Northwest that can hybridize with the globally commercialized red raspberry (R. idaeus). Here we report a 243 Mb draft genome of black raspberry that will serve as a useful reference for the Rosaceae and Rubus fruit crops (raspberry, blackberry, and their hybrids). The black raspberry genome is largely collinear to the diploid woodland strawberry (Fragaria vesca) with a conserved karyotype and few notable structural rearrangements. Centromeric satellite repeats are widely dispersed across the black raspberry genome, in contrast to the tight association with the centromere observed in most plants. Among the 28 005 predicted protein‐coding genes, we identified 290 very recent small‐scale gene duplicates enriched for sugar metabolism, fruit development, and anthocyanin related genes which may be related to key agronomic traits during black raspberry domestication. This contrasts patterns of recent duplications in the wild woodland strawberry F. vesca, which show no patterns of enrichment, suggesting gene duplications contributed to domestication traits. Expression profiles from a fruit ripening series and roots exposed to Verticillium dahliae shed insight into fruit development and disease response, respectively. The resources presented here will expedite the development of improved black and red raspberry, blackberry and other Rubus cultivars. 相似文献
6.
Kemp G Bergmann CW Clay R Van der Westhuizen AJ Pretorius ZA 《Molecular plant-microbe interactions : MPMI》2003,16(11):955-961
Evidence for the presence of a polygalacturonase-inhibiting protein (PGIP) from a monocotyledonous cereal is presented. A 40.3-kDa PGIP that was closely associated with the cell wall was acetone-extracted and purified from wheat (Triticum aestivum L.) leaves and stems. Wheat PGIP exhibited a highly selective inhibitory activity against endopolygalacturonase (EPG) from various fungi. Of nine EPG tested, wheat PGIP only inhibited EPG from Cochliobolus sativus, a pathogen of the tribe Poaceae. A short N-terminal amino acid sequence of wheat PGIP shows no similarity to any other characterized PGIP. 相似文献
7.
8.
Chris S. Jones Pietro P. M. Iannetta Mary Woodhead Howard V. Davies Ronnie J. McNicol Mark A. Taylor 《Molecular biotechnology》1997,8(3):219-221
Previous attempts to extract high-quality, total RNA from raspberry (Rubus idaeus) fruits using published protocols have proven to be unsuccessful. Even the use of protocols developed for the extraction
of RNA from other fruit tissue has resulted in low yields (1) or the isolation of degraded RNA (2). Here, we report on the development of a quick and simple method of extracting total RNA from raspberry fruit. Using this
method, high yields of good quality, undegraded RNA were obtained from fruit at all stages of ripening. The RNA is of sufficient
quality for northern analysis and cDNA library construction. 相似文献
9.
10.
The production of ethylene by red raspberry (Rubus idaeus L.cv. Glen Clova) fruit increased climacterically during development.The concentration of ethylene within green fruit was low butincreased substantially as fruit abscission and ripening commenced.The receptacle contained higher concentrations than the drupeletsat all stages measured. In the mature ripening fruit the ethyleneconcentrations were found to be physiologically significant,and would accelerate the abscission of large green non-abscisingfruit if supplied as a fumigant. The addition of ethylene toripe fruit did not accelerate abscission, probably because saturatinglevels occurred naturally within these fruit. Reduction of ethylenesynthesis rates using the inhibitor of ethylene production aminoethoxyvinylglycine(AVG) reduced the rate of abscission zone weakening which occursin detached large green fruit. The rate of ethylene productionwas found to be dependent on the supply of the precursor l-aminocyclopropane-l-carboxylicacid (ACC). This only accumulated to any extent in those ripefruit with high rates of ethylene production. Rubus idaeus, raspberry, abscission, fruit ripening, ethylene, aminocyclopropane-l-carboxylic acid 相似文献
11.
SEXTON ROY; PALMER JANE M.; WHYTE NICHOLA A.; LITTLEJOHNS SUSAN 《Annals of botany》1997,80(3):371-376
The ripening of raspberry fruit (Rubus ideausL. cv Glen Clova)is associated with a climacteric rise in ethylene production.As the fruit pigments change from green to red there is a progressivesoftening, loss of skin strength and a breakdown of cell wallsin the mesocarp. An increase in cellulase (endo-1,4-ß-D-glucanase)in both drupelets and receptacles accompanies these changes.The localization of cellulase in the regions of the fruit associatedwith abscission zones suggest the enzyme may be involved infruit separation as well as softening. Rubus idaeusL; raspberry; fruit ripening; ethylene; abscission; cell wall breakdown; cellulase; endo-1,4-ß-D-glucanase 相似文献
12.
