030577   KURNIA R S, INDRAWATI A, MAYASARI N L P I, PRIADI A (Animal Disease and Veterinary Health Dep, Bogor Agricultural Univ (IPB), Indonesia, Email: ryanseptakurnia@gmail.com) : Molecular detection of genes encoding resistance to tetracycline and determination of plasmid-mediated resistance to quinolones in avian pathogenic Escherichia coli in Sukabumi, Indonesia. Vet World 2018, 11(11), 1581-6.

This study aimed to identify genes encoding resistance to tetracycline (TE) and plasmid-mediated resistance to quinolones in Escherichia coli isolates from clinical cases of avian colibacillosis in Sukabumi, Indonesia. A total of 25 E. coli archive isolates were collected in 2013-2017 from clinical cases of avian colibacillosis in Sukabumi, Indonesia. All isolates were tested for TE and quinolone resistance using the disk diffusion method. TE -resistant E. coli isolates were screened for the presence of tet(A) and tet(B) genes by single polymerase chain reaction (PCR). The qnr(A), qnr(B), and qnr(S) genes were detected by multiplex PCR in quinolone-resistant E. coli isolates. Result of this study shows that 19 of 25 (76 %) E. coli isolates are resistant to oxytetracycline and 64 % are resistant to TE; among them, 63.2 % and 31.5 % were positive tet(A) and tet(B), respectively. 13 out of 25 (52 %) are resistant to ciprofloxacin and 36 % are resistant to enrofloxacin either norfloxacin; among them, 61.6 % were positive qnr(A), 7.7 % were positive qnr(B), 23 % were positive qnr(S), and 7.7 % were positive both of qnr(A) and qnr(S). This study shows that a few pathogens of E. coli are resistant to TE and quinolone. The frequency of tet and qnr genes that are responsible for this resistance among avian pathogenic E. coli isolates in Sukabumi, Indonesia, was high.

2 illus, 5 tables, 26 ref

030583   NABIH A M, HUSSEIN H A, EL-WAKEEL S A, EL-RAZIK K A A, GOMAA A M (Animal Reproduction and AI Dep, National Research Centre, Dokki, Giza, Egypt, Email: hnyhussein2@yahoo.com) : Corynebacterium pseudotuberculosis mastitis in Egyptian dairy goats. Vet World 2018, 11(11), 1574-80.

Mastitis is an important threat facing goat milk industry and is the most common cause of culling. Efficient control of mastitis, based on efficient diagnosis of diseased animals, would improve milk production and reproductive efficiency. In subclinical mastitis (SCM), infected goats demonstrate neither udder symptoms nor abnormal milk. Corynebacterium pseudotuberculosis is an infectious causative agent of mastitis, mostly results as an extension of infection from the supramammary lymph node, and causes financial losses in the goat industry. This study aimed to estimate the prevalence of SCM with emphasis on C. pseudotuberculosis mastitis in Egyptian dairy goats in the selected farms. A total of 336 half milk samples were collected from 177 dairy goats of various crossbreeds, in mid-to-late lactation period, after clinical examination. All samples were examined bacteriologically, while somatic cell count (SCC) was determined only in 180 half milk samples of the clinically healthy milk samples. The isolated and identified C. pseudotuberculosis was examined for evidence of virulence genes (Phospholipase D [pld] and β-subunit of RNA polymerase [rpoB]) by polymerase chain reaction (PCR). The prevalence of clinical mastitis was 30.5 %, while 69.5 % of animals were apparently healthy and secreted milk was normal. Of those 180 clinically healthy half milk samples, 96 milk samples (53.33 %) showed SCM as detected by SCC (SCC ≥1,000,000 cells/ml). Coagulase-negative staphylococci were the most prevalent bacteria (41.96 %), then Staphylococcus aureus (37.5 %) and C. pseudotuberculosis (7.14 %). Molecular diagnosis of virulence genes revealed evidence of pld gene in 16 isolates (66.66 %), and rpoB gene in 6 samples (25 %) of the 24 bacteriologically isolated C. pseudotuberculosis. Here, we describe, for the 1^st time, isolation and identification of C. pseudotuberculosis from milk of does suffering from SCM in Egypt. C. pseudotuberculosis must be considered for routine bacteriological examination of milk from dairy goats, particularly herds with a history of caseous lymphadenitis. Pld gene-based PCR is more reliable than rpoB gene-based ones for the diagnosis of C. pseudotuberculosis.

3 illus, 6 tables, 61 ref

029205   WANG X, ZHANG Y, WANG D, G, JIN G, LI B, XU F, CHENG J (Institute of Animal Husbandry and Veterinary, Shanxi, 030 032, China) : Transcriptome analysis to characterize genes related to the marbling trait on meat quality of Aberdeen angus. Indian J Anim Health 2018, 57(2), 141-52.

We compared the intramuscular fat deposition in ribeye and subcutaneous fat muscle of Aberdeen Angus beef cattle. Eight samples of the best and worst muscle marbling grade of Angus cattle were collected from 120 Angus cattle after slaughtering. High-quality RNA was subsequently extracted from the beef ribeye and subcutaneous fat muscle and the transcriptome was sequenced using RNAseq technology. Approximately 37.22 GByte of clean sequencing reads were obtained in this study. A total of 15 differentially expressed genes (DEGs) and 16 DEGs were detected in the ribeye and subcutaneous fat muscle tissues respectively. In the beef ribeye, six genes were up-regulated (CYP1A1, RRAD, RETRG1, LOC787269, XIRP1, OTUD1) and nine genes down-regulated (Bos_taurus_newGene_1359, Bos_taurus_newGene_4387, KRT18, MYL9, ACTA2, PTGDS, DSTN, CD5L, MYL6B), whereas the subcutaneous fat muscle, characterized by good marble meat quality, had 14 genes up-regulated (CYP1A1, SLC25A13, S100A1, KLHL40, THRSP, MYLK4, TNFRSF12A, RRAD, HMOX1, AMPD3, DNAJA4, PTPN3, LOC782922, IFITM10) and two down-regulated genes (Bos_taurus_newGene_996, FST). Gene analysis revealed that the differentially expressed genes were mostly associated with myocyte differentiation, for example MYL9, ACTA2 and MYLK4 genes. Gene ontology (GO) was then used to perform a term enrichment analysis and significant GO terms included the vascular smooth muscle contraction pathway and arachidonic acid (AA) metabolism. Taken together, we observed that the DEGs play an important role in marble texture and intramuscular fat deposition of the different parts of Angus cattle. Hence, a strong basis for improving the meat quality of Aberdeen Angus was considered.

5 illus, 2 tables, 34 ref

029207   YUVARAJ N, ARUL V (Biotechnology Dep, Achariya Arts and Science Coll, Puducherry - 607 402, Email: yuvaraj.aasc@ achariya.org) : Sulfated polysaccharides of seagrass Halophila ovalis suppresses tumor necrosis factor-induced chemokine interleukin-8 secretion in HT-29 cell line. Indian J Pharmacol 2018, 50(6), 336-43.

