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Comprehensive Network Analysis of Anther-Expressed
Genes in Rice by the Combination of 33 Laser
Microdissection and 143 Spatiotemporal Microarrays
Koichiro Aya1., Go Suzuki2., Keita Suwabe3,4., Tokunori Hobo1., Hirokazu Takahashi5, Katsuhiro
Shiono5, Kentaro Yano6, Nobuhiro Tsutsumi5, Mikio Nakazono5,7, Yoshiaki Nagamura8, Makoto
Matsuoka1*, Masao Watanabe3,9*
1Bioscience and Biotechnology Center, Nagoya University, Nagoya, Japan, 2Division of Natural Science, Osaka Kyoiku University, Kashiwara, Japan, 3Graduate School of
Life Sciences, Tohoku University, Sendai, Japan, 4Graduate School of Bioresources, Mie University, Tsu, Japan, 5Graduate School of Agricultural and Life Sciences, The
University of Tokyo, Tokyo, Japan, 6School of Agriculture, Meiji University, Kawasaki, Japan, 7Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya,
Japan, 8National Institute of Agrobiological Sciences, Tsukuba, Japan, 9Faculty of Science, Tohoku University, Sendai, Japan
Co-expression networks systematically constructed from large-scale transcriptome data reflect the interactions and
functions of genes with similar expression patterns and are a powerful tool for the comprehensive understanding of
biological events and mining of novel genes. In Arabidopsis (a model dicot plant), high-resolution co-expression networks
have been constructed from very large microarray datasets and these are publicly available as online information resources.
However, the available transcriptome data of rice (a model monocot plant) have been limited so far, making it difficult for
rice researchers to achieve reliable co-expression analysis. In this study, we performed co-expression network analysis by
using combined 44 K agilent microarray datasets of rice, which consisted of 33 laser microdissection (LM)-microarray
datasets of anthers, and 143 spatiotemporal transcriptome datasets deposited in RicexPro. The entire data of the rice co-
expression network, which was generated from the 176 microarray datasets by the Pearson correlation coefficient (PCC)
method with the mutual rank (MR)-based cut-off, contained 24,258 genes and 60,441 genes pairs. Using these datasets, we
constructed high-resolution co-expression subnetworks of two specific biological events in the anther, ''meiosis'' and
''pollen wall synthesis''. The meiosis network contained many known or putative meiotic genes, including genes related to
meiosis initiation and recombination. In the pollen wall synthesis network, several candidate genes involved in the
sporopollenin biosynthesis pathway were efficiently identified. Hence, these two subnetworks are important
demonstrations of the efficiency of co-expression network analysis in rice. Our co-expression analysis included the
separated transcriptomes of pollen and tapetum cells in the anther, which are able to provide precise information on
transcriptional regulation during male gametophyte development in rice. The co-expression network data presented here is
a useful resource for rice researchers to elucidate important and complex biological events.
Citation: Aya K, Suzuki G, Suwabe K, Hobo T, Takahashi H, et al. (2011) Comprehensive Network Analysis of Anther-Expressed Genes in Rice by the Combination
of 33 Laser Microdissection and 143 Spatiotemporal Microarrays. PLoS ONE 6(10): e26162. doi:10.1371/journal.pone.0026162
Editor: Abidur Rahman, Iwate University, Japan
Received June 29, 2011; Accepted September 21, 2011; Published October 26, 2011
Copyright: ? 2011 Aya et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: K. Suwabe was partially supported by Young Researchers Fellowships from the Japan Society for the Promotion of Science. This work was funded in
part by a MEXT Grant (21-COE, 18075003 and 1807512 to MW, 18075005 to NT, G-COE, 18075006 to MM), by a MAFF Grant (IPG-0003 to MM, IPG-0012 to MN, and
IPG-0019 to MW), and by Grants-in-Aid for Scientific Research on Innovative Areas (Nos. 23113006 to GS, 23113005 to MM, and 23113001 to MM, and GS) from
MEXT. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing Interests: The authors have declared that no competing interests exist.
* E-mail: email@example.com (MM); firstname.lastname@example.com (MW)
. These authors contributed equally to this work.
Recent developments in high-throughput microarray, next-
generation sequencing, proteomic analysis, and the accumulated
functional genomics data across species have enabled us to utilize
integrated large-scale data of gene-expression, protein-protein
interaction, and phenotype. There is now an increasing need for
integrated analysis at a system biology level, to gain an
understanding of the complex relationships between gene-product
interactions and biological events (e.g. phenotype). An in-silico-
derived co-expression network is constructed from large-scale gene
expression profiles, and is based on the assumption that genes with
similar expression patterns are likely to interact with each other at
the molecular or physiological level. In some model plants as well
as animals, this strategy has been broadly used to predict
integrated networks in association with protein-protein interaction
data [1], the structural information on metabolic pathways [2–4],
and the functions of gene products [5].
To date, a model plant, Arabidopsis thaliana, has been subjected
to thousands of microarray experiments and the results have been
deposited in publicly-available online databases [6]. By using
these very large expression datasets, high-resolution co-expression
networks have been constructed and these are available as online
information resources in Arabidopsis. Such useful information
resources have enabled Arabidopsis researchers to identify novel
factors involved in cell wall synthesis [3,4] and the aliphatic
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Page 2
glucosinolate biosynthesis pathway [5]. On the other hand, in a
model crop for the grass family, rice (Oryza sativa), there are few
resources for co-expression network analysis, because of inade-
quate proliferation of available rice microarray datasets derived
from various different platforms and a low number of datasets
with few examples of publicly available data [7]. Moreover, in
rice, there have been fewer case studies to estimate the
effectiveness of co-expression networks, compared to those in
Arabidopsis. Thus, a reliable large-scale study of co-expression
network analysis is required to drive forward rice research at this
Male gametophyte development might be a good target for co-
expression network analysis in rice, because pollen transcriptome
can be easily and precisely analyzed using larger rice flowers than
those in Arabidopsis. In flowering plants, pollen development takes
place within a male reproductive organ, the anther, and is
controlled precisely by four sporophytic cell layers of the anther
(tapetum, middle layer, endothecium and epidermis), which
surround the gametophytic pollen grains [8]. After differentiation
of the male germline, pollen mother cells form tetrads of haploid
microspores via meiosis, and the tetrad microspores are connected
to each other by a callose wall. Tapetum cells play an important
role in degradation of the callose wall, which allows the
microspores to be released into the anther locule. The released
microspores subsequently mature into pollen grains through cell
division and pollen wall formation. During the course of pollen
maturation, the tapetum starts to degenerate and provides the
various materials for pollen wall formation on the surface of pollen
grains. So far, by using in situ hybridization [9–15] or microarrays
[16,17], many genes expressed in anthers have been identified in
Arabidopsis [9,13,17], Brassica napus [10], rice [11,12,14], and Lotus
japonicus [15,16]. These studies have revealed the complex patterns
of gene expression in both the gametophytic pollen/microspore
and sporophytic tapetum cells, which are different but influence
each other; consequently, it has been difficult to analyze the
precise regulatory interactions of gene expression between pollen
and tapetum by examination of the whole anther transcriptome In
this context, transcriptome data from separated pollen and
tapetum cells are necessary to compare with the large sets of
other microarray data in order to achieve co-expression network
analysis of male gametophyte development in plants. Recently, we
have performed transcriptome analysis using 44 K microarrays
with RNAs extracted independently from pollen and tapetum cells
within the rice anther by laser microdissection (LM) technology
By using our LM-microarray [18,19] and other publicly
available microarray datasets [7], in this study, we have
conducted comprehensive co-expression analysis and constructed
the co-expression subnetworks responsible for two important
biological events during anther development, meiosis and pollen
wall synthesis. The meiosis network contained many putative
meiotic genes as well as known meiotic genes, which could play
meiotic roles such as meiosis initiation and recombination.
Furthermore, from the pollen wall synthesis network, we
efficiently identified several candidate genes involved in the
sporopollenin biosynthesis pathway. Taken together, our co-
expression network has the potential to be a powerful resource to
dissect various important biological events and/or to isolate novel
genetic factors regulating development and differentiation of the
male reproductive organ in plants at a system biology level. In
addition, this study is an important development as it is one of the
few demonstrations of the efficiency of co-expression network
analysis in rice.
Dataset content
We have previously conducted 44 K agilent LM-microarray
analysis of the microspore/pollen and tapetum cells of japonica rice,
Oryza sativa cv. Nipponbare, during anther development from
meiosis to tricellular pollen stages [18,19]. In addition to the above
study, a LM sample in premeiosis stage was prepared in this study,
for more precise gene expression profiling during anther
development. The overall LM-microarray data, including three
novel microarray datasets, has been deposited at the Gene
Expression Omnibus (GEO; http://www.ncbi.nlm.nih.gov/geo/;
accession number GSE 29217). A detailed procedure for the LM-
microarray is described in our previous reports [18,19].
Recently, co-expression analysis datasets have become a
powerful tool to extract the transcriptional networks of genes
involved in various biological events in plants [20]. Therefore, we
conducted microarray-based co-expression analysis using our LM-
microarray data to gain biological insights into gene expression
networks mediating rice anther development. Because popular
methods that derive regulatory networks from gene expression
data, in general, require large amounts of gene expression data,
here we used 143 spatiotemporal expression profiles of 48 various
vegetative, and reproductive organs and tissues of rice deposited in
RicexPro [7], in addition to our 33 LM-microarray data of rice
anthers [18,19] (Table S1).
A 44K agilent microarray slide contains 29,864 distinct rice
genes. Using the 176 microarray, first, we examined the
spatiotemporal expression pattern by identifying the highest signal
intensity for each gene from different stages and organs (Figure 1,
Tables S2, S3, S4, S5, S6, S7, S8, S9, S10, S11, S12, S13, S14,
S15, S16, S17, S18, S19, S20, S21, S22, S23, S24, S25). Among
the 29,864 genes analyzed, about 10,600 genes were found to be
expressed preferentially in at least one of the anther developmental
stages or tissues (Figure 1, Tables S7, S8, S9, S10, S11, S12, S13,
S14, S15, S16, S17, S18, and S19 for array data of genes
preferentially expressed in anther). The remaining approximately
19,300 genes were expressed in the other 44 vegetative or
reproductive tissues (Tables S2, S3, S4, S5, S6, S20, S21, S22,
S23, S24, S25). Such a large proportion of anther preferentially
expressed genes indicates that many genes are involved in anther
development and, consequently, analysis of co-expression net-
works of anther preferentially expressed genes could potentially
provide good examples to demonstrate the application of
microarray datasets.
