Comparative Physical Mapping of Bph3 with BAC-FISH in Oryza officinalis and O. sativa

A FISH procedure was adopted to physical mapping rice RFLP marker RZ69 and the BAC clone 38J9 screened by RZ69 which is linked to gene Bph3 in Oryza saliva L. and O. officinalis Well ex: Watt. The FISH results showed that both 38J9 and RZ69 were located in the middle of 4S in O. officinalis and the centromere area of 4S in O. sativa. In O. officinalis the percentage distances from the centromere to the hybridization sites were 20.00 +/- 5.40 and 22.12 +/- 3.44, the detection rates were 50.00% and 6.14%, but in O. sativa they were 0 and 0, 56.10% and 6.29% correspondingly. The results obtained from the BAC and RFLP clone were almost the same in the cultivated rice and O. officinalis. It was suggested that die marker RZ69 of the cultivated rice and its homologous sequence in O. officinalis were in the same BAC clone, the homologous sequence of Bph3 in O. officinalis should be located at the sites hybridized by both RZ69 and 38J9. Many signals on different chromosomes of O. officinalis were observed under no blocking with Cot-1 DNA, showing that the repetitive sequences were also homologous between O. sativa and O. officinalis. The identification of chromosome 4 of O. officinalis is based on comparative map with RG214 and BAC clone screened by RG214. The feasibility of comparative physical mapping performed between O. sativa and O. officinalis with rice BAC clones was discussed.

An Integrated Map of Soybean Physical Map and Genetic Map

Soybean is a major crop in the world, and it is a main source of plant proteins and oil. A lot of soybean genetic maps and physical maps have been constructed, but there are no integrated map between soybean physical map and genetic map. In this study, soybean genome sequence data, released by JGI （US Department of Energy＇s Joint Genome Institute）, had been downloaded. With the software Blast 2.2.16, a total of 161 super sequences were mapped on the soybean public genetic map to construct an integrated map. The length of these super sequences accounted for 73.08% of all the genome sequence. This integrated map could be used for gene cloning, gene mining, and comparative genome of legume.

Identification and Physical Mapping of New PCR-Based Markers Specific for the Long Arm of Rye(Secale cereale L.) Chromosome 6

To effectively use elite genes on the long arm of rye chromosome 6（the 6RL arm） in wheat breeding programs,precise and fast identification of 6RL chromatin in wheat backgrounds is necessary.PCR-based 6RL-specific markers can facilitate the detection of elite genes on 6RL in wheat breeding.However,only a limited number of 6RL-specific markers have been developed.In the present study.300 new PCR-based 6RL-specific markers were identified using specific length amplified fragment sequencing（SLAF-seq） technology,and were further physically mapped to four regions on the 6RL arm using 6R and 6RL deletion lines.Interestingly,127 of the 300 markers were physically localized to a region from the site between 2.3 and 2.5 to the telomere,the same region where the powdery mildew resistance gene was mapped.In addition,95 of the 300 markers exhibit polymorphisms,which can be used to investigate the diversity of rye 6RL arms.The markers developed in this study can be used to identify given segments of 6RL in wheat backgrounds and accelerate the utilization of elite genes on 6RL in wheat breeding.

Construction of the Primary Physical Map of Rice Chromosome 12

A primary physical map of rice chromosome 12 was constructed using marker-based chromosome landing and chromosome walking. A BAC library from IR64 was screened using 84 RFLP markers, 4 STS markers and 6 microsatellite markers on chromosome 12 by colony hybridization and polymerase chain reaction (PCR) amplification. A total of 59 contigs consisting of 419 BAC clones including 5 single-clones were physically aligned on rice chromosome 12 with the largest BAC contig covering 855 kb. The whole physical map had a size of ～16 Mb and covered about 52% of rice chromosome 12. This physical map will be certainly helpful for map-based gene cloning of agronomically and biological important genes and understanding the genome structure of the chromosome.

