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1.
Identification,isolation and characterization of a CC-NBS-LRR candidate disease resistance gene family in grapevine 总被引:2,自引:0,他引:2
Andreas Kortekamp Leocir Welter Sarah Vogt Alexander Knoll Florian Schwander Reinhard Töpfer Eva Zyprian 《Molecular breeding : new strategies in plant improvement》2008,22(3):421-432
2.
To achieve higher level expression of Interferon α2b (IFN-α2b) in methylotrophic yeast (Pichia pastoris), a cDNA fragment coding for the mature IFN-α2b was designed and synthesized based on the synonymous codon bias of P. pastoris and optimized G+C content. The synthetic IFN-α2b was inserted into the secreted expression vector pPICZαA, and then integrated
into P. pastoris GS115 genome by electroporation. Multi-copy integrants in the Mut+ recombinant P. pastoris strain were screened by high concentrations of Zeocin. 120 hours culturing allowed expression of the IFN-α2b transformant
up to 810 mg/L as detected by SDS-PAGE and quantitative methods. In addition, Western blot analysis showed that the recombinant
proteins had immunogenicity. The significant antiviral activity of the recombinant IFN-α2b protein was verified by WISH/ VSV
system, which was 3.3×105 IU/mL.
Foundation items: The National ‘973’ Basic Research Program (2002CB111302); The National Natural Science Foundation of China
(30370807) 相似文献
3.
J. G. Roddick G. Melchers 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1985,70(6):655-660
Summary Analyses of leaves and ‘tubers’ from somatic hybrids of potato and tomato (‘pomato’ with plastids of potato, ‘topato’ with
plastids of tomato) produced by fusion of protoplasts from liquid cultures of dihaploid potato and mesophyll of tomato revealed
the presence of the two major potato glycoalkaloids (α-solanine and α-chaconine) as well as the tomato glycoalkaloid (αtomatine). The total alkaloid content of leaves was greater than that of ‘tubers’ and similar to levels in the foliage of
parent plants. However, glycoalkaloids were more abundant in hybrid ‘tubers’ than in normal potato tubers by a factor of 5–15.
In hybrid foliage, approximately 98% of the alkaloid present was of potato origin whereas in ‘tubers’ the reverse was the
case, with tomatine comprising 60–70% of the total alkaloid. The similarities in alkaloid content and ratios between the pomato
and the topato lines indicate that plastomes do not influence the biosynthesis and distribution of these alkaloids. The results
indicate that major secondary metabolites may prove useful for assessing the hybrid nature of such plants. 相似文献
4.
Peroxidase activity of red raspberry canes was dependent on the cultivar and influenced the subsequent lignification. After
inoculation with Didymella applanata, responsible for the spur blight cane disease, the activity of soluble cytoplasmic enzyme increased in the moderately resistant
‘Latham’ and susceptible ‘Malling Promise’, similarly for syringaldazine and guaiacol as hydrogen donors. Systemic induction
found in ‘Latham’ was recognized as a symptom of defence mechanism responsible for fungal restriction. Locally enhanced peroxidase
activity in the ‘M.Promise’ tissues was related to the local lignification and/or may be associated with the loss of cell
integrity caused by pathogen penetration. Pathogen-induced changes of cell wall peroxidases were similar in both cultivars
mentioned above. No influence of the infection was found in the high susceptible Zeva cultivar.
Using native-PAGE analysis and horizontal starch electrophoresis of soluble fraction five constitutive acidic isoperoxidases
were detected in ‘Latham’ and three in ‘M. Promise’. The infection process was accompanied by the appearance of two new anodic
isoforms. 相似文献
5.
Q. Li X. M. Chen D. Li W. D. Zhang J. C. Tian 《Russian Journal of Plant Physiology》2011,58(4):686-695
Wheat powdery mildew is caused by Blumeria graminis f. sp. tritici (Bgt). Pm21 is an effective broad-spectrum powdery mildew resistance gene, which shows a considerable promise in wheat breeding. We report
here a proteomic approach to investigate the resistance response proteins after fungal infection and emphasize the resistance
changes induced by Pm21. Two wheat (Triticum aestivum L.) near-isogenic lines (NILs), recurrent parent ‘Bainong,’ which is susceptible to powdery mildew, and its near-isogenic
line ‘W2132’ carrying resistance gene Pm21) were used to investigate some changes in their proteomes after being infected. Proteins were extracted from the leaves sampled
in 48 h after inoculation, separated by two-dimensional electrophoresis, and stained with Coomassie brilliant blue. Among
these proteins, a total of 56 spots differentially expressed after Bgt infection were detected. Sixteen proteins, identified
by MALDI-TOF-MS, exhibited more than a 1.5-fold increase upon fungal infection. Unfortunately, three spots were not identified
successfully. The predicted functions of identified proteins were related to energy metabolism and defensive responses; they
were involved in many physiological resistance responses, including enhancing energy metabolism, proteins synthesis and stabilization,
antioxidant reactions, cell-wall reinforcement, and lignification. Interestingly that the expression of two proteins related
to the cell-wall reinforcement was enhanced in the resistant line and one protein related to photosynthesis was lost in a
susceptible line. By transmission electronic microscopy, the corresponding physiological characteristics were also observed.