Biological significance of expression similarity
We first conducted an expression coherence (EC) analysis using
gene ontology (GO) terms to examine the biological significance of
expression similarity in the 176 microarray datasets used in this
study. EC analysis is a statistical analysis to determine whether
gene members belonging to a predefined functional gene set are
correlated with each other at the transcriptional level [21]. The
EC score is a measure of the expression similarity within a set of
predefined functional genes [21]; consequently, a higher EC score
is obtained, when most of genes with the same GO term are co-
expressed with each other. In our EC analysis, the EC scores of all
three GO groups (Biological Process, BP; Molecular Function,
MF; and Cellular Component, CC) were higher than that
expected by random sampling (Figure 2). Among the three GO
groups, CC showed the highest expression similarity, in which
approximately 17% of categories exhibited higher EC scores than
random sampling at threshold EC score of 0.05, while approx-
imately 12% and 8% did so in BP and MF, respectively (dotted
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line in Figure 2). For genes annotated according to the BP group,
the ''photosynthesis, light harvesting'' category showed the highest
EC scores (EC=0.868, 17 genes), whereas the scores of the
categories ''small ribosomal subunit'' (EC=0.433, 21 genes) and
''threonine-type endopeptidase activity'' (EC=0.322, 24 genes)
were highest in the CC and MF groups (Table S26). These results
confirm that the levels of expression similarity estimated from the
microarray datasets in this study are good reflections of functional
Construction of the meiosis-specific network
By using the Pearson correlation coefficient (PCC) method with
the mutual rank (MR)-based cut-off (see Materials and Methods),
we identified the final dataset of rice co-expression networks
generated from all the 176 microarray data used in this study,
which contains 24,259 genes (nodes) and 60,449 genes pairs
(edges). From this entire dataset of the rice co-expression network,
we are able to construct a subnetwork around gene(s) of interest
input as guide(s).
To examine whether our co-expression network analysis can
identify useful transcriptional networks related to anther develop-
ment, we first constructed a co-expression subnetwork centered on
9 known meiotic genes (guide genes; red circles in Figure 3A),
PAIR1 [22], PAIR2 [23], PAIR3 [24], MEL1 [25], OsRAD21–4
[26], DMC1A and 1B [27], OsMER3/OsRCK [28], and OsSDS
[28], whose functions in meiosis have been experimentally
demonstrated in rice. The expression profiles of these nine genes
after normalized by the VSN algorism (Figure S1A) are well
corresponding to the previous results of their expression patterns:
indicating that the VSN normalization was a suitable method for
the construction of co-expression network. The meiosis-specific
network consisted of a large main cluster with the 7 guide genes,
and two independent minor clusters of OsMER3 and PAIR1
(Figure 3A), which in total contained 187 genes and 346 gene
interactions (Table S27). The main cluster also contained ZEP1
(rice ZYP1 ortholog) and MEL2 (blue circles in Figure 3A), which
have most recently been reported as rice meiotic genes [29,30].
Moreover, PAIR2 and ZEP1 [23,29], whose products are both
constituents of the synaptonemal complex, were directly connected
in the network (red bold lines in Figure 3A). Good association of
the 7 meiotic guide genes with each other and with the additional
2 meiotic genes in the main expression cluster indicates that genes
involved in meiosis could be strictly co-regulated at the expression
level, and thus the meiotic events are potentially good subjects for
co-expression analysis. This is also supported by the results of GO
analysis (Figures 3B and 3C, Table S28), in which DNA-repair-
related GO terms (such as ''chromosome organization,'' ''DNA
unwinding involved in replication,'' ''Mismatch repair,'' and
Figure 1. Distribution of rice genes with tissue-preferential expression in the 176 microarray datasets. The 29,864 distinct rice genes on
a 44K microarray were classified into 24 spatiotemporal categories (different stages and tissues including LM-separated anther tissues) according to
the highest signal intensity among the 176 microarray datasets. Gene number indicates the number of genes in each category. The spatiotemporal
categories restricted to the anther are indicated in red. MEI, meiosis; TET, tetrad; UN, uninuclear; BC, bicellular; TC, tricellular.
Figure 2. EC analysis of the 176 microarray data of rice. BP, MF,
and CC represent Biological Process, Molecular Function, and Cellular
Component groups of GO, respectively. The total number of categories
in each GO group is indicated in parentheses. In the three GO groups
(BP, blue; MF, red; and CC, green), the EC score was calculated for each
GO category and compared with that of random sampling (purple) to
estimate the statistical significant level. Fraction of categories (y axis)
indicates the ratio of the number of GO categories with higher EC score
than each threshold EC score (x axis) to the total number of GO
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''Mismatched DNA binding'') appear frequently in genes in the
meiosis-specific network.
Known or putative meiotic genes in the subnetwork are listed in
Table 1. The meiosis-specific network contained cyclin genes and
other potential rice orthologs of meiotic genes (Figure 3A). In the
network, genes related to meiotic recombination (e.g. DMC1s and
OsMSH4/5) are located near a core of the network, and genes
functioning in other meiotic events (e.g. MEL1, PAIR3, OsRAD21–
4, and OsSDS) surround them with the small independent cluster of
PAIR1 (Figure 3A). The independent small cluster of OsMER3 (a
meiotic recombination-related gene) is due to its higher vegetative
expression, which could indicate a bifunctional role in meiosis and
other biological events.
Among genes related to ''transition from mitosis into meiosis'' in
Table 1, MEL1 and MEL2 encode ARGONAUTE and RNA-
recognition-motif protein, respectively [24,30], and putative
OsAML1 (Os01g0907900) also encodes an RNA-binding protein
[31], which is highly similar to MEI2 of Schizosaccharomyces pombe
[32]. Together with PWWP- and SET-domain proteins (Table 1),
MEL1, MEL2 and OsAML1 might regulate the initiation of meiosis
in rice through molecular mechanisms related to RNA metabo-
lism, chromatin remodeling and/or transcriptional regulation. On
the other hand, genes related to ''meiotic recombination'' are well
integrated in the network (Figure 3A, Table 1). Co-expression of
recombination-related genes in the network is consistent with
previous reports that DMC1A and DMC1B co-ordinate with
RAD17 [33], and that MSH4 and MSH5 function as a complex
In addition, it can be concluded from the GO analysis
(Figure 3B and 3C, Table S28) that various additional meiotic
genes could be included in the network as unknown genes (open
circles in Figure 3A). This indicates that novel meiotic genes could
be mined from the present data, and that novel interactions
between known meiotic genes could also be identified from the
unknown-gene-mediated connections in the network. Meiosis is an
extremely specialized biological event, thus co-expression network
analysis has the potential to achieve maximum success in the
identification of gene interactions.
Construction of the pollen wall synthesis network
As a second example system, we analyzed the pollen wall
synthesis network to systemically identify the genes that modulate
the formation of the outer pollen wall, the exine. Previous
biochemical analyses have shown that the exine layer consists
mainly of sporopollenin, a polymer of phenylpropanoid and lipidic
monomers covalently linked by ether and ester linkages [35–37].
When three genes for rice sporopollenin biosynthesis or transport
(CYP703A3, CYP704B2, and Osc6) were selected as guide genes
[38–40], a major subnetwork covering all guide genes was
identified, which consisted of 108 genes and 278 gene interactions
(Figure 4A, Table S29). Again, the expression profiles of these
three genes after normalized by the VSN algorism (Figure S1B) are
well corresponding to the previous results of their expression
patterns. Classification according to GO terms showed that this
subnetwork was significantly enriched with fatty acid and several
metabolic related terms, including ''phospholipid metabolic
process'', ''lipid metabolic process'', ''3-oxoacyl-[acyl-carrier-
protein] reductase (NADPH) activity'', and ''phospholipase A2
activity'' (Figure 4B and 4C, Table S30). This is clearly consistent
with previous knowledge of sporopollenin biochemistry [35–37],
indicating that this co-expression network reflects the underlying
molecular mechanism for exine formation.
In addition to the above three guide genes, this network
contained several fatty acid metabolic genes, such as OsMS2,
OsACOS5 (ACYL-COA SYNTHETASE5), OsPKSs (POLYKETIDE
SYNTHASE), and OsTKPRs (TETRAKETIDE a-PYRONE REDUC-
TASE), homologs of which have been demonstrated to be
important for sporopollenin biosynthesis in Arabidopsis [41–44]
(Figure 4A; Table 2). In the latest model of the sporopollenin
biosynthetic pathway [44,45], fatty acid precursors are esterified to
CoA by ACOS5, and then hydroxylated by CYP703A and
CYP704B to produce substrates of the subsequent ACOS5
reaction (Figure 5). Subsequently, ACOS5, PKSs, TKPRs, and
MS2 mediate the biochemical reactions to produce sporopollenin
precursors via fatty alcohols (Figure 5). Consistently, our pollen
wall specific network confirmed significant gene interactions
between these molecular players (red bold lines in Figure 4A).
Furthermore, in order to identify an as-yet-unknown specific
thioesterase producing the CYP703A/CYP704B substrates, we
compared the expression patterns of 15 rice thioesterase genes
with those of the sporopollenin biosynthetic genes, and detected
one thioestrase gene (Os09g0517700) that was co-expressed with
OsPKSB and OsTKPR2 at the significant PCC level (Table 3).
Although Os09g0517700 was not found on the pollen wall
synthesis subnetwork (Figure 4A) because of higher MR values, it
might play an important role in the sporopollenin biosynthetic
In addition to the previously characterized genes, several novel
genes, whose products participate in fatty acid metabolism,
secondary metabolism, and lipid transport, were identified within
this network (Figure 4A and Table 2). It is probable that these
genes are associated with novel metabolic processes related to
sporopollenin biosynthesis or sporopollenin transfer from tapetal
cells to anther locules (Figure 5).
It has also been reported that some MYB and bHLH
transcriptional factors, including GAMYB, AMS, UDT1, and
AtMYB103, regulate the expression of sporopollenin biosynthetic
genes [38,46–48]. The 108 co-expressed genes in the pollen wall
synthesis network included the two candidates (Os01g0293100
and Os03g0296000) for the transcriptional regulation for sporo-
pollenin biosynthesis (Figure 4A and Table 2). Os03g0296000 is a
rice homolog of Arabidopsis MYB35 (TDF1) [49] responsible for
tapetum development, whereas Os01g0293100 is a novel bHLH
type transcription factor, not previously studied. In this network,
Os01g0293100 is directly connected to CYP704B2 (blue line in
Figure 4A), suggesting that Os01g0293100 could be involved in
sporopollenin biosynthesis via modulating CYP704B2 expression
(Figure 5). Thus, the pollen wall synthesis network in the present
co-expression analysis efficiently provides us with useful informa-
tion to identify novel gene functions and interactions.
