028110   CHITTORA M (Biotechnology Dep, Mohanlal Sukhadia Univ, Udaipur-313002, Email: mnshchittora@yahoo.co.in) : Assessment of genetic fidelity of long term micropropagated shoot cultures of Achras sapota L. var. 'Cricket Ball' as assessed by RAPD and ISSR markers. Indian J Biotechnol 2018, 17(3), 492-5.

RAPD and ISSR molecular markers were employed to evaluate genetic fidelity of shoots of Achras sapota L. var. ‘Cricket Ball’ raised through nodal bud cultures. Out of 53 RAPD and 60 ISSR primers screened, 34 RAPD and 20 ISSR primers generated a total of 270 and 120 clear and scorable amplification products, respectively. All the shoot cultures analyzed at different culture passages showed similar RAPD and ISSR profiles as compared to that of mother plant. No genetic variation was observed in in vitro shoot cultures. The results indicated the genetic stability of the tissue culture raised shoot cultures of A. sapota and corroborate the assumption that axillary multiplication is the safest mode for multiplication of true to type plants without any somaclonal variation.

2 illus, 2 tables, 14 ref

028138   SHARMA P, SHARMA R (Biotechnology Dep, Dr Y S Parmar University of Horticulture & Forestry, Solan-173230, Email: rajnish.sharma@yahoo.co.in) : DNA fingerprinting of peach (Prunus persica) germplasm in accessing genetic variation using arbitrary oligonucleotide markers system. Indian J Biotechnol 2018, 17(3), 484-91.

Molecular characterization of 45 peach (Prunus persica) accessions was carried out using 48 RAPD and 46 ISSR molecular markers to assess the value and magnitude of genetic divergence. The RAPD primers revealed 84.20 % polymorphism and ISSR markers generated 89.00% polymorphism. Pooled RAPD and ISSR along with UPGMA clustering based on Jaccard’s coefficient were estimated with a view to assess efficiency of the marker system in Prunus persica. Polymorphic information conten (PIC) values varied from 0.13 to 0.50 in RAPD and 0.12 to 0.49 in ISSR with the mean values for all loci were 0.33 and 0.35, respectively. Jaccard's similarity coefficient among peach accessions with respect to RAPD and ISSR markers ranged from 0.37 to 0.95 and 0.43 to 0.95 which indicated a broad genetic base. Pooled analysis of both molecular markers concluded that genotypes ‘Darli’ and ‘IC-2’ are most distantly related to each other. The use of arbitrary oligonucleotide primers in the amplification reaction facilitated the study of uncharacterized genomes. In the present study, high level of polymorphism indicates their applicability in framing more extensive studies in development of superior progenies, quantitative trait loci (QTL) mapping, molecular breeding, investigation of population genetic diversity, comparative mapping, selection of the parents etc. among various peach crop improvement programmes.

4 illus, 3 tables, 22 ref

028109   BORSE V, ZANAN R, NADAF A (Botany Dep, Savitribai Phule Pune Univ, Pune-411007, Email: abnadaf@unipune.ac.in) : Development of ISSR derived SCAR marker for economically important Benstonea thwaitesii (Martelli) Callm. and Buerki. Indian J Biotechnol 2018, 17(3), 480-3.

Benstonea thawaitesii (Martelli) Callm. and Buerki, an economically important plant of Pandanaceae family, is having fragrant male inflorescence. In vegetative growth phase, it is difficult to distinguish B. thwaitesii from other Pandanaceae species due to similar morphological characters. In the present study, we have developed inter simple sequence repeat (ISSR) derived sequence characterized amplified  region  (SCAR)  marker  for identification  of  B.  thwaitesii.  A  set  of  15 Indian Pandanaceae species was screened using 15 ISSR primers. Among 15 primers, UBC-857 generated unique 1 kb band specific to B. thwaitesii. It was further cloned, sequenced and used to design SCAR primer that amplified the expected amplicon of 750 bp in B. thwaitesii only. This is the first report of developing species-specific SCAR marker in B. thwaitesii using ISSR primers.

3 illus, 2 tables, 12 ref

028121   JAN S A, ALI G M, ALI S, SHAH S H, AHMAD N (National Agricultural Research Centre (NARC), Islamabad -45500, Pakistan, Email: drgmails@gmail.com) : Genetic improvement in tomato (Solanum lycopersicum) against salt stress. Indian J Biotechnol 2018, 17(3), 459-65.

Salinity and drought are the two major abiotic stresses that affect the quality and quantity of tomato. The dehydration response transcription element binding factor (DREB) gene enhances tolerance in plants against salinity. The present study was conducted to transformed DREB1A gene through Agrobacterium in tomato cultivar Roma against salt stresses. A binary vector containing DREB1A gene under the control of inducible promoter Lip 9 along with hygromycin screening marker  was used. The few major factors which directly affect transformation efficiency were optimized such as acetosyringone levels, cefotaxime concentrations, pre-selection time period and hygromycin levels. Four different concentrations (50, 100,150 and 200 mM) of NaCl were applied on both transgenic and non transgenic plants. The addition of 200 µM acetosyringone in co-cultivation media, 200 mg/L cefotaxime in pre-culturing media and 25 mg/L hygromycin  concentration significantly increased transformation efficiency. Maximum transformation efficiency 16.6 % was obtained in tomato cultivar Roma. The transgenic plants were confirmed with PCR and the expected band size ~630 bp was amplified. The transgene express in transgenic tomatoes was confirmed by reverse transcriptase- polymerase chain reaction (RT-PCR). The resulted transgenic plants showed enhanced tolerance against high salt concentration (200 mM) while the non transgenic plants showed the symptoms of wilting and eventually die at that concentration. The DREB1A gene was successfully transformed into local tomato cultivar Roma. The resulted transgenic tomato cultivar Roma will be recommended against extreme salt stresses.

5 illus, 3 tables, 2732 ref

028118   GULERIA S, WALIA A, CHAUHAN A, SHIRKOT C K (Microbiology Dep, Lovely Professional Univ, Jalandhar, Email: shg1988@gmail.com) : Production and eco friendly application of alkaline protease from Bacillus amyloliquifaciens sp1. Indian J Biotechnol 2018, 17(3), 448-58.

Alkaline protease from Bacillus amyloliquefaciens SP1 has been characterized in detail for its ecofriendly application of release of silver particles from gelatin layers of used X-ray films. It exhibited optimum activity at broad temperature range and maximum at 60 ⁰C under alkaline pH environment (8-12). Thermal inactivation of the crude enzyme followed first order kinetics. The half-life of the enzyme at 50, 60 and 65 ⁰C was 70, 15 and 12.6 min, respectively and the denaturation energy was 114.87 kJ/mol. Enzyme retained 53.83 and 108.33 % of its initial activity after heating for 15 min at pH 8.0 and temperature 60 ⁰C, in presence and absence of 10 mM MnSO₄, respectively. Enzymatic decomposition of gelatin layers was enhanced by increase of enzyme concentration from 38 to 3630 µg/ml/min, at 60 ⁰C and pH 8.0. This study reported the shortest time of 1.30 min at 3630 µg/ml/min and 4 : 30 min at 74 µg/ml/min of enzyme concentration for hydrolysis of gelatin layers. Keeping in mind that, nowadays recycling is needed and imperative, this is the first study to report that after addition of Mn₂⁺ ions, thermal stability of enzyme increased and it could be effectively reused for 8 cycles as compared with enzyme without protective agents which also increase its yield of silver recovery i.e. 19.56 ± 0.78 % by eight times.

11 illus, 5 tables, 26 ref

028143   SINGH G, KUMAR V, DUBEY A, AGRAWAL S, VERMA A K (Biochemistry Dep, G B Pant University of Agriculture and Technology, Pantnagar -263145, Email: akv72@rediffmail.com) : Cloning, sequencing and in silico analysis of β-glucosidase from Bacillus subtilis strain PS. Indian J Biotechnol 2018, 17(3), 431-40.

Glycosyl hydrolases are group of enzymes classified into 132 known families based on sequence similarity. Glycosyl hydrolase family 1 comprises of number of enzymes with a known activities including β-glucosidases, which have wide diversity and known to have important applications. In the present study, the β-glucosidase gene of glycosyl hydrolase family 1 from Bacillus subtilis strain PS was successfully cloned and expressed in E. coli BL21 (DE3) by using pET28 (a) vector system. Sequence characterization revealed a 1407 bp long nucleotide sequence with an open reading frame (ORF) encoding 469 amino acids and a calculated molecular mass of 51.6 kDa. Its translated protein sequence was used for comparative analysis to reference protein sequences of β-glucosidases from different organisms for multiple sequence alignment, phylogenetic tree construction, variation in biochemical features and distribution of motifs using various bioinformatics tools. The analysis revealed important sequence features and properties of this important family which would be helpful in further classification, diversity study and genetic engineering of glycosyl hydrolase family 1 β-glucosidases.

3 illus, 5 tables, 30 ref

028116   GOUVEIA M, RODRIGUES M, TEIXEIRA L, CORDEIRO N (Madeira univ, Portugal, Email: mgouveia@uma.p) : Molecular cloning and characterization of cDNAs encoding cytosolic malate dehydrogenase and vacuolar (H⁺)-ATPase in Annona cherimola and their expression during postharvest ripening. Indian J Biotechnol 2018, 17(3), 422-30.

This study aims to investigate the expression of two cherimoya genes putatively related to fruit ripening. Two full-length cDNAs encoding cytosolic NAD-dependent malate dehydrogenase (AccytMDH) and vacuolar (H⁺)-ATPase c subunit (AcVHA-c) were isolated from Annona cherimola using the reverse transcriptase polymerase chain reaction (RT- PCR) and rapid amplification of cDNA ends (RACE). The AccytMDH codes for a 332 amino acid polypeptide with a predicted molecular mass of 35.6 kDa. The deduced amino acid sequence for AccytMDH shared high identity with other plant homologous malate dehydrogenase proteins. The AcVHA-c encodes a proteolipid subunit of the V-type proton ATPase with 166 amino acids (16.7 kDa). Comparison of the deduced amino acid sequence from AcVHA-c revealed four transmembrane domains highly conserved among plant counterparts. The expression of AccytMDH and AcVHA-c, assessed by semi- quantitative RT-PCR showed that there is an increase in the accumulation of transcripts during postharvest ripening, although not correlated by a significant upsurge of titratable acidity they might contribute to organic acid accumulation and translocation during postharvest ripening of cherimoya in association with other enzymes and carriers. By using AccytMDH and AcVHA-c as molecular targets new strategies can be exploited to get a clear picture in the ripening of cherimoya.

