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Anand Agricultural University, Anand
Anand Agricultural University (AAU) was established in 2004 at Anand with the support of the Government of Gujarat, Act No.(Guj 5 of 2004) dated April 29, 2004. Caved out of the erstwhile Gujarat Agricultural University (GAU), the dream institution of Sardar Vallabhbhai Patel and Dr. K. M. Munshi, the AAU was set up to provide support to the farming community in three facets namely education, research and extension activities in Agriculture, Horticulture Engineering, product Processing and Home Science. At present there seven Colleges, seventeen Research Centers and six Extension Education Institute working in nine districts of Gujarat namely Ahmedabad, Anand, Dahod, Kheda, Panchmahal, Vadodara, Mahisagar, Botad and Chhotaudepur
AAU's activities have expanded to span newer commodity sectors such as soil health card, bio-diesel, medicinal plants apart from the mandatory ones like rice, maize, tobacco, vegetable crops, fruit crops, forage crops, animal breeding, nutrition and dairy products etc. the core of AAU's operating philosophy however, continues to create the partnership between the rural people and committed academic as the basic for sustainable rural development. In pursuing its various programmes AAU's overall mission is to promote sustainable growth and economic independence in rural society. AAU aims to do this through education, research and extension education. Thus, AAU works towards the empowerment of the farmers.
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ThesisItem Open Access Study of Pituitary transcription factor, Insulin like growth factor, Leptin, Oxidized low density lipoprotein receptor 1 and Proteases inhibitor gene loci in Mehsana buffaloes(AAU, Anand, 2009) Deshpande, Manisha Ramesh; Rank, D. N.The present work was carried out to study the polymorphism in Pituitary transcription factor (PITl exon 6), Insulin like growth factor (IGFl exon 4), Leptin (exon 2), Oxidized low density lipoprotein receptor 1 (OLRl 3'UTR) and proteases inhibitor gene (PI exon 2) loci in Mehsana buffaloes through PCR-RFLP and sequencing in Mehsana buffaloes. DNA extraction was carried out by John's method from sixty blood samples of non related Mehsana buffaloes from the animals registered under progeny testing programme of Dudhsagar Research and Development Association (DURDA), Mehsana. Locus specific primers were used for PCR amplification for each of five loci. The fragment of 451bp of Pit-1 was amplified by PCR, using the primers reported by Renaville et al. (1997) and digested with restriction enzyme Hinfl. All the samples showed identical restriction pattern consisting of one site at 207 bp obtaining two fiiagments of 207bp and 244 bp. All the animals revealed monomorphic pattern with BB genotype. This result indicated no polymorphism existing in Mehsana buffalo at Pitl exon 6 locus as revealed by Hinfl RFLP. The study revealed only one allele B fixed in Mehsana buffalo with allele frequency 1.0. PCR products of representative samples were purified and cloned in pTZ57R/T vector of InsT/Aclone TM kit. Ligated recombinant vector was transformed in competent E. coli (DH5-a) cells. Recombinant plasmids were obtained and used for cycle sequencing. Sequencing revealed G—>T variation between cattle and buffalo at 283 nucleotide position. The fragment of 320 bp of IGF Iwas amplified by polymerase chain reaction, using the primers reported byDierkes e^a/.(1999) and digested with Ecol30I. On screening the IGF/Ecol30I polymorphism in Mehsana buffalo, all the animals revealed monomorphic pattern with three bands of 34, 127, and 159 bp (B allele) indicating two RE sites at 127 and 161 bp position. The investigation revealed B allele fixed in Mehsana buffaloes. As there is no polymorphism, association analysis is not warranted. PCR products of 320bp of IGF-1 (exon 4) from representative samples were purified and cloned in pTZ57R/T vector. Recombinant plasmids were obtained and used for cycle sequencing. The nucleotide sequence variation between cattle and buffalo was present at four nucleotide positions i.e. 35bp, 102bp, 132bp, 257 bp. The fragment of 331 bp of leptin gene loci was amplified by PCR, using the primers reported by Haegeman et al. (2000). Amplified products were digested with HphI and electrophoresed on 2% agarose. All the samples showed identical restriction pattern with 331 bp fiugment only. On screening the Leptia/HphI polymorphism in Mehsana buffalo one genotype AA was observed indicating allele