The present study aims to investigate the anti‑oxidant and anti‑inflammatory properties of seagrass Halophila ovalis sulfated polysaccharide on HT‑29 cell line. Monosaccharides composition was identified using liquid chromatography‑mass spectrometry (LC‑MS) and the functional groups were analyzed using Fourier transform‑infrared (FT‑IR) spectroscopy. The antioxidant and anti‑inflammatory potential of crude extract and purified fractions was investigated in vitro. FT‑IR spectra revealed that the presence of different functional groups and the presence of galactose (82.4 %), xylose (7.6 %), fructose (4.0 %), mannose (2.0 %), fucose (1.6 %), glucose (1.2 %), and arabinose (1.0 %) was observed using LC‑MS. Ho‑SP and its fractions showed radical scavenging activity in hydroxyl, 2‑azinobis‑3‑ethylbenzothiazoline‑6‑sulfonic acid, and ferric reducing antioxidant power assay in a dose‑dependent manner. Noticeable anti‑inflammatory activity of purified fraction Ho FrIV (IC₅₀ = 43.85 μg/ml) was observed in a noncytotoxic range of concentrations and inhibited the tumor necrosis factor‑α (TNF‑α)‑induced interleukin‑8 (IL‑8) secretion (0.27 ng/ml) in HT‑29 cell line. Overall, the results presented in this study suggest that purified fraction Ho FrIV of Ho‑SP could suppress the TNF‑α‑induced secretion of IL‑8 in HT‑29 and thus could be used as a promising antioxidant and anti‑inflammatory candidate with potential benefits.

6 illus, 27 ref

029162   FENG J, LI H, XU X, LIN R, WANG F, XU S (Chinese Academy of Agricultural Sciences, Beijing 100193, People’s Republic of China, Email: jingfeng@ippcaas.cn.) : Genetic analysis and location of a resistance gene for Puccinia striiformis f. sp. tritici in wheat cultivar Zhengmai 7698. J Genet 2018, 97(4), 931–7.

Wheat stripe (yellow) rust, caused by Puccinia striiformis West. f. sp. tritici (Pst), is one of the most destructive diseases in many wheat-growing countries, especially in China, the largest stripe rust epidemic area in the world. Growing the resistant cultivars is an effective, economic and environmentally friendly way to control this disease. Wheat cultivar Zhengmai 7698 has shown a high-level resistance to wheat stripe rust. To elucidate its genetic characteristics and location of the resistance gene, Zhengmai 7698 was crossed with susceptible variety Taichung 29 to produce F1, F2 and BC1 progeny generations. The genetic analysis showed that the stripe rust resistance in Zhengmai 7698 to Pst predominant race CYR32 was controlled by a single-dominant gene, namedYrZM. Bulked segregant analysis and simple sequence repeat (SSR) markers were used to map the gene. Four SSR markers, Xbarc198, Xwmc179, Xwmc786 and Xwmc398 on chromosome 6BL were polymorphic between the parents and resistance, and susceptible bulks. A linkage genetic map was constructed using 212 F₂ plants in the sequential order of Xwmc398, Xwmc179, YrZM, Xbarc198, Xwmc786. As this gene is effective against predominant race CYR32, it is useful in combination with other resistance genes for developing new wheat cultivars with resistance to stripe rust.

3 illus, 5 tables, 37 ref

029164   GAO Z, TIAN G, WANG Y, LI Y, CAO Q, HAN M, SHI Z (Shijiazhuang Academy of Agricultural and Forestry Sciences, Shijiazhuang 050041, Hebei, People’s Republic of China, Email: zhenxiangao@163.com) : Allelic variation of high molecular weight glutenin subunits of bread wheat in Hebei province of China. J Genet 2018, 97(4), 905–10.

In common wheat (Triticum aestivum L.), allelic variations of Glu-1 loci have important influences on grain end-use quality. The allelic variations in high molecular weight glutenin subunits (HMW-GSs) were identified in 151 hexaploid wheat varieties representing a historical trend in the cultivars introduced or released in Hebei province of China from the years 1970s to 2010s. Thirteen distinct alleles were detected for Glu-1. At Glu-A1, Glu-B1 and Glu-D1, we found that the most frequent alleles were the 1 (43.0 %), 7+8 (64.9 %), 2+12 (74.8 %) alleles, respectively, in wheat varieties. Twenty two different HMW-GS compositions were observed in wheat. Twenty-five (16.6 %) genotypes possessed the combination of subunits 1, 7+8, 2+12, 25 (16.6 %) genotypes had subunit composition of 2*, 7+8, 2+12; 20 (13.2%) genotypes had subunit composition of null, 7+8, 2+12. The frequency of other subunit composition was less than 10 %. The Glu-1 quality score greater than or equal to 9 accounted for 20.6 % of the wheat varieties. The percentage of superior subunits (1 or 2* subunit at Glu-A1 locus; 7+8, 14+15 or 17+18 at Glu-B1 locus; 5+10 or 5+12 at Glu-D1 locus) was an upward trend over the last 40 years. The more different superior alleles correlated with good bread-making quality should be introduced for their usage in wheat improvement efforts.

1 illus, 3 tables, 42 ref

029158   BOZKAYA F, GURLER S (Genetics Dep, Harran Univ, 63200 Eyyubiye, Sanliurfa, Turkey, Email: farukbozkaya@yahoo.com.) : Sequence diversity of MHC class-II DRB gene in gazelles (Gazella subgutturosa) raised in Sanliurfa of Turkey. J Genet 2018, 97(4), 897–903.

In this study, we aimed to assess the sequence diversity of major histocompatibility complex (MHC) class-II DRB gene at exon 2 in gazelles raised in Sanliurfa Province of Turkey. Twenty DNA samples isolated from gazelles (Gazella subgutturosa) were used for sequencing exon 2 of MHC class-II DRB gene. Target region was amplified by polymerase chain reaction (PCR) and their products were directly sequenced. Nine of these 20 samples yielded unambiguously readable sequences. Three of the nine samples were homozygotes and each showed different sequences. A 262-bp sequence obtained from the three homozygote samples were submitted to GenBank (accession numbers: KC309405, KC309406 and KC309407). Using an allele specific PCR, we detected 10 additional haplotypes. Among 13 haplotypes, 45 nucleotide positions were polymorphic and most of the polymorphic nucleotide positions localized at peptide-binding region (PBR). Rates of nonsynonymous substitutions were significantly higher than synonymous substitutions at PBR. Phylogenetic analysis of the haplotypes showed that 10 haplotypes of the gazelles were clustered together while three were clustered with ovine and bovine haplotypes. The results indicated that at least 13 haplotypes at exon 2 of MHC class-II DRB gene were showing high degree of nucleotide and amino acid diversity, and certain haplotypes of G. subgutturosa were more similar to haplotypes from sheep or cattle than to each other. Rates of synonymous and nonsynonymous substitutions suggested that positive selection was a driving force for diversity at this locus in G. subgutturosa.

2 illus, 2 tables, 41 ref

029204   WAN L, DONG L, XIAO S, HAN Z, WANG X, WANG Z (Jimei Univ, Xiamen- 361 021, People’s Republic of China, Email: zywang@jmu.edu.cn.) : Genomewide association study for economic traits in the large yellow croaker with different numbers of extreme phenotypes. J Genet 2018, 97(4), 887–95.

A traditional genomewide association study (GWAS) detects genotype–phenotype associations by the vast number of genotyped individuals. This method requires large-scale samples and considerable sequencing costs. Extreme phenotypic sampling proposes make GWAS more cost-efficient and are applied more widely. With extreme phenotypic sampling, we performed a GWAS for n-3 highly unsaturated fatty acids (HUFA) and eviscerated weight (EW) traits in the large yellow croaker population. Of the 32,249 and 29,748 detected SNPs for the two traits, three candidate regions were found in each trait. Three candidate regions associated with HUFA were known near genes on chromosomes 4 and 11, and three candidate regions were on chromosome 6, and 15 for the EW trait. By combing through our GWAS results and the biological functional analysis of the genes, we suggest that the FABP, DGAT, ATP8B1, FAF2 and CERS2 genes, as well as the IGF2, BORA, CYP1A1, GRTP1 and HOX genes are promising candidate genes for n-3 HUFA and EW, respectively, in the large yellow croaker. Moreover, compared with the different numbers of the extreme phenotypic sampling, we conclude that 60 % of the extreme phenotypic subsample can obtain a similar result as GWAS with whole phenotypes. Thus, extreme phenotypic sampling could save 40 % of the cost for genotyping and DNA extraction without loss of the candidate regions and functional genes. Our study may provide a basis for further genomic breeding and a reference for others who want to perform GWAS with extreme phenotypes.

