Beruflich Dokumente
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ISBN : 978-1-63315-205-2
Short Views on Insect Biochemistry and Molecular Biology Vol.(1), October 2014
2014
Section IV
Insect Molecular Genetics
NAL B OO
IO
TERNA
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SION
MIS
Short
on Insect
Biochemistry
and Molecular
Biology and
Vol.(1),
2014 Biology
Views Short
Views
on Insect
Biochemistry
Molecular
Vol. (1), 00 00, 2009
Invited Review
Review
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Chapter 12
Sex chromosomes of the silkworm, Bombyx mori, are designated as ZW (XY) for females and ZZ
(XX) for males. The female mode of development is determined by the presence of a single W
chromosome. Therefore, it is presumed that the female-determining gene (Fem) is present on the
W chromosome. Until date, 12 W-specific random amplified polymorphic DNA (RAPD) markers
have been identified on the normal W chromosome. We have compared the W-translocation
chromosomes with normal W chromosome. The T(W;3)Ze (the sex-limited Zebra strain)
chromosome lacked 2 W-specific RAPD markers, indicating that the region containing these two
W-specific RAPD markers has been deleted. Additionally, we have investigated T(W;2)Y (the
sex-limited Yellow-cocoon strain). The T(W;2)Y chromosome lacked 11 such markers. These
results indicate that the W chromosomes of the sex-limited strains are the products of reciprocal
translocation accompanied by deletion, and an extremely limited region is required to determine
femaleness. No genes for morphological traits have been mapped to the W chromosome. Contrary,
Z chromosome is rich in genes. Many long terminal repeat (LTR) and non-LTR retrotransposons,
retroposons, DNA transposons, and their derivatives, had accumulated as strata on the W
chromosome. It is notable that some of these transposable elements contained the Bombyx short
interspersed element (Bm1) sequences in the elements. On the other hand, the transposable
elements on the Z chromosome were excluded by unequal crossing over or intra-element
homologous recombination between LTRs.
Key words: Bombyx mori, female-determining gene, transposable elements, single nucleotide, polymorphism, random amplified polymorphic DNA, W chromosomes, Z chromosomes
*For Correspondence (email: wfem@cc.tuat.ac.jp)
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Over view
1. Overview of W chromosome in Bombyx mori
2. The beginning of the molecular biological studies of the W chromosome
3. Presence of the W-markers in the silkworm strains maintained in Japan
4. Mapping of the W-markers on the W chromosome
5. Nested structure of transposable elements on the W chromosome
6. The structure of W chromosome and novel transposable elements
6.1. Structure of W chromosome
6.2. Structure of transposable elements the W chromosome
6.2a. BMC1
6.2b. Kendo
6.2c. Bm1 group
6.2d. Supermite-BMC1
6.2e. Other transposable elements
7. Recognition of W chromosome by FISH
8. piRNA derived from the W chromosome
9. The relations of the Z chromosome with W chromosome
9.1. Z-linked genes involved in the sexual differentiation
9.2. Transposable elements on the Z chromosome
10. Analyses of W chromosome of Lepidopteran insects except B. mori
11. Conclusion
12. References
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B. mori. Thus, based on genetic studies, the existence of the W chromosome was
almost certain, but the true characteristic of the W chromosome was unclear.
