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1.
The wheat line H960642 is a homozygous wheat- Thinopyrum intermedium translocation line with resistance to BYDV by genomic in situ hybridization (GISH) and RFLP analysis. The genomic DNA of Th. intermedium was used as a probe, and common wheat genomic DNA as a blocking in GISH experiment. The results showed that the chromosome
segments of Th. intermedium were transferred to the distal end of a pair of wheat chromosomes. RFLP analysis indicated that the translocation line H960642
is a T7DS-7DL-7XL translocation by using 8 probes mapped on the homoeologous group 7 in wheat. The translocation breakpoint
is located between Xpsr680 and Xpsr965 about 90–99 cM from the centromere. The RFLP markers psr680 and psr687 were closely
linked with the BYDV resistance gene. The gene is located on the distal end of 7XL around Xpsr680 and Xpsr687.
Project supported by the 863 program and the National Natural Science Foundation of China (Grant No. 39680027). 相似文献
2.
Thinopyrum intermedium is a useful source of resistance genes for Barley Yellow Dwarf Virus (BYDV), one of the most damaging wheat diseases. In this study, wheat/ Th. intermedium translocation lines with a BYDV resistance gene were developed using the Th. intermedium 7Ai-1 chromosome. Genomic in situ hybridization (GISH), using a Th. intermedium total genomic DNA probe, enabled detection of 7Ai-1-derived small chro-matins containing a BYDV resistance gene, which were translocated onto the end of wheat chromosomes in the lines Y95011 and Y960843. Random amplified polymorphic DNA (RAPD) analyses using 120 random 10-mer primers were conducted to compare the BYDV-resistant translocation lines with susceptible lines. Two primers amplified the DNA fragments specific to the resistant line that would be useful as molecular markers to identify 7Ai-1-derived BYDV resistance chromatin in the wheat genome. Additionally, the isolated Th. intermedium-specific retrotransposon-like sequence pTi28 can be used to identify Th. intermedium chromatin transferred to the wheat genome. 相似文献
3.
以小麦-中间偃麦草二体附加系L1衍生抗病系PP9-1为抗源,与小麦推广品种陕7859.丰抗8号杂交并自交,在F6代中选到农艺性状优良的高抗黄矮病小麦新品系YW443。对YW443及其亲本进行抗病性鉴定。结果表明:YW443高抗大麦黄矮病毒GPV、GAV株系。利用基因组原位杂交,RFLP分析和RAPD分析,研究诉遗传构成及其抗病基因染色体归属。结果表明:YW443(2n=43)的遗传构成了40条(2 相似文献
4.
Barley yellow dwarf virus (BYDV) can cause significant losses of wheat worldwide. The long arm segment of Thinopyrum intermedium chromosome 7Ai#1 carrying the BYDV resistance gene Bdv2 was translocated to the distal region of the long arm of wheat chromosome 7D in translocation line Yw642. In this study,
40 wheat EST sequences located in the distal region of 7DL were explored to identify specific PCR markers for the Bdv2 region on the basis of the homoeologous relationship between wheat chromosome 7D and Th. intermedium chromosome 7Ai# 1. Our results revealed 8 novel EST-PCR markers specific to the Bdv2 region, including 5 EST-STS markers of BE404744, BE498985, BE591497, BG606695 and BQ161842, and 3 EST-SSCP markers of BE404953,
BG312663 and BE498985. These EST-PCR markers could distinguish Bdv2 from another BYDV-resistance gene located on Th.intermedium chromosome 2Ai-2. These specific bands for the Bdv2 region were further cloned and sequenced. The sequencing analysis indicated that the specific sequences for the Bdv2 region were highly homologous with the original wheat EST sequences that were used to design primers, and encode respectively
a protein kinase, P450, centrin, transducin, and a hypothetical protein. This study created a starting point for eventual
cloning of the Bdv2 gene and understanding the defense mechanism. 相似文献
5.
小偃麦附加系Z1和Z2中外源染色体2Ai-2的结构组成@张增燕$中国农业科学院作物育种栽培研究所!北京100081@辛志勇$中国农业科学院作物育种栽培研究所!北京100081@陈孝$中国农业科学院作物育种栽培研究所!北京100081小偃麦;;附加系;;染色体 相似文献
6.