2 illus, 6 tables, 41 ref

029167   GUNDA P, MANNE M, ADEEL S S, KONDAREDDY R K R, TIRUNILAI P (Genetics Dep, Osmania Univ, Hyderabad - 500 007, Email: padmatirunilai@gmail.com) : Detection of c.139G>A (D47N) mutation in GJA8 gene in an extended family with inheritance of autosomal dominant zonular cataract without pulverulent opacities by exome sequencing. J Genet 2018, 97(4), 879–85.

The aim of this study was to identify the gene causing bilateral autosomal dominant zonular congenital cataract (ADZCC) without pulverulent opacities in an extended Muslim family by exome sequencing and subsequent analysis. An extended family of 37 members (14 affected and 23 unaffected) who belong to different nuclear families was screened for causative gene. Proband and her unaffected son were screened for causative variant by exome sequencing followed by Sanger sequencing of the proband’s entire nuclear family. The rest of the members were further screened for variants detected, by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS PCR). Review of exome sequencing data of the proband and her unaffected son for 40 known genes causing congenital nonsyndromic cataracts revealed two variants, namely c.139G > A (p.Asp47Asn; D47N) in the GJA8 gene and c.2036C > T in the FYCO1 gene to be potentially pathogenic. Further, validation of these two variants in the entire family showed cosegregation of c.139G > A variant in GJA8 with ADZCC without pulverulent opacities. Variation of c.2036C > T in FYCO1 was not associated with disease in the family. The mutation c.139G > A in the GJA8 gene detected in the present study was also previously reported in Caucasian and Chinese families but with different phenotypes, i.e. nuclear and nuclear pulverulent cataracts. Thus, the mutation c.139G > A in GJA8 appears to exhibit marked interfamilial phenotypic variability.

2 illus, 4 tables, 24 ref

029206   XIA X, WAN R, HUO W, ZHANG L, XIA X, CHANG Z (Henan Normal Univ, Xinxiang 453007, Henan, People’s Republic of China, Email: 13837331530@163.com) : Molecular cloning and mRNA expression pattern of Sox4 in Misgurnus anguillicaudatus. J Genet 2018, 97(4), 869–77.

Sox4, a member of the SoxC subfamily which of the Sox family, plays important roles in the development of the vertebrate gonad and nervous system.We have cloned a Sox4 homologue from the brain of Misgurnus anguillicaudatus using homologous cloning and rapid amplification of cDNA ends. We named the cloned gene as MaSox4. The full-length cDNA was 2122 bp, containing a 718 bp 5 -untranslated region and a 267 bp 3 -untranslated region. The open-reading frame of the cloned gene encoded 378 amino acids and contained a characteristic HMG-box DNA-binding domain with the specific motif (RPMNAFMVW). Phylogenetic analysis indicated that MaSox4 is highly homologous to Sox4 in different species. Protein sequence analysis showed that MaSox4 is a nonsecretory hydrophilic protein. Quantitative real-time reverse transcription polymerase chain reaction and in situ hybridization assay revealed that MaSox4 was ubiquitously expressed during embryogenesis and is present in various adult tissues, especially in the central nervous system. Our study suggests that MaSox4 is highly conserved among vertebrates’ evolution and might be involved in developmental processes such as embryogenesis, neurogenesis and gonad development.

5 illus, 1 table, 57 ref

029175   KAZEMI H, NAJAFI M, GHASEMIAN E, RAHIMI-MIANJI G, PIRSARAEI Z A (Animal Breeding and Genetics, SANRU, Sari 4818168984, Iran, Email: mojtaba_najafy@yahoo.com.) : Polymorphism detection of promoter region of IFN-γ and IL-2 genes and their association with productive traits in Mazandaran native breeder fowls. J Genet 2018, 97(4), 843–51.

To identify polymorphism in interferon gamma (IFN-γ) and interleukin-2 (IL-2) genes, blood samples were collected from 380 breeder hens of the Mazandaran native fowls breeding station. DNA extraction was performed through a modified saltingout method and fragments of 670 and 659 bp from the promoter regions of IFN-γ and IL-2 genes were amplified by using specific primers, respectively. Following genotyping in the IFN-γ gene using the Tsp509I restriction enzyme, two alleles of A and G with the frequencies of 0.55 and 0.45 and three genotypes of AA, AG and GG were observed with the frequencies of 0.32, 0.46 and 0.22, respectively. For the IL-2 gene, two alleles of A and G were also detected using the MnlI restriction enzyme with the frequencies of 0.58 and 0.42 and three genotypes of AA, AG and GG with the frequencies of 0.33, 0.50 and 0.17, respectively. Statistical analysis revealed significant associations between IL-2 gene single-nucleotide polymorphism and productive traits including the average egg weight (EW) at 345–375 days of age, egg number (EN) at 345–375 days of age and body weight (BW) at 8 weeks of age traits (P < 0.05). Further, in a mean comparison analysis, there were also significant differences between different genotypes of the IL-2 gene in average EW at 28 and 30 weeks of age, in which AG genotypes showed higher performance. Additionally, for the IFN-γ gene, a significant difference was found between the genotypes in average EW at 28 weeks of age trait. Therefore, it can be concluded that the above-mentioned polymorphisms could be considered as the pivotal genetic makers to improve Mazandaran native fowl breeding programmes to achieve the optimum performance in productive traits more efficiently.

1 illus, 5 tables, 70 ref

029155   ANTONY A C, ALONE P V (Homi Bhabha National Institute, Mumbai - 400 094, Email: pankaj@niser.ac.in.) : Fidelity of HIS4 start codon selection influences 3-amino-1,2,4-triazole sensitivity in GTPase activating protein function defective eIF5. J Genet 2018, 97(4), 953–64.

The eIF5 protein plays an important role in the fidelity of AUG start codon selection. However, the hyper GTPase eIF5^G31R mutation in yeast causes preferential utilization of UUG as initiation codon and is termed as suppressor of initiation codon (Sui⁻) phenotype. The eIF5^G31R mutant recognizes upUUG initiation codon from the 5' regulatory leader region of GCN4 transcript and dominantly represses GCN4 expression thereby conferring sensitivity to 3-amino-1,2,4-triazole (3AT)-induced starvation. The 3AT sensitivity was rescued by supplementing HIS4^UUG allele. The eIF5^G31R mutant has a better efficiency of UUG codon recognition from the HIS4^UUG allele under starvation conditions. Moreover, the expression of HIS4^UUG allele was significantly lower than the critical level causing additional derepression of GCN4 expression in eIF5^G31R mutant to rescue its 3AT sensitivity. The overexpression of eIF1 improved expression of HIS4^AUG allele and GCN4 transcript causing 3AT resistance, whereas overexpression of eIF1 resulted in diminished UUG codon recognition of HIS4^UUG allele causing 3AT sensitivity, despite having higher GCN4 expression. This paper reports the critical role of HIS4 expression necessary in response to 3AT-induced starvation in the eIF5^G31R mutant which is ostensibly not a direct target of 3AT inhibition.