These results provide us with the information to further reveal the resistance mechanism of Pm21 action and comprehensively investigate the physiological response to powdery mildew at the protein level. 相似文献
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‘Galia’ muskmelon (Cucumis melo L. var. reticulatus Ser.) has been recalcitrant to transformation by Agrobacterium tumefaciens. Transformation of the ‘Galia’ male parental line, ‘Krymka’, with an ACC oxidase (CMACO-1) gene in antisense orientation
is described herein. Explants were transformed using A. tumefaciens strain ABI, which contained a vector pCmACO1-AS plasmid, bearing an antisense gene of CMACO-1 and the CP4 syn gene (glyphosate-tolerance). Both CMACO-1 and CP4 syn genes were assessed by a polymerase chain reaction method. Flow cytometry analysis was performed to determine plant ploidy
level of primary transformants. Two completely diploid independent transgenic plants were obtained. Southern blot and segregation
analysis in the T1 generation determined that each independent transgenic line had one single insertion of the transgene. These transgenic muskmelon
male parental lines have potential for use in the production of ‘Galia’ F1 hybrids with improved shelf life. 相似文献
10.
Reported crystallographic data and calculated molecular models indicated that chlorophyll (Chl) a and bacteriochlorophyll (BChl) a tend to bind the fifth ligand on the side of the macrocycle where the C132-(R)-methoxycarbonyl moiety protrudes (denoting the ‘back’ side). The crystal structures of 34 photosynthetic proteins possessing
(B)Chl cofactors revealed that most of Chl a and BChl a (and b) are coordinated by any peptidyl residue (e.g., histydyl-imidazolyl group), peptidyl backbone or water from the ‘back’ side.
Almost all the cofactors that bind a water molecule as the fifth ligand in these proteins have a ‘back’ configuration. Theoretical
model calculations for methyl chlorophyllide a (MeChlid a) and methyl bacteriochlorophyllide a (MeBChlid a) bound to an imidazole molecule indicated that the ‘back’ side is energetically favored for the ligand binding. These results
are consistent with the fact that ethyl chlorophyllide a (EtChlid a) dihydrate crystal consists of the ‘back’ complex. The modeling also showed that both removal and stereochemical inverse
of the C132-methoxycarbonyl group affect the relative stability between the ‘back’ and ‘face’ complexes. The effect of the C132-moiety on the choice of the macrocycle side for the ligand binding is discussed in relation to the function of P700.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
11.