Genomic in situhybridization (GISH) to root-tip cells at mitotic metaphase, using genomic DNA probes from Thinopyrum intermedium and Pseudoroegneria strigosa, was used to examine the genomic constitution of Th. intermedium, the 56-chromosome partial amphiploid to wheat called Zhong 5 and disease-resistant derivatives of Zhong 5, in a wheat background.
Evidence from GISH indicated that Th. intermedium contained seven pairs of St, seven J S and 21 J chromosomes; three pairs of Th. intermedium chromosomes with satellites in their short arms belonging to the St, J, J genomes and homoeologous groups 1, 1, and 5 respectively.
GISH results using different materials and different probes showed that seven pairs of added Th. intermedium chromosomes in Zhong 5 included three pairs of St chromosomes, two pairs of J S chromosomes and two pairs of St-J S reciprocal tanslocation chromosomes. A pair of chromosomes, which substituted a pair of wheat chromosomes in Yi 4212 and
in HG 295 and was added to 21 pairs of wheat chromosomes in the disomic additions Z1, Z2 and Z6, conferred BYDV-resistance
and was identical to a pair of St-J S tanslocation chromosomes (StJ S) in Zhong 5. The StJ S chromosome had a special GISH signal pattern and could be easily distinguished from other added chromosomes in Zhong 5; it
has not yet been possible to locate the BYDV-resistant gene(s) of this translocated chromosome either in the St chromosome
portion belonging to homoeologous group 2 or in the J S chromosome portion whose homoeologous group relationship is still uncertain. Among 22 chromosome pairs in disomic addition
line Z3, the added chromosome pair had satellites and belonged to the St genome and homoeologous group 1. Disomic addition
line Z4 carried a pair of added chromosomes which was composed of a group-7 J S chromosome translocated with a wheat chromosome; this chromosome was different to 7 Ai-1, but was identical to 7 Ai-2. The
leaf rust and stem rust resistance genes were located in the distal region of the long arm, whereas the stripe rust resistance
gene(s) was located in the short arm or in the proximal region of the long arm of 7 Ai-2. A pair of J S-wheat translocation chromosomes, which originated from the WJ S chromosomes in Z4, was added to the disomic addition line Z5; the added chromosomes of Z5 carried leaf and stem rust resistance
but not stripe rust resistance; Z5 is a potentially useful source for rust resistance genes in wheat breeding and for cloning
these novel rust-resistant genes. GISH analysis using the St genome as a probe has proved advantageous in identifying alien
Th. intermedium in wheat.
Received: 17 May 1999 / Accepted: 22 June 1999 相似文献
7.
Genomic in situ hybridization (GISH) and multicolor GISH (mcGISH) methodology were used to establish the cytogenetic constitution of five partial amphiploid lines obtained from wheat × Thinopyrum intermedium hybridizations. Line Zhong 1, 2 n=52, contained 14 chromosomes from each of the wheat genomes plus ten Th. intermedium chromosomes, with one pair of A-genome chromosomes having a Th. intermedium chromosomal segment translocated to the short arm. Line Zhong 2, 2 n=54, had intact ABD wheat genome chromosomes plus 12 Th. intermedium chromosomes. The multicolor GISH results, using different fluorochrome labeled Th. intermedium and the various diploid wheat genomic DNAs as probes, indicated that both Zhong 1 and Zhong 2 contained one pair of Th. intermedium chromosomes with a significant homology to the wheat D genome. High-molecular-weight (HMW) glutenin and gliadin analysis revealed that Zhong 1 and Zhong 2 had identical banding patterns that contained all of the wheat bands and a specific HMW band from Th. intermedium. Zhong 1 and Zhong 2 had good HMW subunits for wheat breeding. Zhong 3 and Zhong 5, both 2 n=56, possessed no gross chromosomal aberrations or translocations that were detectable at the GISH level. Zhong 4 also had a chromosome number of 2 n=56 and contained the complete wheat ABD-genome chromosomes plus 14 Th. intermedium chromosomes, with one pair of Th. intermedium chromosomes being markedly smaller. Multicolor GISH results indicated that Zhong 4 also contained two pairs of reciprocally translocated chromosomes involving the A and D genomes. Zhong 3, Zhong 4 and Zhong 5 contained a specific gliadin band from Th. intermedium. Based on the above data, it was concluded that inter-genomic transfer of chromosomal segments and/or sequence introgression had occurred in these newly synthesized partial amphiploids despite their diploid-like meiotic behavior and disomic inheritance. 相似文献
8.