5 illus, 2 tables, 26 ref

029186   MENON R, SARAO N K, PATHAK M (Vegetable Science Dep, Punjab Agricultural Univ, Ludhiana - 141 004, Punjab, Email: navraj-soab@pau.edu) : In planta Agrobacterium-mediated genetic transformation in okra {Abelmoschus esculentus (L.) Moench}. Appl Biol Res 2018, 20(3), 221-7.

In Planta methods have been developed in several plant species to overcome the problems associated with their tissue culture-based transformation. In present study, a method for genetic transformation in okra (Abelmoschus esculentus) was established using in planta Agrobacterium-mediated genetic transformation. The embryos of imbibed seeds pricked from meristematic region of plumule were used as explants for transformation. Agrobacterium tumefaciens strain EHA105 was carrying cry1Ac gene against fruit and shoot borer, CaMV 35S as promoter and nptII as plant selectable marker gene. The putative transgenic plants were confirmed by amplifying transgene through polymerase chain reaction.

7 illus, 1 table, 22 ref

029198   SRINARANG P, NAGANVONPANIT K, PRADIT W, BUDDACHAT K, SIENGDEE P, SOONTORNVIPART K, CHOMDEJ S (Biology Dep, Chiang Mai Univ, Chiang Mai 50200, Thailand, Email: siriwadee.submission@gmail.com) : Dystroglycan 1: A new candidate gene for patellar luxation in chihuahua dogs. Vet World 2018, 11(9), 1277-84.

The objective of this study was to uncover new candidate genes related to patellar luxation (PL) in dogs to select for those with low susceptibility for breeding purposes. The inter simple sequence repeat (ISSR) technique was performed to construct DNA fingerprints of 61 Chihuahua dogs with PL and 30 healthy Chihuahua dogs. DNA polymorphisms were detected by comparing the sequences between the affected and unaffected dogs, using the pairwise alignments in MultAlin. Genotyping was performed using allele-specific polymerase chain reaction (AS-PCR). The association analysis of ISSR DNA fingerprints and genotypes or phenotypes was performed using the Chi-square (χ²) model and generalized linear model (GLM), respectively. Two single nucleotide polymorphisms (SNPs), namely SNP1UBC811 (g.91175C>G) and SNP2UBC811 (g.92259T>C), were found in the intron of the Dystroglycan 1 (DAG1) gene, which was obtained using the PL-related marker UBC811 primer (p=0.02), and genotyped by AS-PCR. When investigated using the GLM, g.91175C>G had a significant association with PL (p=0.0424), whereas g.92259T>C did not have such an association (p=0.0959). DAG1 might be one of the genes related to PL in Chihuahuas and could aid the process of marker-assisted selection in genetic breeding for Chihuahua dogs without PL.

5 illus, 6 tables, 19 ref

029170   HIDAYATI D N, UNTARI T, WIBOWO M H, AKIYAMA K, ASMARA W (Microbiology Dep, Gadjah Mada Univ, Daerah Istimewa Yogyakarta 55281, Indonesia, Email: wied_as@ugm.ac.id) : Cloning and sequencing gB, gD, and gM genes to perform the genetic variability of bovine herpesvirus-1 from Indonesia. Vet World 2018, 11(9), 1255-61.

Previous research has shown thatbovine herpesvirus-1 (BHV-1) in Indonesia was closely related to subtype-1 based on glycoprotein D genes. This study aimed to analyze the genetic variability of the BHV-1 isolated from the recent case in Indonesia not only based on gD but also other genes such as gB and gM and to study the homology and similarity of the sample to other BHV-1 isolated in other countries or regions. Samples were drawn from the tracheal organ in recent field case and prepared for DNA extraction. The gB, gD, and gM were amplified usingnested polymerase chain reaction (nPCR) with our specifically designed primer pair and based on the specified bands of 350 bp gB, 325 bp gD, and 734 bp gM confirmed as BHV-1. The PCR product was ligated into pGEM-T and transformed into competent Escherichia coli. The purified plasmid was subsequently sequenced. The virus sample isolated from the recent field case of infectious bovine rhinotracheitis (IBR) from Indonesia showed variability based on the gB, gD, and gM sequences. However, all of the genes had high similarity (98-100 %) to BHV-1.2.

1 illus, 4 tables, 43 ref

029185   MANSOUR K A, NASER H H, HUSSAIN M H (Internal and Preventive Veterinary Medicine Dep, Al-Qadisiyah Univ, Iraq, Email: muthanna.hussain@qu.edu.iq) : Clinical, molecular detection and phylogenetic analysis study of local foot-and-mouth disease virus in Al-Qadisiyah province of Iraq. Vet World 2018, 11(9), 1210-3.

This study was directed during an outbreak of suspected foot-and-mouth disease (FMD) in cattle in Al-Qadisiyah province, Iraq 2016. The disease has made a huge economic loss in livestock. It was suspected that the vaccination has failed to protect the animals from the infection because of the difference in the strains. Consequently, we designed the study to make the diagnosis and detect the strain of the causative virus. The extraction of the DNA was done on 73 samples and  Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) was used in the detection of FMD virus (FMDV) for primary diagnosis, and serotype-specific diagnosis was done with universal primer sets 1F/1R, A-1C612, and O-ARS4 with the expected band of 329, 865, and 1301 bp, respectively. Universal primer pair 1F/1R detected FMD in 55 of 73 (75.3 %); of these, 37 (67.3 %) were females and 18 (32.7 %) were males, with high significance (p < 0.01) between males and females in the PCR positivity ratio. The tested samples with positive universal primer were amplified with specific primers A-IC612 with no reaction for serotype O-ARS4. The products of RT-PCR were sent for RNA sequencing, and the results were 100 % positive to serotype A which means that it is the predominant type in Iraq. It may help in the importing or production of the vaccine to make a preventive plan for the disease. The virus of FMD is contagious and dangerous due to its role in the huge economic loses. The detection of this virus is widely explained in lots of articles, but it is more specific and sensitive in RT-PCR and sequencing. Consequently, the authorities responsible for importing and/or production vaccines have to avoid the importing of other serotypes because it will be losing money and more outbreaks will explode.

3 illus, 3 tables, 19 ref

029196   SALIM S A (Al-Furat Al-Awsat Technical Univ, Iraq, Email: dr.sihamabdalrazzaq@yahoo.com) : In vitro induction of callus from different explants of Terminalia arjuna (Roxb.) Wight and Arn. and detection of its active secondary metabolites using GC-MS analysis. Plant Arch 2018, 18(2), 2519-27.

Terminalis arjuna is one of the most important medicinal plants that used in folk medicine in different countries. The current study was achieved to identify the active phytochemical compounds in the callus using GC-MS technique. In order to induce callus, the leaves, petioles and internodes used as explants were taken from 10 years old trees and cultured in MS medium supplemented with different concentrations(0.1 - 4.0 mg.l^-1) of auxin 2, 4-D. Callus was extracted with hexane and analyzed with GC-MS for detection of its phytochemical components. The results displayed that internodal explants were the best for callus induction and proliferation under the concentration 3.0 mg.l^-1 of 2, 4-D. The main compounds obtained from callus extract were Benzo[h] quinolone, 2, 4-dimethyl, ¹H-Indole,5-methyl-2-phenyl-, 1, 2, 4-Oxadiazole,3- (1, 3-benzodioxol-5-yl) -5- [(4- iodo-1H-pyrazol-1-yl) methyl]-, alpha.-Amyrin; Urs-12-en-24-oic acid,3-oxo-, methyl ester, (+)-. This study focused on the detection of phytochemical active compounds in vitro from callus cultures and it is useful for the production of these compounds in the future in large quantities for pharmaceutical and commercial purposes.