In Japan, sericulture is stable in the spring season. However, it becomes
unstable in summer and autumn because the quality of mulberry leaves deteriorates
after spring and because of unfavorable weather conditions, namely high temperature
and humidity. Therefore, establishment of more vigorous silkworm strains was
desired. In 1906, Toyama (3), one of the founders of silkworm genetics, rediscovered
Mendels laws in a study of inheritance of cocoon color. He recognized heterosis in
hybrids between Japanese and Thai strains (3,4). Following his advice, the Japanese
government decided to encourage the rearing of hybrid silkworms for general
sericultural farms and began distributing hybrid eggs in 1914. Thereafter, almost all
the silkworm strains reared on Japanese sericultural farms have been F1 hybrids. At
the same time, for the production of F1 eggs, specialized farms for F1 eggs were
necessary, and males and females must be separated at or before the cocoon stage. In
the absence of separation, just after emergence, sibling moths copulate mutually and
females cannot be used for the production of F1 hybrid eggs. Therefore, determining
the sex of silkworms for hybridization became very critical to silkworm egg
producers. The traditional separation method was inspecting Ishiwatas germal discs
in the female and Herolds gland in the male on the post-ventral surface of the final
(5th) instar larvae. This method was generally performed by a professional
discriminator, but it was inconvenient. Therefore, development of a convenient
method (visible marker) of discriminating males from females that even an amateur
was capable of was desired. Tazima, during the investigations of chromosomal
aberrations induced by X-rays, proposed, If we can translocate artificially any one
autosome that carries dominant genes for noticeable traits to the W chromosome, we
will be able to discriminate females and males based on these traits easily. For these
reasons, Tazima (5) attempted to achieve translocation between the W chromosome
and the 2nd chromosome, which carries the pSa gene (sable marking) for larval trait
by X-ray irradiation and achieved wonderful success at last! Stimulated by Tazimas
successful experiment, several researchers undertook the challenge of inducing
translocation between the W chromosome and an autosome carrying the dominant
genes for noticeable traits (egg color, larval marking, and cocoon color Fig. 1), and
were also successful (5-9). These traits were inherited from mother to daughter
(sex-limited inheritance). These translocations were also used to verify and conclude
that the W chromosome does exist and femaleness is determined solely based on the
presence of the W chromosome in B. mori. Tazima (6) also demonstrated that the
determiner of femaleness (female determining gene(s), Fem) is localized in a
particular region of the W chromosome not diffusely distributed.
Molecular biological techniques have been introduced to the biological studies of
B. mori, various genes have been cloned as the DNA nucleotide sequence data. In
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However, although 400 or more visible mutations have been placed on linkage
maps in B. mori (10-12), no gene for morphological character has been found on the
normal W chromosome. Thus, we were interested in the following question: What is
the nature of the nucleotide sequence of the nonrecombinant W chromosome, which
contains no gene except for the Fem gene? Therefore, the research group led by Abe,
who was one of the authors, tried to find a DNA marker as a beginning of the W
chromosome analysis. We continue analyzing the W and Z chromosomes of B. mori
now.
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In subsequent analyses, Abe et al. (24, 25) used genomic DNA from several
strains with the normal W chromosomes (for example p50T strain) as templates.
Fortunately, they obtained 12 W-specific RAPD markers from the normal W
chromosome (24, 25). Then, the conversion of these female-specific RAPD markers
(W-markers) into Sequence Characterized Amplified Region (SCAR) markers using
longer primers was successful. Thus, the existence of the W chromosome of B. mori
became clear a molecular biological level.
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containing the dominant genes for visible traits have been translocated to the
W chromosome by using X-ray or gamma-ray irradiation (5-7, 9), resulting in the
strains having mutated W chromosomes (28 and 29). Abe et al (26) investigated the
presence or absence of the W-markers in these strains and cleared that there were
differences at each W chromosome. The T(W;2)+p chromosome of larval-marking-W
strain contained all 12 W-markers, same as the normal W chromosome. The
T(W;10)+w-2 chromoosme of the Black-egg-W strain (7) did not contain the W-Mikan
marker. The T(W;3)Ze chromosome of the Zebra-W strain (8) did not contain the
W-Samurai and W-Mikan markers. Moreover, in the newly established strains of the
W-translocation strains, a part of the W chromosome inherited to male (25-29).
Furthermore, the T(W;2)Y-Abe chromosome (sex-limited yellow cocoon strain)
contained only the W-Rikishi marker (26). From these results, it is possible to map the
positions of the W-markers relative to the putative Fem gene. The order of W-Kabuki,
W-Kamikaze, W-Musashi, W-Sasuke, W-Sakura, and BMC1-Kabuki markers could
not be determined, but Abe et al. (26) concluded that the W-Rikishi marker is distal to
the end of the W chromosome (Fig. 2). This positional information should be of
significant help towards the goal of cloning the putative Fem gene.
In B. mori, efficient construction methods for transgenic silkworm have been
developed (30,31). Uchino et al. (32) constructed 105 enhancer trap lines in which the
BmA3-GAL4 gene is inserted into different positions of the chromosomes. However,
no line shows W-linkage of the transgene. Recently, insertion of the transgene into the
W chromosome has been reported (33) One of 160 newly established transgenic line
shows W-linkage of the transgene. In the strain, EGFP is expressed stably under the
control of the 3 x P3 promoter. Interestingly, insertion position of sex-limited EGPF
strain is mapped to the BAC clone containing W-Rikishi marker. This result suggests
that a small region of the W chromosome containing the W-Rikishi marker is
transcrptionally active compared to the rest of the heterochromatic W chromosome.