7 illus, 39 ref

028144   SUDHAMANI M, BATISH V K, HELLER K J (Biotechnology Dep, K L Univ, Guntur-522502, Email: sudhamani1@rediffmail.com) : Application of mobilization gene promoter for heterologous expression and curing of plasmid pSMA23. Indian J Biotechnol 2018, 17(3), 416-21.

Lactobacilli have been recognized as key members of the group of probiotic bacteria by virtue of expressing a wide spectrum of physiological functions beneficial to human health. Genetic modification of this genus requires the availability of tools for gene expression like promoter and suitable host vector systems. In the context, plasmid pSMA23 was cured from its native host Lactobacillus casei A23 by electroporation with a construct based on the pSMA23 replicon. Cured strain A23 can be employed as host for vectors or constructs based on the replicon of pSMA23 and for heterologous expression of genes. The promoter less gene cat194 coding for chloramphenicol resistance was cloned under the control of the putative mobilisation gene promoter present on the native plasmid pSMA23 of Lactobacillus casei A23. Constructs were electroporated into Lactobacillus casei LK1 and transformants were obtained on media containing chloramphenicol, indicating expression of the cat194 gene. The putative promoter is thus active and can be recruited for gene expression.

3 illus, 2 tables, 32 ref

028115   GOMAA E Z (Biological and Geological Sciences Dep, Ain Shams Univ, Egypt, Email: emann7778@yahoo.com) : β-galactosidase from Lactobacillus delbrueckii and Lactobacillus reuteri: Optimization, characterization and formation of galactooligosaccharides. Indian J Biotechnol 2018, 17(3), 407-415.

Beta-galactosidase (β-gal) is one of the important commercial enzymes having several applications in food, pharmaceutical industry and in the synthesis of galactooligosaccharides known for their prebiotic properties. The production of β-gal from Lactobacillus delbrueckii and Lactobacillus reuteri was studied. The highest yield of β-gal was achieved using wheat flour as carbon source. Beef extract gave the highest value of enzymatic production by L. delbrueckii reached 24.25 U/ml, while L. reuteri produced the highest β-gal (20.20 U/ml) in the presence of peptone. The optimum β-gal activity was recorded on 15 mM O-nitrophenyl β-D-galactopyranoside (ONPG) at 40 °C and pH 7. Further enzyme activity was enhanced in the presence of Mg²⁺, Mn²⁺and Fe²⁺, while it was decreased by Ag⁺ and Ni⁺ for both strains. The production of galactooligosaccharides (GalOS) from lactose using the produced β-gal was investigated. The maximum yield of 40 and 35 % (w/w) GalOS could be achieved with β-gal of L. delbrueckii and L. reuteri, respectively from 40 % lactose solution at 40 °C and pH 7. These characteristics of produced β-gal showed that it could be a promising candidate for various industrial as well as biotechnological applications.

7 illus, 3 tables, 32 ref

028141   SINGH A, BANERJEE R (Agricultural and Food Engineering Dep, Indian Institute of Technology, West Bengal-721302, Email: rb@agfe.iitkgp.ernet.in) : Value addition to soybean whey through microbial and enzymatic intervention. Indian J Biotechnol 2018, 17(3), 397-401.

The wide range of functional properties of soy protein and its high nutritive value makes it base of a novel food platform. Various products are in the market like soy protein isolate, soy protein concentrate, tofu, soymilk which are well accepted by the consumers. Nevertheless, soybean processing operations generate a large proportion of liquid effluent termed as soybean whey. This yellowish liquid waste can be functionally and nutritionally valuable because of their nutrient composition. Discarded whey is not only accountable for pollution problem, but also represents an economic and nutritional penalty in this era. Till now, there are only few reports on effective use of soybean whey, for this reason the present article emphasis to summarize all the extensive research developed for its utilization, so that soybean whey can be well recognized as a potential feedstock both for the microbial and enzymatic intervention.

1 illus, 1 table, 28 ref

026680   KAUSHIK N, BHATT A K, CHAKRABARTI S K (Crop Improvement Div, Central Potato Research Institute (CPRI), Shimla- 171 001, Email: aneelamkaushik@gmail.com) : Plastid genome engineering and its potential applications: A review. Int J Agric Environ Biotechnol 2018, 11(4), 615-21.

Plastid genome engineering is a credible tool for the basic biotechnological research and various innovative techniques have led to the better understanding of the complex processes involved in the plastid transformation. Plastids in higher plants are the major biosynthetic centers for photosynthesis which is the main source of energy requirement. Plastids have their own genome i.e. plastome which is maternally inherited in most angiospermic plant species. Although production of transgenic plants has traditionally been through expression of transgene in the nucleus, but plastid transformation is considered more attractive and efficient target for genetic engineering due to several advantages over nuclear transformation including high level of foreign protein, eliminating the risk of cross pollination with weeds, absence of silencing mechanism and ability to engineer multiple genes rather than a single gene. The potential utility of plastid genome engineering has been explored in development of crops with various agronomic traits, development of vaccine, biopharmaceuticals, therapeutic proteins, biomaterials and industrial enzymes, which will definitely prove beneficial in near future. Plastid transformation is still to be fully utilized for product commercialization, because of the problems associated with protein purification and expression level control. This review article highlights the various possibilities and potential applications of plastid genome engineering for generation of marker free transplastomic plants, improvement in agronomic traits and role of plastids in the production of cost effective biopharmaceuticals and biomaterials.

62 ref

026679   YADAV S K, MAURYA S K, YADAV A K, KUMAR A, YADAV K (Narendra Deva Univ of Agriculture & Technology, Faizabad - 224 229, Email: dr.shivkumarjnp@gmail.com) : Study of prolactin receptor gene (PRLR5) polymorphisms and its association with egg production in Kadaknath hens. Indian J Anim Res 2018, 52(8), 1232-5.

The present study was conducted to investigate the polymorphisms of prolactin receptor (PRLR5) gene and its association with egg production in Kadaknath hens. Egg production is a polygenic inheritance trait.  Study was conducted on twenty female birds of Kadaknath kept for laying. Egg production performances were recorded as age at first laying (AFE), Body Weight at First Egg (WFE), Mean Egg Weight (MEW) and Total No. of Eggs at 90 days of laying (TEN). Genomic DNA isolated from 2- 3 of blood collected from wing vein of each bird was amplified for prolactin receptor (PRLR5) gene with specific primer by standardizing and optimising the PCR protocols.  PCR was performed in a final volume of 20 µl. The amplified PCR products were resolved on the gels to generate polymorphisms. PRLR5 was digested with BamHI. Retriction digested products were run on 2 % agarose gel electrophoresis. PRLR5 showed two alleles & two genotypes. The frequency of AA genotype at this locus was 0.75 & BB genotype was 0.25. The AFE (d), WFE (Kg), MEW (g) and TEN of Kadaknath hens in the present study were found to be 188.00 ± 0.71, 1.26 ± 0.03, 42.83 ± 0.21 & 37.75 ± 0.59 respectively. Birds with AA genotype of PRLR5 had a significantly (P < 0.05) better WFE & AFE than BB genotype. Prolactin receptor (PRLR5) genes produced polymorphism in Kadaknath and were associated with egg production traits.

1 illus, 2 tables, 23 ref

026678   DIALLO T, SINGLA L D, SUMBRIA D, KAUR P, BAL M S (Veterinary Parasitology Dep, Guru Angad Dev Veterinary and Animal Sciences Univ, Ludhiana- 141 004, Email: ldsingla@gmail.com) : Conventional and molecular diagnosis of haemo-protozoan infections in cattle and equids from Republic of Guinea and India. Indian J Anim Res 2018, 52(8), 1206-11.

A cross-sectional study was carried out on 50 N’ Dama cattle and 35 equids blood samples from Republic of Guinea and Punjab, India, respectively to assess the level of exposure to obligatory haemoprotozoa Theileria equi and Trypanosoma evansi by 18S rRNA gene based primary and nested polymerase chain reaction (nPCR) and RoTat1.2 based card agglutination test (CATT), and to investigate risk factors associated with the infection. Blood smear examination revealed the prevalence rate of 6 % (95 % CI = 2.06-16.22) for Trypanosoma spp. in N’dama cattle and 5.7 % for T. equi in equids. In equids, 17.14 % (95 % CI = 8.10-32.68) samples showed positive titer by CATT. Primary PCR showed 5.71 % (95 % CI = 1.58-18.8) infection and on the other hand nested PCR depicted 20 % (95 % CI = 10.04-35.89) T. equi infection. Moreover, only 8.57 % (95 % CI = 2.96-22.38) prevalence of T. evansi was recorded by T. evansi based multiplex PCR. PCRs revealed higher risk of infection of both T. equi (OR = 5.28, 95 % CI = 0.68-49.81) and T. evansi (OR = 3.33, 95 % CI = 0.20-104.83) in the farms where proper deworming/vaccination schedule was not followed. The risk factor associated with the type of host species had an odds ratio (OR) of 5 (95 % CI = 3.90-74.33) for donkeys/mules versus horses for T. equi infection. This group was also at higher risk of infection with OR of 4.8 (95 % CI = 0.12-124.47) for T. evansi. The present exploration brings out a variety of commodities at risk of infectivity pertaining to trypanosomosis and theileriosis calculated by different PCRs assay.

2 tables, 23 ref

026677   CHANG W, WANG J, ZHANG Y, WU J (Tarim Univ, Xinjiang 843 300, China, Email: wjyn-w@126.com) : Discovery of two novel miRNAs from the Ovis aries by a combinatorial approach of experiments and bioinformatics. Indian J Anim Res 2018, 52(8), 1155-61.