A fixed in Mehsana buffalo. Representative sample PCR products of 331bp of leptin (exon 2) was purified and cloned in pTZ57R/T vector. Recombinant plasmids were obtained and used for cycle sequencing. The nucleotide sequence variation between cattle and buffalo was presentat six nucleotide positions i.e. 100,112,119,135,185,321. OLRl 3' UTR region was explored for Pst 1 RFLP in Mehsana buffalo. The primers reported by Khatib et al. (2006) for Bos taunts could not amplify the region in Mehsana buffaloes. Hence, new primer sets were designed using Bioinformatic tools, Primer 3.0 and Primer Express softwares (http://www.genome.wi.mit.edu/cgi bin/primer/primer3_www.cgi) on the basis of gene sequence available in the data base NW_174132.2. A 288 bp fragment of OLR gene loci was amplified by PCR, using the designed primers and digested with Pst 1. It has aPst 1 site at 215 bp and produced two fragments of 215 and 73 bp. Representative sample PCR products of 288 bp (3'UTR) was purified and cloned in pTZ57R/T vector. Recombinant plasmids were obtained and used for cycle sequencing. The nucleotide sequence variation between cattle and buffalo was present at nine nucleotide positions i.e. 85,91,116,129,151,168,171,217,240. The fragment of 448 bp of PI gene loci was amplified by PCR, using the primers reported by Khatib et al. (2005) and digested with restriction enzyme SfaNI. SfaNI digestion of PI axon 2 gene fragments revealed monomorphic pattern with appearance of three bands of 242bp, 124bp, 82 bp (B allele) in all the samples. Representative sample PCR products of 448 bp of PI (exon 2) was purified and cloned in pTZ57R/T vector. Recombinant plasmids were obtained and used for cycle sequencing. The nucleotide sequence variation between cattle and buffalo was present at ten nucleotide positions i.e. 110,141,146,170,236,257,277,306,350,396. None of the polymorphic site reported in cattle for PITl, IGFl, Leptin, OLRl and PI could be verified in buffalo. Genomic nucleotide sequences of PITl, IGFl, Leptin, OLRl and PI was submitted to Genbank of NCBI database. Bankit online sequence submission tool was used for submission of sequences to Genbank. 1. GQ385224: Bubalus bubalis POU domain class 1 transcription factor 1 (PITl) gene, exon 6 and partial cds. 2. GQ385225 : Bubalus bubalis serpin peptidase inhibitor clade A member 1 (SERPINAl) gene, partial cds. 3. GQ385226: Bubalus bubalis oxidized low density lipoprotein receptor 1 (OLRl)gene,3'UTR. 4. GQ385227: Bubalus bubalis insulin-like growth factor 1 (IGFl) gene, exon 4 and partial cds. 5. GQ385228: Bubalus bubalis leptin gene, exon 2 and partial cds.ThesisItem Open Access RESPONSE TO SELECTION FOR EGG PRODUCTION IN IWN PURE LINE WHITE LEGHORN CHICKEN(AAU, Anand, 2007) PALEJA, H. I.; SOLANKI, J. V.A total of 961 (S3), 1219 (S4), 2049 (S5), and 1309 (S6) progenies of four consecutive generations of IWN pure line White Leghorn were considered for the study. In each generation 49, 50 and 60 sires of S3, S4 and S5 respectively produced progenies in S4, Ss and Sg generations. Corresponding number of dams were 197, 220 and 279. The selection differentials for generation S3, S4 and S5 were 17.17, 28.94 and 18.12 respectively. The corresponding selection intensity were 1.63, 1.81 and 1.93. The females were selected based on 64 week egg number (EN64) and independent culling level for egg weight at 40 week (EW40). The male progenies were selected based on the production performance of the fuUsib and halfsib sister progenies. The least squares means for AFE (day) were 148.75±0.50, 149.60 ±0.45, 140.34±0.39 and 150.46±0.42 in S3 to S6 generations respectively. There were significant differences between the generations except for S3 and S4 as well as S4 and S6 (P>0.05). The LSMs for egg production (no.) upto 64 weeks of age were 252.23±1.13, 255.00±1.02, 268.91±0.87, 254.71±0.96 in Se generation, respectively. There were significant differences between generations for egg production upto different age between two or more generations. The egg weights (g) at 40 weeks of age were 47.86±0.17, 51.65±0.16, 50.6110.14 and 50.49±0.15 in consecutive four generations under study. The differences for EW40 were significant between generations except between S4 and S5. The age at first egg was moderate to highly heritable trait. The differences between generations for heritability estimates of age at first egg were non significant. The heritability estimates for egg number upto different stages showed no significant differences between generations. The heritability estimates were 0.361±0.104, 