The wheat-Thinopyrum intermedium addition lines Z1,Z2 contain a pair of Th. intermedium chromosomes 2Ai-2 carrying the gene with resistance to barley yellow dwarf virus (BYDV). Genomic in situ hybridization (GISH) was used to analyze the chromosome constitution of Z1,Z2 by using genomic DNA probes from Th. intermedium and Pseudoroegneria strigosa. The results showed that the chromosome constitution of either Z1 or Z2 composes of 42 wheat chromosomes and two Th. intermedium chromosomes (2Ai-2). The 2Ai-2 chromosome is St-E intercalary translocation, in which the E genomic chromosome segment translocated into the middle region of the long arm of chromosome belonging to St genome. With the genomic DNA probe of Ps. strigosa, the GISH pattern specific to the 2Ai-2 chromosome may be used as a molecular cytogenetic marker. A detailed RFLP analysis on Z1, Z2 and their parents was carried out by using 12 probes on the wheat group 2 chromosomes. Twenty RFLP markers specific to the 2Ai-2 chromosome were identified. Two RAPD markers of OPR16 –350 and OPH09 -1580, specific to the 2Ai-2 chromosome, were identified from 280 RAPD primers. These molecular markers could be used to assisted-select translocation lines with small segment of the 2Ai-2 chromosome and provide tools to localize the BYDV resistance. 相似文献
9.
Thinopyrum intermedium is a promising source of resistance to wheat streak mosaic virus (WSMV), a devastating disease of wheat. Three wheat germplasm lines possessing resistance to WSMV, derived from Triticum aestivum× Th. intermedium crosses, are analyzed by C-banding and genomic in situ hybridization (GISH) to determine the amount and location of alien chromatin in the transfer lines. Line CI15092 was confirmed as a disomic substitution line in which wheat chromosome 4A was replaced by Th. intermedium chromosome 4Ai?2. The other two lines, CI17766 and A29-13-3, carry an identical Robertsonian translocation chromosome in which the complete short arm of chromosome 4Ai?2 was transferred to the long arm of wheat chromosome 4A. Fluorescence in situ hybridization (FISH) using ABD genomic DNA from wheat as a probe and S genomic DNA from Pseudoroegneria stipifolia as the blocker, and vice versa, revealed that the entire short arm of the translocation was derived from the short arm of chromosome 4Ai?2 and the breakpoint was located at the centromere. Chromosomal arm ratios (L/S) of 2.12 in CI17766 and 2.15 in A29-13-3 showed that the translocated chromosome is submetacentric. This translocated chromosome is designated as T4AL? ? 4Ai?2S as suggested by Friebe et al. (1991). 相似文献
10.
小麦-中间偃麦草二体异附加系Z1、Z2具有一对携带抗黄矮病基因的中间偃麦草染色体2Ai-2。利用中间偃麦草(Thinopyrum intermedium (Host) Bakwoth and Dewey)和拟鹅冠草(Pseudoroegneia strigosa)基因组DNA作探针,对Z1、Z2进行基因组原位杂交分析。结果表明,Z1、Z2附加的一对中间偃麦草染色体2Ai-2为St-E染色体,E组染 相似文献
11.
Seedlings of a series of addition or substitution lines of wheat containing different Thinopyrum intermedium chromosomes were inoculated with the PAV and RPV serotypes of barley yellow dwarf virus (BYDV). Reduced virus titres in infected plants were ascribed to a single pair of homoeologous group 7 chromosomes from Th. intermedium in the disomic addition lines L1 and TAF 2. The group 7 chromosome is associated with red pigmentation of coleoptiles, which was also observed in two lines ditelosomic for the α arm of the chromosome. However, when infected with the PAV serotype of BYDV, the ditelosomic lines had normal virus titres and it is concluded that potential determinants of BYDV resistance are located on the β arm of the Group 7 chromosome. 相似文献
12.