4 illus, 2 tables, 36 ref

029173   JASSIM E H (Baghdad Univ, Iraq, Email: emadJassim081@gmail.com) : Micro propagation of Carissa macrocarpa L. plant in vitro. Plant Arch 2018, 18(2), 2412-6.

Carissa macrocarpa L. is an important medicinal plant are used for various medical treatments. An efficient micro propagation protocol was developed for using nodal segment as the explants. After sterilization of explants, no contaminations were recorded. Explants were cultured on MS medium supplemented with various concentrations of BA and 0.5 mg/l NAA. The higher average of shoots Number 4.30 per explant and mean length of shoots 1.25 cm and maximum fresh and dry weight of shoots induction 442.0, 38.4 mg respectively in nutrient medium supplemented with 1 mg/l of BA + 0.5 mg/l of NAA. Results of multiplication stage showed that the best multiplication was on the medium supplemented with 3mg/l of BA + 0.5 mg/l of GA₃ , induced large number of shoots reached 5.70 and the highest length of multiple shoots 1.64 cm with maximum fresh and dry weight of shoots 539.7, 51.20 mg, respectively. Rooting was readily achieved upon transferring the shoots on to halfstrength MS medium the highest average number of roots was 2.80 while the highest average of root length was 1.50 cm at the concentration of 4.5 mg/l of NAA. Rooted plants were successfully acclimatized to greenhouse conditions.

3 illus, 3 tables, 22 ref

029172   ISSA F H, NAJIALHASNAWI A, SABAH S S (Al Muthanna Univ, Samawa, 66001, Iraq, Email: falah70hasan@gmail.com) : Influence of gamma radiation on in vitro growth microtubersation and hormonal content of some potato (Solanum tuberosum L.) cultivars. Plant Arch 2018, 18(2), 2317-23.

Gamma irradiation, which is a mutagen used in vegetable and crop plant breeding is expected to induce novel mutations and alteration agents in developing a genetic variety. The objective of this study was to assess the vegetative growth, plant hormonal parameters and yield of three cultivars of potato (Solanum tuberosum L. cv Arnova, Lizeta and Safari) which were irradiated with different doses of gamma rays (0, 2.5, 5, 10, 15 and 20 Gy) respectively, using in vitro techniques. The results of the experiment showed that Arnova was significantly superior in plant height, number of leaves, microtubers fresh weight, IAA and GA hormones content. Furthermore, the microtubers reached (7.25 cm, 10.5 leaf, 0.4 g, 0.673 µg.g^-1 and 1.3 µg.g^-1 in microtubers) respectively per plant. The total yield per plant and the percentage representing the number of tubers with the size 1.5-2 cm, reached (9.3, 26.56, 6.34 % and 47 %) respectively. However, Lizeta significantly differed from Arnova and Safari in the total yield which reached (0.72, 0.55, and 0.62 g) respectively. The gamma radiation (2.5 Gy) increased the average plant height significantly, and the number of shoots and tubers were determined to have a significant increment ratio (1.86 %, 19.04 %) which was the average compared to the control treatment. The interaction of the Arnova and 2.5 Gy was significantly superior in the total yield reaching (0.9 g per explant), while Lizeta at 2.5 Gy significantly superior in a number of tubers reaching 2.8 tubers per explant. From the previous results, the best growth was observed in the total yield of microtubers at the dose 2.5 Gy in Arnova or Lizeta cultivar.

4 illus, 10 tables, 22 ref

029154   AL-ZAYDI A M J, MITAB H H (Al-Muthanna Univ, Iraq, Email: abiralward420@gmail.com) : Real time PCR detection multidrug-resistance Mycobacterium tuberculosis in Al-Sammawa City. Plant Arch 2018, 18(2), 2102-8.

Multidrug-resistant tuberculosis (MDR-TB) is a global problem that many countries are challenged. Rapid and accurate detection of MDR-TB is critical for appropriate treatment and controlling of TB. The aims of study at using repaid real time PCR based assay to detection multidrug-resistant tuberculosis from culture samples, mutation in rpoB and katG genes Mycobacterium tuberculosis that responsible resistance to rifampicin and isoniazid, respectively. A total of 30 M. tuberculosis isolates from cases with diagnosed TB By GeneXpert, AFB and Culture on L.J media. DNA extraction from bacteria colonies. Resistant isolates were tested for characterization of mutations in the rpoB, katG genes by Real Time PCR. The test showed positive results for resistance genes (20 %, 10 %, respectively) as well as note that the values of Ct for this test ranged from (12 - 38.25) and the melting points of the genes were between (85-88.5 ºC). Real time PCR results identified three mutations of MDR (rifampicin and isoniazide) resistance genes, whereas there was one MDR mutation of molecular diagnostic results with the GeneXpert MTB / RIF test for rifampicin. When comparing the results of the Real time PCR and GeneXpert tests at the level of the genetic mutation with rifampicin, the real time PCR test showed four resistance mutations for the rpoB gene for both new cases and relapse tuberculosis as well as one rpoB mutant for under treatment patient. Both molecular tests have agreed to identify one rpoB mutant in the case of failure TB treatment.

5 illus, 2 tables, 19 ref

029180   KOUADRIA R, BOUZOUINA M, LOTMANI B (Ain Shams Univ, Egypt, Email: rabia-rk@hotmail.com) : Isolation and identification of endophytic fungi on plants growing in salt environments, using ITS and 18S molecular methods. Plant Arch 2018, 18(2), 1835-40.

Endophytic fungi seem to play particle roles for the survival of plants inhabiting stressful habitats. This study focused on the identification of fungal endophytic community, associated with roots plants growing in salt environments by sequencing ITS and 18S rDNA regions. Based on the culture characteristics and growth morphology of colonies, 6 fungi species obtained from roots were collected from three plots with different salinities (< 4 dS/m, 4 to 8 dS/m and 8 to 16 dS/m). ITS sequences and 18S rDNA gene were compared with those available in the GenBank databases, to identify the following species: Alternaria chlamydospora and Chaetomium coarctatum (salinity < 4 dS/m), Alternaria chlamydospora, Embellisia phragmospora, Phoma betae, Fusarium equseti, Chaetomium coarctatum and Fusarium graminearum (4 to 8 dS/m); and Chaetomium coarctatum (8 to 16 dS/m). Results indicate that Chaetomium coarctatum was considered as the most dominant fungus in studied plots. The fungal root endophytic community in natural vegetation under abiotic salt stress opens up possibilities for further investigations on the role of endophytes.

2 illus, 2 tables, 35 ref

029171   IBRAHIM S K, SALEH N M (Industrial Research and Development Dep, Ministry of Science and Technology, Iraq) : Study of the optimal nutritional conditions for the production of Kojic acid from the local isolation of the Aspergillus oryzae. Plant Arch 2018, 18(2), 1801-8.

The cultivation conditions for respective ability to produce kojic acid from local Aspergillus oryzae isolates were studied. The obtained results showed that the best cultivation medium to produce kojic acid from local Aspergillus oryzae isolates included a modified molasses medium with a concentration of 5 % reduced sugars, 0.5 % sodium nitrate, 4.5 pH, fertilization (3.2 × 10⁷ spore/ml), 0.1 % potassium dihydrogen phosphate and 0.05 % of magnesium hydroxide sulfate.