Figure 3. The meiosis-specific co-expression network in rice. (A) The co-expression subnetwork constructed using 9 reported rice meiotic
genes as guide genes (red circles). The network contains the most-recently reported meiotic genes (blue circles), putative meiotic genes (green
circles) and other genes (open circles). A red bold line indicates the direct connection of PAIR2 and ZEP1 in the co-expression network. A link between
two nodes indicates direct interaction with PCC .0.64 and MR ,10. The subnetwork vicinity is extracted by taking 2 steps out from a guide gene. (B,
C) Percentage of genes classified in enriched GO terms of BP (B) or MF (C) for the genes in the meiosis-specific network (red) relative to for 11,456
genes (blue) that had at least one connection at the PCC (.0.64) and MR (,10) thresholds. Enrichment analysis is performed using the
hypergeometric distribution (P value ,0.05).
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Co-expression network analysis in plants
In Arabidopsis, useful databases for co-expression analysis are
being constructed, and several novel genes have been identified
from these data. Aoki et al. [3] and Obayashi et al. [50] listed the
available co-expression databases in Arabidopsis, including CressEx-
press [51], CoP [52], ACT [53], GeneCAT [54], and ATTED-II
[55]. As described in these review articles, novel genes for enzymes
involved in metabolic pathways, signal factors of biological events
(including transcription factors), and components of protein
complexes have been identified by using the publicly available
databases or analyzing a laboratory's own co-expression networks.
For example, irregular xylem8 (IRX8) and irregular xylem13 (IRX13),
which are involved in secondary cell wall formation, were
identified from co-expression analysis guided by members of a
cellulose synthase (CESA) family [4]. IRX8 encodes glycosyltrans-
ferase (GT8), which is involved in pectin synthesis, whereas IRX13
Table 1. Known or putative meiotic genes in the meiosis-specific subnetwork.
RAP-ID Gene_name DescriptionCategory
Transition from mitosis into meiosis
Os03g0800200 MEL1PAZ domain containing proteinGuide gene
Os12g0572800MEL2RNA recognition motif family proteinKnown meiotic gene
Os01g0907900OsAML1 AML1, putative, expressed Known meiotic gene
Os05g0129900RNA polymeraseRNA polymerase II-associated protein 3 Putative
Os08g0399500SET domain YDG/SRA domain containing proteinPutative
Os05g0122500PWWP domain PWWP domain containing proteinPutative
Os08g0384100 Double-stranded RNA binding double-stranded RNA binding motif containing proteinPutative
Os02g0730100Ribonucleoproteinpre-mRNA processing ribonucleoprotein, binding region Putative
Homologus pairing
Os03g0106300PAIR1 PAIR1Guide gene
Os09g0506800PAIR2 retrotransposon protein, putative, SINE subclass Guide gene
Os04g0452500 ZEP1synaptonemal complex protein 2 Known meiotic gene
Os10g0405500PAIR3expressed proteinGuide gene
Meiotic replication and chrmosome structure control
Os02g0511900OsPOLE1POLE1 - Putative DNA polymerase epsilon catalytic subunitPutative
Os09g0521900 Flap endonucleaseflap endonucleasePutative
Os05g0580500OsRAD21-4Rad21 / Rec8 like protein, putativeGuide gene
Os01g0904400OsSMC2 chromosome segregation proteinPutative
Os06g0693300 OsRPA2C RPA2CPutative
Meiotic recombination
Os12g0143800 DMC1ADNA repair protein Rad51 Guide gene
Os11g0146800DMC1BDNA repair protein Rad51 Guide gene
Os04g0648500BRCA1-associated protein BRCA1-associated proteinPutative
Os03g0242100OsRAD17 cell cycle checkpoint protein RAD17Putative
Os05g0389800RNA helicaseATP-dependent RNA helicasePutative
Os07g0636200SNF2SNF2 family N-terminal domain containing protein Putative
Os02g0617500OsMER3 DEAD/DEAH box helicase domain containing protein Guide gene
Os07g0486000OsMSH4mutS family domain IV containing protein Known meiotic gene
Os05g0498300OsMSH5 mutS domain V family proteinKnown meiotic gene
Meiotic progression
Os03g0225200 OsSDScyclin Guide gene
Os01g0233500 CycA (Cyclin A)cyclin-A1 Putative
Os01g0281200CycB (Cyclin B)cyclinPutative
Os07g0556000CycC (Cyclin D) cyclinPutative
Os07g0624900SKP1BSKP1-like protein 1BPutative
Os03g0716200OsMMD1PHD-finger domain containing proteinKnown meiotic gene
Os09g0242300OsCDC20 WD domain, G-beta repeat domain containing proteinPutative
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encodes an unknown protein similar to an arabinogalactan
protein. Involvement of IRX8 and IRX13 in secondary cell wall
formation was demonstrated by T-DNA insertion mutagenesis.
Similarly, Oikawa and co-workers [56] conducted a comparative
co-expression analysis between Arabidopsis and rice, and proposed a
model for secondary cell wall formation of Arabidopsis and rice,
based on the combined approach of co-expression and predicted
cellular localization. More recently, by using Arabidopsis co-
expression networks, UGP3, a gene coding for UDP-glucose
pyrophosphorylase 3 implicated in sulfolipid biosynthesis [57],
NAI2 implicated in endoplasmic reticulum body formation [58],
BTS, a BRUTUS gene coding for bHLH-type transcription factor
Figure 4. The pollen wall synthesis co-expression network in rice. (A) The co-expression subnetwork constructed using 3 reported rice genes
for sporopollenin biosynthesis or transport as guide genes (red circles). The network contains putative genes related to pollen wall synthesis (green
circles). Red bold lines indicate the direct connection of CYP703A3, CYP704B2, OsACOS5, OsPKSs, and OsTKPRs in the co-expression network. A blue
bold line indicates the direct connection between Os01g0293100 (a bHLH-type transcription factor) and CYP704B2. A link between two nodes
indicates direct interaction with PCC .0.64 and MR ,10. The subnetwork vicinity is extracted by taking 2 steps out from a guide gene. (B, C)
Percentage of genes classified in enriched GO terms of BP (B) or MF (C) for the genes in the meiosis-specific network (red) relative to for 11,456 genes
(blue) that had at least one connection at the PCC (.0.64) and MR (,10) thresholds. Enrichment analysis is performed using the hypergeometric
distribution (P value ,0.05).
Table 2. Known or putative rice genes involved in pollen wall synthesis in the pollen wall synthesis subnetwork.
RAP-ID Gene_name DescriptionCategory
The synthesis of Fatty acids precursers
Os12g0242700 KAR 3-oxoacyl-reductase, chloroplast precursorPutative
Os02g0504800 Os02g0504800 PE-PGRS family proteinPutative
Sporopollenin biosynthesis
Os04g0310800OsACOS5 AMP-binding domain containing proteinKnown pollen wall synthesis gene
Os08g0131100CYP703A3 cytochrome P450Guide gene
Os03g0168600CYP704B2 cytochrome P450Guide gene
Os10g0484800OsPKSA stilbene synthase Known pollen wall synthesis gene
Os07g0411300 OsPKSBchalcone and stilbene synthases Known pollen wall synthesis gene
Os08g0515900OsTKPR1dihydroflavonol-4-reductaseKnown pollen wall synthesis gene
Os01g0127500OsTKPR2dihydroflavonol-4-reductaseKnown pollen wall synthesis gene
Os03g0167600OsMS2male sterility protein Known pollen wall synthesis gene
Os090517700Os090517700 acyl-coenzyme A thioesterase 10, mitochondrial precursorPutative
The transcriptional regulation of sporopollenin biosynthetic genes
Os01g0293100 bHLHhelix-loop-helix DNA-binding domain containing protein Putative
Os03g0296000OsTDF1MYB family transcription factorKnown pollen wall synthesis gene
The secondary metabolism
Os01g0933900 Os01g0933900glutathione S-transferasePutative
Os01g0651500Os01g0651500wax synthase isoform 3Putative
Os03g0263600Os03g0263600strictosidine synthase Putative
Os05g0498200Os05g0498200glycine-rich cell wall proteinPutative
Os04g0449800 Os04g0449800alpha/beta hydrolase foldPutative
The transport of Sporopollenin
Os11g0582500Osc6LTPL68 - Protease inhibitor/seed storage/LTP family protein precursorGuide gene
Os07g0244900LTPLTPL54 - Protease inhibitor/seed storage/LTP family protein precursorPutative
Os09g0525500 LTPLTPL45 - Protease inhibitor/seed storage/LTP family protein precursor Putative
Os03g0357500LTP LTPL1 - Protease inhibitor/seed storage/LTP family protein precursorPutative
Os08g0546300LTPLTPL44 - Protease inhibitor/seed storage/LTP family protein precursorPutative
Os08g0545800LTP LTPL97 - Protease inhibitor/seed storage/LTP family protein precursorPutative
Os01g0691300 LTPLTPL150 - Protease inhibitor/seed storage/LTP family protein precursorPutative
Os08g0496800 OsRAFTINBURP domain containing proteinKnown pollen wall synthesis gene
Os09g0480900 BURPBURP domain containing proteinPutative
Os06g0607700 OsWBC27ABC-2 type transporter domain containing proteinKnown pollen wall synthesis gene
Network Analysis of Anther-Expressed Genes in Rice
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Page 9
co-expressed with POPEYE in iron homeostasis [59], and
STOMAGEN, encoding secretory peptide regulating stomatal
density [60], were successfully identified. Thus, co-expression
network analysis is a powerful tool for the identification of novel
Arabidopsis genes, and such an approach is undoubtedly also
applicable to rice research.
In the present study, we performed co-expression network
analysis of rice genes by using 176 datasets of 44 K agilent
microarrays (including 33 LM-microarray datasets of the male
reproductive organs), and constructed two different co-expression
subnetworks (meiosis-specific and pollen-wall-synthesis networks)
to demonstrate the efficiency of this technique. Integrative
application of such comprehensive microarray data enabled us
to construct high-resolution networks (Figures 3A and 4A), which
included useful information for the dissection of gene regulation in
biological processes. This demonstrates that we can now perform
co-expression screening to pursue our interests in specific genes or
biological events in rice, as well as in Arabidopsis, by the use of our
reliable network datasets and the other effective rice databases
reported to date [61–64].
To the best of our knowledge, this is one of the few reports of
the application of co-expression network analysis to specific
biological processes in rice. Four rice co-expression databases
are now publicly available. In the most recent version of ATTED-
II, the co-expression network can be constructed by using the
mutual rank of the Pearson's correlation coefficient (PCC) in rice
as well as in Arabidopsis [61]. Ficklin et al. [62] demonstrated
application of the Gene Co-expression Network Browser, a
website providing a rice co-expression network constructed by
random matrix theory and weighted correlation network analysis.