In addition, these W-translocation results indicate that the W chromosomes of the
sex-limited strains are not the result of simple fusion of the autosomal fragment to the
end of the unaltered W chromosome but, rather, the product of reciprocal
translocation accompanied by the breakage of the W chromosome. The intention was
to isolate translocations of autosomal fragments to the W chromosome, rather than to
delete the W chromosome. However, in the process of generating these strains,
deletions of the W chromosome were unconsciously selected. From these strains, we
can conclude that even large deletions of the W chromosome or large translocations to
W do not affect the female sex determination.
In addition, the pSa+pW chromosome, in which two 2nd chromosome fragments
(pSa and +p, respectively) were contained, was generated by translocation of a
fragment of chromosome 2 (pSa connected with +p) to the left end of the W
chromosome (4, 6). Subsequently, the pSa+pW+od chromosome was produced by
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connecting the short fragment of the Z chromosome having the +od gene with the
pSa+pW chromosome (4, 34). Through a dissociation experiment on the pSa+pW+od
chromosome, Tazima reached the conclusion that the +od gene was translocated to the
right end of the pSa+pW chromosome as a result of exceptional crossover between the
Z chromosome and the pSa+pW chromosome (right-end model). However, based on
the results of the presence or absence of W-markers on the W chromosome variants,
left-end model could be supported over the right-end model (26,28). The generation
of the Df(pSa+pW+od)Fem chromosome connected with the deleted Z chromosome can
be easily explained by the translocation between the pSa+pW+od chromosome and the
Z chromosome. Therefore, Fujii et al. (28,35) designated the pSa+pW+od
chromosome and the Df(pSa+pW+od)Fem chromosome as +odpSa+pW and
Df(+odpSa+pW)Fem, respectively.
Fig. 2. Higher-resolution mapping of W-specific RAPD markers and the putative Fem gene on the
W chromosome using W chromosome variants. The order of six W-specific RAPD markers
(W-Kabuki, W-Kamikaze, W-Musashi, W-Sasuke, W-Sakura and BMC1-Kabuki) and two W-specific
RAPD markers (W-Yukemuri-L and W-Yukemuri-S) could not be determined. Mapping of the other
markers is shown. For the T(W;3)Ze, T(W;10)+w-2 and DfZ-DfW chromosomes, see Abe et al. (25) and
Fujii et al. (28). Adapted from Abe and Fujii (74).
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Fig. 3. Nested transposable elements in the BAC clone that includes the W-Kabuki RAPD marker
sequence. See Abe et al. (38). This figure is adapted based on the recent DNA sequence information
from Abe and Fujii (74).
Based on the BLASTX search, except for the W-Musashi, all of the deduced
amino acid sequences of W-markers show similar to previously reported amino acid
sequences of retrotransposable elements. Moreover, almost all amino acid sequences
of W-markers contain boundaries of two or three retrotransposable elements (24, 25).
To analyze the W chromosome in detail, genomic DNA lambda phage libraries have
been constructed (36). Then, two lambda phage clones were obtained containing the
W-Kabuki and W-Samurai RAPD sequences. The lambda clone containing the
W-Kabuki RAPD sequence (18.1 kb) comprises a nested structure of at least six
transposable elements (36). The lambda clone containing the W-Samurai RAPD
sequence (14.7 kb) comprises a nested structure of six transposable elements (25).
Three of these six transposable elements are non-LTR retrotransposon BMC1
(37, 41). Moreover, to analyze the flanking regions of W-Kabuki RAPD marker in
detail, Abe et al. (25, 38) obtained a BAC (bacterial artificial chromosome) clone
containing a W-Kabuki RAPD marker and subjected this clone to shotgun sequencing.
The resulting assembly of sequences did not produce a single contiguous sequence
due to the presence of many repetitive DNA elements, in particular the non-LTR
retrotransposon BMC1 (37) and Kendo (39). As shown in Fig. 3, the structural
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features of the W chromosome of B. mori are quite different from structures typical of
other chromosomes. The W chromosome comprises the nested structures of many
transposable elements (38-40).