10 illus, 2 tables, 32 ref

029208   ZIEDAN E S H, ATTALLAH A G, ABD-EL-AAL S K, SAHAB A F (Plant Pathology Dep, National Research Centre, Egypt, Email: ziedanehe@yahoo.com) : Molecular identification and pathogenic potential of Botrytis cinerea isolates causing fruit blight of cucumber under protective greenhouse in Egypt. Plant Arch 2018, 18(2), 1563-9.

Survey of fruits blight (grey mould disease) on fruits of cucumber plants cultivations during growing winter season 2016 - 2017 in protective plastic houses of some Governorates in Egypt indicated that fruits blight is the most epidemic foliar disease of cucumber. Botrytis cinerea was the common fungi of diseased fruits at El-Beheira followed by El- Gharbeia Governorates respectively. Pathogenicity test of fungal isolates revealed that Botrytis cinerea isolates (No. 5 and 2) respectively which isolated from El- Gharbeia caused highly incidence of fruit rot of cucumber followed by isolate (No.7) from E Beheira Governorate. Fungal isolates were identified as Botrytis cinerea according to cultural, morphological and molecular characterizations based on sequencing of internal transcribed space (ITS). Some nucleotides sequences were registries of fungi in Gene Bank under accession number of MF996362, MF996363, MF996364, MF996365, MF996366, MF996367 and MF996368.

4 illus, 2 tables, 42 ref

029153   AL-AMIRY A H A (Horticulture Dep, Univ of Basrah, Iraq, Email: aqeelhadi6@gmail.com) : Comparison between three different methods for isolation genomic dna from dry date palm leaves without liquid nitrogen. Plant Arch 2018, 18(1), 1011-4.

Three different methods for isolation genomic DNA from Date palm leaves is described. The leaves was dried and ground before DNA extraction by CTAB development method [100 mM Tris- HCl (pH 8.0), 20 mM EDTA, 4M NaCl, 6 % CTAB, 2 % Polyvinylpyrrolidone, 0.2 % B-mercaptoethanol and 15 µl of proteinase K] and compared with two other methods, the results shows the developing method gives highly quality of DNA (A260/280 = 1.6 to 1.9) in all three Date palm cultivars under tests. This study found the development method is very suitable to DNA extraction from dried leaves without liquid nitrogen.

3 illus,16 ref

029197   SHARAN S, MUKHOPADHYAY K, SARIN N B (Jawaharlal Nehru Univ, New Delhi-110 067, Email: neerasarin@rediffmail.com) : Establishment of in vitro callus cultures and comparative phytochemical study of in vitro callus cultures and field grown plants of Ocimum tenuiflorum L.. Plant Arch 2018, 18(2), 1251-7.

The present study deals with callus induction from explants along with the comparative analysis of content of eugenol and ursolic acid present in aerial parts of field grown plants and in vitro callus cultures of Ocimum tenuiflorum L. Leaf explants exhibited higher callus induction, followed by stem explants when inoculated onto MS medium containing α-naphthalene acetic acid (0.25 mg/l) and 6-benzyl amino purine (0.5 mg/l). The content of eugenol and ursolic acid was found to be higher in the aerial parts (leaf and stem) of the field grown plants in comparison with that of in vitro callus cultures.

6 illus, 1 table, 23 ref

029156   ARA H, KHAN J A (Biosciences Dep, Jamia Millia Islamia (Central Univ), New Delhi -110 025, Email: jkhan1@jmi.ac.in) : Biotechnological approaches for engineering resistance against viruses in plants. Plant Arch 2018, 18(2), 1191-208.

Plant viruses, generally cause diseases on wide varieties of agronomically important crop species and hence bring a serious threat to the food security and global economy as well. Introduction of crop immunity against viruses has been a major challenging task. The infection of viruses is difficult to control as they are strict intracellular pathogens. Moreover, their chemical control is not usually advised in practice for long as it not only affects the environment but also is supposed to lessen quality of the crops. Using biotechnological approaches for genetic engineering and molecular biology, induction of defence mechanism against viruses in crop plants is considered as one of the powerful alternative strategy. Over the past few decades tremendous progresses have been made to unravel our knowledge in the area of plant immunity against viruses. Present review describes existing strategies for developing resistance in plants against viruses. Role of protein- and nucleic acid- mediated resistance to generate pathogen-derived resistance is described. In addition, importance of the genes of host plant origin, plant’s hormone and ribosome inactivating proteins for virus resistance is also discussed.

3 tables, 128 ref

029179   KORUKAPPILLIL P K, SHERIF R, SARAYU L, SIMI (Microbiology Dep, Azeezia Institute of Medical Sciences and Research, Kollam, Kerala, Email: prasobh13@gmail.com) : Identification of BENr gene from Candida albicans in sputum samples. J Evolution Med Dent Sci 2018, 7(37), 4088-90.

Candidiasis is caused by infection with species of the genus Candida, predominantly with Candida albicans. The growing problem of mucosal and systemic candidiasis reflects the enormous increase in the number of patients at risk and the increased opportunity that exists for Candida species to invade tissues that are normally resistant to invasion with the help of the Candida virulence markers such as adhesin, Germ tube, Pseudohyphae, slime production, exoenzymes etc. The respiratory tract is frequently colonized with Candida species, especially in hospitalised patients. The drug resistance shown by Candida is increasing day-by-day. The most common gene responsible for multi-drug resistance was noted as BENr. Candida albicans was isolated from the sputum by conventional methods. Antifungal susceptibility testing was done, and multidrug-resistant strains were subjected to genotyping to identify BENr gene by multiplex PCR. From a total of 561 Candida albicans strains, 68 of them were multi-drug resistant and four strains have BENr gene. BENr gene, which is responsible for multidrug resistance in Candida was present in our region.

4 illus, 1 table, 9 ref

028113   ELSAYED E A, EL ENSHASY H A (Chemistry of Natural and Microbial Products Dep, National Research Centre, Dokki, 12622 Cairo, Egypt, Email: eaelsayed@ksu.edu.sa) : Effects of different aeration rates and feeding strategies on cell growth and invertase production kinetics by Saccharomyces boulardii. J Sci Ind Res 2018, 77(10), 575-82.

Invertase (β-D-fructofuranoside fructohydrolase; EC 3.2.1.26) constitutes an improtant microbial enzyme with wide applications in differnt food and pharmaceutical sectors. The present work used the biotherapeutic yeast Saccharomyces boulardii to produce invertase under different aeration rates in stirred tank bioreatcr. Our results showed that an aeration rate of 1 v v^-1 m^-1 was the most suitable in terms of cell growth and invertase productivity. Highest enzyme production was recorded 14950 U L^-1 after 50 h of cultivation. The production process was further optimized using different feeding strategties to overcome substrate limitation side effects encountered during batch cultivation. Sucrose feeding enhanced cell growth and enzyme productivity over batch cultivation by about 56.5 % (from 7.35 to 11.5 g L^-1) and 35.5 % (from 14950 to 20250 U L^-1), respectively. Furthermore, during feeding phase, invertase prodcution rate was improved by about 62.5 % (from 299 to 486 U L^-1 h^-1), while growth rate remained constant. Additionally, the improved invertase production was mainly due to increased biomass and not cell productivity, since both batch and fed-batch cultivations have nearly similar specific growth values (2243.9 and 2194.1 U g^-1, respectively). On the other hand, feeding complete medium greatly enhanced process parameters. Cell growth and invertase production increased from the batch cultivation by about 143.8 and 120.1 % (17.92 g L^-1 and 32900 U L^-1, respectively), and from the sucrose-feeding cultivation by about 55.8 and 62.5 %, respectively.