In the study, we successful constructed a library from the ovine testis using next-generation sequencing technology, and identified 219 novel miRNAs by bioinformatics. Two of the novel miRNAs (ovis_aries_testis-m0052_5p and ovis_aries_testis-m0165_5p), which were expressed in the sheep testis and ovary, and were confirmed by real-time PCR and northern blotting. Ovis_aries_testis-m0052_5p was 23 nucleotides in length, was located on chromosome 15, and had 100 % similarity to mmu-miR-34c-5p, hsa-miR-34c-5p, gga-miR-34c-5p, and cfa-miR-34c. Ovis_aries_testis-m0165_5p was 21 nucleotides in length, located on chromosome 5, and had 100 % similarity to mmu-miR-145a-5p, hsa-miR-145-5p, ssc-miR-145-5p, and bta-miR-145. The pre-miRNAs for Ovis_aries_testis-m0052_5p and Ovis_aries_testis-m0165_5p were 75 and 81 nucleotides in length, and both had a standard hairpin stem-loop structure. From the consistency of the sequence and structure, we speculated that ovis_aries_testis-m0052_5p had a function similar to hsa-miR-34c-5p, mmumiR-34c-5p, and ovis_aries_testis-m0165_5p had a function similar to hsa-miR-145-5p, which were involved in the fine regulation of cell survival, spermatozoon generate, breeding activities. Therefore, we defined the identified miRNAs as oar-miR-34c-5p and oar-miR-145-5p. The results will enrich the miRNA database, and provide the basis for research into the regulatory mechanisms of miRNA in relation to breeding activities.

6 illus, 2 tables, 30 ref

026676   RUBAN S W, BABU R N, ROBINSON J J A, KUMAR T M A S, KUMARASAMY P, PORTEEN K, RAJA P (Livestock Products and Technology Dep, Veterinary Coll, Hassan- 573 202, Email: rubanlpt@gmail..com) : Molecular detection of enterotoxigenic Staphylococcus aureus isolated from mutton marketed in retail outlets of Chennai, India. Indian J Anim Res 2018, 52(7), 1048-52.

The present study was aimed at detection of enterotoxigenic S. aureus in mutton marketed in retail outlets of Chennai. A total of 120 meat samples were collected from across Chennai for isolation of S. aureus and it was observed that 66.28 per cent of the samples were contaminated with S. aureus. The S. aureus count in mutton samples ranged from 1.8 x 10² to 4.9 x 10⁴ CFU/ g with an overall average of 1.30 x 10⁴ CFU/ g. All the isolates presumptively identified as S. aureus biochemically, amplified 181 bp product specific for nuc gene by PCR, which is species specific marker for S. aureus. Enterotoxin gene profiles (multiplex PCR) results revealed that 70.17 percent of the isolates were enterotoxigenic carrying only six genes (seb, sed, seg, seh, sei and sej) either alone or in combination, whereas none of these isolates harbored sea, sec and see. It was clear that seb (72.5 %) was the predominant enterotoxin gene followed by seg and sei, seh, sej and sed. Six different toxin gene profiles were exhibited by different isolates and majority of the isolates (55 %) carried two or more genes as compared to only one toxin gene.

2 illus, 2 tables, 26 ref

026675   KUMAR V, MAAN S, KUMAR A, BATRA K, CHAUDHARY D, DALAL A, GUPTA A K, BANSAL N, SHEORAN N, MAAN N S (Animal Biotechnology Dep, Lala Lajpat Rai Univ of Veterinary and Animal Sciences, Hisar- 125 004, Email: sushilamaan105@gmail.com) : Real time PCR assay for differentiation of Brucella abortus and Brucella melitensis. Indian J Anim Res 2018, 52(7), 1037-42.

Brucellosis is one of the zoonotic diseases of major concern and can cause huge economic losses to livestock industry. Serological tests and bacterial isolation are considered as the gold standard assay for diagnosis of Brucella spp. but they are time-consuming, hazardous and lack specificity. To control and eradicate a disease, a confirmatory diagnostic method which is sensitive, quick and specific is the foremost requirement. Therefore in this study, we evaluated the performances of two newly designed TaqMan real-time PCR assays targeting the BruAB_0168 gene and BMEII0466 gene for Brucella abortus and Brucella melitensis (respectively). Both the assays were found to be highly specific in differentiation of respective species. Both the assays can detect as low as 0.02 fg of DNA and there was no detectable difference found in sensitivity of these two tests. R2 value and efficiency of these tests ranged from 0.992 - 0.998 and 100- 106 %, respectively showing that these assays are highly efficient.  Compared to conventional PCR assays these qPCR assays were 100 times higher sensitive. In conclusion, the present study showed that the developed real-time PCR assays are more sensitive, specific, have high reproducibility and repeatability and are faster than serological and conventional PCR methods for differentiation of Brucella abortus and Brucella melitensis.

4 illus, 1 table, 23 ref

026674   NEUPANE S, MA Q, MATHEW F M, VARENHORST A J, ANDERSEN E J, NEPAL M P (Biology and Microbiology Dep, South Dakota State Univ, Brookings, United states of America, Email: madhav.nepal@sdstate.edu) : Evolutionary divergence of TNL disease-resistant proteins in soybean (Glycine max) and common bean (Phaseolus vulgaris). Biochem Genet 2018, 56(4), 397–422.

Disease-resistant genes (R genes) encode proteins that are involved in protecting plants from their pathogens and pests. Availability of complete genome sequences from soybean and common bean allowed us to perform a genome-wide identification and analysis of the Toll interleukin-1 receptor-like nucleotide-binding site leucine-rich repeat (TNL) proteins. Hidden Markov model (HMM) profiling of all protein sequences resulted in the identification of 117 and 77 regular TNL genes in soybean and common bean, respectively. We also identified TNL gene homologs with unique domains, and signal peptides as well as nuclear localization signals. The TNL genes in soybean formed 28 clusters located on 10 of the 20 chromosomes, with the majority found on chromosome 3, 6 and 16. Similarly, the TNL genes in common bean formed 14 clusters located on five of the 11 chromosomes, with the majority found on chromosome 10. Phylogenetic analyses of the TNL genes from Arabidopsis, soybean and common bean revealed less divergence within legumes relative to the divergence between legumes and Arabidopsis. Syntenic blocks were found between chromosomes Pv10 and Gm03, Pv07 and Gm10, as well as Pv01 and Gm14. The gene expression data revealed basal level expression and tissue specificity, while analysis of available microRNA data showed 37 predicted microRNA families involved in targeting the identified TNL genes in soybean and common bean.

4 illus, 1 table, 13 ref

026673   LAWANIA S, SINGH N, BEHERA D, SHARMA S (Biotechnology Dep, Thapar Univ, Punjab - 147 002, Email: siddharthsharma.phd@thapar.edu) : Association of XPA polymorphisms towards lung cancer susceptibility and its predictive role in overall survival of north Indians. Biochem Genet 2018, 56(4), 375–96.

The present study investigated the role of Xeroderma pigmentosum group A (XPA) polymorphism (A23G and G709A) with lung cancer risk and its association with overall survival in North Indians. 370 cases and 370 controls were investigated to evaluate association between XPA polymorphism (A23G and G709A) with lung cancer risk using logistic regression analysis. A follow-up study was also conducted for 291 lung cancer cases illustrating correlation between overall survival in lung cancer patients and XPA variants. GG genotype showed an increased lung cancer risk (p = 0.0007) for A23G polymorphism whereas G709A polymorphism was associated with significant protective effect in heterozygous (AG) subjects (p = 0.001). When stratified according to smoking status an increased risk for lung cancer was observed for GG genotype in A23G polymorphism (p = 0.0002). A poor survival in females carrying variant genotype (GG) was observed (p = 0.001; MST = 4.16 months) for A23G polymorphism. Adenocarcinoma patients with heterozygous genotype showed an increased hazard ratio (p = 0.02) for A23G polymorphism. G709A was associated with a reduced hazard ratio marking a better survival among mutant females (HR 0.17; p = 0.05; MST = 18.63 months). It can be concluded that A23G polymorphism might contribute to increased lung cancer risk in North Indian population emphasizing on poor survival among females. G709A polymorphism might result in protective effect in lung cancer subjects. The present study had a low sample size but it could act as reference for the large sample studies in future.

2 illus, 5 tables, 30 ref

026672   BO GAO B, WANG W, WU H, CHEN C, SHEN D, WANG S, CHEN W, ZHANG L, CHAN S, SONG C (Yangzhou Univ, Yangzhou 225009, China, Email: cysong@yzu.edu.cn) : Changes in skeletal muscle and body weight on sleeping beauty transposon-mediated transgenic mice overexpressing pig mIGF-1. Biochem Genet 2018, 56(4), 341–55.

Insulin-like growth factor (IGF-I) is an important growth factor in mammals, but the functions of the local muscle-specific isoform of insulin-like growth factor 1 (mIGF-1) to skeletal muscle development have rarely been reported. To determine the effect of pig mIGF-1 on body development and muscle deposition in vivo and to investigate the molecular mechanisms, the transgenic mouse model was generated which can also provide experimental data for making transgenic pigs with pig endogenous IGF1 gene. We constructed a skeletal muscle-specific expression vector using 5′- and 3′-regulatory regions of porcine skeletal α-actin gene. The expression cassette was flanked with Sleeping Beauty transposon (SB)-inverted terminal repeats. The recombinant vector could strongly drive enhanced green fluorescence protein (EGFP) reporter gene expression specifically in mouse myoblast cells and porcine fetal fibroblast cells, but not in porcine kidney cells. The EGFP level driven by α-actin regulators was significantly stronger than that driven by cytomegalovirus promoters. These results indicated that the cloned α-actin regulators could effectively drive specific expression of foreign genes in myoblasts, and the skeletal muscle-specific expression vector mediated with SB transposon was successfully constructed. To validate the effect of pig mIGF-1 on skeletal muscle growth, transgenic mice were generated by pronuclear microinjection of SB-mediated mIGF-1 skeletal expression vector and SB transposase-expressing plasmid. The transgenepositive rates of founder mice and the next-generation F1 mice were 30 % (54/180) and 90.1 % (64/71), respectively. The mIGF-1 gene could be expressed in skeletal muscle specifically. The levels of mRNA and protein in transgenic mice were 15 and 3.5 times higher, respectively, than in wild-type mice. The body weights of F1 transgenic mice were significantly heavier than wild-type mice from the age of 8 weeks onwards. The paraffin-embedded sections of gastrocnemius from 16-week-old transgenic male mice showed that the numbers of myofibers per unit were increased in comparison with those in the wild-type mice. mIGF-1 overexpression in mice skeletal muscle may promote myofibers hypertrophy and muscle production, and increased the average body weight of adult mice. Transgenic mice models can be generated by the mediation of SB transposon with high transgene efficiency.