Thus, in the process of the W chromosome analysis, its become clear that it is
very difficult to use the normal molecular biological method for analysis of the W
chromosome due to the presence of many interspersed transposable elements
(repetitive sequences). For example in the case of non-LTR BMC1, there are many
copies on the autosomes but almost all open reading frames (ORFs) of BMC1 copies
are incomplete, and the full-length and complete ORF BMC1 are not found (41).
However, almost all BMC1 copies on the W chromosome are full-length and have
complete ORFs (37). When there are multiple BMC1 elements in one phage or BAC
clone of the W chromosome, the shotgun sequence analysis is very difficult.
Moreover, when a computer performs an assembly operation, often a false result is
given. It seems to be an extremely difficult jigsaw puzzle that includes a lot of
resembling pieces. There is not a completely connected, single contig BAC clone of
W chromosome so far.
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two intact ORFs, and a 3-UTR sequence, which terminates in a poly(A) tail. The
ORF1 of BMC1 encodes three putative cysteine histidine (Cys) motifs, while ORF2
encodes a protein containing an endonuclease (EN) domain and a reverse transcriptase
(RT) domain (37). Why does a complete full-length BMC1 element exist on the
W chromosome? One possible explanation is that because crossing over is restricted
to males in B. mori and the W chromosome is recombinationally isolated from the
Z chromosome, the BMC1 elements inserted into the W chromosome would be
expected to change more slowly than those inserted into other chromosomes.
Moreover, higher mutation rates are predicted for Z chromosomes or autosomes
relative to the W chromosome because of the greater number of cell divisions required
for spermatogenesis relative to oogenesis. Unfortunately, the presence of many
complete full-length BMC1 elements on the W chromosome is a main cause making
the assembly of the shotgun clone sequence extremely difficult.
Fig. 4. Structure of six retrotransposons and Supermite-BMC1 of Bombyx mori. Each box with an
arrow except in Supermite-BMC1 indicates one LTR (long terminal repeat). Each box with an arrow in
Supermite-BMC1 indicates an IR (inverted repeat). PBS, primer-binding site; PPT, polypurine tract;
Pro, protease domain; RT, reverse transcriptase domain; RH, RNase H domain; Int, integrase domain;
EN, endonuclease domain; 5-UTR, 5untranslated region; 3-UTR, 3untranslated region; poly(A),
poly(A) tail. Cys, cysteine and histidine motif are indicated solid bar in the box of ORF.
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6.2b. Kendo
Kendo is the most abundant non-LTR retrotransposon next to the BMC1 in B.
mori genome (39). Despite numerous sequences of B. mori, a complete full-length
Kendo had not been found. As mentioned below, the longest Kendo has been found in
the Z-specific BAC clone (47). However, in the WGS sequence data (18), we could
not find longer Kendo than that in the Z chromosome. Therefore, we think that the
Kendo in the Z chromosome (47) is the complete full-length element. The
full-length Kendo is about 3670 bp long and contains a single ORF encoding an EN
and an RT domains (Fig.4f).
6.2c. Bm1 group
The Bm1 element is a major short interspersed element (SINE) in the genome of
B. mori (48). Therefore, the Bm1 element is thought to be analogous to the human Alu
element. The longest Bm1 element is about 450 bp containing no coding region and
having a poly(A) tail at the 3 end (48). It is thought that the enzyme required for
mobility of the Bm1 element is supplied by other elements. The Bm1 element is often
found on the W chromosome of B. mori (36). We thought that if Bm1 element
insertions occurred randomly in the genome, the Bm1 elements on the W chromosome
would be useful for the analysis of the W chromosome as landmarks. In other words,
even if there are identical nucleotide sequences before the Bm1 element was inserted
on the autosomes, it will be possible to discriminate the PCR product amplified from
the W chromosome based on the size. However, when we carried out such a PCR
experiment, the same size PCR products were amplified from both the male genome
and female genome. We were not able to understand the meaning of these results.
Then, when the same sequences containing Bm1 elements on the W chromosome
were found in the WGS data of males, we suspected that these sequences resulted
from contamination of male genomic DNA female DNA during WGS analyses. If
this were so, we would have been able to find the W chromosome-specific sequences
(25,26,38) in the WGS data. However, no W-specific sequences, for example, the
nested structure of retrotransposable elements, were found in the WGS data.