3 illus, 1 table, 41 ref

028108   BASAKA S (Pharmaceutical Chemistry Div, Dr. B.C. Roy Coll of Pharmacy & Allied Health Sciences, Durgapur, WB, Email: souvik_basak1@yahoo.com) : Scaling up asymmetric biocatalysis with cofactor regeneration by heterologous expression of a supra-active carbonyl reductase from Candida glabrata. J Sci Ind Res 2018, 77(9), 537-41.

A carbonyl reductase (cr) gene from Candida glabrata CBS138 has been cloned, over-expressed, characterised and subsequently employed in biotransformation of a prochiral keto ester (COBE) to a chiral alcohol (ethyl-4-chloro-3-hydroxybutanoate or CHBE). Using NADPH as cofactor and as substrate, the isolated enzyme (CR) exhibited a towering specific activity of 173.49 ± 6.08 Umin^-1mg^-1 with K_m and K_cat as 0.45 ± 0.02 mM and 112.77 ± 3.95 s^-1 respectively. Unlike other proteins of this class which usually show substrate inhibition at high substrate concentration (≥ 230 mM), the CR enzyme exhibited marked velocity at substrate concentration as high as 363 mM with highest turnover number (112.77 ± 3.95 s^-1). This advocated utility of the enzyme in a batch reactor where maximum COBE conversion has been achieved (161.04 g.L^-1 CHBE per g of dry cell weight) compared to the reported so far (1.51~ 149 g.L^-1 CHBE per g of dry cell weight). The reaction yielded sparingly available yet greatly important (R) isomer in over 99 % enantiomeric excess (e.e) with 88.30 % molar bioconversion. Although numerous proteins have been investigated to accomplish the prochiral COBE to chiral CHBE bioconversion, we present our finding as a highly efficient choice for conversion of COBE into CHBE through an efficient batch reaction system.

3 illus, 18 ref

028128   MOHANRAJ R, HAIDAR S, NOBRE M, ANANTHAN R (Medical Biotechnology Dep, MGM Institute of Health Sciences, Kamboth, Navi Mumbai-410 209, Email: remyam@gmail.com) : Anti HIV-1 activity, anti bacterial activity and phytochemical analysis of leaf extracts from Cleistanthus collinus (Roxb.) Benth. ex Hook.f.. Indian J Tradit Know 2018, 17(4), 770-5.

This study is an attempt to identify new chemical entities with anti HIV1 activity. In vitro anti-HIV activity of Cleistanthus collinus extracts was evaluated using the p24 antigen assay. A dose-dependent inhibition of the p24 antigen expression was observed and the extract was found to be efficient against HIV-1. Antibacterial activity was tested against four different bacteria employing the agar well diffusion method and the highest activity was exhibited by the chloroform and methanolic extracts of C. collinus. HPLC analysis of the crude extract revealed the presence of phytochemicals like Gallic acid, Chlorogenic acid, 3,4 Dihydroxy B-acid, Diadizin, p-Coumaric acid, Epi-GC Gallate, Ellagic acid, Luteoline, Hesperitin and Quercetin. This report on the anti HIV 1 activity of C. collinus and the results obtained from the present study suggest that the extracts could serve as a source for providing potential lead compound for drug discovery against HIV-1.

1 illus, 3 table, 27 ref

028120   JAIN P, DANWRA K, SHARMA H P, MAHATO D (Botany Dep, Ranchi Univ, Ranchi- 834 008, Email: paras.jain42@yahoo.in) : In vitro tissue culture studies and synthetic seed formation from Plumbago zeylanica L. Indian J Exp Biol 2018, 56(10), 769-73.

Synthetic seeds and in vitro propagation are the need of the hour, especially for conservation of medicinal plants which are under the threat of extinction due to extensive exploitation. The plumbagin, Plumbago zeylanica L., is one such highly exploited medicinal plant. Here, we attempted in vitro propagation of its roots by tissue culture and also synthetic seed development towards conservation of this plant. Its leaves were used as the explant. Surface sterilized explants were aseptically cultured on MS medium supplemented with different plant growth hormones. The embryoid callus produced from tissue culture was then used to produce synthetic seed for large scale production of the plants and to reduce the risk of maintenance, storage and transportation of the cultured plants. For the production of synthetic seed, the embryoid callus were chopped aseptically and were encapsulated with sodium alginate and liquid MS medium without CaCl₂ supplemented with growth hormones of similar concentration as used in tissue culture. Best result of callus induction and root regeneration was observed on MS medium supplemented by 2 ppm NAA (naphthalene acetic acid).

3 illus, 2 tables, 21 ref

028111   DASH P K, GUPTA P, JAILANI A K, RAI R (ICAR-NRC on Plant Biotechnology, New Delhi, Email: pdas@nrcpb.org) : Hydropenia induces expression of drought responsive genes (DRGs) erd1, hat, plD-δ, and zfa in Linum usitatissimum L. Indian J Exp Biol 2018, 56(10), 743-9.

Globally, abiotic stresses affect growth and yield of an economically and industrially important field crop flax, Linum usitatissimum. Being genomically unmapped, the molecular details of abiotic stress signaling in flax has not been elucidated. One such important and most damaging abiotic stress in flax is hydropenia or drought that inhibits growth and development of the plant. With the release of its genome sequence, there is a renewed interest in functional genomics study in flax. In an endeavour to get insights into the molecular events of hydropenia i.e., drought stress, an in-depth study of four marker genes induced by drought stress was carried out in flax. Expression profiling of these four drought responsive genes viz. erd1, hat, plD δ and zfa in flax were correlated to the expression profile in model crops such as Arabidopsis and rice. Based on phenotypic expression, relative water content and semi-quantitative PCR expression data, we confirmed the applicability of these four genes in screening drought tolerant varieties of an industrially important crop like flax.

3 illus, 1 table, 54 ref

028149   ZOTHANPUIA, PASSARI A K, YADAV M K, SINGH B P (Biotechnology Dep, Mizoram Univ, Mizoram-796 004, Email: bhimpratap@gmail.com) : In vitro evaluation of antimicrobial activities and antibiotic susceptibility profiling of culturable actinobacteria from fresh water streams. Indian J Exp Biol 2018, 56(9), 665-73.

Actinobacteria are major producers of antibiotics, industrially significant enzymes and many pharmaceutically important biologically active compounds. Twenty two actinobacterial strains were isolated from fresh water stream sediment samples of Murlen National Park, Mizoram, India. The actinobacterial strains were screened against antifungal pathogens (Fusarium oxysporum, Fusarium udum, Fusarium proliferatum, Fusarium oxysporum ciceri and Fusarium graminearum), and antibacterial activities against five bacterial pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Micrococcus luteus, Bacillus subtilis and Escherichia coli) and a yeast pathogen Candida albicans. All strains showed antibacterial activity against E. coli and F. proliferatum. Based on the results of antagonistic, antibacterial and anti-yeast, two most potent strains Kocuria sp. and Streptomyces intermidus were further evaluated for their antibiotics susceptibility activity against 21 different antibiotics. Kocuria sp. showed resistance to 10 antibiotics whereas Streptomyces intermidus was resistance to 15 antibiotics. Modular genes Polyketide Synthase (PKS II) and Nonribosomal Peptide Synthetase (NRPS) were also detected in these two strains, which might be responsible for the production of secondary metabolites. Two volatile compounds, Di-N-octyl phthalate and 1-Bromo-3, 7-Dimethyloctane were identified from the extract of Streptomyces intermidus BPSWAC29 strain using Gas chromatography Mass spectrometry (GC-MS). This study highlights the promise of discovering novel actinobacteria with antimicrobial activity from underexplored niche biotopes such as fresh water stream sediments.