0.132 ± 0.058, 0.231±0.060 and 0.180±0.084 for EN64 in S3 to S6 generations, respectively. The heritability estimates of egg weight at 40 weeks of age were 0.496 ± 0.124, 0.682 ±0.140, 0.432 ± 0.089 and 0.248±0.074 in S3 to Se generations, respectively. The genetic and phenotypic correlations between egg number and AFE and EW40 were negative. The egg production has shown phenotypic response of 2.13 ± 3.86 eggs per generation for ENe4. As a correlated response to selection for EN64, an increase was observed in all the traits under consideration except BW40 and AFE which declined over generations. The direct response and correlated response to selection for egg were however, non significant. All the traits except AFE have shown increasing trend for LSMs over the generations. Heritability estimates for AFE, EN64 and EW40 showed decline. The genetic and phenotypic correlations between EN64 and AFE showed declining trend whereas between EN64 and EW40 showed increasing trend over generations. However, the trend for LSMs of traits, their heritability estimates, genetic and phenotypic correlations were non significant (P>0.05). Out of total 25 different model of curve attempted to fit the egg production curve, model Y=A+B / X + C / X*X, second order hyperbola fitted to the three (S3. S4 and Be) generations. The curve fit model Y=A+B*X + C/ X, which was linear and reciprocal observed to be fitting S5 generation.ThesisItem Open Access MOLECULAR GENETIC CHARACTERIZATION OF BANTAM, BANTAMISED WHITE LEGHORN, AND WHITE LEGHORN CHICKENS USING PCR-RAPD, PCR-RFLP & MICROSATELLITE TECHNIQUES(AAU, Anand, 2004) Pipalia, Devendrakumar L.; Solanki, J. V.The molecular genetic techniques RAPD, PCR-RFLP and microsatellite markers studies were used for investigation of genetic composition and genetic distances among BNT, BWLH and WLH genetic groups of chicken at molecular level. Out of 13 RAPD primers, 9 primers gave amplifications and amongst them 3 primers (BG-1, PLANTAGO-01 and PLANTAGO-06) showed variation between pooled DNA samples from each sex of BNT, BWLH and WLH groups. The BG-1 produced 4 to 7 scorable bands from sexwise pooled DNA samples within 550 to 1550 bp range and amplified approximately 1235 bp female specific band and 625 & 550 bp fragments specific to BWLH and WLH. The band sharing (BS) value was highest between BWLH and BNT group (0.4454) as compared to BWLH and WLH group (0.3880). The PLANTAGO-01 produced 7 to 8 scorable bands from sexwise pooled DNA samples within 724 to 2038 bp range and amplified approximately 1014 bp size fragment specific to BWLH and WLH group. While 724 bp fragment was only found in BNT group. The PLANTAGO-06 primer produced 5 to 8 scorable bands from sexwise pooled DNA samples within 696 to 2530 bp range and amplified approximately 790 and 832 bp fragments specific to BNT group. These primers, PLANTAGO-01 and 06, showed higher BS values between BWLH and WLH (0.6235 and 0.5235) as compared to BWLH and BNT groups (0.5346 and 0.4637). Phylogenetic tree using data revealed that distance between BWLH was less with WLH than BNT group. The PCR products amplified by cGH1 and cGH2 primers specific to chicken growth hormone gene showed size variation as compared to literature i.e. 1216 bp instead of 1163 bp (53 bp difference) for cGHI and 770 bp instead of 756 bp (14 bp difference) for cGH2 region. PCR-RFLP of growth hormone showed restriction enzyme (RE) sites at 628 and 1074 bp at cGHI/Sac / locus and RE sites at 373 and 529 bp at cGH2//Wsp / locus. The new RE sites found at 628 bp for cGHI/ Sac I locus and at 529 bp for cGH2/Msp / locus. The exact size, sequence and position of restriction sites can be more precisely known only by sequencing. For cGHI/Sac / locus, alleles A2 and A3 were not observed in BNT and WLH groups, while only allele A3 was absent in BWLH group. The allele A was found in highest frequency in BNT (0.5357) and WLH (0.5968) groups. While allele A1 was at highest frequency in BWLH (0.7766) group. The allele A2 was only found in BWLH group. It may happen due to insertion of particular restriction site (628 bp) at the time of gametic recombination. The WLH population significantly (P<0.05) deviated from genetic equilibrium at this locus. For cGH2IMsp I locus, the allele frequencies for A1 and A3 alleles were found intermediate in BNT genetic group. For BWLH group, A1 allele showed higher frequency (0.8878), while WLH group showed lack of polymorphism with only A1 allele present. This may be due to mutation or deletion of restriction site at 373 bp in WLH. BNT and BWLH