Moreover, we analyzed the WGS data to find out if there were ancestral states of these
sequences that did not contain the Bm1 sequence. However, the ancestral states were
not found in the WGS data. Therefore, we could not but change a thought
fundamentally as follows. The Bm1 sequences found in these sequences were not
inserted, but the flanking sequences of Bm1 sequence and Bm1 are one unit of
transposition! We got possible to explain the relationship of the Bm1 sequence data of
the W chromosome and the WGS data without contradiction. We designated these
transposable elements as secondary-Bm1 transposable elements (SBTEs) (49).
There are several types of SBTEs as shown in Fig. 5, long SBTEs more than 5kb
long (ChoBm1), short SBTEs about 1kb long (Bm1modoki), and the LTR types
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(Yaocho, Rinne-L and Rinne-S). We initially thought that Bm1 sequences of SBTEs
were carried on each SBTE merely as a package. For example, in ChoBm1, we
thought that the Bm1 sequence was inserted fortuitously into this position of ChoBm1.
If this were so, the putative ancestral ChoBm1 not containing the Bm1 sequence would
be found in the WGS data. However, we could not find the ancestral ChoBm1
sequence in the WGS data. These results strongly indicate that the ChoBm1 acquired
the ability to propagate after this structure containing the Bm1 sequence had been
constructed. This applies to all SBTEs. The generation processes of SBTEs in the
genome of B. mori are unknown. The role of Bm1 sequences in SBTEs for
transcription or translation is also unknown. It is likely that the Bm1 sequence has a
specific role for transcription or transposition because the sequences of the flanking
regions of Bm1 sequences in each SBTE are not homologous to each other.
In the case of human Alu elements, there is no report of a transposable element
containing an Alu sequence [International Human Genome Sequencing Consortium
(52)]. In Drosophila, there is no SINE in the genome (51). A unified classification
system for eukaryotic transposable elements was proposed (42). However, SBTEs
cannot be classified into the known groups. Therefore, SBTEs should be classified as
a new group of transposable elements. It seems likely that the SBTEs are unique
transposable elements found in the Bombyx genome. However, our idea was not
accepted at first. It took a long time until our idea was understood and accepted.
Because mysterious sequences will appear in the data of the W chromosome in the
future, we have to analyze the data without having a preconception.
6.2d. Supermite-BMC1
Previously, we found an amino acid coding sequence that is a part of the
non-LTR retrotransposon BMC1 in 45J23 BAC clone (Accession no. AB126052; 38).
Moreover, this coding sequence was flanked with a relatively longer inverted repeat
(500 bp). Initially, we thought that this sequence was the result of the insertion of
5-truncated BMC1 into the DNA transposon-like sequence or into MITE (miniature
inverted-repeat transposable element) like sequence. If this were so, the putative
ancestral DNA transposon-like sequence would be found in the WGS and Buil2 data.
However, we could not find the putative ancestral DNA transposon-like sequence.
Therefore, we consider that this sequence is the unit of transposition. We designated
this sequence (nucleotide position 13270-15260 in AB126052) as Supermite-BMC1
(Fig.5g).
6.2e. Other transposable elements
In the processes of the analyses of the W chromosome, typical LTR and
non-LTR retrotransposons, and DNA transposons have been found. Additionally,
many SINE-like sequences have been found. For these include Nagisa (390 bp),
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Sayama (952 bp), Shirigonomi (279 bp), Kohichi (1302 bp), Kusanagi (421 bp),
Chikuri (712 bp), Minichikuri (413 bp), and Amanoya (834 bp). The characteristics of
these transposable elements can be seen in Abe et al. (49).
Fig. 5. Schematic diagrams of SBTEs (secondary-Bm1 transposable elements). (a) ChoBm1 (5412
bp). (b) Neet (4046 bp). (c) Bm1modoki (1178 bp). (d) Ins11 (1088 bp). (e) Tenshi (1360 bp). (f)
Hikikomori (1740 bp). (g) Yaocho (5851 bp). (h) Rinne-L (3695 bp). (i) Rinne-S (3437 bp). Each box
with an arrow in Yaocho, Rinne-L and Rinne-S indicates one LTR or LTR-like sequence. PBS,
primer-binding site. Adapted from Abe et al. (49).