Twenty-five partial amphiploids (2n=8x=56), which were derived from hybrids of wheat ( Triticum aestivum L.) with either Thinopyrum ponticum (Podpera) Liu & Wang, Th. intermedium (Host) Barkworth & D. Dewey, or Th. junceum (L.) A. Löve, were assayed for resistance to BYDV serotype PAV by slot-blot hybridization with viral cDNA of a partial coat protein gene. Three immune lines were found among seven partial amphiploids involving Th. ponticum. Seven highly resistant lines were found in ten partial amphiploids involving Th. intermedium. None of eight partial amphiploids or 13 addition lines of Chinese Spring — Th. junceum were resistant to BYDV. Genomic in situ hybridization demonstrated that all of the resistant partial amphiploids, except TAF46, carried an alien genome most closely related to St, whether it was derived from Th. ponticum or Th. intermedium. The two partial amphiploids carrying an intact E genome of Th. ponticum are very susceptible to BYDV-PAV. In TAF46, which contains three pairs of St- and four pairs of E-genome chromo somes, the gene for BYDV resistance has been located to a modified 7 St chromosome in the addition line L1. This indicates that BYDV resistance in perennial polyploid parents, i.e., Th. ponticum and Th. intermedium, of these partial amphiploids is probably controlled by a gene(s) located on the St-genome chromosome(s). 相似文献
13.
Summary The Agropyron intermedium chromosome 7Ai #2 is the source of the leaf rust resistance gene Lr38 which was transferred to wheat by irradiation. The chromosomal constitutions of eight radiation-induced rust-resistant wheat- Agropyron intermedium derivatives were analyzed by C-banding and genomic in-situ hybridization (GISH). Five lines were identified as wheat Ag. intermedium chromosome translocation lines with the translocation chromosomes T2AS·2AL-7Ai#2L, T5AL · 5AS-7Ai # 2L, T1DS · 1DL-7Ai # 2L, T3DL · 3DS-7Ai#2L, and T6DS · 6DL-7Ai#2L. The sizes of the 7Ai#2L segments in mitotic metaphases of these translocations are 2.42 m, 4.20 m, 2.55 m, 2.78 m, and 4.19 m, respectively. One line was identified as a wheat- Ag. intermedium chromosome addition line. The added Ag. intermedium chromosome in this line is different from 7Ai # 2. This line has resistance to leaf rust and stem rust. Based on the rust reactions, and the C-banding and GISH results, the remaining two lines do not contain any Ag. intermedium-derived chromatin. 相似文献
14.
Thinopyrum intermedium translocations derived from the wheat ( Triticum aestivum L.) substitution line P-29 were previously characterized by RFLP. We have further analyzed these lines and additional related
germplasm with publicly available STS and SSRs. Primers which showed a polymorphism between wheat and P-29, were tested in
all recombinant and nulli-tetrasomic lines confirming their position on chromosome 7D. The resulting 7D/7E chromosome maps
appeared as a mosaic of wheat and Th. intermedium chromatin sections. To verify the composition of the translocation lines suggested by the RFLP-PCR map, F 2 progeny of two crosses (CS/216-1 and CS/260-1) were analyzed with molecular markers. Both populations gave an unexpectedly
diverse number of recombinant individuals, suggesting that interstitial translocations occur more frequently than previously
thought. This analysis also showed that there is a wide range in the number and position of the interstitial translocations
within a given line such as the mosaic chromosome in recombinant line 260-1/CS-26, which has four Th. intermedium chromosome segments. Phenotypic data of the two populations suggested the presence of one gene which we have called Bdv3 to differentiate it from the previously reported orthologous gene Bdv2. Using the PCR-based molecular markers identified in this study, 5 out of 12 elite lines that showed good yields and no YDV
symptoms contained Th. intermedium chromatin. Due to the multiple components involved in the YDV disease complex, combining selection for YDV resistance with
the molecular markers and maps identified in this study will increase the efficiency of introgressing Th. intermedium chromatin containing YDV resistance or other beneficial traits into elite wheat germplasm. 相似文献
15.