Gerald Kastberger Ronald Thenius Anton Stabentheiner Randall Hepburn 《Journal of Insect Behavior》2009,22(1):65-85
Colony defence in Apis mellifera involves a variety of traits ranging from ‘aggressive’ (e.g. entrance guarding, recruitment
of flying guards) to ‘docile’ (e.g. retreating into the nest) expression. We tested 11 colonies of three subspecies (capensis, scutellata, carnica) regarding their defensiveness. Each colony was selected as reportedly ‘aggressive’, ‘intermediate’ or ‘docile’ and consisted
of about 10,000 bees. We applied three stimulation regimes (mechanical disturbance, exposure to alarm pheromones, and the
combination of both) and measured their behaviours by tracking the rates of outflying bees at the entrance sites of the test
hives. We provided evidence that for mechanical disturbances the test colonies resolved into two response types, if the ‘immediate’
defence response, assessed in the first minute of stimulation, was taken as a function of foraging: ‘releaser’ colonies allocated flying guards, ‘retreater’ colonies reduced the outside-hive activities. This division was observed irrespective
of the subspecies membership and maintained in even roughly changing environmental conditions. However, if pheromone and mechanical
stimulation were combined, the variety of colony defensiveness restricted to two further types irrespective of the subspecies
membership: six of nine colonies degraded their rate of flying defenders with increasing foraging level, three of the colonies
extended their ‘aggressiveness’ by increasing the defender rate with the foraging level. Such ‘super-aggressive’ colonies
obviously are able to allocate two separate recruitment pools for foragers and flying defenders. 相似文献
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Maria Luiza Peixoto de Oliveira Vicente J. Febres Marcio Gilberto Cardoso Costa Gloria A. Moore Wagner Campos Otoni 《Plant cell reports》2009,28(3):387-395
An improved method for the Agrobacterium infiltration of epicotyl segments of ‘Pineapple’ sweet orange [Citrus sinensis (L.) Osbeck] and ‘Swingle’ citrumelo [Citrus paradisi Macf. X Poncirus trifoliata (L.) Raf.] was developed in order to increase transformation frequency. Sonication-assisted Agrobacterium-mediated transformation (SAAT), vacuum infiltration, and a combination of the two procedures were compared with conventional
Agrobacterium-mediated inoculation method (‘dipping’ method). It was observed that the morphogenic potential of the epicotyl segments decreased
as the duration of SAAT and vacuum treatments increased. Transient GUS expression was not affected by the different SAAT treatments,
but it was significantly enhanced by the vacuum infiltration treatments. The highest transformation efficiencies were obtained
when the explants were subjected to a combination of SAAT for 2 s followed by 10 min of vacuum infiltration. PCR and Southern
blot analysis of the uidA gene were used to confirm the integration of the transgenes. The transformation frequencies achieved in this study (8.4%
for ‘Pineapple’ sweet orange and 11.2% for ‘Swingle’ citrumelo) are the highest ones reported for both cultivars. 相似文献
14.
V. R. Bommineni P. P. Jauhar T. S. Peterson R. N. Chibbar A. B. Almouslem 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,95(5-6):757-763
The objective of this study was to detect the presence of alien chromatin in intergeneric hybrids of durum wheat (Triticum turgidum, 2n=4x=28; AABB genomes) with the perennial grass Thinopyrum junceiforme (2n=4x=28; J1J1J2J2) using RAPD markers. The first step was to identify amplification of species-specific DNA markers in the parental grass species
and durum wheat cultivars. Initially, the genomic DNA of five grass species (Thinopyrum junceiforme, Th. bessarabicum, Lophopyrum elongatum, Leymus karataviensis and Elytrigia pycnantha) and selected durum cultivars (‘Langdon’, ‘Durox’, ‘Lloyd’, ‘Monroe’, and ‘Medora’) was screened with 40 oligonucleotide
primers (nano-mers). Three oligonucleotides that amplified DNA fragments specific to a grass species or to a durum cultivar
were identified. Primer PR21 amplified DNA fragments specific to each of the five durum cultivars, and primers PR22 and PR23
amplified fragments specific to each of the grass species. Intergeneric hybrids between the durum cultivars ‘Langdon’, ‘Lloyd’
and ‘Durox’ and Th. junceiforme, and their backcross (BC) progeny were screened with all 40 primers. Six primers amplified parent-specific DNA fragments
in the F1 hybrids and their BC1 progeny. Three primers, PR22, PR23 and PR41, that amplified Th. junceiforme DNA fragments in both F1 and BC1 were further analyzed. The presence of an amplified 1.7-kb Th. junceiforme DNA fragment in the F1 hybrids and BC1 progeny was confirmed using Southern analysis by hybridization with both Th. junceiforme genomic DNA and Th. junceiforme DNA amplified with primer PR41. With the exception of line BC1F2 no. 5, five selfed progeny of BC1 and a BC2 of line 3 (BC1F2 no. 3בLloyd’) from a cross of ‘Lloyd’×Th. junceiforme showed the presence of the 1.7-kb DNA fragment. All selfed BC1 and BC2 lines retained the 600-bp fragment that was confirmed after hybridization with Th. junceiforme DNA amplified with primer PR22. Other experiments using RFLP markers also showed the presence of up to seven Th. junceiforme DNA fragments in the F1 hybrids and their BC progeny after hybridization with Th. junceiforme DNA amplified with primer PR41. These studies show the usefulness of molecular markers in detecting alien chromatin/DNA fragments
in intergeneric hybrids with durum wheat.
Received: 21 November 1996 / Accepted: 21 March 1997 相似文献
15.
