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Appukuttan Retnabhavan Pradeep,Arvind Awasthi,Raje Urs 한국분자세포생물학회 2008 Molecules and cells Vol.25 No.4
Simple sequence repeats (SSRs) and interSSR (ISSR) marker systems were used in this study to reveal genetic changes induced by artificial selection for short/long larval duration in the tropical strain Nistari of the silkworm Bombyx mori. Artificial selection separated longer larval duration (LLD) (29.428 ± 0.723 days) and shorter larval duration (SLD) (22.573 ± 0.839 days) lines from a base, inbred population of Nistari (larval span of 23.143 ± 0.35 days). SSR polymorphism was observed between the LLD and SLD lines at one microsatellite locus, Bmsat106 (CA7) and at two loci of 1074 bp and 823 bp generated with the ISSR primer UBC873. Each of these loci was present only in the LLD line. The loci segregated in the third generation of selection and were fixed in opposite directions. In the F2 generation of the LLD × SLD lines, the alleles of Bmsat106 and UBC8731074bp segregated in a 1:1 ratio and the loci were present only in the LLD individuals. UBC873823bp was homozygous. Single factor ANOVA showed a significant association between the segregating loci and longer larval duration. Together, the two alleles contributed to an 18% increase in larval duration. The nucleotide sequences of the UBC8731074bp and UBC873823bp loci had 67% A/T content and consisted of direct, reverse, complementary and palindromic repeats. The repeats appeared to be “nested” (59%) in larger repeats or as clustered elements adjacent to other repeats. Of 203 microsatellites identified, dinucleotides (67.8%) predominated and were rich in A/T and T/A motifs. The sequences of the UBC8731074bp and UBC873823bp loci showed similarity (E = 0.0) to contigs located in Scaffold 010774 and Scaffold 000139, respectively, of the B. mori genome. BLASTN analysis of the UBC8731074bp sequence showed significant homology of (nt.) 45–122 with upstream region of three exons from Bombyx. The complete sequence of this locus showed ~49% nucleotide conservation with transposon 412 of Drosophila melanogaster and the Ikirara insertions of Anopheles gambiae. The A + T richness and lack of coding potential of these small loci, and their absence in the SLD line, reflect the active process of genetic change associated with the switch to short larval duration as an adaptation to the tropics.
Kumaresan P.,Somasundaram P.,Kumar K. Ashok,Urs S. Raje Korean Society of Sericultural Science 2006 International Journal of Industrial Entomology Vol.13 No.2
Heterosis was studied involving two multivoltine silkworm breeds viz, APM1 and SLKSPM through rearing and isozyme analysis. A positive significant heterotic effect was observed in fecundity, hatching % and survivability. The heterobeltiosis was observed only in fecundity and hatching %. Isozyme analysis of ${\alpha}-esterase$ showed variation in loci and allelic expression. The allele with heterozygosity $(Est-2^{12})$ was observed at the Est-2 locus in $F_1$ progeny. Est-3 was observed in $F_1$ progeny, whereas it was completely absent in both parental lines. The present study suggests that the markers ($Est-2^{12}$ and Est-3) targeted for introgression may be useful for the improvement of fecundity and survivability as the phenomenon of heterosis was observed only in $F_1$ progeny.
Maji, M.D.,Sengupta, T.,Das, C.,Urs, S.Raje Korean Society of Sericultural Science 2004 International Journal of Industrial Entomology Vol.9 No.2
Post-infectional biochemical changes due to Xanthomonas campestris pv. mori (Xcm) infection in five elite mulberry varieties viz., $S_1$, $S_{1635}$, $V_1$, RF $S_{175}$ and JRH was studied under inoculated condition. It was revealed that total soluble sugar and protein content was significantly declined in all the varieties due to X. campestris infection. Total phenol content was at par prior to inoculation in all varieties, but it was significantly increased in $S_1$, RF $S_{175}$, $S_{1635}$ and JRH 7 days after inoculation. The correlation coefficient (r) between total soluble sugar and total phenol content was found positive (r = 0.825) and statistically significant. Similarly, correlation coefficient (r) between total soluble protein and phenol content was found positive (r = 0.897) and statistically significant. The present study indicates that X. campestris infected leaves are nutritionally inferior in quality and the duration of phenol production in a mulberry variety play decisive role on disease resistance.nce.