7 illus, 4 tables, 36 ref

026671   SWELLAM M, HASHIM M, MAHMOUD M S, RAMADAN A, HASSAN N M (Biochemistry Dep, National Research Centre, Giza 12622, Egypt, Email: menhamswellam@gmail.com) : Aberrant expression of some circulating miRNAs in childhood acute lymphoblastic leukemia. Biochem Genet 2018, 56(4), 283–94.

Acute lymphoblastic leukemia (ALL) is a heterogeneous cancer commonly affecting children due to dysregulation of miRNA expression. In the current study, authors investigated the expression profile for miRNA-125b-1 and miRNA203 among childhood ALL. Blood samples were collected from newly diagnosed childhood ALL and healthy control children. The expression profile for candidate miRNAs was detected using quantitative RT-PCR analysis. Statistical analysis were performed using receiver operating characteristic curve (ROC) to examine the diagnostic efficacy of the two miRNA and their levels among ALL clinicopathological factors and phenotypes. The median expression level for miRNA-125b-1 was significantly high in childhood ALL; while miRNA-203 level was significantly low in childhood ALL as compared to control ones. MiRNA-125-1 reported significant increase in T-ALL as compared to other ALL phenotypes. Median miRNA203 level was high in T-ALL followed by pre-B-ALL although no significant difference was reported. Clinicopathological factors did not emphasize significance with either detected miRNAs. Using ROC curve the diagnostic efficacy was significant with an area under the curve 0.858 for miRNA-125b-1 (83.72, 100 %) and 0.878 for miRNA-203 (97.67, 86.96 %). The combination of the two key miRNAs revealed absolute sensitivity (100 %). MiRNA-125b-1 and miRNA-203 can be useful molecular markers for diagnosis of ALL. Further studies with large cohort are warranted to validate these results.

3 illus, 3 tables, 34 ref

026670   YADAV S, CHANDRA A, KUMAR A, MITTAL B (Biotechnology Dep, Babasaheb Bhimrao Ambedkar Univ, Uttar Pradesh - 226 025, Email: balrajmittal@gmail.com) : Association of TERT-CLPTM1L and 8q24 common genetic variants with gallbladder cancer susceptibility and prognosis in north Indian population. Biochem Genet 2018, 56(4), 267–82.

Gallbladder carcinoma (GBC) is one of the common malignancy of the biliary tract. Several genome wide and candidate gene studies have reported associations between multiple cancer types and single-nucleotide polymorphisms on 5p15.33 and 8q24.21 loci. However, predisposition potential of these genetic variants has not been assessed in GBC. We performed the present study to assess the potential of five polymorphisms on 5p15.33 and one on 8q24.21 locus in GBC risk and treatment response in patients undergoing chemoradiotherapy. We extracted genomic DNA from peripheral blood and genotyped selected SNPs using TaqMan allelic discrimination assays in 523 GBC cases and 274 controls from the north-Indian population. Statistical tests were performed to assess the association of selected common genetic variants with gallbladder cancer susceptibility and prognosis. Binary logistic regression analysis showed significant association of TERT rs2736100C > A [OR(CI) = 0.690(0.515–0.924), p value = 0.013], CLPTM1L rs401681C > T [OR(CI) = 0.586(0.405–0.847), p value = 0.004], and CASC8 rs6983267G > T [OR(CI) = 1.629(1.215–2.186), p value = 0.001] with GBC risk. Further, using multivariate logistic regression, we observed that haplotype CLPTM1L  C_rs401681C_rs31489 TERT  T_rs2853676A_rs2736100 MIR4457  G_rs4635969 [OR(CI) = 7.52 (1.79–31.52), p value = 0.0064] is significantly associated with poor treatment response. In survival analysis, Kaplan–Meier survival curves showed significantly poor survival and COX regression suggested significantly higher hazard ratio in TT genotype carriers of CASC8 rs6983267 [OR(CI) = 4.28(1. 07–17.10), p value = 0.040] as compared to major allele and heterozygous (GG+GT) genotypes in metastatic GBC cases. The study revealed that 5p15.33 and 8q24.21 genetic variants significantly influence GBC risk and treatment response in north-Indian population. 

1 illus, 4 tables, 46 ref

026684   KANNAKI T R, VERMA P C, REDDY M R, SHANMUGAM M (Directorate of Poultry Research, Hyderabad-500 030, Email: trkannaki@gmail.com) : Molecular characterization of duck (Anas platyrynhos) Toll-like receptors, mRNA expressions profile in day-old duckling's tissues and cytokine response to in vitro TLR agonsists stimulation. Indian J Anim Res 2018, 52(6), 851-7.

TLR repertoire of duck, profiling of their mRNA expression in a range of duckling tissues and cytokine gene expressions upon TLR agonists stimulation in in vitro assay have been investigated. All ten TLR genes orthologous to chicken TLR repertoire were found in duck. Duck TLR genes showed 77-83 % similarity at amino acid level to their chicken counterparts. All ten TLRs-TLR1LA, 1LB, 2A, 2B, 3, 4, 5, 7, 15 and 21 mRNA expressions were significantly higher in bursa than other tissues studied, whereas in muscle all TLRs mRNA expressions were significantly lower except for TLR15 (P < 0.01). TLR7 gene expression was significantly higher in spleen, bursa and also in lung tissues (P < 0.01). The cytokine gene expression levels in duck PBMCs upon LPS and poly I:C stimulation have been quantified. IL-1γ gene expression level in LPS stimulated duck PBMC culture was significantly higher at both 12 h and 24 h time intervals (P < 0.05). However, there were no significant changes in IFN-γ gene expression levels in poly I:C stimulated duck PBMC culture at both the intervals. TLR gene expression in young ducklings together with cytokine response upon LPS stimulation demonstrates the innate preparedness of younger birds to encounter pathogens and their functional ability to respond to their ligands.

2 illus, 3 tables, 36 ref

026683   BORAH B, SINGH A P, GOGOI H, PHUKAN A J, SARKHEL B C (Nanaji Deshmukh Veterinary Science Univ, Jabalpur- 482 004, Email: borah.drbiswajyoti@gmail.com) : Myostatin silencing effect on basic helix-loop-helix transcription factors in caprine foetal fibroblast cells. Indian J Anim Res 2018, 52(6), 843-50.

Transgenic food animal production is one of the potential and need oriented research to mitigate the food crises of the world. In vitro gene silenced animal cells and making use of these cells for transgenesis one of the suitable way to produce productive animals. Myostatin is a negative regulator of muscle growth, has the potential to increase the muscle mass upon its silencing. Four Hush 29-mer anti- myostatin (MSTN) shRNA constructs were checked for myostatin gene silencing in caprine foetal fibroblast cells and its subsequent effect on basic helix– loop–helix (bHLH) transcription factors. These factors are necessary for the terminal differentiation, proliferation, and homeostasis of muscle development. Different shRNA constructs displayed 55.1 to 91.5 % (p < 0.01) of myostatin silencing in caprine foetal fibroblast cells and upregulation of myogenic gene. Upregulation of 7.97 to 111.67 % for MyoD, 77.0 % to 319.47 % for myogenin, 16.67 % to 138.0 % for Myf5 were observed . The Pearson correlation established a negative correlation between myostatin and genes under study. Result suggests that knockdown of MSTN a potential approach to improve caprine musculatures.

8 illus, 2 tables, 20 ref

026682   BASANG W, AN T, LUO X, ZHU Y, DANJIU L, HE S, E G (Southwest Univ, Lasa 850009, China, Email: eguangxin@swu.edu.cn) : Identification and characterization of microRNAs in white and black coated yak. Indian J Anim Res 2018, 52(6), 816-9.

In this study, we used high-throughput technology to provide the first transcriptome dataset for differentially expressed miRNA in mixed pools of dermis tissue from black- and white-coated yak to research the possible molecular mechanisms of yak coat pigmentation. In this study, 92,636,002 and 95,917,842 clear reads were generated through Illumina paired-end sequencing. A total of 78 differentially expressed miRNAs (DEMs) were identified, including 59 upregulated and 19 downregulated miRNAs in the mixed pools of white-coated yak compared with the mixed pools of black-coated yak. In addition, 3634 genes were predicted as putative targets of DEMs. These DEGs related to 59 GO categories and were enriched in 216 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, including melanogenesis and the Wnt signaling pathway. The results of the current study indicated that the coat color of the yak involved the transcriptional regulation process of miRNAs. These results provide helpful data to understand the molecular mechanisms of yak coat pigmentation.

1 illus, 1 table, 18 ref

026681   DARWISH A M, NADY G H E, ALI N I, ABDELSALAM A Z E (Cell Biology Dep, National Research Centre, Giza, Egypt, Email: am.darwish@nrc.sci.eg) : Evaluation of β-casein variants in Egyptian goat, sheep and cattle by allele specific PCR and relevance to β- casomorphin. Indian J Anim Res 2018, 52(6), 799-804.

The beta casein gene (CSNS2) has 12 genetic variants divided into two groups: the first group (A1, B, and G) which differ from the second group (A2, A3 C, D, E, F, H, H2 and I) where A base replaces C base, this leads to potential liberation of a bioactive peptide, $\mathrm{\beta }$-casomorphin, upon digestion where a histidine replaces a proline at position 67. The allele specific polymerase chain reaction (AS-PCR) was evaluated to distinguish between the beta casomorphin releasing variants (A1 and B) and the non-releasing variants. The sequence analysis was used to determine these variants and confirm it in goat, sheep and cattle. The results showed that cattle carrying allele A1 either homozygous or heterozygous more than sheep and goats. The allele frequency of A1 and A2 is 0.44, 0.56 in goats, 0.43, 0.57 in sheep and 0.54, 0.46 in cattle, respectively. The sequence results reported changing of C base to A base in goat, sheep and cattle. Therefore, this study reported that goat and sheep milk was more safe than cattle milk.

3 illus, 3 tables, 23 ref

026669   NEGI Y K, PANDEY C, SAXENA N, SHARMA S, GARG F C, GARG S K (Basic Sciences Dep, Forestry Coll (VCSG UUHF), Tehri Garhwal, Uttarakhand, Email: yknegi@rediffmail.com) : Isolation of antibacterial protein from Lactobacillus spp. and preparation of probiotic curd. J Food Sci Technol 2018, 55(6), 2011-20.