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In the antenna of moth, two genes encoding sex pheromone receptors, Or1 and
Or2, are expressed specifically in males, which are tuned to bombykol and bombykal
respectively (62,63). Both of the genes are linked to the Z chromosome (The
International Silkworm Genome Consortium, 60). The neurons expressing pheromone
receptors are projected into a macroglomerular complex (MGC) in the male brain
where pheromone information is further integrated to elicit the mating behavior (64).
spli (soft and pliable) is a Z-linked mutant showing an extremely soft and pliable
larval body (65). Moreover, sex-pheromone preference of spli male moths is different
from normal male moths. In the spli strain, the behavioral responsiveness of male
moths to bombykol was markedly reduced, whereas bombykal alone evoked full
courtship behavior (66).
In the spli strain, the Z chromosome is broken into two pieces. A Bombyx
homolog of Drosophila acj6 is broken by the breakage event of the Z chromosome
(67). Drosophila acj6 is a transcriptional factor involved in the olfactory receptor
gene choice and axon targeting of olfactory receptor neurons (ORNs) (68, 69).
Quantitative RT-PCR analysis of olfactory receptors in male moth antenna revealed
that expression levels of Or3 in the spli males was almost the same as those in normal
males. However, about 1000-fold reduction in Or1 expression was observed in the
spli males. (1) These results suggest that expression of the Or1 depends on Bmacj6
while that of the Or3 is independent of Bmacj6 (2). The reduced response of spli
males to bombykol was caused by the paucity of bombykol receptors on the male
antennae.
The MGC of B. mori consists of three subdivisions: the toroid, cumulus, and
horseshoe. ORNs responding to bombykol and bombykal send their axons to the
toroid and cumulus, respectively (70). It was also found that in the spli males, neurons
projecting to the toroid responded strongly to bombykal. A possible explanation for
this phenomenon is mistargeting of the neuron expressing Or1 caused by the lack of
Bmacj6 (67).
9.2. Transposable elements on the Z chromosome
A 320-kb region of the Z chromosome of B. mori containing a kettin gene has
been analyzed (47). This 320-kb sequence contains many transposable-like sequences,
47 non-LTR retrotransposons, 50 retroposons, 10 DNA-type transposons, and other
uncharacterized repetitive sequences. The non-LTR retrotransposons BMC1, the
DNA-type transposon mariner (71,72), and the retroposon Bm1 (48) are dispersed
throughout the region of 320-kb on the Z chromosome. These transposable elements
were found to be incomplete. However, as mentioned above, we think that the kendo
in this region (AB090307, nucleotide positions: 89135-85464 from 5 to 3; Koike et
al.(47) is the complete full-length element (74). The nested structure of many
transposable elements found on the W chromosome is not found in this 320-kb region
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11. Conclusion
The W chromosome of B. mori has been artificially modified for practical use by
irradiation with X-rays of gamma ray (5-9, 34). In these processes, the intention was
to isolate translocations of autosomal fragments to the W chromosome, rather than to
delete the W chromosome. However, deletions of the W chromosome were selected
unconsciously in the processes of generating sex-limited strains, and these modified
W chromosomes became very useful materials for molecular biological analysis.
From our results using the W chromosomes of these W-translocation strains, we can
conclude that even large deletions of the W chromosome or large translocations to W
do not affect the female sex determination. Therefore, we think that a full-length
normal W chromosome is not essential for femaleness. Instead, we conclude from
translocation tests and deletion strains that putative Fem genes is not distributed
evenly over the W chromosome. Provably, a putative Fem gene exists in an extremely
limited region around the W-Rikishi RAPD marker (26). Although the
W chromosome continues to be analyzed, nobody has successfully identified the Fem
gene.
In the genomic sequence of Drosophila melanogaster, there are 1572 different
transposable elements belonging to many families (51). In contrast, although the
accurate number is not determined, there are many more transposable elements in the
genomic sequence of B. mori. It is surprising that the numerous copy numbers of
transposable elements in addition to a great many kinds including unpublished
elements. It is very likely that the SBTEs containing the Bm1 sequence that cannot be
classified into the conventional classification are unique transposable elements only in
the genome of B. mori (49). In the nucleotide sequences of the Z chromosome, a trace
of the W chromosome could not be found. It is widely accepted that mammal
(including human) X and Y chromosomes originated from the same ancestral
autosomes (non-sex chromosomes) during the evolution of sex determination (79).