A bacterial artificial chromosome-based physical map of Manihot esculenta ssp. flabellifolia

Cassava(Manihot esculenta) is known as the third most important food crop in the tropics and also used for industrial feedstock for biofuels. Two new bacterial artificial chromosome(BAC) libraries were constructed for W14(M. Esculenta ssp. flabellifolia), a wild ancestor of domesticated cassava. The libraries were constructed with EcoR I and Hind III insertion vectors, respectively.The EcoR I library has 29952 clones with an average insert size of 115 kb, while the Hind III library consists of 29952 clones with an average insert of 129 kb. The combined libraries contain a total of 59904 clones with an average insert size of 125 kb, representing approximately 10 ?haploid genome equivalents. A total of 29952 clones were fingerprinted and resulted in a cassava physical map composed of 2485 contigs with an average physical length of 336 kb and 2909 singletons, representing approximately762 Mb of the cassava genome. 5000 clones located at the ends of BAC contigs were selected and sequenced. A total of 6077 SNPs and 231 indels were identified, that covered459 gene sequences, of which 6 genes were associated with starch and sucrose metabolism. This BAC-based physical map provides valuable tools to understand the genetics and evolution of cassava.

PHYSICAL VALUE MAPPING ALGORITHM OF MESH IN FINE CAE ANALYSIS OF AUTOMOBILE BODY STRUCTURE

A physical value mapping （PVM） algorithm based on finite element mesh from the stamped part in stamping process to the product is presented, In order to improve the efficiency of the PVM algorithm, a search way from the mesh of the product to the mesh of the stamped part will be adopted. At the same time, the search process is divided into two steps： entire search （ES） and local search （LS）, which improve the searching efficiency. The searching area is enlarged to avoid missing projection elements in ES process. An arc-length method is introduced in LS process. The validity is confirmed by the results of the complex industry-forming product.

An Integrated Genetic,Physical and Transcript Map of Homoeologous Chromosomes 12 and 26 in Upland Cotton(Gossypium hirsutum L.)

While Upland cotton(Gossypium hirsutum L.) represents 95% of the world production,its genetic improvement is hindered by the shortage of effective genomic tools and resources.The

Comparative Physical Localization of Rice Pib Gene and Its Linked RFLP Markers in Oryza sativa, O. officinalis and Zea mays

Comparative genetic studies have shown that there are widespread synteny and colinearity of the genes among different species within grass family. Rice ( Oryza sativa L.) is a model plant, and analysis of its genome allows us to reveal the common features and the evolutionary rules of the gramineous genomes and accumulate the data for establishment of a common genetic system in the Poaceae. In this study, a rice gene Pib ( 10.3 kb), a map-based cloned gene, and RFLP markers linked with it are used as the tested probes to investigate their homology and physical location among the tested species. Southern blotting analysis showed that there were sequences homologous to Pib in maize genome. Further, Pib was localized onto the chromosomes of O. sativa ssp. indica cv. Guangluai 4, O. officinalis Wall ex Watt and the inbred line of Zea mays cv. Huangzao 4. The results of fluorescence in situ hybridization (FISH) and double-color FISH indicated that a synteny of Pib and RFLP markers linked with Pib existed among the genomes of the three tested species.

Mapping of Microsatellite SW943 to Porcine Chromosome 12p11-(2/3p13) Using Primed in situ Synthesis and Somatic Cell Hybrid Panel

The porcine microsatellite SW943 was regionally localized on 12p11-(2/3p13) by the two methods: the Primed in situ (PRINS) labelling on the pachytene bivalents of pigs using the Dig-11-dUTP as the report molecule and pig X rodent Somatic Cell Hybrid Panel (SCHP) which contains 27 cell lines through PCR amplification. Advantages and disadvantages of the two methods for physical mapping of microsatellites were also discussed.