Evaluation of ISSR and RAPD Markers for the Detection of Genetic Diversity in Mulberry (Morus spp.)
Venkateswarlu, M.,Nath, B.Surendra,Saratchandra, B.,Urs, S.Raje Korean Society of Sericultural Science 2004 International Journal of Industrial Entomology Vol.9 No.2
The present study was carried out to evaluate the ISSR and RAPD markers for their efficiency as genetic marker systems to establish the relationships between 18 mulberry genotypes. A total of 36 from 56 (64%) RAPD primers and 12 from 48 (25%) ISSR primers produced reproducible amplification patterns. A high proportion of polymorphic bands ranging from 44 to 91% was observed respectively with RAPD and ISSR markers. The average Resolving Power (Rp) of ISSR primers was higher than RAPD primers. The ISSR primers, UBC 825, 868 and 873, and RAPD primers, UBC 712, 720 and 729, possessed the highest Rp values and could in each instance distinguish all the 18 genotypes. Similarity matrix values were estimated based on Jaccards coefficient, considering 109 polymorphic ISSR and 212 polymorphic RAPD bands and two dendrograms were constructed. The dendrograms obtained with ISSR and RAPD markers distinguished the eight exotic genotypes from the ten indigenous (Indian) genotypes. A significant correlation value (r=0.959; p=0.001) for the cophenetic matrix between the RAPD and ISSR matrices was observed. The results indicated that the ISSR and RAPD markers could assist in the differentiation of genotypes and permit the determination of genetic distances that might be exploited by mulberry breeders in improvement programs.
Moorthy, S. M.,Das, S. K.,Kar, N. B.,Urs, S. Raje Korean Society of Sericultural Science 2007 International Journal of Industrial Entomology Vol.14 No.2
The success of rearing with presently available conventional bivoltine is unpredictable in some seasons of the tropical regions due to highly fluctuating adverse climatic conditions. Thus, in order to popularize bivoltine breeds in tropical parts of India, it is very much essential to have a bivoltine breed(s), which can give stable cocoon crop under variable environments. With this objective a breeding programme was undertaken to improve the survival trait in bivoltine silkworm by introducing multivoltine genes into bivoltine through back crossing. Resultant bivoltine lines showed significantly higher survival in compared to the receptor (Bivoltine) parent and control bivoltine breed. Esterase isozyme analysis revealed similar banding pattern in the developed bivoltine and in the donor multivoltine, which predicts the introgression of multivoltine character into evolved bivoltine.
Moorthy, S.M.,Das, S.K.,Mukhopadhyay, S.K.,Mandal, K.,Urs, S. Raje Korean Society of Sericultural Science 2007 International Journal of Industrial Entomology Vol.15 No.1
An indigenous multivoltine silkworm, Nistari was evaluated for their thermo tolerance by exposing the larvae to various temperature regimes for eight hours. Among different temperature exposed, this strain has significant tolerance at $32^{\circ}C$. Analysis of heat shock protein revealed the expression of 70 kDa and 64 kDa polypeptides in fat body and midgut tissues. Interestingly esterase isozyme pattern in midgut showed characteristic expression of Est-1 and Est-3 at different temperatures signifying role in heat and cold shock.
( S. M. Moorthy ),( S. K. Das ),( P. R. T. Rao ),( S. Raje Urs ),( A. Sarkar ) 한국잠사학회 2007 International Journal of Industrial Entomology Vol.14 No.1
In order to find out the appropriate parents for the breeding programme, twelve bivoltine and three multivoltine silkworm breeds were evaluated on the basis of multivariate selection index and isozyme analysis. Of which, four [CSR2, D6 (P), SK3, SK4] bivoltine and two multivoltine (Nistari, Cambodge) breeds were selected and breeding initiated to develop higher survival bivoltine silkworm breed suitable for tropical conditions. Among two isozyme (Esterase and acid phosphatase) analyzed, only esterase exhibited polymorphism among the bivoltine breeds. No polymorphism was observed among multivoltine in respect of esterase as well as acid phosphatase.