In addition, RiceArrayNet [63], based on PCC, and OryzaExpress
[64], based on correspondence analysis, are available as rice co-
expression databases. These four recent papers [61–64] reported
the availability of the rice co-expression databases, but did not
provide a demonstration of the identification of genes in relation to
specific biological events. Therefore, our present report is a
valuable addition to the understanding of the utility of rice co-
expression network analysis, together with discussion of specific
examples of successful construction of meiosis-specific and pollen-
wall-synthesis networks.
Application of co-expression network analysis in sexual
reproduction research
Our co-expression analysis included the LM microarray data of
male reproductive organs, which can discriminate transcriptomes
of gametophytic pollen/microspore cells and sporophytic tapetum
cells in the anther, and will provide more precise information on
transcriptional regulation of anther genes than has been available
previously. None of the examples of gene identification using
Arabidopsis co-expression networks described above focus on the
microarray databases of reproductive organs, conditions or
developmental stages [4,56–60]. Hence, our co-expression net-
work datasets, focusing on male gametophyte development, shed
light on a new and interesting aspect of the potential of co-
expression analysis. It may be interesting to apply co-expression
network analysis to other LM microarray datasets targeted at
specific tissues and cells.
Our meiosis-specific and pollen-wall-synthesis networks reveal
that many previously unidentified genes are co-expressed in these
specific reproductive events. A reverse genetics approach to the
study of these genes will lead to identification of novel meiosis-
specific and pollen-wall-synthesis genes. Because there are many
interesting and specific biological events during male gametophyte
development, the very large co-expression datasets presented here
will provide an excellent opportunity for plant researchers to study
the molecular biology of sexual reproduction in rice.
Materials and Methods
Plant materials
Rice (O. sativa L. ssp. japonica cv. Nipponbare) plants were grown
in a greenhouse under normal conditions. Anthers at the
premeiosis stage were collected after their developmental stages
were confirmed by DAPI staining using one of the six anthers of
each flower.
Anthers were fixed in Farmer's fixative (ethanol: acetate=3:1)
overnight at 4uC. Dehydration and paraffin embedding were
performed using a microwave processor [31]. Paraffin-embedded
sections were cut to a thickness of 16 mm and mounted on PEN
membrane glass slides (Molecular Devices, Sunnyvale, CA, USA)
for LM. To remove paraffin, slides were immersed in 100% xylene
(twice), 50% xylene/50% ethanol, and 100% ethanol (v/v), for
5 min at each step and then air-dried completely at room
temperature. Three or four individual flowers were used for each
LM experiment. LM was performed using the Veritas Laser
Microdissection System LCC1704 (Molecular Devices). Selected
areas were captured by an infrared (IR) laser onto CapSure Macro
Figure 5. The current model of the sporopollenin biosynthetic
Network Analysis of Anther-Expressed Genes in Rice
PLoS ONE | www.plosone.org9October 2011 | Volume 6 | Issue 10 | e26162
Page 10
LCM Caps (Molecular Devices) and subsequently cut by a UV
laser. The target cells that fused to the LCM cap were collected by
removing the cap from the tissue section.
Microarray analysis
Total RNAs were extracted from the LM cells with a
PicoPureTMRNA isolation kit (Molecular Devices), quantified
with a Quant-iTTMRiboGreen RNA reagent and kit (Invitrogen,
Carlsbad, CA, USA), and subjected to the rice 44 K oligo
microarray (Agilent Technologies, Santa Clara, CA, USA), which
contains ,42,000 oligonucleotides based on the nucleotide
sequence and full-length cDNA of the Rice Annotation Project
Database (RAP-DB) [65]. Fluorescent probe labeling, hybridiza-
tion, and scanning were performed according to the manufactur-
er's instructions (Agilent Technologies), with slight modifications.
Each experiment was performed three times using independently
isolated samples (three biological replicates). Feature extraction
software (Agilent Technologies) was used to delineate and measure
the signal intensity of each spot in the array. All microarray data,
including our microarray data and publically available data, were
statistically normalized by variance stabilization normalization
(VSN) using R software (http://www.r-project.org/). The nor-
malized signal intensities were then transformed to log base 2. All
microarray data is MIAME compliant and the raw data from this
study have been deposited in Gene Expression Omnibus (GEO;
GO and EC analyses
GO terms were obtained for rice genes to classify their
biological functions [66]. GO terms for GO enrichment analysis
and EC analysis were retrieved from the RAP-DB. EC analysis
was performed as previously described [21,67], with the following
modifications: genes with the same GO term were used as a set of
predefined functional genes (each set contained 10 or more genes).
EC reports the fraction of gene pairs per GO category that show
elevated co-expression. Here, the PCC was utilized as a measure
for expression similarity, and we used a PCC threshold of 0.64,
corresponding to the 99thpercentile of random PCC distribution
derived for 1,000 random genes (approximately 1,000*999*0.5
gene pairs). To calculate the random EC for GO categories,
random gene sets were sampled with the same size as the category
under investigation. For GO enrichment analysis, the statistical
significance of GO enrichment within co-expressed genes was
evaluated against a background set consisting of 11,456 genes with
at least one connection from the dataset of rice co-expression
networks using the hypergeometric distribution without a multiple-
testing correlation, and P values ,0.05 were set as the significant
Co-expression analysis
To construct co-expression networks, we calculated the PCC
values for all combinations of unique 29,864 probes present on the
rice 44 K oligo microarray. We estimated two PCC thresholds of
0.48 and 0.65, corresponding to the 95thand 99thpercentile of
random PCC distribution derived for 1,000 random genes
(approximately 1,000*999*0.5 gene pairs). In some previous co-
expression studies [3,68], any two genes with a PCC value greater
than 0.6 between their expression profiles were considered as co-
expressed genes. Therefore, we set a PCC threshold of 0.64
corresponding to the 99thpercentile of random PCC distribution,
as described above. For all gene pairs with a significant PCC value,
we also calculated MR values between them as another value of
co-expression measure to further reduce the number of false
positives, according to a previous report [69]. Finally, if the
absolute value of MR was lower than 10, the gene pair was
considered as a significant connection for the co-expression
network. These calculations were conducted using the R/
Bioconductor. In extracting meiosis or pollen-wall subnetwork
datasets, the network vicinity was extracted by taking 2 steps out
from a guide gene, as described previously by Mutwil et al. [70].
The network was illustrated using the program Cytoscape.
Table 3. Expression similarities between 15 putative thioesterase genes and 7 sporopollenin biosynthetic genes in rice.
RAP-ID OsTKPR2CYP704B2 OsACOS5OsPKSBCYP703A3OsTKPR1 OsPKSA
20.136 0.090
Os05g01377000.3150.0390.286 0.2820.1180.319 0.110
Os02g0660800 N.T.N.T.N.T.N.T.N.T.N.T.N.T.
The PCC value of each gene pair was used as a measure of expression similarity.
Values with asterisk indicate a higher PCC value than the threshold value, 0.64.
N.T., Not tested because of no probe in microarray slide.
Network Analysis of Anther-Expressed Genes in Rice
PLoS ONE | www.plosone.org10October 2011 | Volume 6 | Issue 10 | e26162
Page 11
Supporting Information
Figure S1
the guide genes after normalization by VSN algorism.
Nine meiosis guide genes (A) are preferentially expressed in anther
and microspore at meiosis (MEI) stage, whereas three pollen wall
guide genes (B) are expressed in microspore and tapetum at tetrad
(TET) or uninuclear (UNI) stage. The bar at top represents the
average signal intensity (log2). Details on the individual samples
can be found in Table S1. All 176 microarray data used for co-
expression analysis show a similar expression pattern to those of
these guide genes, respectively (Tables S2, S3, S4, S5, S6, S7, S8,
Representative tissue expression profile of
Table S1
GEO IDs of the 176 microarray profiles in this
Table S2
expressed genes.
Microarray data of Leaf Blade preferentially
Table S3
expressed genes.
Microarray data of Leaf Sheath preferentially
Table S4
pressed genes.
Microarray data of Root preferentially ex-
Table S5
pressed genes.
Microarray data of Stem preferentially ex-
Table S6
tially expressed genes.
Microarray data of Inflorescence preferen-
Table S7
erentially expressed genes.
Microarray data of Anther_0.3–0.6 mm pref-
Table S8
entially expressed genes.
Microarray data of Premeiosis anther prefer-
Table S9
preferentially expressed genes.
Microarray data of Meiosis (MEI)-microspore
Table S10
preferentially expressed genes.
Microarray data of Meiosis (MEI)-tapetum
Table S11
erentially expressed genes.
Microarray data of Anther_0.7–1.0 mm pref-
Table S12
preferentially expressed genes.
Microarray data of Tetrad (TET)-microspore
Table S13
preferentially expressed genes.
Microarray data of Tetrad (TET)-tapetum
Table S14
erentially expressed genes.
Microarray data of Anther_1.2–1.5 mm pref-
Table S15
spore preferentially expressed genes.
Microarray data of Uninuclear (UN)-micro
Table S16
preferentially expressed genes.
Microarray data of Uninuclear (UN)-tapetum
Table S17
erentially expressed genes.
Microarray data of Anther_1.6–2.0 mm pref-
Table S18
preferentially expressed genes.
Microarray data of Bicellular (BC)-pollen
Table S19
preferentially expressed genes.
Microarray data of Tricellular (TC)-pollen
Table S20
expressed genes.
Microarray data of Pistil preferentially
Table S21
expressed genes.
Microarray data of Lemma preferentially
Table S22
expressed genes.
Microarray data of Palea preferentially
Table S23
expressed genes.
Microarray data of Ovary preferentially
Table S24
expressed genes.
Microarray data of Embryo preferentially
Table S25
ly expressed genes.
Microarray data of Endosperm preferential-
Table S26
EC scores for different GO categories.
Table S27
meiotic subnetwork.
PCC and MR values of all gene pairs in the
Table S28
meiotic subnetwork.
Frequency of GO categories for genes in the
Table S29
pollen wall synthesis subnetwork.
PCC and MR values of all gene pairs in the
Table S30
pollen wall synthesis subnetwork.
Frequency of GO categories for genes in the
We thank M. Miyano (Tohoku University), S. Tashiro (Soma Agricultural
School), and H. Kamakura (The University of Tokyo) for their helpful
assistance. We are also grateful to K. Yano (Meiji University), M.
Kawagishi-Kobayashi (National Institute of Crop Science) and A.
Higashitani (Tohoku University) for their useful discussion.