Xingguo Zhang Chaozhi Ma Tingdong Fu Yuanyuan Li Tonghua Wang Qingfang Chen Jinxing Tu Jinxiong Shen 《Molecular breeding : new strategies in plant improvement》2008,21(3):305-315
‘SI1300’ is a self-incompatible Brassica napus line generated by introgressing an S haplotype from B. rapa ‘Xishuibai’ into a rapeseed cultivar ‘Huayou No. 1’. Five S-locus specific primer pairs were employed to develop cleaved amplified polymorphic sequences (CAPS) markers linked the S haplotype of ‘SI1300’. Two segregating populations (F2 and BC1) from the cross between ‘SI1300’ and self-compatible European spring cultivar ‘Defender’, were generated to verify the molecular
markers. CAPS analysis revealed no desirable polymorphism between self-incompatible and self-compatible plants. Twenty primer
pairs were designed based on the homology-based candidate gene method, and six dominant sequence characterized amplified region
(SCAR) markers linked with the S-locus were developed. Of the six markers, three were derived from the SRK and SP11 alleles of class II B. rapa
S haplotypes and linked with S haplotype of ‘SI1300’. The other three markers were designed from the SLG-A10 and co-segregated with S haplotype of ‘Defender’. We successfully combined two pairs of them and characterized two multiplex PCR markers which could
discriminate the homozygous and heterozygous genotypes. These markers were further validated in 24 F3 and 22 BC1F2 lines of ‘SI1300 × Defender’ and another two segregating populations from the cross ‘SI1300 × Yu No. 9’. Nucleotide sequences
of fragments linked with S-locus of ‘SI1300’ showed 99% identity to B. rapa class II S-60 haplotype, and fragments from ‘Defender’ were 97% and 94% identical to SLG and SRK of B. rapa class I S-47 haplotype, respectively. ‘SI1300’ was considered to carry two class II S haplotypes and the S haplotype on the A-genome derived from B. rapa ‘Xishuibai’ determines the SI phenotype, while ‘Defender’ carry a class I S haplotype derived from B. rapa and a class II S haplotype from B. oleracea. SCAR markers developed in this study will be helpful for improving SI lines and accelerating marker-assisted selection process
in rapeseed SI hybrid breeding program. 相似文献
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Guohua Xu Shaoling Zhang Yonghua Yang Caiping Zhao Joseph N. Wolukau 《Acta Physiologiae Plantarum》2008,30(2):233-241
The objective of this study was to examine whether S-RNase plays a specific role in the pre-germinated Pyrus pollen. Effects of exogenous RNase and endogenous S-RNase on concentration of cytosolic-free calcium ([Ca2+]i) variation of pre-germinated Pyrus pollen were studied. [Ca2+]i variation caused by different RNases were complex. In 1 h after being cultured, exogenous RNase, RNase T1 and RNase A, and endogenous incompatible ‘Hohsui’ RNase promoted the [Ca2+]i of ‘Hohsui’ pollen. Acid proteins of ‘Hohsui’ had no remarkable influence on the [Ca2+]i of self-pollen. Endogenous compatible ‘Kohsui’ RNase reduced the [Ca2+]i of ‘Hohsui’ pollen, but compatible ‘Hohsui’ RNase can stimulate the [Ca2+]i of ‘Kohsui’ pollen. RNase T1, RNase A and incompatible ‘Kohsui’ S-RNase can also make ‘Kohsui’ pollen [Ca2+]i increase. Different from ‘Hohsui’ pollen, acid proteins of ‘Hohsui’ pull down the ‘Kohsui’ pollen [Ca2+]i remarkably. Conclusion can be made that during the prophase of pollen germination, endogenous S-RNase has no specific effect
on pollen [Ca2+]i changes. 相似文献
19.
Two genes that encode proteins which share 30–35% sequence identity with yeast OYE (Old Yellow Enzyme, an NAD(P)H FMN-oxidoreductase),
the well-studied archetype of the OYE protein family, have been identified in Gluconobacter oxydans M5. The two genes are localized in the chromosome and plasmid, respectively. Comparison of the deduced amino acid sequences
of the enzymes with database entries revealed 75.1% similarity and 64.9% identity to that of the Pseudomonas syringae pv. glycinea NAD(P)H-dependent 2-cyclohexen-1-one reductase. The two proteins were expressed as His-tag fusion proteins in Escherichia coli and purified. The ability of the purified proteins to hydrogenate citral was identified. The results showed that the α,β-double
bond of citral cis-isomer ‘neral’ could be stereoselectively reduced to produce citronellal by the purified OYE homologues. 相似文献