genetic groups were in genetic equilibrium (P<0.01). The phylogenetic trees constructed from cGH1 and cGH2 motifs showed that WLH was at equidistance from BWLH and BNT. The five microsatellite markers i.e. ADL-102, ADL-136, ADL-158, ADL-171 and ADL-172 have shown presence of genetic polymorphism in all genetic groups. The number of alleles ranged from 2 to 6. The marker ADL- 171 was found to be highly polymorphic than other four microsatellite markers. BNT and WLH genetic groups deviated significantly (P<0.01) from genetic equilibrium at ADL-102 locus. All three genetic groups deviated significantly (P<0.01) from genetic equilibrium at ADL-136 locus and ADL-158. The genotypic frequencies of BWLH and WLH populations significantly deviated (P<0.01) from equilibrium at ADL-171 locus. The genetic groups BNT and BWLH were in genetic equilibrium while genetic group WLH significantly (P<0.01) deviated from equilibrium at ADL-172. The heterozygosity for five microsatellite markers ranged from 0.48 for ADL-102 to 0.78 for ADL-171. The PIC values ranged from 0.463 for ADL-102 to 0.683 for ADL-171. The microsatellite marker ADL-171 was more polymorphic and informative than other four microsatellite markers. A consensus tree obtained by bootstrapping multiple data set analysis from all 5 microsatellite markers revealed that WLH and BWLH were forming one cluster in cent percent cases, where as BNT branched out separately from the cluster. The statistical analysis revealed non-significant effect of genotypes at cGH1/Sac / and cGH2/Wsp / locus on BW-8, BW-20, BW-40, TEN-40, EW- 40 and TFC-40 traits. The statistical analysis revealed that various genotypes of all 5 microsatellite markers exerted no significant effect on different production traits under study except for BW-8 in WLH at Adl-102; BW-40 in BNT at ADL- 171; and BW-20, BW-40, TEN-40 and EW-40 in BNT at ADL-172.ThesisItem Open Access GENETIC ANALYSIS OF TWO STRAINS OF WHITE LEGHORN UNDER RECIPROCAL RECURRENT SELECTION PROGRAMME(AAU, Anand, 2000) MISHRA, RAM KAILASH; PATEL, M. M.The present study was carried out to study the inheritance pattern in terms of magnitude of genetic, phenotypic variances and covariances for economic traits in the two strains and their reciprocal crosses (viz. IWD, IWK, DK and KD) under reciprocal recurrent selection (RRS) for five successive generations under All India Coordinated Research Project on Poultry (AICRP) at Poultry Complex, Gujarat Agricultural University, Anand Campus, Anand. Number of pullets from 1^^ to 5 generation under different genetic groups and crosses utilized as experimental materials were 879, 1551, 1395, 2080 and 1911. Over the generations, number of pullets utilized were 1819, 2048, 1650 and 2229 in IWD, IWK, DK and KD genetic groups respectively. The data obtained on these birds were analyzed to obtain the estimates of the means, heritability, genetic and phenotypic correlations and genetic correlation between crossbreds and purebreds for different economic traits. For computation of the estimates least squares analysis technique using LSMLMW and MIXED MODEL computer , programme (Harvey, 1990) was utilized. The least squares means for growth traits(BW8, BW20 and BW40), Egg production traits (EN40) and egg weight traits (EW32 and EW40) were computed in DD, KK, DK and KD genetic groups separately in each generation. In general, crosses performed better than pures (P<0.05) for EN40 over the generation, (DD Vs DK and KK Vs KD). The heritability estimates for DD and KK, pure lines for BWg were ranging from low to moderate across the generations. However, the heritability estimates for BWg for crosses ranged from medium to high across the generation. The estimates of h2 for BW20 and BW40 were moderate to high in magnitude in all the genetic groups across the generations. The estimates of heritability for EN40 pooled over generations for DD, DK, KK and KD genetic groups were 0.107 ± 0.054, 0.115 ± 0.049, 0.292 ± 0.073 and 0.131 ± 0.048 respectively. The estimates observed in KK line were higher in all the generations as compared to DD line except in third generation. The estimates in crosses were higher than the pures. Selection based on crossbreds performance can bring about genetic improvement in EN40 at fairly good rate. The heritability estimates for EW32 and EW40 in pure lines were low to high in magnitude. The genetic correlations amongst body weight traits (BW8, BW20 and BW40) were moderate to high in magnitude and in desired direction. All phenotypic correlations were statistically highly significant (P<0.01).