However in B. mori, it is hard to think that the W chromosome was originated from
the ancestral Z chromosome degenerated by insertion of numerous transposable
elements. It is thought that the origin of the W chromosome of B. mori is different
from the Z chromosome.
In Drosophila genome projects, it was impossible to assemble Y chromosome
sequence data into a single contig due to the presence of many repetitive sequences
(73). Therefore, to identify new genes on the Y chromosome of D. melanogaster,
another strategy, TBLAST search, was developed (77, 80). In the Africa malaria
mosquito Anopheles gambiae, numerous Y-linked repetitive sequences in the Y
chromosome have been obtained (81, 82). However, no genes on the Y chromosome
were identified based on a TBLASTN search (83). Thus, both in Drosophila and
Anopheles, molecular biological analyses of the Y chromosome are made very
difficult by the presence of unusual chromosomal features. However, the W
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Acknowledgment
This work was supported by the Grants-in-Aid for Scientific Research, JSPS/MEXT
(No. 26450464).
12. References
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_____________________________________________________________________________________
Article History: Received on 15th December 2013; Revised on 15th Feburary 2014; Accepted on 2nd May 2014;
Publsihed 30th October 2014.
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Table Contents
SION
MIS
TERNA
IN
T
N AL B OO
IO
Page No.
Preface
Forward message
Contributors
Reviewers
Acknolwedgement
i
ii
iii
iv
v
Volume1
Section I: Insect Biochemical approaches
Raman Chandrasekar, P.G., Brintha, Enoch Y.Park, Paolo Pelsoi, Fei Liu,
Marian Goldsmith, Anthony Ejiofor, B.R., Pittendrigh, Y.S., Han,
Fernando G. Noriega, Manickam Sugumaran, B.K., Tyagi, Zhong Zheng Gui,
Fang Zhu, Bharath Bhusan Patnaik, and P. Michailova
2.
57
Sahayaraj, K.
3.
75
4.
99
5.
127
xvii
149
Manickam Sugumaran
185
217
Insect Immunity
233
253
271
291
317
331
Paraskeva V. Michailova
355
Dhanenjeyan, K. J., Paramasivam, R., Thanmozhi, V., Chandrasekar,R., and Tyagi, B.K.
Index
363
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Volume2
Section V:
373
385
in Lepidoptera.
409
Section VI:
429
449
473
497
509
Ronald J. Nachman
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Section VII:
533
549
Usha Rani, P.
575
595
Section VIII:
Insect Bioinformatics
621
633
685
Jitrayut Jitonnom
Index
709
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Book Mission Project # 2: Initiated on June 2010; Completed on March 2014 and Published on Oct. 2014.
PREFACE
Entomology as a science of inter-depended branches like biochemistry, molecular entomology, insect
biotechnology; has made rapid progress in its attributes in the light of modern discoveries. This also
implies that there is an urgent need to manage the available resources scientifically for the good of man.
In the past five decades, entomology in the world/country has taken giant steps ahead. Continued
research has evolved better pest management through molecular approaches. The aim of the Short
Views on Insect Biochemistry and Molecular Biology book is to integrate perspectives across
biochemistry and molecular biology, physiology, immunology, molecular evolution, genetics,
developmental biology and reproduction of insects. This century is proclaimed as the Era of
Biotechnology and its consists of all types of Mol-Bio applications, which is an essential component for
a through understanding of the Insect Biology. This volume 1 & 2 (8 section with 30 chapters)
establishes a thorough understanding of physiological and biochemical functions of proteins, genes in
insects life processes; the topics dealt with in the individual chapters include chemistry of the insect
cuticle, hormone and growth regulators; biochemical defenses of insects; the biochemistry of the toxic
and detoxification action; modern molecular genetics and evolution; inter- and intra-specific chemical
communication and behavior; insect pheromone and molecular architecture, phylogeny and chemical
control of insect by using insect pheromones biotechnology; insect modern biology and novel plant
chemical and microbial insecticides for insect control, followed by a discussion of the various
mechanisms of resistance (both behavioral and physiological) and resistance management; modern insect
pest management through biochemical and molecular approaches; Mimetic analogs of insect
neuropeptide for pest management; entomo-informatics and computer-aided pesticide designing. In short
this book provides comprehensive reviews of recent research from various geographic areas around the
world and contributing authors area recognized experts (leading entomologist/scientist) in their
respective filed of molecular entomology. We will miss this collaboration now it has ended, but will feel
rewarded if this book is appreciated by our team/colleagues and remarkable mile stone in entomology
field.