( P. Kumaresan ),( P. Somasundaram ),( K. Ashok Kumar ),( S. Raje Urs ) 한국잠사학회 2006 International Journal of Industrial Entomology Vol.13 No.2
Heterosis was studied involving two multivoltine silkworm breeds viz, APM1 and SLKSPM through rearing and isozyme analysis. A positive significant heterotic effect was observed in fecundity, hatching % and survivability. The heterobeltiosis was observed only in fecundity and hatching %. Isozyme analysis of α-esterase showed variation in loci and allelic expression. The allele with heterozygosity (Est-212) was observed at the Est-2 locus in F1 progeny. Est-3 was observed in F1 progeny, whereas it was completely absent in both parental lines. The present study suggests that the markers (Est-212 and Est-3) targeted for introgression may be useful for the improvement of fecundity and survivability as the phenomenon of heterosis was observed only in F1 progeny.
$\alpha$- and $\beta$-Amylase Isozyme Expresser Native Proteins in Tropical Silkworm Bombyx mori L.
Chattopadhyay, G.K.,Verma, A.K.,Sengupta, A.K.,Das, S.K.,Urs, S.Raje Korean Society of Sericultural Science 2004 International Journal of Industrial Entomology Vol.8 No.2
Amylase isozyme based three multivoltine viz., N+p, Np, N+ $p^{cho}$ and two bivoltine-D6+p, D6p syngenic lines (Syn. L) were developed from germplasm (GP) stocks Nistari (N) and D6 respectively. haemolymph isozyme pattern at pH 7.0 and 8.5 depicted a total 11 number (Am $y_{1 to 6}$ at pH 7.0 and Am $y^{l to 5}$ at pH 8.5) of native proteins (NP) of various sizes are amylase isozyme expressers. Among eleven NPs, two NPs of 770 kDa (Am $y^{6}$ at pH 7.0) and 376 kDa (Am $y^3$ at pH 8.5) are $\alpha$-amylase expressers and remaining NPs of 370, 364, 350, 329 and 274 kDa at pH 7.0 and 206, 292, 416, 725 kDa at pH 8.5 are $\beta$-amylase expressers. Accordingly, digestive juice amylase isozyme pattern at aforesaid pH also depicted a total number of 10 NPs (Am $y^{1 to 5}$) at each pH 7.0 and 8.5 are amylase expressers of which NP of 387 kDa (Am $y^4$ at pH 7.0) and 780 kDa (Am $y^{5}$ at pH 8.5) are a-amylase expresser. Remaining NPs of 338,297 & 216 kDa at pH 7.0 and 370, 341, 329 &302 kDa at pH 8.5 are $\beta$-amylase expresser. Recurrent backcross lines (RBL) viz., N+pRBL and NpRBL were developed through introgression of high shell weight character (a multigenic trait) to be used further for congenic line (Con. L) development and to understand any association with introgressed character. Isozyme pattern in haemolymph of RBLs depicted only one $\alpha$-amylase of 770 kDa at pH 7.0 and 376 kDa at pH 8.0 with three and four respective $\beta$-amylase bands but in bivoltine lines numbers of $\beta$-amylase bands vary between 1 to 2 at aforesaid pH. Variability was also observed in digestive juice of multivolitine and its RBLs but bivoltine lines express null activity at both pH except appearance of one very week $\alpha$-amylase band D6+p at pH 8.5. Overall study suggests that not a single NP at both pH is common for expression of any band of amylase isozyme i.e., a totally different set of proteins are the amylase isozyme expresser at specific pH and no molecular factor of amylase is associated in developed RBLs which showed improvement on survival, single cocoon shell weight (SCSW) and single filament length over receptor parents.s.s.s.