The study was aimed to isolate antagonistic lactobacilli and the molecules responsible for their antagonistic ability from curd. Preparation of probiotic curd and the ability of the selected lactobacilli to suppress the pathogen therein was also assessed. All the 116 isolates were identified as Lactobacillus spp. based on morphological, biochemical and curdling assays. Five of these lactobacilli (Lb-17, Lb-33, Lb-108, Lb-112, and Lb-N3) were found most promising to inhibit all test pathogens (Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Salmonella typhi and Shigella sonnei). The cell-free culture supernatants of these five lactobacilli were recorded as thermo-tolerant when subjected to heat treatment at 100 C for 20 min. The loss in the activity after protease treatment indicated the proteinaceous nature of the antimicrobial molecule present in the culture supernatants. Active protein (19 kDa) produced by lactobacilli was confirmed by SDS-PAGE followed by agar-overlay method. Antibiotic sensitivity assay revealed that the selected Lactobacillus spp. isolates were resistant to methicillin and vancomycin. Probiotic curd prepared by using Lb-108 and Lb-N3 was found to be superior to rest of the three isolates based on organoleptic tests and shelf-life. Complete inhibition of all the test pathogens in curd was shown by Lb-108 and Lb-N3. Inhibition spectrum, production of thermostable protein and preparation of quality curd suggest Lb-108 and Lb-N3 as promising candidates to prepare probiotic curd.

1 illus, 4 tables, 41 ref

026668   VAZQUEZ M A D, GARCIA-UGLDE C R, ALVAREZ B E, VILLEGAS L M, GARICA-ALMENDAREZ B E, ROSADO J L, REGALADO C (Autonoma de Queretaro Univ, 76010 Queretaro, Mexico, Email: carlosr@uaq.mx) : Use of urea-polyacrylamide electrophoresis for discrimination of A1 and A2 beta casein variants in raw cow's milk. J Food Sci Technol 2018, 55(5), 1942-7.

Beta-casein (BC) in cow’s milk occurs in several genetic variants, where BC A1 (BCA1) and BC A2 (BCA2) are the most frequent. This work deals with a method based on modified polyacrylamide gel electrophoresis using urea PAGE to discriminate BCA1 and BCA2 variants from Holstein Friesian (HF) and genetically selected Jersey A2/A2 (JA2) cow’s milk. Two well defined bands were obtained from BC fraction of HF milk, while that of JA2 showed a single band. Proteins from these bands were sequenced by HPLC-quadrupole linear ion trap/mass spectrometry, resulting in BCA1 and BCA2 separation from the BC fraction of HF milk, whereas BCA2 was the only constituent of JA2 fraction. This method represents a feasible and useful tool to on site phenotyping of BC fraction of cow’s milk for pharmaceutical and food industries applications.

3 illus, 28 ref

026667   LIU Z J, LI F, WANG L G, LIU R Z, MA J J, FU M C (Cotton Research Center, Jinan 250100, People’s Republic of China, Email: scrcliuzhanji@sina.com) : Molecular characterization of a stress-induced NAC gene, GhSNAC3, from Gossypium hirsutum. J Genet 2018, 97(2), 539-48.

NAC genes, specific to plants, play important roles in plant development as well as in response to biotic and abiotic stresses. Here, a novel gene encoding a NAC domain, named as GhSNAC3, was isolated from upland cotton (Gossypium hirsutum L.). Sequence analyses showed that GhSNAC3 encodes a protein of 346 amino acids with an estimated molecular mass of 38.4 kDa and pI of 8.87. Transient localization assays in onion epidermal cells confirmed GhSNAC3 is a nuclear protein. Transactivation studies using a yeast system revealed that GhSNAC3 functions as a transcription activator. Quantitative real-time polymerase chain reaction analysis indicated that GhSNAC3 was induced by high salinity, drought and abscisic acid treatments. We overexpressed GhSNAC3 in tobacco by using Agrobacterium-mediated transformation. Transgenic lines produced longer primary roots and more fresh weight under salt and drought stresses as compared to wild-type plants. Collectively, our results indicated that overexpression of GhSNAC3 in tobacco can enhance drought and salt tolerances.

6 illus, 2 tables, 45 ref

026666   SEYOUM M, DU X M, HE S P, JIA Y H, PAN Z, SUN J L (Jimma Univ, Jimma, Ethiopia, Email: mulugeta.seyoum@hotmail.com) : Analysis of genetic diversity and population structure in upland cotton (Gossypium hirsutum L.) germplasm using simple sequence repeats. J Genet 2018, 97(2), 513-22.

Improvement of cotton fibre yield and quality is challenging due to the narrow genetic base of modern cotton cultivars, which emphasizes the great need to effectively explore the existing germplasm resources. With major objective to assess the genetic diversity and population structure at DNA level, 302 elite upland cotton germplasm accessions (253 Chinese and 49 different exotic origins), were genotyped using 198 simple sequence repeats (SSRs) markers. Each of the 198 markers differed greatly in its ability to detect variations in the panel of cotton germplasm. The SSRs amplified 897 alleles, of which 77.7 % were polymorphic. The number of alleles varied from 2 to 12 (mean 4.53). Gene diversity ranged from 0.020 to 0.492 with a mean of 0.279. The polymorphic information content (PIC) values ranged from 0.371 to 0.019 (mean 0.225). Genetic distances in the whole cotton germplasm ranged from 0.451 to 0.052 (mean 0.270), demonstrating relatively wider genetic diversity range. Chinese-origin cotton germplasm showed the highest level of SSR polymorphisms (gene diversity = 0.268, PIC = 0.218), whereas American-origin revealed the highest mean genetic distance (0.274). Model-based Bayesian analysis clustered the whole cotton germplasm into three subpopulations, and the highest molecular variation ws revealed between subpopulations (4 %, P < 0.001). The SSRs revealed moderate level of genetic diversity at DNA level, identified three structured subpopulations, suggesting a potential use of these markers for genomewide association mapping studies and for identifying and conserving useful alleles in upland cotton germplasm.

3 illus, 4 tables, 54 ref

026665   YANG N, MU L, ZHAO B, WANG M, HU S, ZHAO B, CHEN Y, WU X (Yangzhou Univ, Jiangsu, People’s Republic of China, Email: xswu@yzu.edu.cn) : RNAi-mediated SLC7A11 knockdown inhibits melanogenesis-related genes expression in rabbit skin fibroblasts. J Genet 2018, 97(2), 463-8.

Solute carrier family 7 member 11 (SLC7A11) is a cystine/glutamate exchanger, also known as xCT, has been found to play an important role in pheomelanin synthesis. Adjusting the cystine content of cells to influence pheomelanin synthesis affects the proportion of total melanin, changing mammalian coat colour. In our previous study, we used RNA-seq to show that SLC7A11 was involved in coat colour regulation in Rex rabbits. However, the precise role of SLC7A11 in rabbit coat colour formation has not been investigated. To better understand the functions of SLC7A11 in rabbits, we used RNA interference (RNAi) to explore the effects of small interfering RNA (siRNA)-mediated downregulation of SLC7A11 gene expression on the expression of melanogenesis-related genes in rabbit skin fibroblasts (RAB-9). The effects of siRNA treatment were measured by quantitative real-time polymerase chain reaction and the efficiency of RNAi was calculated. The expression levels of melanogenesis-related genes, including MITF, MC1R, Agouti, CREB1 and SLC24A5 were detected at 24 h after RNAi transfection. The results showed that MITF, MC1R, SLC24A5, Agouti and CREB1 expression was significantly downregulated after SLC7A11 inhibition. This suggested that changes in SLC7A11 gene expression could directly affect the transcription of genes related to melanin production. This provides a scientific basis for further study of the role of SLC7A11 in the formation of coat colour.

3 illus, 3 tables, 23 ref

025536   AI D, CHENG S, CHANG H, YANG T, WANG G, YU C (China Univ of Mining and Technology, Beijing-100 083, China, Email: caihongyu2013@126.com) : Gene cloning, prokaryotic expression, and biochemical characterization of a soluble Trehalase in Helicoverpa armigera H?bner?(Lepidoptera: Noctuidae). J Insect Sci 2018, 18(3), 1-8.

Trehalase is an indispensable component of insect hemolymph that plays important role in energy metabolism and stress resistance. In this study, we cloned and expressed the gene encoding soluble trehalase (HaTreh-1) of Helicoverpa armigera (cotton bollworm) and characterized the enzyme. HaTreh-1 had a full-length open reading frame encoding a protein of 571 amino acids. Sequence comparison indicated that HaTreh-1 was similar to some known insect trehalases. Two essential active sites (D321 and E519) and three essential residues (R168, R221, and R286) were conserved in HaTreh-1. The recombinant trehalase was expressed in Escherichia coli and purified by nickel exchange chromatography. Molecular weight of the recombinant protein was about 71 kDa, and the optimum HaTreh-1 enzyme activity is at 55 °C with pH 6.0. Enzymatic assays showed a K_m value of 72.8 mmol/liter and a V_max value of 0.608 mmol/(liter·min). Inhibition assays in vitro indicated that castanospermine, a polyhydroxylated alkaloid, was an effective competitive inhibitor of trehalase with a K_i value of 6.7 μmol/liter. The inhibitor action of castanospermine was linked to its modification effect on trehalase structure. The circular dichroism spectrum showed that the percentage of α-helix increased under the presence of castanospermine. Results of our study will aid in developing effective trehalase inhibitors for controlling H. armigera in the future.

7 illus, 2 tables, 64 ref

025535   SUGANTHI M, ARVINTH S, CHANDRASHEKARA K N, RAJKUMAR R, SENTHILKUMAR P (Genetic Engineering Dep, SRM Univ, Kattankulathur-603 203, Email: sugenthi88@gmail.com) : Molecular characterization of bacterial biocontrol agents and their chitinase genes from tea soil. Indian J Exp Biol 2018, 56(6), 395-401.

In tea, bacterial biocontrol agents viz. Bacillus and Pseudomonas and an enzyme like chitinase from these bacterial strains are used to control tea pests and pathogens. However, literature on molecular identification of the same is quite scarce. In this study, Bacillus and Pseudomonas strains isolated from tea soil samples, were systematically identified by 16S rRNA sequencing. Molecular characterization of bacteria was carried out to identify the species of different level chitinase producing bacteria and diversity among them. Further, chitinase gene was characterized from these bacteria to understand the gene diversity among different bacterial chitinase that has potential application in controlling the plant pests and pathogens. Sequence analysis of 16S rRNA and chitinase gene sequences was made among thirteen Bacillus and five Pseudomonas species submitted in NCBI Genbank

5 illus, 1 table, 31 ref

025534   VERMA B, THAKUR Y, TRIPATHI M, PARDHI M, KHILARI R, PANDE R (Pt. Ravishankar Shukla Univ, Raipur- 492 010, Email: rama.pande11@gmail.com) : N-arylhydroxamic acids as a drug like molecule: A motif of binding mode with calf thymus DNA. Indian J Biochem Biophys 2018, 55(3), 215-21.