This book emphasizes upon the need for and relevance of studying molecular aspects of entomology in
Universities, Agricultural Universities and other centers of molecular research. To encompass this
knowledge and, particularly disseminate it to the scientific community free of cost, was the major
inspiring force behind the launch of Short Views on Insect Biochemistry and Molecular Biology.
Editors
Raman Chandrasekar
Brij Kishore Tyagi
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vi
ShortViewson
InsectBiochemistryand
MolecularBiology
Editedby
Raman Chandrasekar, Ph.D.,
Kansas State University, USA.
B.K.Tyagi, Ph.D.,
Centre for Research in Medical Entomology (ICMR), India.
Zhong Zheng Gui, Ph.D.,
Jiangsu University of Science and Technology,
Sericultural Research Institute, Chinese Academy of
Agricultural Sciences, China.
Gerald R. Reeck, Ph.D.,
Kansas State University, USA.
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Contributing Authors
Dr. B.K.Tyagi
Prof.Fernando G. Noriega
Prof. K. Sahayaraj
Prof.Yanyuan Bao
Institute of Insect Science,
Zhejiang University, China.
Prof. PatriciaY.Scaraffia
Department of Tropical Medicine,
Tulane University, New Orleans,
LA 70112, USA.
Dr. P. Somasundaram
Central Sericultural Germplasm Resources Centre,
P.B.No.44, Thally Road,
Hosur-635109,
Tamilnadu, India.
College of Forestry,
Northwest A & F University
Yangling, Shaanxi 712100, China
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Dr. R. Srinivasan
School of Biotechnology,
Trident Academy of Creative Technology
(TACT), Bhubaneswar 751013 Odisha, India.
School of Science
University of Phayao, Thailand.
Department of Entomology,
University of Illinois, Urbana-Champaign, IL,
61801, USA
.
Prof. K. Murugan
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Acknowledgements
Writing and publishing a book requires the assistance of individuals who are
creative, talented, and hard-working. All of these qualities were present in the
individuals assembled to produce this book volume. I would like to express my
heartfelt gratitude to my former teacher Prof. Seo Sook Jae, (GSNU, South Korea),
Prof. Subba Reddy Palli (University of Kentucky, USA), and other external mentors
Prof. Marian R. Goldsmith (University of Rhode Island, USA), Prof. Enoch Y. Park
(Shizuoka University, Japan), Prof. M. Kobayashi (Nagoya University, Japan), Prof.
CHU Jang Hann (National University of Singapore, Singapore), Prof. Thomas W.
Sappington (USDA-ARS, USA), Prof. Fernando G. Noriega (Florida International
University, USA), Dr. Srinivasan Ramasamy, AVRDC, The World Vegetable
Center, Taiwan), Dr. H.C. Sharam (ICRISAT, India), who inspiration and
supported me at many ways for the commencement of this International Book
Mission Program. The book mission program was initiated on May 2010,
completed on March 2014 and published on October 2014. I have no words to
express my feeling for all those who provided valuable contributions from USA,
South Korea, Japan, China, India, Thailand, Taiwan, Bulgaria, France, Iseral, and
Portugal (Contributors name list, see page no. v) and made the completion of this
book possible. We express our appreciation to the following people (Reviewer
name list, see page no. vii) who reviewed various part of the manuscript as it was
being developed and improved quality of each chapter. I thank the ICMR, New
Delhi, and Chinese Academy of Agricultural, China, and Kansas State University for
support from several aspects. Many others (scientists and publishers) have also
allowed us to use their materials in the various chapters, their color image have then
been converted to gray color/BW. Iam especially indebted to International Book
Mission Organization, Academic Publishing Services for the production of book. I
thank my Co-Editors for their continuous vigilance over the book project and for
always giving advance notice of the editing and proofreading schedules. I thank also
my Brintha, P.G., (my wife), who in all possible way, encouragement helped
transform our original efforts into an acceptable final form. I apologize to those
whose work could not be cited owing to space considerations limitation. Further, I
wish to recognize the moral support extended by colleagues and friends. I hope that
this volume will inspire interest on the diverse aspects of insect biochemistry and
molecular biology in aspiring and established scientists.
Raman Chandrasekar
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Book Series
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