Network Analysis of Anther-Expressed Genes in Rice
PLoS ONE | www.plosone.org11 October 2011 | Volume 6 | Issue 10 | e26162
Page 12
Author Contributions
Conceived and directed the project: MW MM. Designed and performed
laser microdissection experiment: HT K. Shiono NT MN. Designed and
performed microarray analysis: YN. Analyzed microarray data and
performed GO, EC and co-expression network analyses: KA GS K.
Suwabe KH KY TH. Discussed the results and commented on the
manuscript: KA GS K. Suwabe TH HT K. Shiono KY NT MN YN MW.
Wrote the manuscript: KA GS K. Suwabe MW MM.
1. Lee HK, Hsu AK, Sajdak J, Qin J, Pavlidis P (2004) Coexpression analysis of
human genes across many microarray data sets. Genome Res 14: 1085–
2. Ihmels J, Levy R, Barkai N (2004) Principles of transcriptional control in the
metabolic network of Saccharomyces cerevisiae. Nat Biotechnol 22: 86–92.
3. Aoki K, Ogata Y, Shibata D (2007) Approaches for extracting practical
information from gene co-expression networks in plant biology. Plant Cell
Physiol 48: 381–390.
4. Persson S, Wei H, Milne J, Page GP, Somerville CR (2005) Identification of
genes required for cellulose synthesis by regression analysis of public microarray
data sets. Proc Natl Acad Sci U S A 102: 8633–9638.
5. Hirai MY, Sugiyama K, Sawada Y, Tohge T, Obayashi T, et al. (2007) Omics-
based identification of Arabidopsis Myb transcription factors regulating aliphatic
glucosinolate biosynthesis. Proc Natl Acad Sci U S A 104: 6478–6483.
6. Swarbreck D, Wilks C, Lamesch P, Berardini TZ, Garcia-Hernandez M, et al.
(2008) The Arabidopsis Information Resource (TAIR): gene structure and
function annotation. Nucleic Acids Res 36: D1009–1014.
7. Sato Y, Antonio BA, Namiki N, Takehisa H, Minami H, et al. (2011) RiceXPro:
a platform for monitoring gene expression in japonica rice grown under natural
field conditions. Nucleic Acids Res 39: D1141–1148.
8. Suzuki G (2009) Recent progress in plant reproduction research: the story of the
male gametophyte through to successful fertilization. Plant Cell Physiol 50:
9. Koltunow AM, Truettner J, Cox KH, Wallroth M, Goldberg RB (1990)
Different temporal and spatial gene expression patterns occur during anther
development. Plant Cell 2: 1201–1224.
10. Scott R, Dagless E, Hodge R, Paul W, Soufleri I, et al. (1991) Patterns of gene
expression in developing anthers of Brassica napus. Plant Mol Biol 17: 195–207.
11. Tsuchiya T, Toriyama K, Ejiri S, Hinata K (1994) Molecular characterization of
rice genes specifically expressed in the anther tapetum. Plant Mol Biol 26:
12. Hihara Y, Hara C, Uchimiya H (1996) Isolation and characterization of two
cDNA clones for mRNAs that are abundantly expressed in immature anthers of
rice (Oryza sativa L.). Plant Mol Biol 30: 1181–1193.
13. Rubinelli P, Hu Y, Ma H (1998) Identification, sequence analysis and expression
studies of novel anther-specific genes of Arabidopsis thaliana. Plant Mol Biol 37:
14. Jeon J-S, Chung Y-Y, Lee S, Yi G-H, Oh B-G, et al. (1999) Isolation and
characterization of an anther-specific gene, RA8, from rice (Oryza sativa L.). Plant
Mol Biol 39: 35–44.
15. Masuko H, Endo M, Saito H, Hakozaki H, Park J-I, et al. (2006) Anther-specific
genes, which expressed through microsporogenesis, are temporally and spatially
regulated in model legume, Lotus japonicus. Genes Genet Syst 81: 57–62.
16. Endo M, Matsubara H, Kokubun T, Masuko H, Takahata Y, et al. (2002) The
advantages of cDNA microarray as an effective tool for identification of
reproductive organ-specific genes in a model legume, Lotus japonicus. FEBS Lett
514: 229–237.
17. Amagai M, Ariizumi T, Endo M, Hatakeyama K, Kuwata C, et al. (2003)
Identification of anther-specific genes in a cruciferous model plant, Arabidopsis
thaliana, by using a combination of Arabidopsis macroarray and mRNA derived
from Brassica oleracea. Sex Plant Reprod 15: 213–222.
18. Suwabe K, Suzuki G, Takahashi H, Shiono K, Endo M, et al. (2008) Separated
transcriptomes of male gametophyte and tapetum in rice: validity of a laser
microdissection (LM) microarray. Plant Cell Physiol 49: 1407–1416.
19. Hobo T, Suwabe K, Aya K, Suzuki G, Yano K, et al. (2008) Various
spatiotemporal expression profiles of anther-expressed genes in rice. Plant Cell
Physiol 49: 1417–1428.
20. Usadel B, Obayashi T, Mutwil M, Giorgi FM, Bassel GW, et al. (2009) Co-
expression tools for plant biology: opportunities for hypothesis generation and
caveats. Plant Cell Environ 32: 1633–1651.
21. Pilpel Y, Sudarsanam P, Church GM (2001) Identifying regulatory networks by
combinatorial analysis of promoter elements. Nat Genet 29: 153–159.
22. Nonomura K-I, Nakano M, Fukuda T, Eiguchi M, Miyao A, et al. (2004) The
novel gene HOMOLOGOUS PAIRING ABERRATION IN RICE MEIOSIS1 of rice
encodes a putative coiled-coil protein required for homologous chromosome
pairing in meiosis. Plant Cell 16: 1008–1020.
23. Nonomura K-I, Nakano M, Eiguchi M, Suzuki T, Kurata N (2006) PAIR2 is
essential for homologous chromosome synapsis in rice meiosis I. J Cell Sci 119:
24. Yuan W, Li X, Chang Y, Wen R, Chen G, et al. (2009) Mutation of the rice
gene PAIR3 results in lack of bivalent formation in meiosis. Plant J 59: 303–315.
25. Nonomura K-I, Morohoshi A, Nakano M, Eiguchi M, Miyao A, et al. (2007) A
germ cell specific gene of the ARGONAUTE family is essential for the progression
of premeiotic mitosis and meiosis during sporogenesis in rice. Plant Cell 19:
26. Zhang L, Tao J, Wang S, Chong K, Wang T (2006) The rice OsRad21-4, an
orthologue of yeast Rec8 protein, is required for efficient meiosis. Plant Mol Biol
60: 533–554.
27. Deng ZY, Wang T (2007) OsDMC1 is required for homologous pairing in Oryza
sativa. Plant Mol Biol 65: 31–42.
28. Chang L, Ma H, Xue HW (2009) Functional conservation of the meiotic genes
SDS and RCK in male meiosis in the monocot rice. Cell Res 19: 768–
29. Wang M, Wang K, Tang D, Wei C, Li M, et al. (2010) The central element
protein ZEP1 of the synaptonemal complex regulates the number of crossovers
during meiosis in rice. Plant Cell 22: 417–430.
30. Nonomura K-I, Eiguchi M, Nakano M, Takashima K, Komeda N, et al. (2011)
A novel RNA-recognition-motif protein is required for premeiotic G1/S-phase
transition in rice (Oryza sativa L.). PLoS Genet 7: e1001265.
31. Kaur J, Sebastian J, Siddiqi I (2006) The Arabidopsis-mei2-like genes play a role in
meiosis and vegetative growth in Arabidopsis. Plant Cell 18: 545–559.
32. Watanabe Y, Shinozaki-Yabana S, Chikashige Y, Hiraoka Y, Yamamoto M
(1997) Phosphorylation of RNA-binding protein controls cell cycle switch from
mitotic to meiotic in fission yeast. Nature 386: 187–190.
33. Shinohara M, Sakai K, Ogawa T, Shinohara A (2003) The mitotic DNA
damage checkpoint proteins Rad17 and Rad24 are required for repair of
double-strand breaks during meiosis in yeast. Genetics 164: 855–865.
34. Higgins JD, Vignard J, Mercier R, Pugh AG, Franklin FC, et al. (2008) AtMSH5
partners AtMSH4 in the class I meiotic crossover pathway in Arabidopsis thaliana,
but is not required for synapsis. Plant J. 55: 28–39.
35. Rozema J, Broekman RA, Blokker P, Meijkamp BB, de Bakker N, et al. (2001)
UV-B absorbance and UV-B absorbing compounds (para-coumaric acid) in
pollen and sporopollenin: the perspective to track historic UV-B levels.
J Photochem Photobiol B 62: 108–117.
36. Bubert H, Lambert J, Steuernagel S, Ahlers F, Wiermann R (2002) Continuous
decomposition of sporopollenin from pollen of Typha angustifolia L. by acidic
methanolysis. Z Naturforsch C 57: 1035–1041.
37. Ahlers F, Lambert J, Wiermann R (2003) Acetylation and silylation of piperidine
solubilized sporopollenin from pollen of Typha angustifolia L. Z Naturforsch C 58:
38. Aya K, Ueguchi-Tanaka M, Kondo M, Hamada K, Yano K, et al. (2009)
Gibberellin modulates anther development in rice via the transcriptional
regulation of GAMYB. Plant Cell. 21: 1453–1472.
39. Li H, Pinot F, Sauveplane V, Werck-Reichhart D, Diehl P, et al. (2010)
Cytochrome P450 family member CYP704B2 catalyzes the {omega}-hydrox-
ylation of fatty acids and is required for anther cutin biosynthesis and pollen
exine formation in rice. Plant Cell. 22: 173–190.
40. Zhang D, Liang W, Yin C, Zong J, Gu F, et al. (2010) OsC6, encoding a lipid
transfer protein, is required for postmeiotic anther development in rice. Plant
Physiol. 154: 149–162.
41. Aarts MG, Hodge R, Kalantidis K, Florack D, Wilson ZA, et al. (1997) The
Arabidopsis MALE STERILITY 2 protein shares similarity with reductases in
elongation/condensation complexes. Plant J 12: 615–623.
42. de Azevedo Souza C, Kim SS, Koch S, Kienow L, Schneider K, et al. (2009) A
novel fatty Acyl-CoA Synthetase is required for pollen development and
sporopollenin biosynthesis in Arabidopsis. Plant Cell 21: 507–525.
43. Kim SS, Grienenberger E, Lallemand B, Colpitts CC, Kim SY, et al. (2010)
LAP6/POLYKETIDE SYNTHASE A and LAP5/POLYKETIDE SYNTHASE B
encode hydroxyalkyl a-pyrone synthases required for pollen development and
sporopollenin biosynthesis in Arabidopsis thaliana. Plant Cell 22: 4045–4066.