A drug-like molecule, which has a propensity of binding with DNA play a vital role in drug designing mechanism. In this paper, we tried to find out the DNA binding affinity of two derivatives of N-arylhydroxamic acids: (i) N-p-Chlorophenyl-2- methoxybenzohydroxamic Acid (Cl-2-MBHA) and (ii) N-p-Chlorophenyl-3-methoxybenzohydroxamic Acid (Cl-3-MBHA) with calf thymus DNA (ct-DNA) by applying techniques such as UV-visible spectroscopy, Fluorescence spectroscopy, and Viscometry measurements. The findings concluded with experimental techniques were verified with theoretical calculation using computer-based method, Molecular Docking. Absorption spectra revealed that both the hydroxamic acids derivatives bind to ct-DNA, among two, Cl-2-MBHA exhibits the higher value of binding affinity Kb (9.52 × 103±0.08 M-1). Fluorescence spectra showed that ct-DNA successfully quenches the emission spectra of N-arylhydroxamic acid. Ethidium bromide displacement method was used as a standard for analyzing the mode of binding. Both the hydroxamic acids were found to be groove binders. The Stern–Volmer Constant was found to be 2.05 × 10-2 ± 0.001 M-1 and 3.35 × 10-2 ± 0.002 M-1 for Cl-2-MBHA and Cl-3-MBHA respectively. Theoretical analysis molecular docking was done using Hex software for validating the experimental findings. Hence, it was observed that both experimental and computational method complimented the results and deduces groove binding as the mode of interaction.

9 illus, 1 table, 43 ref

025533   SANKARGANESH D, RAMCHANDRAN R, ASHOK R, SARVANAKUMAR V R, SUKRITA R, ARCHUNAN G, ACHIRAMAN S (Animal Science Dep, Bharathidasan Univ, Tiruchirappalli- 620 024, Email: achiramans@gmail.com) : Buck odor production in the cornual gland of the male goat, Capra hircus? validation with histoarchitecture, volatile and proteomic analysis. Indian J Biochem Biophys 2018, 55(3), 183-90.

In many animals, glandular secretions or pheromones that possess biological moieties contain messages encoded by the intrinsic smell. In male goats, the cornual gland (a sebaceous gland), may synthesize and excrete relevant chemical components that are responsible for the ‘buck effect’. To test this, cornual glands from freshly-slaughtered male goats (N=6) were subjected to histoarchitecture analysis, to infer about the structural alignment, to the GC–MS analysis for volatile compounds and to SDS–PAGE for protein profiling followed by MALDI-TOF to characterize specific protein bands. The gland possesses sebum, vacuoles and hair follicles inferring its capability to synthesize and extrude the scent. We found 14 volatiles in GC–MS analysis, in which 1-octadecanol might be a putative pheromone of buck odor. We identified seven different proteins in SDS-PAGE. Two proteins, 28 and 33 kDa, were highly matched with DNA mismatch repair protein and Abietadiene synthase, respectively, as inferred from MALDI-TOF. Conclusively, the volatiles identified in the cornual gland suggest that the structural microelements of the gland may synthesize (sebum and vacuoles) and release the key volatiles through the hair follicles. The volatile(s) thus produced in male goats either solely or synergistically may confer the buck odor.

5 illus, 1 table, 33 ref

025531   TOMY M J, SHARANYA C S, MAHAPATRA D K, SUREH K J, SABU A, HARIDAS M (Biotechnology and Microbiology Dep, Kannur Univ, Palayad– 670 661, Email: mharidasm@rediffmail.com) : In vitro assessment of selected benzoic acid derivatives as anti-inflammatory compounds. J Sci Ind Res 2018, 77(6), 330-6.

Inflammation is a major reason of pathophysiology of several diseases, causing a number of disarrays. The present research involved exploring inhibition of soybean 5-lipoxygenase (5-LOX) by various benzoic acid derivatives (eudesmic acid, veratric acid, cumic acid, and syringic acid) and their kinetics studies, comparing with vanillin as a standard reference. Isothermal titration calorimetry (ITC) was performed to determine kinetics of the biomolecular interactions. Further, molecular docking studies were performed using GLIDE module of Schrodinger software to determine the inter-molecular interactions between the ligand and the target. Result underlined that four of the investigated derivatives exhibited potent inhibition of 5-LOX in a competitive manner with IC50 values <60 µM. ITC study demonstrated that all derivatives bind effectively with a single domain of the target. Molecular docking studies revealed that all the four candidates displayed good interaction with the target. As found from Glide scores, veratric acid interacted stronger than other derivatives (GLIDE Score of -10.72 kcal/mol), forming two hydrogen bonds with residues GLN514 and GLN716, quite similar to the standard reference. ITC and GLIDE score analyses showed difference in the binding strength of the derivatives. This study would help in developing novel inhibitors, anti-inflammation as well as understanding mechanism(s) of ligand-enzyme interaction.

3 illus, 3 tables, 10 ref

025554   WAKEKAR R S, JADHAV P V, KALE P B, MOHARIL M P, NANDANWAR R S, MANE S S, DESHMUKH A G, MANJAYA J, DANI R G (Agricultural Biotechnology Dep, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Maharashtra - 444 104, Email: jpraveen26@yahoo.co.in) : Pollen dysfunction causes 'Floral Bud Distortion' in Indian soybean (Glycine max). Agric Res 2018, 7(1), 10-24.

Floral bud distortion (FBD) is a peculiar disorder limiting yield of soybean in central India. Investigations were conducted to understand tissue-specific cytological, biochemical and molecular alterations associated with FBD. The conspicuous morphological symptom seen was extended vegetative phase, which remained green even after R7 (beginning maturity) stage of growth and failure to produce pods leading to complete yield loss in affected plants. Reduced numbers of pollen grains as well as pollens with high percent of sterility were recorded in symptomatic plants. Although the stigma was found to be receptive in the symptomatic plants, pollens showed irregular shape and had a thicker exine wall. Carbohydrate (2.21 times), protein (2.38 times) and chlorophyll (2.17 times) contents were found to be significantly higher in symptomatic plants. Tissue-specific differentially expressed TDFs (transcript-derived fragments) were generated through cDNA-RAPD using 40 primers. Seven primers were found polymorphic and size ranged from 160 to 1100 bp. Re-amplified TDFs were sequenced, and they showed partial homology with characterized (ARF, XM_003529058) and uncharacterized (LOC 100815325) transcription factors. This indicates investigating differentially expressed genes are needed to understand the insights of molecular alterations associated with floral bud distortion.

7 illus, 5 tables, 61 ref

025553   KUMARI N, BATRA N G, SHARMA V (Bioscience and Biotechnology Dep, Banasthali Univ, Rajasthan- 304 022, Email: vinaysharma30@yahoo.co.uk) : Photosynthetic performance and drought-induced changes in activity of antioxidative enzymes in different varieties of Vigna radiata. Agric Res 2018, 7(1), 1-9.

In the present study, we analysed the photosynthetic performance in five varieties of Vigna radiata, viz. vars RMG 268, K-851, RMG 492, RMG 975 and Anand using chlorophyll fluorescence parameters. We observed that var. RMG 268 tended to reach highest effective quantum yield of PSII [$∆$F/F_m$\text{'}$], maximum apparent electron transport rate [ETR_max] and saturating photosynthetically active photon flux density [PPFDsat], followed by var. K-851. Thus as judged by its photosynthetic performance, ecophysiologically var. RMG 268 seems to be better adapted to the semi-arid environment of the state of Rajasthan, India. On the contrary, var. Anand was least adapted to its environment as indicated by lowest ETR_max, PPFDsat and $∆$F/F_m$\text{'}$ values. The activities of certain antioxidant enzymes of Vigna radiata in response to drought were also examined in var. RMG 268 and var. Anand. The increased activities of antioxidant enzymes, presumed to limit cellular damage, were observed in var. RMG 268. Cellular malondialdehyde content signal indicators of lipid peroxidation were much higher in var. Anand compared to var. RMG 268. These data revealed that var. RMG 268 had high resistance to environmental and drought conditions and thus substantiated our results obtained on the basis of plant performance.

5 illus, 2 tables, 36 ref

025552   TSIARA C G, NIKOLOPOULOS G K, DIMOU N L, PANTAVOU K G, BAGOS P G, MENSAH B, TALIAS M, BRALIOU G G, PARASKEVA D, BONOVAS S, ET AL. (Cyprus Univ, Cyprus, Email: gknikolopoulos@gmail.com) : Interleukin gene polymorphisms and susceptibility to HIV-1 infection: A meta-analysis. J Genet 2018, 97(1), 235-51.

Some subjects are repeatedly exposed to human immunodeficiency virus (HIV), yet they remain uninfected. This suggests the existence of host-resistance mechanisms. The current study synthesizes the evidence regarding the association between interleukin (IL) gene polymorphisms and HIV susceptibility. Medline, Scopus and the Web of Science databases were systematically searched, and a meta-analysis of case–control studies was conducted. Univariate and bivariate methods were used. The literature search identified 42 eligible studies involving 15,727 subjects. Evidence was obtained on eight single-nucleotide polymorphisms (SNPs): IL1A −889 C>T (rs1800587), IL1B +3953/4 C>T (rs1143634), IL4 −589/90 C>T (rs2243250), IL6 −174 G>C (rs1800795), IL10 −592 C>A (rs1800872), IL10−1082 A>G (rs1800896), IL12B −1188 A>C (rs3212227) and IL28B C>T (rs12979860). The IL1B +3953/4 C>T variant appears to increase the risk of HIV acquisition, under the assumption of a recessive genetic model (odds ratio (OR): 4.47, 95 % CI: 2.35–8.52). The AA homozygotes of the IL10 -592 C>A SNP had an increased, marginally nonsignificant, risk (OR: 1.39, 95 % CI: 0.97–2.01). It reached, however, significance in subanalyses (OR: 1.49, 95 % CI: 1.04–2.12). Finally, the well-studied hepatitis C virus (HCV) infection IL28B (rs12979860) CT/TT genotypes were associated with a 27 % decrease in HIV infection risk, especially in populations infected with HCV (OR: 0.73, 95 % CI: 0.57–0.95). Interleukin signalling is perhaps important in HIV infection and some interleukin genetic variants may affect the risk of HIV acquisition. Approaches targeting specific genes and genomewide association studies should be conducted to decipher the effect of these polymorphisms.