The wheat--Thinopyrum intermedium addition lines Z1 and Z2 carry 21 pairs of wheat chromosomes and one pair of Th. intermedium chromosomes (2Ai-2) conferring resistance to barley yellow dwarf virus (BYDV). GISH results using the genomic DNA of Pseudoroegneria strigosa (S genome) as the probe indicated that the 2Ai-2 chromosome in Z1 and Z2 is an S-J intercalary translocation. Most of the 2Ai-2 chromosome belongs to the S genome, except for about one third in the middle region of the long arm that belongs to the J genome. The results of detailed RFLP analyses confirmed that the 2Ai-2 chromosome is extensively homoeologous to wheat group 2 chromosomes. Some new RFLP markers specific to the 2Ai-2 chromosome were identified. A RAPD marker, OP-R16(340), specific to the 2Ai-2 chromosome, was screened. We converted the RAPD marker into a sequence-characterized amplified region (SCAR) marker (designated SC-R16). The study establishes the basis for selecting translocation lines with small segments of the 2Ai-2 chromosome and localizing the BYDV resistance gene when introgressed into a wheat background. 相似文献
16.
Fluorescence and genomic in situ hybridization (FISH and GISH) were used to establish the cytogenetic constitution of two
wheat × Thinopyrum intermedium partial amphiploids H95 and 55(1-57). Both partial amphiploids are high-protein lines having resistance to leaf rust, yellow rust and powdery mildew and have
in total 56 chromosomes per cell. Repetitive DNA probes (pTa71, Afa family and pSc119.2) were used to identify the individual
wheat chromosomes and to reveal the distribution of these probes within the alien chromosomes. FISH detected 6B tetrasomy
in H95 and a null (1D) -tetrasomy (1B) in 55(1-57). GISH was carried out using biotin labeled Th. intermedium DNA and digoxigenin labeled Pseudoroegneria spicata DNA as probes, subsequently. GISH results revealed 44 wheat chromosomes and four Thinopyrum chromosome pairs, including three S and one J chromosome pairs in line H95. Line 55(1-57), contained 42 wheat chromosomes and six Th. intermedium pairs, including two S and one J S pairs. Additionally, two identical translocated chromosome pairs with diminished affinity to the alien chromatin were detected
in both amphiploids. Another two translocations were found in 55(1-57), with satellite sections from the Thinopyrum J genome. 相似文献
17.
Homology-based gene/gene-analog cloning method has been extensively applied in isolation of RGAs (resistance gene analogs)
in various plant species. However, serious interference of sequences on homoeologous chromosomes in polyploidy species usually
occurred when cloning RGAs in a specific chromosome. In this research, the techniques of chromosome microdissection combined
with homology-based cloning were used to clone RGAs from a specific chromosome of Wheat-Thinopyrum alien addition line TAi-27,
which was derived from common wheat and Thinopyrum
intermedium with a pair of chromosomes from Th. intermedium. The alien chromosomes carry genes for resistance to BYDV. The alien chromosome in TAi-27 was isolated by a glass needle
and digested with proteinase K. The DNA of the alien chromosome was amplified by two rounds of Sau3A linker adaptor-mediated
PCR. RGAs were amplified by PCR with the degenerated primers designed based on conserved domains of published resistance genes
(R genes) by using the alien chromosome DNA, genomic DNA and cDNA of Th. intermedium, TAi-27 and 3B-2 (a parent of TAi-27) as templates. A total of seven RGAs were obtained and sequenced. Of which, a constitutively
expressed single-copy NBS-LRR type RGA ACR3 was amplified from the dissected alien chromosome of TAi-27, TcDR2 and TcDR3 were
from cDNA of Th. intermedium, AcDR3 was from cDNA of TAi-27, FcDR2 was from cDNA of 3B-2, AR2 was from genomic DNA of TAi-27 and TR2 was from genomic
DNA of Th. intermedium. Sequence homology analyses showed that the above RGAs were highly homologous with known resistance genes or resistance gene
analogs and belonged to NBS-LRR type of R genes. ACR3 was recovered by PCR from genomic DNA and cDNA of Th. intermedium and TAi-27, but not from 3B-2. Southern hybridization using the digested genomic DNA of Th. intermedium, TAi-27 and 3B-2 as the template and ACR3 as the probe showed that there is only one copy of ACR3 in the genome of Th. intermedium and TAi-27, but it is absent in 3B-2. The ACR3 could be used as a specific probe of the R gene on the alien chromosome of
TAi-27. Results of Northern hybridization suggested that ACR3 was constitutively expressed in Th. intermedium and TAi-27, but not 3B-2, and expressed higher in leaves than in roots. This research demonstrated a new way to clone RGAs
located on a specific chromosome. The information reported here should be useful to understand the resistance mechanism of,
and to clone resistant genes from, the alien chromosome in TAi-27. 相似文献
18.