44. Grienenberger E, Kim SS, Lallemand B, Geoffroy P, Heintz D, et al. (2010)
Analysis of TETRAKETIDE a-PYRONE REDUCTASE function in Arabidopsis
thaliana reveals a previously unknown, but conserved, biochemical pathway in
sporopollenin monomer biosynthesis. Plant Cell 22: 4067–4083.
45. Ariizumi T, Toriyama K (2011) Genetic regulation of sporopollenin synthesis
and pollen exine development. Annu Rev Plant Biol 62: 437–460.
46. Sorensen AM, Kro ¨ber S, Unte US, Huijser P, Dekker K, et al. (2003) The
Arabidopsis ABORTED MICROSPORES (AMS) gene encodes a MYC class
transcription factor. Plant J 33: 413–423.
47. Jung KH, Han MJ, Lee YS, Kim YW, Hwang I, et al. (2005) Rice Undeveloped
Tapetum1 is a major regulator of early tapetum development. Plant Cell 17:
48. Zhang ZB, Zhu J, Gao JF, Wang C, Li H, et al. (2007) Transcription factor
AtMYB103 is required for anther development by regulating tapetum
development, callose dissolution and exine formation in Arabidopsis. Plant J 52:
49. Zhu J, Chen H, Li H, Gao JF, Jiang H, et al. (2008) Defective in Tapetal development
and function 1 is essential for anther development and tapetal function for
microspore maturation in Arabidopsis. Plant J 55: 266–277.
Network Analysis of Anther-Expressed Genes in Rice
PLoS ONE | www.plosone.org12 October 2011 | Volume 6 | Issue 10 | e26162
Page 13
50. Obayashi T, Kinoshita K (2010) Coexpression landscape in ATTED-II: usage of
gene list and gene network for various types of pathways. J Plant Res. 123:
51. Srinivasasainagendra V, Page GP, Mehta T, Coulibaly I, Loraine AE (2008)
CressExpress: a tool for large-scale mining of expression data from Arabidopsis.
Plant Physiol. 147: 1004–1016.
52. Ogata Y, Suzuki H, Sakurai N, Shibata D (2010) CoP: a database for
characterizing co-expressed gene modules with biological information in plants.
Bioinformatics. 26: 1267–1268.
53. Manfield IW, Jen CH, Pinney JW, Michalopoulos I, Bradford JR, et al. (2006)
Arabidopsis Co-expression Tool (ACT): web server tools for microarray-based
gene expression analysis. Nucleic Acids Res. 34: W504–W509.
54. Mutwil M, Obro J, Willats WG, Persson S (2008) GeneCAT--novel webtools
that combine BLAST and co-expression analyses. Nucleic Acids Res. 36:
55. Obayashi T, Kinoshita K, Nakai K, Shibaoka M, Hayashi S, et al. (2007)
ATTED-II: a database of co-expressed genes and cis elements for identifying co-
regulated gene groups in Arabidopsis. Nucleic Acids Res. 35: D863–D869.
56. Oikawa A, Joshi HJ, Rennie EA, Ebert B, Manisseri C, et al. (2010) An
integrative approach to the identification of Arabidopsis and rice genes involved in
xylan and secondary wall development. PLoS One 5: e15481.
57. Okazaki Y, Shimojima M, Sawada Y, Toyooka K, Narisawa T, et al. (2009) A
chloroplastic UDP-glucose pyrophosphorylase from Arabidopsis is the committed
enzyme for the first step of sulfolipid biosynthesis. Plant Cell 21: 892–909.
58. Yamada K, Nagano AJ, Nishina M, Hara-Nishimura I, Nishimura M (2008)
NAI2 is an endoplasmic reticulum body component that enables ER body
formation in Arabidopsis thaliana. Plant Cell 20: 2529–2540.
59. Long TA, Tsukagoshi H, Busch W, Lahner B, Salt DE, et al. (2010) The bHLH
transcription factor POPEYE regulates response to iron deficiency in Arabidopsis
roots. Plant Cell 22: 2219–2236.
60. Sugano SS, Shimada T, Imai Y, Okawa K, Tamai A, et al. (2010) Stomagen
positively regulates stomatal density in Arabidopsis. Nature 463: 241–244.
61. Obayashi T, Nishida K, Kasahara K, Kinoshita K (2011) ATTED-II updates:
condition-specific gene coexpression to extend coexpression analyses and
applications to a broad range of flowering plants. Plant Cell Physiol. 52:
62. Ficklin SP, Luo F, Feltus FA (2010) The association of multiple interacting genes
with specific phenotypes in rice using gene coexpression networks. Plant Physiol.
154: 13–24.
63. Lee TH, Kim YK, Pham TT, Song SI, Kim JK, et al. (2009) RiceArrayNet: a
database for correlating gene expression from transcriptome profiling, and its
application to the analysis of coexpressed genes in rice. Plant Physiol. 151:
64. Hamada K, Hongo K, Suwabe K, Shimizu A, Nagayama T, et al. (2011)
OryzaExpress: an integrated database of gene expression networks and omics
annotations in rice. Plant Cell Physiol. 52: 220–229.
65. Rice Annotation Project (2008) The Rice Annotation Project Database (RAP-
DB): 2008 update. Nucleic Acids Res 36: D1028–1033.
66. The Gene Ontology project in 2008 (2008) Gene Ontology Consortium. Nucleic
Acids Res 36: D440–444.
67. Vandepoele K, Quimbaya M, Casneuf T, De Veylder L, Van de Peer Y (2009)
Unraveling transcriptional control in Arabidopsis using cis-regulatory elements
and coexpression networks. Plant Physiol 150: 535–546.
68. Fu FF, Xue HW (2010) Coexpression analysis identifies Rice Starch Regulator1,
a rice AP2/EREBP family transcription factor, as a novel rice starch biosynthesis
regulator. Plant Physiol. 154: 927–938.
69. Obayashi T, Kinoshita K (2009) Rank of correlation coefficient as a comparable
measure for biological significance of gene coexpression. DNA Research 16:
70. Mutwil M, Ruprecht C, Giorgi FM, Bringmann M, Usadel B, et al. (2009)
Transcriptional wiring of cell wall-related genes in Arabidopsis. Mol Plant 2:
Network Analysis of Anther-Expressed Genes in Rice
PLoS ONE | www.plosone.org13October 2011 | Volume 6 | Issue 10 | e26162


text:
I currently have a leak on my hose, but I cant replace it yet, I'm sure my shocks are gone to...
Questions for specific Mercedes-Benz S500 years:
Mercedes-Benz Q&AAsk Your Question
All Mercedes-Benz S500 questions
What kind of fluid for the steering pump, and brakes do you use for a 97 S 500
Is there a recall on the instrument cluster panel?
Mine does not light up
How do you replace the front brake pads
I am trying to save $$ by doing some of the repairs myself to my 1998 S500 Mercedes.
I have been...
heater will not get hot nor will my remote unlock the doors now.
My car ran hot back in October so I replace the thermostat,water pump and radiator. Now the car d...
my car makes a whining noise when i crank it.
when I crank it this whining sound comes from under the hood....everything else seems to be worki...
car wont idle, runs rough but smoothes out with added throttle.
No Engine Codes, no vacuum leaks, changed spark plugs twice (first autolite plat, than mercedes ...
How to remove coolant temperature sensor?
i need to replace the coolant temperature sensor but don't know how to remove it.
My check engine light came on and then my stalled, what does that mean?
My batter cable was not completely clamped on, could that be the problem?
Instrument cluster malfunctioning.
S500 MB yr.2000 having malfunction...
part number 220-820-06021
the right side door mirror ha a broken part, how do I repair it
Radiator Fan Assembly
I've being told that I need a new radiator fan assembly for my 2001 S500. What is the level ...
What has caused the gear shift to lock in park?
Initially when the car ws purchased this occured. It was stated that because the car had sat undr...
The steps for replacing the rearveiw ext mirror gaskets, the easiest safest way?
My S500 - 2000 MBENZ is - Eleven years old orginal rubber seal gaskets worn out falling apart - ...
ticking noise
I get a ticking sound when i start my car and as im driving. i need to know what the problem is a...
reset remote on a 1996 s500
I already changed the batteries and sometime it works< but most the time it don't
engine is mising
What steps are necessary to change spark plugsin
I need to change spark plugs in mbz500 coupe.
How do I change spark plugs in mbz s-500 coupe?
how do i remove the air conditioer unit
how do i remove the air condtioner unit
car fire
2001 s500 85,000 miles, just service by dealer. bought new
and always service by dealer.... fire...
oil filter leak
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The dashboard control buttons that work the emergency flashers, suspension options, head rests an...
trunk motor
trunk will not close tight, motor does not seem to be pulling it snug
how many quarts of oil
Regular oil & Filter change
the car stalls and cools off and starts agian
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i drive for 10mins and the car turns off
Can the rear suspension stablizing pump be unplugged?
Whenever someone sits in the rear of the car the pump starts screaming and will not stop until th...
how to atabilize the proper hight on the rear suspension?
Suddenly i noticed that my rear suspension rised noticibly.I am not sure if it was due to an elec...
engine will crank but not start some times after setting
The car will run normal but after setting some time it will crank normally but will not start.
clear service A"
how you clear service A'

subset: commoncrawl


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This pattern includes both a left-hand and right-hand version - they can be combined to make a full rainbow. Not as awesome as a Double Rainbow, but still pretty ;-)The pattern is full-color and clearly numbered/lettered.Basic paper-piecing instructions are not included. There are many great print and video tutorials available online!This pattern is for personal and non-profit use only.For each pattern sold I am donating $1 to Blood:Water Mission, a grassroots organization that works alongside African communities to improve access to clean water and medical care. You can read more about them at http://www.bloodwatermission.com/
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Three New eBooks Feature Easy to Make Christmas Crafts
FaveCrafts sister sites AllFreeKnitting, AllFreeCrochet and AllFreeSewing have come out with three new fabulous eBooks featuring easy to make Christmas crafts including knit Christmas stockings, crochet Christmas wreaths and easy to make Christmas ornaments.
November 11, 2010 - Northbrook, IL -Readers looking for fabulous Christmas ideas will be amazed at the great projects included in these three new eBooks. They can find anything from knit Christmas stockings to crochet Christmas wreaths to easy to make Christmas ornaments to hang on the tree. These projects are for anyone no matter what the skill level.
In the first eBook, 5 Knit Christmas Stocking Patterns eBook, you will find 5 patterns for Christmas Knit stockings that are great for the holidays, including some that are perfect beginner knitting patterns. You will see a classic knit stocking as well as a fun snowman stocking amongst others.