4 illus, 3 tables, 124 ref

025551   CALVELLO R, CIANCIULLI A, PANARO M A (Biosciences, Biotechnologies and Biopharmaceutics Dep, Bari Univ, Bari, Italy, Email: mariaantonietta.panaro@uniba.it) : Unusual structure and splicing pattern of the vertebrate mitochondrial solute carrier SLC25A3 gene. J Genet 2018, 97(1), 225-33.

The DNA sequence corresponding to the second exon of the SLC25A3 gene is duplicated in vertebrates. The second exon codes for the first transmembrane segment and parts of the immediately adjoining intermembrane and mitochondrial matrix segments. The two genomic exon 2 sequences are 84 % similar in zebrafish (slc25a3b gene), 70 % in chicken, 66 % in mouse and 67 % in human. The amino acid identity is 86 % in zebrafish, 77 % in chicken and 70 % in mouse and human. The two copies of exon 2 are separated by an intronic interval. Translation of both exon 2 sequences would alter the reading frame of the downstream sequence, generating a modified aa sequence which would soon be truncated by a stop codon. As a matter of fact the splicing machinery is tuned in such a way that in some species only one of the two copies is expressed and the other is spliced out, while in other species both copies are expressed but only one at a time, generating two alternative protein products.

5 illus, 1 table, 20 ref

025550   ZHANG J, CHEN J, MA T, GUO H, YANG B (Gastroenterology Dep, Jiangsu Taizhou People’s Hospital, Taizhou, People’s Republic of China, Email: yangbintaizhou@126.com) : Genetic variants of FOXP1 and FOXF1 are associated with the susceptibility of Oesophageal adenocarcinoma in Chinese population. J Genet 2018, 97(1), 213-8.

This study aimed to investigate whether the genetic variants of CRTC1, BARX1, FOXP1 and FOXF1 are associated with the development of oesophageal adenocarcinoma (OA) in Chinese population. A total of 744 OA patients and 1138 controls were included in this study. Here we genotyped four SNPs, rs10419226 of CRTC1, rs11789015 of BARX1, rs2687201 of FOXP1 and rs3111601 of FOXF1. The chi-square test was used to compare the genotype and allele frequencies between the patients and controls. The student’s t-test was used to compare FOXP1 expression in the tumour and the adjacent normal tissues. The relationship between genotypes of rs2687201 and FOXP1 expression was investigated by one-way analysis of variance test. Patients were found to have significantly higher frequency of allele A of rs2687201 and allele C of rs3111601 when compared with the controls (49.2 vs 43.4 %, P = 0.0008 for rs2687201; 29.1 vs 24.0 %, P = 0.0003 for rs3111601). There was a significantly higher expression level of FOXP1 in the tumour than in the adjacent normal tissue (0.0052 ± 0.0021 vs 0.0027 ± 0.0018, P < 0.001). Patients with genotype AA were found to have remarkably higher FOXP1 expression in the tumour than those with genotype CC (P = 0.01). To conclude, the varients of FOXP1 and FOXF1 genes are functionally associated with OA in Chinese population. With the identification of more susceptible loci, the combined effect of these markers may be helpful for the surveillance of OA.

1 illus, 3 tables, 28 ref

025549   ALI S G, DARVISHZADEH R, EBRAHIMI A, BIHAMTA M R (Plant Breeding and Biotechnology Dep, Urmia Univ, Iran, Email: r.darvishzadeh@urmia.ac.ir) : Identification of SSR and retrotransposon-based molecular markers linked to morphological characters in oily sunflower (Helianthus annuus L.) under natural and water-limited states. J Genet 2018, 97(1), 189-203.

Sunflower is an important source of edible oil. Drought is known as an important factor limiting the growth and productivity of field crops in most parts of the world. Agricultural biotechnology mainly aims at developing crops with higher tolerance to the challenging environmental conditions, such as drought. This study examined a number of morphological characters, along with relative water content (RWC) in 100 inbred sunflower lines. A 10 × 10 simple lattice design with two replications was employed to measure the mentioned parameters under natural and water-limited states during two successive years. In molecular trial, 30 simple sequence repeat (SSR) primer pairs, as well as 14 inter-retrotransposon amplified polymorphism (IRAP) and 14 retrotransposon-microsatellite amplified polymorphism (REMAP) primer combinations were used for DNA fingerprinting of the lines. Most of the examined characters had lower average values under water-limited than natural states. Maximum and minimum reductions were observed in the cases of yield and oil percentage, respectively. The broad-sense heritabilities for all the examined characters were 0.20–0.73 and 0.10–0.34 under natural and water-limited states, respectively. In the studied samples, 8.97% of the 435 possible locus pairs of the SSRs represented significant linkage disequilibrium (LD) levels. In the association analysis using SSR markers, 22 and 21 markers were identified (P ≤ 0.05) for the studied characters under natural and water-limited states, respectively. The corresponding values were 50 and 37 using retrotransposon-based molecular markers. Some detected markers were communal between the characters under water-limited and natural states. This was in line with the phenotypic correlations detected between the characters. Communal markers facilitate the simultaneous selection of several characters and can thus improve the efficacy of selection based on markers in the plant-breeding activities.

5 illus, 4 tables, 53 ref

025548   GOPALAKRISHNAN A, VINEESH N, ISMAIL S, MENON M, AKHILESH K V, JEENA N S, PAULTON M P, VIJAYAGOPAL P (ICAR-Central Marine Fisheries Research Institute, Ernakulam - 682 018, Email: vineeshnedumpally@gmail.com) : Mitochondrial signatures revealed panmixia in Lutjanus argentimaculatus (Forsskal 1775). J Genet 2018, 97(1), 179-87.

Mangrove red snapper, Lutjanus argentimaculatus is a commercially important fish. The genetic stock structure of L. argentimaculatus from Indian waters was identified using mitochondrial ATPase 6 and ATPase 8, and cytochrome b (Cytb) genes. A 842 bp region of ATPase 6/8 genes and 1105 bp region of Cytb gene were amplified in 120 samples from six different locations along the Indian coast and obtained 58 and 66 haplotypes, respectively. The high haplotype and low nucleotide diversity values along with mismatch distribution, Tajima’s D and Fu’s F_s analysis suggested a genetic bottleneck events or founder effect, with subsequent population expansion in L. argentimaculatus. Coefficient of genetic differentiation (F_ST) values was low and nonsignificant for both ATPase 6/8 gene and Cytb genes indicating low genetic differentiation in L. argentimaculatus which can be managed as a unit stock in Indian waters.

5 illus, 1 table, 48 ref

025547   LIANG T, JIA Y, ZHANG R, DU Q, CHANG Z (Henan Normal Univ, China, Email: 041019@htu.edu.cn) : Identification, molecular characterization and analysis of the expression pattern of soxf subgroup genes the yellow river carp, Cyprinus carpio. J Genet 2018, 97(1), 157-72.

Sox7, Sox17 and Sox18 are the members of the Sry-related high-mobility group box family (SoxF) of transcription factors. SoxF factors regulate endothelial cell fate as well as development and differentiation of blood cells and lymphatic vessels. There is very less information about the functions of these genes in fish. We obtained the full-length cDNA sequence of SoxF genes including Sox7, Sox17 and Sox18 in Cyprinus carpio, where Sox7 and Sox18 had two copies. The construction of a phylogenetic tree showed that these genes were homologous to the genes in other species. Chromosome synteny analysis indicated that the gene order of Sox7 and Sox18 was highly conserved in fish. However, immense change in genomic sequences around Sox17 had taken place. Numerous putative transcription factor binding sites were identified in the 5$\text{'}$ flanking regions of SoxF genes which may be involved in the regulation of the nervous system, vascular epidermal differentiation and embryonic development. The expression levels of SoxF genes were highest in gastrula, and was abundantly expressed in the adult brain. We investigated the expression levels of SoxF genes in five specific parts of the brain. The expression levels of Sox7 and Sox18 were highest in the mesencephalon, while the expression level of Sox17 was highest in the epencephalon. In carp, the expression patterns of SoxF genes indicated a potential function of these genes in neurogenesis and in vascular development. These results provide new information for further studies on the potential functions of SoxF genes in carp.

8 illus, 3 tables, 52 ref

025546   ZHENG H, LI H, TAN W, XU C, JIA L, WANG D, LI Z, SUN G, KANG X, YAN F, LIU X (Henan Agricultural Univ, Zhengzhou 450 002, People’s Republic of China, Email: xjliu2008@hotmail.com) : Oestrogen regulates the expression of cathepsin E-A-like gene through ERβ in liver of chicken (Gallus gallus). J Genet 2018, 97(1), 145-55.

The cathepsin E-A-like, also known as ‘similar to nothepsin’, is a new member of the aspartic protease family, which may take part in processing of egg yolk macromolecules, due to it was identified in the chicken egg-yolk. Previously, studies have suggested that the expression of cathepsin E-A-like increased gradually during sexual maturation of pullets, but the exact regulation mechanism is poorly understood. In this study, to gain insight into the function and regulation mechanism of the gene in egg-laying hen, we cloned the cathepsin E-A-like gene and evaluated its evolutionary origin by using both phylogenetic and syntenic methods. The mode of the gene expression regulation was analysed through stimulating juvenile hens with 17 β-estradiol and chicken embryo hepatocytes with 17 β-estradiol combined with oestrogen receptor antagonists including MPP, ICI 182,780 and tamoxifen. Our results showed that cathepsin E-A-like was an orthologoues gene with nothepsin, which is present in birds but not in mammals. The expression of cathepsin E-A-like significantly increased in a dose-dependent manner after the juvenile hens were treated with 17 β-estradiol (P < 0.05). Compared with the 17 β-estradiol treatment group, the expression of cathepsin E-A-like was not significantly changed when the hepatocytes were treated with 17 β-estradiol combined with MPP (P < 0.05). In contrast, compared with the 17 β-estradiol combined with MPP treatment group, the expression of cathepsin E-A-like was significantly downregulated when the hepatocytes were treated with 17 β-estradiol combined with tamoxifen or ICI 182,780 (P < 0.05). These results demonstrated that cathepsin E-A-like shared the same evolutionary origin with nothepsin. The expression of cathepsin E-A-like was regulated by oestrogen, and the regulative effect was predominantly mediated through ER-β in liver of chicken.