To introduce useful genes from the wild species Thinopyrum intermedium into cultivated wheat, a wheat- Th. intermedium disomic addition line (TAI27) was used as source material for tissue culture. TAI27 contains, beside the 42 wheat chromosomes,
a pair of smaller chromosomes that is cytologically discernible. Based on restriction fragment length polymorphism (RFLP)
analysis, this chromosome pair was determined to be a recombinant one, comprising segments with homoeology to at least two
chromosome groups of wheat, i.e., group 2 and 7. Sixty-eight green plants were regenerated from six month-old embryogenic
calli initiated from immature embryos of TAI27. Four of the plants were found to have only 42 cytologically normal-looking
chromosomes. Southern blot analysis using a Th. intermedium-enriched repetitive probe showed that one of the plants had hybridization fragments specific to Th. intermedium, indicating introgression of chromatin during the cell culture process.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
19.
The genetic constitutions of chromosome 2M of Aegilops comosa and the derived wheat- Ae. comosa translocations were analyzed by molecular cytogenetic techniques. Hybridization of 15 RFLP markers covering the entire length of the group-2 chromosomes revealed that chromosome 2M was structurally rearranged compared to the homoeologous chromosomes of wheat by either a pericentric inversion or a terminal intrachromosomal translocation. The breakpoint of the rearrangement was located in a region between the loci Xpsr131 and Xcdo405, resulting in the translocation of 47% of 2MS to 2ML. This aberrant structure of 2M allowed homoeologous recombination between 2M and its wheat counterpart only in the translocated segment on 2ML. C-banding and genomic in situ hybridization analyses confirmed that all translocation chromosomes consisted of the complete 2MS arm, a large part of 2ML, and very small distal segments derived from 2AS or 2DS, as expected from the aberrant structure of chromosome 2M. Thus, the translocation in the line 2A-2M?4/2 can be described as T2AS-2M?1L???2M?1S and the translocations in the lines Compair and 2D-2M?3/8 as T2DS-2M?1L???2M?1S. RFLP analysis determined the breakpoints in these translocation chromosomes to be within the telomeric 16% of the wheat chromosome arms. The breakpoint of the 2A/2M translocation was between Xbcd348 and Xcdo783, and that of the 2D/2M translocation was between Xcdo783 and Xpsr666. Because the translocation chromosomes retain the structural aberration found in chromosome 2M, further exploitation of the wheat- Ae. comosa translocations for cultivar improvement is questionable. 相似文献
20.
Main conclusion
Fluorescence in situ hybridization and molecular markers have confirmed that several chromosomes from
Thinopyrum intermedium
ssp.
trichophorum
have been added to a wheat background, which originated from a cross between a wheat–
Thinopyrum
partial amphiploid and triticale. The lines displayed blue grains and resistance to wheat stripe rust.
Thinopyrum intermedium has been used as a valuable resource for improving the disease resistance and yield potential of wheat. With the aim to transfer novel genetic variation from Th. intermedium species for sustainable wheat breeding, a new trigeneric hybrid was produced by crossing an octoploid wheat–Th. intermedium ssp. trichophorum partial amphiploid with hexaploid triticale. Fluorescence in situ hybridization (FISH) revealed that Thinopyrum chromosomes were transmitted preferably and the number of rye chromosomes tended to decrease gradually in the selfed derivatives of the trigeneric hybrids. Four stable wheat–Th. intermedium chromosome substitution, addition and translocation lines were selected, and a 2JS addition line, two substitution lines of 4JS(4B) and 4J(4B), and a small 4J.4B translocation line were identified by FISH and molecular markers. It was revealed that the gene(s) responsible for blue grains may located on the FL0.60–1.00 of long arm of Th. intermedium-derived 4J chromosome. Disease resistance screenings indicated that chromosomes 4JS and 2JS appear to enhance the resistance to stripe rust in the adult plant stage. The new germplasm with Th. intermedium introgression shows promise for utilization of Thinopyrum chromosome segments in future wheat improvement. 相似文献
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