In the second eBook, 5 Free Christmas Crochet Patterns: Crochet Christmas Wreaths eBook, you will see 5 free wreath crochet patterns including a fun candy cane wreath, a Santa and gifts wreath and a beautiful ornament wreath. Wreaths aren't just for hanging on your door anymore, they can be hung on your tree as well. Whether you're crocheting something for yourself or giving it as a gift, there is nothing more special than a homemade wreath.
In the third eBook, 6 Sew Easy to Make Christmas Ornaments eBook, a collection of 6 Christmas ornaments can be found. Whether you're a hand-sewer or a sewing machine whiz, you can sew your own Christmas ornaments easily by following along with these simple Christmas craft sewing patterns. Included in this eBook are felt ornaments, fabric ornaments, tips on making quilted ornaments and more.
Visit each sister site to download each of their latest free eBooks.
Christine Weiher, Editor
Prime Publishing LLC
About AllFreeCrochet.com:
AllFreeCrochet.com is a website dedicated to the best free crochet patterns, tutorials, tips and articles on crochet. Find all varieties and skills levels, from easy to crochet afghan patterns to complex Tunisian crochet patterns.
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text: I am from Minneapolis, Minnesota and I am a freelance writer and the owner of web sites about travel to Mexico. I have homes in Mexico and Minnesota and travel between the two frequently. I am also a Destination Expert for Mexico and Senior Hotel Reviewer for the (website hidden) travel site.


text:
Release Date: 2013 |
Album Rating: 4.1Listen here:
Tada. I've never reviewed an EP before.
yeah good band name dickheads
great review as usual despite a weak-ass first sentence : )
user ratings (1)
"Even tough it's somewhat baffling that "
one mistake, good review though
Digging: Saosin - Translating the Name
Love that band name
love your pic chesse
Is this anything like Sluts? Probably not I'm guessing, but I thought it'd be mildly humorous to ask.
Ooh these whores are from my area.
Digging: CHON - Grow
Know them?
Digging: Geinoh Yamashirogumi - Ecophony Gaia
I checked out their last EP a while back. It was pretty good from what I remember. I like the album art on this one.
Digging: Kendrick Lamar - To Pimp a Butterfly
nice rev, Baby Bird is infectious
Digging: Different Strings - The Sounds of Silence, Pt. II: The Counterfeits
Album Rating: 4.0...aren't just 2 songs out for now?
Dafuq did I miss?
It's a band name that let's you know they play noise rock.
Good review. They sound pretty gnar. Might make it on my top EPs if the other songs live up to the two I heard.
#lmfao #goodtimes #bitches #trapshit #bitches2013 #ondamoneyz #theywontseeitcomin #allnightbaby
#avicii #sluts #kush #ballaz4lyfe
whores making noise
Digging: Jlin - Dark Energy
Album Rating: 4.1Thanks guys. Yeah, the next 4 tracks are equally good.
Nice. I love noise rock like this.
Nicer Dicer. Cows rule so this too must rule.
Bands: A B C D E F G H I J K L M N O P Q R S T U V W X Y Z


text:
tub with low side???
My mother is determined to get a certain kind of tub that I'm not sure even exists. She wants a standard sized tub that has the side cut very low so that she can step in it easier. She thinks they make them that way. I can't find any on the internet. We can't afford the nice "walk-in" tubs, so that's not an option. She believes that even though the side is cut down low, that in a shower, it will not go over that side and make the floor wet.
Any ideas? Money is definately an issue as well.
You will have a tough time finding this low side wall tub with out paying top dollar. Your Mom is used to all the old tubs of the early 50's and 60's.
They where about 30" wide and had low sides because most homes had small 40 gallon max hot water tanks. These tubs are easier for elderly people to step in and out of and easier still for Dad and Mom to bathe the little ones after supper.
I would suggest you look at your salvage yards for this "Old School" look. My town is full of them and most times they sadly make there way to the dump.
My Nana's is Green....
Oceania Baths make a couple of easy step over tubs.
Here is a link that might be useful: Oceania Baths - Estime Collection
The corrolary to a low tub is that the water level is very shallow and poor for actual bathing. Yes, it's OK for showering, but you're not really gaining any utility for bathing with such a tub. Why not just replace the tub with a shower? If you're at the project level of replacing a bath tub, you'll also have to replace the walls/waterproofing as well, so you're already not doing a small project. If she'd actually prefer a shower only setup, this is the time to do it.
Thanks for all your ideas! Much appreciated. I also think it would be a good time to just remove the tub and put in a stand up shower. Nothing fance, but just a shower. She will not be using it as a tub because she doesn't have the strength to get up and down. The tub she has is a "garden tub" in a mobile home. If you are familiar with them, they are a big pain. There is one step to go up, and then you have to step pretty far down. She has been stepping over the step to get in. She did not like the shower bench and it didn't fit well because of the sides of the tub. The tub is rectangle shape, but inside the tub it's oval. I will look into the Oceania tubs. THANKS AGAIN!!
The Kohler Villager is a cast iron tub with only a 14 inch deep basin. It might be worthwhile taking a look at it.
We're also considering a renovation of an upstairs bathroom. And I'm also looking for a low step over height tub. We never use a tub. Always shower. But the reason I want a tub and not just change over the tub to a shower is that many real estate folks recc. having at least one tub in a house. If we sell our home in the future, it could be to a family with babies or very young children. They would want a bath to bathe their kids. But they would not need a deep tub for that. Just a guess, but a 10-12" tall bathtub would probably work fine for them.
With more and more people going to showers over bathtubs, you would think the mfgs. would be offering a very low height bathtub for situations just like this. But that doesn't seem to be the case. I called Kohler/Sterling and American Standard. They do not offer anything less than a 14" height bathtub.


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Samsung to release 14nm octa, production underway
Eight cores of terror
Samsung has announced an updated version of its Exynos 7 Octa processor. Last year it announced a 64-bit, ARMv8 processor based on a 20nm design, but these new chips will be 14nm processors.
Vaio resurrects Z
Zed is not dead baby
The new independent Vaio has resurrected its Z range. There will be the Vaio Z and the Vaio Z canvas
Apple to kill iPad mini
Not a moment too soon
Apple has decided that its iPad mini is not worth making any more and is about to discontinue the line.
Japanese do not give a flip about smartphones
Smart moves
While the Japanese are famous for their love of brightly coloured gadgets, it seems that they are rejecting their smartphones in favour of traditional flip-phones.
Sharp denies display business sell off
Rumours wrong
A senior Sharp executive has denied that the Japanese company wants to flog off its failing display business, after it warned it will likely book its third annual net loss in four years.
Software engineers do not vaccinate
Cleverest minds are sometimes the dumbest
Figures from the US show that software engineers are more likely to fall for fake science and refuse to vaccinate their kids.
ARM does well because of embedded
ARMv8 does well
Analysts have been looking under the bonnet of ARM's good results and discovered that they might not be down to its traditional mobile stomping ground.
iOS crashes more than Lollipop
Still, you get what you pay for
A survey which is not being widely reported among the Tame Apple Press, shows that Apple's latest iOS software is not as reliable as Android Lollypop.
Microsoft still thinks the pen is mightier than the swipe
Buys Israeli company
Software giant Microsoft has written a cheque for the Israeli tech manufacturer N-trig which specialises in the development and production of digital pens and touchscreens.
South Korean anti-trust watchdog yaps at Qualcomm
Looks like everyone wants a piece
The South Koreans have looked at Qualcomm writing a billion dollar cheque to make the Chinese anti-trust watchdogs go away and thought, "I'll be having something of that."


text:
Early on I veered, then I soul-searched, dug deep like a gopher, figured I should go first, reason why I don't curse, reason why I'm sober, reason I feel younger, even though I'm older
from Homeboy Sandman – The Carpenter Lyrics on Genius
Homeboy Sandman details his life and his wanderings. He started off the path (going to law school, indulging in drugs) but then slowly found his way back and now he's a straight-edge guy who's dedicated to his craft.
To help improve the quality of the lyrics, visit Homeboy Sandman – The Carpenter Lyrics and leave a suggestion at the bottom of the page


text:
This 2004 Harley Davidson Road King Police has a 111 C.I. S & S V-Twin, Baker 6 Speed Transmission, Baker Oil Pan, Harley Davidson... More
GR Auto Gallery is pleased to offer this 2005 Harley Davidson Touring Ultra Classic for purchase. The Harley Davidson Company (HD) was started a... More
2007 Harley-Davidson FLSTN ''Deluxe'', Black, 96 cu in, 6-Speed, Vance & Hines ''True Dual'' exhaust, Road King Classic leather bags, Luggage rack and backrest,... More
Call Rick today @ 888-245-6479 Here is a rare find and deal! 2005 Harley Davidson FLSTSC Softail Heritate Springer Blue and... More
Call Rick @ 888-245-6479... More
2004 Harley Davidson Fat Boy FLSTF Loaded with extras 2 tone paint Vance and Hines Exhaust Custom floorboards and controls... More
This super-clean 5-speed bike is powered by a fuel-injected 88 ci engine. It features a detachable windshield with bag, Hooker True Duals, chrome forks... More
Offered here for sale is a mint 2005 Harley Davidson CVO (Custom Vehicle Operations) bike. Purchased new, this one-owner, lady-driven beauty has been meticulously... More
This frame-up restoration of the famous military motorcycle is a real work of art. It features a 45ci engine. The bike runs strong and... More
Powered by an 88 ci fuel-injected engine, this 5-speed bike is in very good condition. It features HD Nostalgic grips and riding pegs, chrome... More
Thats right! You can lease this 2011 Harley Davidson XL883 low for $192 per month and 0 down with your good credit! UNLIMITED MILES!!!... More
Call Rick @ 888-245-6479... More
2014 Harley Davidson Screamin Eagle CVO Breakout FOR SALE!!! Part of a Private collection. High Metallic Orange with Black finish. Extras include: Vance & Hines Full exhaust system. Chrome... More
JUST IN! AN AWESOME ONE OWNER, ADULT RIDDEN XL1200R HARLEY DAVIDSON SPORTSTER ROADSTER edition. IT HAS FLAWLESS BLACK PAINT AND IS READY FOR A... More
In honor of the historic Sturgis motorcycle rally, Harley-Davidson released the FXSB Sturgis model, employing belt drive, black chrome appointments and 80 cubic inch... More
Harley Davidson celebrated its 90th birthday by producing a few of these rare FLSTN Heritage Softail Specials - only 1500 were made sporting a... More
Ride in style! Unique, flame paint job sets this bike apart from the crowd. Custom, white seat smooths out the miles. Mileage shown is... More
Ready for around the town or long distance cruising with just 257 miles since engine rebuild. Windshield keeps those pesky bugs out of your... More
Only produced for a few years for Harley Davidson by Aermacchi in Italy, these motorcycles are difficult to find in this excellent condition. Collect... More
Yes, you are looking at an awesome Harley Davidson Buell Thunderbolt S2. It has a very low production # and also has very low... More
Lease me today with unlimited mileage!! 2010 Harley Davidson FLHX Street Glide Pearl white with Black Ice insert 2 into 1... More
This loaded 6-speed bike features a 96 ci fuel-injected engine and is decked out in chrome, including the forks, swing arms, timing cover, primary,... More
Pauchgo frame , Rebuilt 1000 cc original sportster engine , Pm quad front caliper, JP hand controls , Ace oil tank, 41mm wide glide front... More
2006 HARLEY DAVIDSON ELECTRA GLIDE CLASSIC : I BOUGHT THIS SHOWROOM CONDITION, BEAUITFUL ORIGINAL GLOSS BLACK PAINT WITH ONLY 5K MILES FROM A GOOD FRIEND... More
I PUT ALOT OF MONEY AND PLENTY OF BIKE MECHANIC'S TIME INTO RESTORING THIS BEAUTIFUL HARLEY DAVIDSON SHOVELHEAD. NO EXPENSES WERE SPARED AND THE BEST... More
custom everything,wheels,vance and hines staggers,seat,backrest,excellent paint,... More
Stk. 1940 2009 Harley-Davidson Dyna Glide Low Rider Ready to ride; this one owner 2009 Harley Davidson Dyna Glide Low Rider has an amazingly low... More
For sale in our Tampa showroom is a truly custom 2002 Harley Davidson Fat Boy. Powered by a 1450 CC engine this Harley can... More
1989 HARLEY DAVIDSON SOFTAIL EVO ENGINE One Owner 16500 MILES . CAPTAIN AMERICA , LOTS OF CHROME AND GOLD PLATED , NEW BATTERY AND... More
For sale in our St. Louis showroom is a beautiful 2006 Harley-Davidson Heritage Softail Classic. Harley-Davidson motorcycles have had a huge following in... More


text:
Utility Applications
Bill Reminder
Why miss the bill due dates and pay extra money. Use this application to keep track of your bill dates. No late fees, simply key in your bill due date and Bill Reminder will remind you in advance about the bill due dates. Recurring bills are also supported, hence reducing extra effort to key in...