5 illus, 5 tables, 45 ref

025545   QIAN Y, ZHANG Y, WEI B, ZHANG M, YANG J, LENG C, GE Z, XU X, SUN M (Neurology Dep, The Second Affiliated Hospital of Soochow Univ, Suzhou- 215 004, People’s Republic of China, Email: xingshunxu@suda.edu.cn) : A novel Alu-mediated microdeletion in the RUNX2 gene in a Chinese patient with Cleidocranial dysplasia. J Genet 2018, 97(1), 137-43.

Cleidocranial dysplasia (CCD; OMIM: 119600) is a rare autosomal dominant skeletal dysplasia caused by RUNX2 gene mutations. The present study described a sporadic case with CCD. The clinical data of the proband with CCD was reported and genetic analysis was performed. The proband presented with typical CCD features including supernumerary impacted teeth, bilateral clavicle dysplasia, delayed closure of cranial sutures, and short stature; while his hands were normal. Sequencing analysis of the entire coding region of the RUNX2 gene revealed no pathogenic changes; however, copy-number analysis with the Affymetrix HD array found $~$500 kb genomic microdeletion. Real-time quantitative PCR validated this microdeletion in the 1–4 exons of the RUNX2 gene. The junction point of the breaking DNA was located in the directly oriented AluSz6 and AluSx repetitive elements, indicating that this microdeletion might be generated through an Alu–Alu mediated mechanism. In addition, this microdeletion existed in 21.8 % of the asymptomatic mother’s peripheral blood cells, demonstrating that the mosaicism was not associated with CCD phenotypes. In summary, a pathogenic microdeletion in the RUNX2 gene located on chromosome 6 was responsible for CCD.

3 illus, 23 ref

025544   HE Y, WANG X, WU X, ZHU Y, YANG D (Chinese Academy of Fishery Sciences, Hubei- 430 223, People’s Republic of China, Email: yangdg@yfi.ac.cn.) : Expression profiles of amh and foxl2 in Schizothorax kozlovi, and their response to temperature during the early developmental stage. J Genet 2018, 97(1), 127-36.

To elucidate the role of amh and foxl2 in sex differentiation of the teleost fish Schizothorax kozlovi, the full-length cDNAs were cloned from the mature testis and ovary by rapid amplification of cDNA ends (RACE), and their relative mRNA expression levels were determined by quantitative real-time polymerase chain reaction among tissues and temperature groups. The complete amh and foxl2 cDNAs of S. kozlovi were 2060 bp and 1750 bp, which encoded 568 and 306 amino acids, respectively. The amh were expressed only in gonads, while foxl2 was expressed in the gills, brain and gonads, both exhibiting relatively high tissue specificity. The amh exhibited sex-specific expression pattern in the gonads. No sex differences in the foxl2 expression were observed in the brain and gonads, but significant sex differences were found in the gills. No significant differences were found in the foxl2 expression, from the larval to the juvenile stage, and also between different temperature groups. However, significant differences were found in the expression levels of amh from the larval (12–63 days posthatching (dph)) to the juvenile stage (190 dph), and also among the 18$°$C and 10$°$C groups at 31 dph. This result suggested that amh plays an important role in male sex differentiation of S. kozlovi during the early developmental stage, but no similar effect was observed in foxl2.

6 illus, 1 table, 43 ref

025543   MEMON S, WANG L, LI G, LIU X, DENG W, XI D (Yunnan Agricultural Univ, China, Email: 2522935343@qq.com) : Isolation and characterization of the major histocompatibility complex DQA1 and DQA2 genes in Gayal (Bos frontalis). J Genet 2018, 97(1), 121-6.

The species origin of Yunnan gayal has been controversial since many years. However, few recent genetic studies have suggested that it has perhaps originated from the hybridization between male Bos frontalis and female B. taurus or B. indicus. Being an important semi-wild bovid species, this has also been listed under the red list of International Union of Conservation of Nature and Natural Resources. However, there is limited information available about the immunogenicity of this precarious species of Bos. Major histocompatibility complex (MHC) plays a pivotal role in immune response to infectious diseases in vertebrates. In the present study, we have investigated the structural and functional characteristics and possible duplication of the MHC-DQA genes in gayal (B. frontalis). Two full-length cDNA clones of the MHC-DQA genes were amplified and designated as Bofr-DQA1 (DQA*0101) and Bofr-DQA2 (DQA*2001) with GenBank accession numbers KT318732 and KT318733, respectively. A comparison between BofrDQA1, Bofr-DQA2 and to other MHC-DQA molecules from different animal species showed that nucleotide and encoded amino acid sequences of these two identified MHC-DQA genes have more similarity to alleles of specific DQA1 and DQA2 molecules from other Ruminantia species than to each other. The phylogenic investigation also demonstrated a large genetic distance between these two genes than to homologous from the other species. The large genetic distance between Bofr-DQA1 and Bofr-DQA2, and the presence of different bovine DQA putative motifs clarify that these sequences are nonallelic type. These results could suggest that duplication of the DQA genes has also occurred in gayal. The findings of the present study have strengthened our understanding to MHC diversity in rare ruminants and mutation of immunological functions, selective and evolutionary forces that affect MHC variation within and between species.

2 illus, 1 table, 36 ref

025542   DUBEY N K, MISHRA D K, IDRIS A, NIGAM D, SINGH P K, SAWANT S V (CSIR-National Botanical Research Institute, Lucknow - 226 001, Email: samirsawant@nbri.res.in) : Whitefly and aphid inducible promoters of Arabidopsis thaliana L.. J Genet 2018, 97(1), 109-19.

Lack of regulated expression and tissue specificity are the major drawbacks of plant and virus-derived constitutive promoters. A precise tissue or site-specific expression, facilitate regulated expression of proteins at the targeted time and site. Publically available microarray data on whitefly and aphid infested Arabidopsis thaliana L. was used to identify whitefly and aphid-inducible genes. The qRT-PCR further validated the inducible behaviour of these genes under artificial infestation. Promoter sequences of genes were retrieved from the Arabidopsis Information Resources database with their corresponding 5$\text{'}$UTR and cloned from the A. thaliana genome. Promoter reporter transcriptional fusions were developed with the beta-glucuronidase (GUS) gusA gene in a binary expression vector to validate the inducible behaviour of these promoters in eight independent transgenic Nicotiana tabaccum lines. Histochemical analysis of the reporter gene in T₂ transgenic tobacco lines confirmed promoter driven expression at the sites of aphid and whitefly infestation. The qRT-PCR and GUS expression analysis of transgenic lines revealed that abscisic acid largely influenced the expression of both aphid and whitefly inducible promoters. Further, whitefly-specific promoter respond to salicylic acid and jasmonic acid (JA), whereas aphid-specific promoters to JA and 1-aminocyclopropane carboxylic acid. The response of promoters to phytohormones correlated to the presence of corresponding conserved cis-regulatory elements.

6 illus, 60 ref

025541   PAUL P, MALAKAR A K, CHAKRABORTY S (Biotechnology Dep, Assam Univ, Silchar - 788 011, Email: supriyoch2008@gmail.com) : Codon usage vis-a-vis start and stop codon context analysis of three dicot species. J Genet 2018, 97(1), 97-107.

To understand the variation in genomic composition and its effect on codon usage, we performed the comparative analysis of codon usage and nucleotide usage in the genes of three dicots, Glycine max, Arabidopsis thaliana and Medicago truncatula. The dicot genes were found to be A/T rich and have predominantly A-ending and/or T-ending codons. GC3s directly mimic the usage pattern of global GC content. Relative synonymous codon usage analysis suggests that the high usage frequency of A/T over G/C mononucleotide containing codons in AT-rich dicot genome is due to compositional constraint as a factor of codon usage bias. Odds ratio analysis identified the dinucleotides TpG, TpC, GpA, CpA and CpT as over-represented, where, CpG and TpA as under-represented dinucleotides. The results of (NcExp−NcObs)/NcExp plot suggests that selection pressure other than mutation played a significant role in influencing the pattern of codon usage in these dicots. PR2 analysis revealed the significant role of selection pressure on codon usage. Analysis of varience on codon usage at start and stop site showed variation in codon selection in these sites. This study provides evidence that the dicot genes were subjected to compositional selection pressure.

5 illus, 3 tables, 55 ref

025540   LI J, ZHU J-L, LOU S-D, WANG P, ZHANG Y-S, WANG L, YIN R-C, ZHANG P-P (Anhui Univ, Anhui, China, Email: eveyin@163.com) : The complete mitochondrial genome of Coptotermes ?suzhouensis? (syn. Coptotermes formosanus) (Isoptera: Rhinotermitidae) and molecular phylogeny analysis. J Insect Sci 2018, 18(2), 26.

Coptotermes suzhouensis (Isoptera: Rhinotermitidae) is a significant subterranean termite pest of wooden structures and is widely distributed in southeastern China. The complete mitochondrial DNA sequence of C. suzhouensis was analyzed in this study. The mitogenome was a circular molecule of 15,764 bp in length, which contained 13 protein-coding genes (PCGs), 22 transfer RNA genes, two ribosomal RNA genes, and an A+T-rich region with a gene arrangement typical of Isoptera mitogenomes. All PCGs were initiated by ATN codons and terminated by complete termination codons (TAA), except COX2, ND5, and Cytb, which ended with an incomplete termination codon T. All tRNAs displayed a typical clover-leaf structure, except for tRNA^Ser(AGN), which did not contain the stemloop structure in the DHU arm. The A+T content (69.23 %) of the A+T-rich region (949 bp) was higher than that of the entire mitogenome (65.60 %), and two different sets of repeat units (A+B) were distributed in this region. Comparison of complete mitogenome sequences with those of Coptotermes formosanus indicated that the two taxa have very high genetic similarity. Forty-one representative termite species were used to construct phylogenetic trees by maximum likelihood, maximum parsimony, and Bayesian inference methods. The phylogenetic analyses also strongly supported (BPP, MLBP, and MPBP = 100 %) that all C. suzhouensis and C. formosanus samples gathered into one clade with genetic distances between 0.000 and 0.002. This study provides molecular evidence for a more robust phylogenetic position of C. suzhouensis and inferrs that C. suzhouensis was the synonymy of C. formosanus.

3 illus, 6 tables, 68 ref