Daniel J Sargent Felicidad Fernández-Fernández Alicja Rys Victoria H Knight David W Simpson Kenneth R Tobutt 《BMC plant biology》2007,7(1):15
Background
Raspberry breeding programmes worldwide aim to produce improved cultivars to satisfy market demands and within these programmes there are many targets, including increased fruit quality, yield and season, and improved pest and disease resistance and plant habit. The large raspberry aphid, Amphorophora idaei, transmits four viruses and vector resistance is an objective in raspberry breeding. The development of molecular tools that discriminate between aphid resistance genes from different sources will allow the pyramiding of such genes and the development of raspberry varieties with superior pest resistance. We have raised a red raspberry (Rubus idaeus) F1 progeny from the cross 'Malling Jewel' × 'Malling Orion' (MJ × MO), which segregates for resistance to biotype 1 of the aphid Amphorophora idaei and for a second phenotypic trait, dwarf habit. These traits are controlled by single genes, denoted (A 1) and (dw) respectively. 相似文献13.
Purification and molecular characterization of a soybean polygalacturonase-inhibiting protein 总被引:14,自引:0,他引:14
A polygalacturonase-inhibiting protein (PGIP) was detected in soybean (Glycine max (L.) Merr.) seedlings. The protein was purified from germinating seeds and appeared to consist of at least three components with very close molecular weights (between 37 and 40 kDa) but each showing a unique N-terminal sequence. Primers specific for N-terminal and C-terminal nucleotide sequences of field bean (Phaseolus vulgaris L.) PGIP were used in a polymerase chain reaction (PCR) on soybean DNA, and only one amplification band was obtained. The amplified product was cloned and one of the PCR clones was sequenced. The nucleotide sequence comprises 942 bp with a single open reading frame which encodes a polypeptide of 313 amino-acid residues with a predicted molecular weight of 33984 Daltons and an isoelectric point of 8.21. Analysis of genome organization showed a single gene copy of PGIP with few related sequences, and wounding of soybean hypocotyls showed a strong induction of expression of the PGIP gene. The PGIP showed different activities toward three purified fungal endo-polygalacturonases (endo-PGs) (two endoPGs from Sclerotinia sclerotiorum and one endo-PG from Aspergillus niger). A possible involvement of soybean PGIP in plant defence against fungal pathogens is discussed. 相似文献
14.
Szankowski I Briviba K Fleschhut J Schönherr J Jacobsen HJ Kiesecker H 《Plant cell reports》2003,22(2):141-149
The objective of the present research was to introduce genes with antifungal potential into the commercially important apple cvs. Elstar
and Holsteiner Cox in order to establish resistance against fungal diseases. The gene encoding the stilbene synthase
(Vst1) from Vitis vinifera L., responsible for the synthesis of the phytoalexin resveratrol in grapevine, and the gene for a polygalacturonase-inhibiting protein (PGIP) from kiwi (Actinidia deliciosa) were transferred into Holsteiner Cox
and Elstar via Agrobacterium tumefaciens-mediated transformation. A total of nine transgenic Holsteiner Cox clones and one transgenic E clone carrying the stilbene-synthase gene as well as three transgenic Holsteiner Cox lines harbouring the polygalacturonase-inhibiting protein from Kiwi were identified via polymerase chain reaction and Southern blot analysis. High performance liquid chromatography analysis revealed the accumulation of a resveratrol-derivate, a glycoside, in transgenic Vst1 plants.Abbreviations BAP
6-Benzylaminopurine
- E
Elstar
- H
Holsteiner Cox
- HPLC
High performance liquid chromatography
- IBA
Indole-3-butyric acid
- NAA
-Naphthaleneacetic acid
- TDZ
Thidiazuron
- YEP
Yeast extract broth
Communicated by H. Lörz 相似文献
15.
Bertazzon N Raiola A Castiglioni C Gardiman M Angelini E Borgo M Ferrari S 《Plant cell reports》2012,31(1):133-143
Grapevine is an economically important crop, and the recent completion of its genome makes it possible to study the function
of specific genes through reverse genetics. However, the analysis of gene function by RNA interference (RNAi) in grapevine
is difficult, because the generation of stable transgenic plants has low efficiency and is time consuming. Recently, transient
expression of genes in grapevine leaves has been obtained by Agrobacterium tumefaciens infiltration (agroinfiltration). We therefore tested the possibility to silence grapevine genes by agroinfiltration of RNAi
constructs. A construct to express a double strand RNA (dsRNA) corresponding to the defense-related gene VvPGIP1, encoding a polygalacturonase-inhibiting protein (PGIP), was obtained and transiently expressed by agroinfiltration in leaves
of grapevine plants grown in vitro. Expression of VvPGIP1 and accumulation of PGIP activity were strongly induced by infiltration with control bacteria, but not with bacteria carrying
the dsRNA construct, indicating that the gene was efficiently silenced. In contrast, expression of another defense-related
gene, VST1, encoding a stilbene synthase, was unaffected by the dsRNA construct. We have therefore demonstrated the possibility of transient
down-regulation of grapevine genes by agroinfiltration of constructs for the expression of dsRNA. This system can be employed
to evaluate the effectiveness of constructs that can be subsequently used to generate stable RNAi transgenic plants. 相似文献
16.