An automated method for mapping physical soil and water conservation structures on cultivated land using GIS and remote sensing techniques

An efficient and reliable automated model that can map physical Soil and Water Conservation（SWC） structures on cultivated land was developed using very high spatial resolution imagery obtained from Google Earth and Arc GIS？ERDAS IMAGINE？and SDC Morphology Toolbox for MATLAB and statistical techniques. The model was developed using the following procedures：（1） a high-pass spatial filter algorithm was applied to detect linear features,（2） morphological processing was used to remove unwanted linear features,（3） the raster format was vectorized,（4） the vectorized linear features were split per hectare（ha） and each line was then classified according to its compass directionand（5） the sum of all vector lengths per class of direction per ha was calculated. Finallythe direction class with the greatest length was selected from each ha to predict the physical SWC structures. The model was calibrated and validated on the Ethiopian Highlands. The model correctly mapped 80% of the existing structures. The developed model was then tested at different sites with different topography. The results show that the developed model is feasible for automated mapping of physical SWC structures. Thereforethe model is useful for predicting and mapping physical SWC structures areas across diverse areas.

Characterization and Genetic Analysis of a Novel Rice Spotted-leaf Mutant HM47 with Broad-spectrum Resistance to Xanthomonas oryzae pv. oryzae

A stable inherited rice spotted-leaf mutant HM47 derived from an EMS-induced IR64 mutant bank was identified. The mutant expressed hypersensitive response （HR）-like symptoms throughout its whole life from the first leaf to the flag leaf, without pathogen invasion. Initiation of the lesions was induced by light under natural summer field conditions. Expression of pathogenesis-related genes including PAL, PO-C1, POX22.3 and PBZ1 was enhanced significantly in association with cell death and accumulation of H2O2 at and around the site of lesions in the mutant in contrast to that in the wild-type （WT）. Disease reaction to Xanthomonas oryzae pv. oryzae from the Philippines and China showed that HM47 is a broad-spectrum disease-resistant mutant with enhanced resistance to multiple races of bacterial blight pathogens tested. An F2 progeny test showed that bacterial blight resistance to race HB-17 was cosegregated with the expression of lesions. Genetic analysis indicated that the spotted-leaf trait was controlled by a single recessive gene, tentatively named spl HM47 , flanked by two insertion/deletion markers in a region of approximately 74 kb on the long arm of chromosome 4. Ten open reading frames are predicted, and all of them are expressed proteins. Isolation and validation of the putative genes are currently underway.

Dynamic Intra-Japonica Subspecies Variation and Resource Application

We constructed a physical map of O. sativa ssp. japonica cv. ZH11 and compared it and its random sample sequences with the Nipponbare RefSeq derived from the same subspecies. This comparison showed that the two japonica genomes were highly syntenic but revealed substantial differences in terms of structural variations, rates of substitutions and indels, and transposable element content. For example, contractions/expansions as large as 450 kb and repeat sequences that were present in high copy numbers only in ZH11 were detected. In tri-alignment regions using the indica variety 93-11 sequence as an outgroup, we found that： （1） the substitution rates of the two japonica-indica inter- subspecies comparison combinations were close but almost a magnitude higher than the substitution rate between the japonica rice varieties ZH11 and Nipponbare; （2） of the substitutions found between ZH11 and Nipponbare, 47.2% occurred in ZH11 and 52.6% in Nipponbare; （3） of the indels found between ZH11 and Nipponbare, the indels that occurred in ZH11 were 15.8 times of those in Nipponbare. Of the indels that occurred in ZH11, 75.67% were insertions and 24.33% deletions. Of the indels that occurred in Nipponbare, 48.23% were insertions and 51.77% were deletions. The ZH11 com- parative map covered four Nipponbare physical gaps, detected assembly errors in the Nipponbare sequence, and was integrated with the FSTs of a large ZH11 T-DNA insertion mutant library. ZH11 BAC clones can be browsed, searched, and obtained at our website, http：//GResource.hzau.edu.cn.