Disco Lights
Disco light effects, party anywhere anytime. Turn your device into a party application. Move your shoes with the rhythm. Rock your body in the disco lights. This app is supported via search. The following will be added to your device once you download the app – Search icon, bookmark link and...
Pnr Status
Check PNR status anytime, store your PNR so you need not check your ticket to find PNR everytime. Stores your berth details and makes it available offline too. Informs about train timings and your journey details.


text:
More "Faces" wanted for popular video series
If you're a full-time faculty member and you haven't yet been filmed for a Faces of UVic Research video, the project team would like to hear from you.
In each video—which is just over one minute long—a researcher gives a short and succinct summary of their work in everyday language that quickly gets to the heart of what they do and why it matters.
President Cassels emphasizes need to sharpen focus and build on strengths
President Cassels emphasizes need to sharpen focus and build on strengths
UVic has made some big strides in the last few years—in terms of national and international recruitment, research, and graduates' success—but that's not to say we're sheltered from demographic change, competition, and the sustainability challenges facing post-secondary institutions, UVic President Jamie Cassels said during an Oct. 7 Campus Update session.
Environmental Studies at 40: learning and growing a greener campus
From the very first plans for the Gordon Head campus, natural features have been valued as a defining force in UVic's learning environment. And the Environmental Studies program, starting small with a single course in 1975, has grown substantially — not just alongside, but as a driver of green initiatives that have helped transform the campus into an internationally recognized leader in sustainability.
Many of the practices that helped UVic become the only Canadian university to make the Princeton Review's 2015 Green Honour Roll sprouted from Environmental Studies course projects, student initiatives and partnerships across the institution.
Vanier Scholar advancing FASD diagnostics
It certainly sounds novel: could a smelling test, easy enough to administer to young children, help diagnose Fetal Alcohol Spectrum Disorder (FASD)?
That possibility is taking shape, thanks to work done by UVic neuroscience PhD student Christine Fontaine.
New HSD centre to support Indigenous student success
Shauna Underwood remembers all too well what it was like to be an Indigenous student at UVic—trying to do well in her studies while also being a single parent of three children and having cultural responsibilities in her community.
"I felt like I didn't have anyone to go to. There were individual instructors who were great, but they are not always on campus or available," said Underwood, now the Indigenous advisor at the newly opened Indigenous Student Support Centre (ISSC) in the Faculty of Human and Social Development.
Two UVic scholars help debut new national college
Two University of Victoria researchers—a psychologist who studies how we age mentally and a computer scientist who explores new ways to sort through large amounts of information—are charter members of a new national "college" for outstanding young scholars.
Ancient technology, meet the future of underwater archaeology
Using some of the newest technologies in underwater archaeology, Quentin Mackie (anthropology) and Alison Proctor (engineering) may have discovered one of the oldest archaeological finds in Canada.
Right now all they know for sure is that they've discovered a line of basketball-sized rocks on the seafloor. Those rocks may well add to the mounting evidence about the ingenuity of an ice age people who lived and thrived on the west coast of Canada while much of Europe was still under ice. It's a tantalizing possibility.
History mystery central to First World War exhibit
It's a history mystery worthy of its own exhibition.
When Dr. Marcus Milwright, of UVic's recently renamed Art History and Visual Studies department, began planning his upcoming exhibit The Arts of World War I, he knew there was one item in the university library that he just had to use: a beautiful two-volume leather diary set illustrated with watercolours and pen-and-ink drawings of life during wartime. There's only one problem: he has no idea to whom it actually belonged.
India field school spotlights intergenerational theatre
The buildings may have been repaired, but two key segments of the population in the southeastern coastal region of India are still struggling to overcome the effects of the 2006 tsunami: seniors and rural youth. Now, a new UVic field school hopes to bring a sense of joy to these marginalized people by creating India's first intergenerational theatre company.
Unite to Change with United Way
It's that time of year again. United Way Greater Victoria's annual fundraising campaign is underway and this year the charitable organization has set its fundraising goal at $6 million. United Way counts on the support of the Greater Victoria community to reach its goal each year.
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- Co-op Students of the Year
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- New research on same-sex marriage
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- Campus Checklist: March 24, 2015
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Event Specialist: Cosmetics
Event Planning - Retail / Marketing / Events - Full Training
The main focus of this position is to plan events and promote our clients' brand names by developing and supporting field marketing programs in retail environments. You will work closely with other Event Marketing ...
Mar 16 -
Personal Trainer - Capital District, NY
Best Fitness is looking for Personal Trainers to staff our clubs in Albany, Schenectady and our newest club in Colonie, NY. If you've committed to a healthy, fit lifestyle and enjoy helping others achieve the same benefits from it that you've experienced, then you're ideally ...
Mar 16 -
Regional Manager - Youth Programs - National - National Sports Brand
MUST HAVE PREVIOUS EXPERIENCE in the Health and Fitness industry! FULL TIME
The Regional Program Manager is responsible for driving revenue through the execution of youth fitness programming in a specific geography of clubs that host Sports Club for Kids (SCFK) ...
Mar 15 -
Retail Sales Representatives
Sales Representatives Needed in Schenectady...Immediate Openings!!
We have a Salon conveniently located at:
11 Saratoga Rd. Schenectady, NY 12302
Zoom Tan is America's fastest growing tanning salon chain. We offer incentive based pay to our associates...The more you sell. ...
Mar 6 -
Feb 2 -


text:
outdoor solar led light
Was $16.98
Your Price: $9.98
shaped solar lights
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string store set
Was $19.98
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flag tree lights
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10 piece lantern with wreath path lights
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candlestick path lights
Was $19.98
Your Price: $14.99
motion activated cordless swivel light
Your Price: $15.99
solar spot light set of two
Your Price: $19.98
led flip light lantern
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Money - Page 6
Gov. Patrick's Daughter Engaged To New Bedford Woman
Gas Prices Continue To Plummet
Scientists Urge Fishing Regulators to Tighten Management Plan
Scientists are urging federal regulators to go further to protect fish habitat in New England waters as the regulators finalize a long-anticipated management plan.
The New England Fishery Management Council has been working on a habitat management plan for several years and is considering a host of options
A-G's Office Investigates Santander Bank's Auto Loan Program
Mass. Gas Prices Drop Again
Alves Proposes Lottery Ticket To Help Pay For Southcoast Rail
New Bedford City Councilor David Alves wants the state to establish a lottery ticket to help raise money to pay for the Southcoast Rail Project.
Alves is proposing that the tickets be sold in Eastern Massachusetts only, but he says if state lawmakers from our region could muster the support for a statewide ticket, he wouldn't be opposed


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Luigi Frusci
Luigi Frusci (16 January 1879 – 1949) was an officer in the Italian Royal Army (Regio Esercito) during World War II.
Frusci fought on the southern front for General Rodolfo Graziani during the Second Italo-Abyssinian War. In April 1936, during the Battle of the Ogaden, Frusci commanded the center column of three columns attacking the Ethiopian "Hindenburg Wall".
Frusci was the deputy commander of the Italian "volunteers" in the Corps of Volunteer Troops (Corpo Truppe Volontarie) during the Spanish Civil War.
During World War II, Frusci was a Lieutenant-General and the Italian Governor of Eritrea. As a Lieutenant-General, he commanded Italian forces fighting in Eritrea during the East African Campaign. In mid-1940, Frusci oversaw the initial Italian attacks into the Sudan Later in 1940, even when ordered to do so, he chose not pull out of the Sudan. Instead, he rebuffed the initial efforts of British and Commonwealth forces to retake the border towns. In November, an assault on Gallabat was stopped short of its goals, the attacking force was hit hard from the air, and the position was re-taken by Italian ground forces.
After the British and Commonwealth forces crossed the border and launched an offensive in January 1941, Frusci also oversaw the defensive actions at Agordat, Keren, and the rest of Eritrea.
With the fall of Eritrea, Frusci became a prisoner of war.
Command history[edit]
- Inspector of Infantry
- Colonel, Ogaden border region command, Italian Somaliland – 1935 to 1936
- Deputy General Officer Commanding, (Corpo Truppe Volontarie), Spain – 1936 to 1937
- General Officer Commanding, 20th Division Friuli, Spain – 1937 to 1938
- General Officer Commanding, XX Corps, Libya – 1938 to 1939
- Governor of Amhara, Ethiopia – 1939 to 1941
- Governor of Eritrea – 1940 to 1941
- General Officer Commanding, Eritrean Army, East Africa – 1940 to 1941
- Prisoner of War – 1941