为了探讨氟虫脲可能的作用靶标及毒性机制, 本研究以重要农业害虫东亚飞蝗Locusta migratoria manilensis (Meyen)和中华稻蝗Oxya chinensis (Thunberg)为材料, 采用简并引物扩增中华稻蝗几丁质合成酶1基因(OcCHS1)的部分cDNA序列; 以氟虫脲浸渍法处理2龄中期中华稻蝗及1, 2和3龄东亚飞蝗若虫为处理组, 丙酮处理为对照组, 使用RT-PCR和实时荧光定量PCR方法分析氟虫脲对蝗虫几丁质合成酶基因mRNA表达的影响。结果获得的OcCHS1部分cDNA序列, 其长度为312 bp, 编码104个氨基酸, GenBank登录号为HM214491, 与东亚飞蝗几丁质合成酶1基因(LmCHS1)在氨基酸水平上相似度达95%。RT-PCR结果显示, 处理组几丁质合成酶1扩增带均强于对照组。实时荧光定量PCR结果表明: 与对照组相比, 处理组中华稻蝗2龄中期若虫OcCHS1 mRNA表达提高了1.02倍, 东亚飞蝗1, 2, 3龄若虫LmCHS1 mRNA表达分别提高了34%, 82%和89%, 差异显著(P<0.05)。分析基因表达提高的原因是几丁质合成受阻后基因表达水平的一种代偿性增加, 由此推测几丁质合成酶可能是氟虫脲作用的靶标之一。 相似文献
17.
Construction,characterization, and screening of a transformation-competent artificial chromosome library of peach 总被引:7,自引:1,他引:6
Feng Shan Liang Kai Chun Zhang Zhan Wang Yu Ji Liang Yang Xiao Ming Zhang Jin De Min Bin Wang 《Plant Molecular Biology Reporter》2004,22(1):37-48
As a genome model of fruit trees, peach (Prunus persica [L.] Batch) has advantages for studying structural and functional genomics. Okubo, a traditional peach variety used as a
parent in Asian peach breeding, displays economically valuable agronomic traits. To develop an efficient platform for peach
gene cloning and genomic research, a large-insert genomic DNA library of Okubo was constructed in a transformation-competent
artificial chromosome (TAC) vector, pYLTAC7, which can accept and stably maintain large genomic DNA fragments in bothEscherichia coli andAgrobacterium tumefaciens. The TAC library contains 41,472 recombinant clones with an average insert size of approximately 42 kb, and it is equivalent
to 6 haploid peach genomes. The TAC library was stored in 2 ways: one copy as frozen cultures in 108 pieces of 384-well plates
and another copy as bulked pools in 36 pieces of 96-well plates, each well containing 12 individual clones. The lack of hybridization
signal to chloroplast and mitochondrial genes indicated that the TAC library had no significant cytoplast organelle DNA contamination.
TAC clones were stable inE. coli cells until generation 100 and stable in bothE. coli andA. tumefaciens. Twenty-one clones containing the polygalacturonase-inhibiting protein (PGIP) gene were detected by using pooled PCR in the
TAC library. Positive clones can be used for peach PGIP gene cloning and functional analysis. The library is well suited for
gene cloning and genetic engineering in peach. 相似文献
18.
19.
Inactivation of 1-aminocyclopropane-1-carboxylate (ACC) oxidase 总被引:1,自引:0,他引:1
Smith Julian J.; Zhang Zhi Hong; Schofield Christopher J.; John Philip; Baldwin Jack E. 《Journal of experimental botany》1994,45(5):521-527
The enzyme 1-aminocyclopropane-1-carboxylate (ACC) oxidase,which catalyses the final step in the biosynthesis of ethylene,showed a non-linear time-course in vitro and activity decayedwith a half-life of around 14 min. This loss of activity wasstudied using tomato ACC oxidase purified from Escherichia coiltransformed with the cDNA clone pTOM13. Inactivation was notdue to end-product inhibition by dehydroascorbic acid or cyanide.Preincubatlon of enzyme in the combined presence of Fe2+ ascorbateand ACC, which together allowed catalytic turnover, resultedin almost total loss of ACC oxidase activity. Enzyme Inactivatedby catalysis could not be reactivated by passage through SephadexG-25 or by treating with combina tions of DTT and CO2 A non-lineartime-course and inactivation in the presence of all substratesand cofactors was also shown for the enzyme assayed in vivowith melon fruit discs. Using the purified tomato enzyme a distinctascorbate-dependent inactivation was also observed, which occurredIn the absence of catalysis and was prevented, although notreversed, by catalase. This ascorbate-dependent inactivationmay thus be due to H2O2 attack on ACC oxidase. Key words: 1-aminocyclopropane-1-carboxylate (ACC) oxidase, catalase, catalytic inactivation, ethylene 相似文献
20.