5 illus, 5 tables, 46 ref

028134   ROHELA G K, SHABNAM A A, SHUKLA P, AURADE R, GANI M, YELUGU S, SHARMA S P (Central Sericultural Research & Training Institute, Pampore-192 121, Jammu & Kashmir, Email: gulab_biotech@yahoo.co.in) : In vitro clonal propagation of PPR-1, a superior temperate mulberry variety. Indian J Biotechnol 2018, 17(4), 619-25.

A protocol was developed and standardized for the in vitro clonal propagation of PPR-1, a superior temperate mulberry variety using nodal explants. When the nodal explants of PPR-1 mulberry variety were inoculated onto various concentrations and combinations of cytokinins supplemented media, maximum axillary bud proliferation was obtained on combinational rather than individually supplemented hormonal media. Over all, maximum axillary shoot length (7.2 ± 0.61 cm) and maximum number of leaves per explant (8.1 ± 0.85) was obtained on combinational media of 6 benzylaminopurine (BAP) (1.5 mg/L) and kinetin (2.0 mg/L) after 20 days of culture. On individually supplemented cytokinins hormonal media maximum axillary shoot length (4.7 ± 0.61 and 3.2 ± 0.22 cm) with more number of leaves (7.4 ± 0.16 and 6.0 ± 0.35) per explant was obtained at 1.5 mg/L and 2.0 mg/L concentration of BAP , respectively after 20 days of culture. The proliferated axillary shoots when transferred on to different concentrations of auxins containing rooting media, good response of rooting (100 %) was observed on Murashige & Skoog (MS) media supplemented with 2.0 mg/L concentration of indole butyric acid (IBA). The raised plantlets were then hardened using 1 : 1 ratio of vermicompost and soil, then gradually they were acclimatized to field conditions. The survival rate of in vitro raised PPR-1 plantlets in field conditions is about 70 %.

5 illus, 2 tables, 39 ref

028123   KUMARI S N, SINGH S P, CHATTOPADHYAY T, KUMAR S, KUMAR M (Plant Breeding and Genetics Dep, Bihar Agricultural Univ, Bhagalpur-813 210, Bihar, Email: mankesh2008@gmail.com) : Diversity analysis of rice landraces of Bihar using linked and functional markers. Indian J Biotechnol 2018, 17(4), 611-8.

In the present study, 45 simple sequence repeat (SSR) markers distributed throughout the 12 chromosomes of rice and a gene-specific co-dominant marker for the fragrance (fgr) gene of rice were used to characterize 13 landraces collected from the adjoining areas of Bhagalpur in Bihar and 3 released rice genotypes. The number of alleles generated by each marker ranged from 2 to 4 with an average of 2.41 alleles per locus. Marker RM 489 generated maximum number of alleles (4). Polymorphic information content (PIC) varied greatly for all SSR loci ranging from 0.08 to 0.49 with average of 0.25. The highest PIC value (0.49) was obtained for RM 431, which suggests the utility of this marker for analyzing genetic diversity in rice. The constructed dendrogram showed two distinct clusters, where the major cluster (cluster I) contained 14 out of the 16 rice genotypes under study. Dice similarity coefficient was maximum between Malida and Burma Bhusi and minimum between Kalanamak and Gutraj. By using gene-specific marker for grn gene, 11 genotypes showed amplification of fragrance allele-specific band. Thus, the present study suggests that there is scope for exploitation of genetic diversity of landraces available in Bihar for future rice breeding programmes.

3 illus, 3 tables, 24 ref

028139   SHARMA R K, BHULLAR M B, SINGH S, JINDAL V (Entomology Dep, Punjab Agricultural Univ, Ludhiana-141 004, Email: rksharma@pau.edu) : Molecular analysis of fenazaquin selected resistant strain of two-spotted spider mite Tetranychus urticae Koch. Indian J Biotechnol 2018, 17(4), 602-10.

The two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae) is an important agricultural pest in a wide range of outdoor and protected crops worldwide. Fenazaquin is mitochondrial electron transport inhibition (METI)-acaricide and its extensive and frequent use for control of this mite has facilitated resistance development. So, present studies were conducted to investigate genetic differences between fenazaquin resistant and susceptible population of T. urticae as no information is available regarding mutation/variability in genes involved in imparting resistance. Fenazaquin selected resistant population was developed in the laboratory by giving selection pressure for up to 15 generations which resulted into 166.49 fold increase in resistance level when compared with susceptible population. Molecular analysis of resistant and susceptible population revealed no changes in genes structure of CYP392A11 and CYP392A12 in the resistant compared to the susceptible population. However, expression profiling of nine different genes associated (CYP392A11, CYP392A12, CYP392A16, CYP392D2, CYP392D3, CYP392D6, CYP392D7, CYP392D8 and CYP392D10p) with METI resistance showed increased mRNA transcripts in CYP392A11, CYP392A12, CYP392D2 and CYP392D10p. The increased MFO activity in the resistant population corresponds to nearly two fold increase in the expression of CYP392A11 and CYP392A12. The target site mutation might not have any role in low to moderate level of fenazaquin resistance in mites. Differences in mRNA expression are a simple, fast and reliable tool for early detection of resistance.

5 illus, 2 tables, 32 ref

028112   DIXIT S J, APPU KUTTAN K K, SHRIVASTAVA R M (Biological Sciences and Engineering Dep, Maulana Azad National Institute of Technology, Bhopal, Madhya Pradesh, Email: manitbiotech@gmail.com) : Genetic characterization of sulphur and iron oxidizing bacteria in manganese mining area of Balaghat and Chhindwara, Madhya Pradesh, India. Indian J Biotechnol 2018, 17(4), 595-601.

The aim of present study was to explore microbiology of manganese mining area of Balaghat and Chhindwara, Madhya Pradesh, India with the objective of reducing load of mine based pollution to support environmental sustainability with help of bacterial isolates. The research involves physicochemical analysis, culture dependent methods, 16S rDNA based sequencing and computational phylogenetic analysis. The 16S rDNA sequence analysis revealed the occurrence of two iron oxidizing bacteria (Staphylococcus hominis and Pseudomonas sp.) and four sulphur oxidizing bacteria (Bacillus cereus HYM74, B. anthracis, B. cereus D42 and Pantoea calida) in the selected sites. All cultures were able to grow on acidic as well as neutral pH medium and at low temperature of 4 °C. Bacterial isolates were also found with heavy metal tolerance for Mn⁺⁷ and Cr⁺⁶ up to the concentration of 1000 ppm. This study assists the idea of biomineralization, bioremediation and future reclamation in the selected mining area with the help of bacteria.

8 illus, 2 tables, 24 ref

028131   PATTNAIK S (Biotechnology and Bioinformatics Dep, Sambalpur Univ, Burla- 768 019, Email: smaranika2010@gmail.com) : Induction of bioactive compounds in a co-cultured strain of Micromonosperma echinospora (DST-4) isolated from secluded Hirakud dyke soil. Indian J Biotechnol 2018, 17(4), 586-94.

An actinomycetes (DST-4) strain was isolated from soil of Hirakud dyke, Western Odisha, India. The isolated actinomycetes strain was subjected to 16S rRNA gene partial sequencing and the sequence in FASTA format was analysed at National Centre Biotechnology Information (NCBI) portal using basic local alignment search tool (BLAST) analysis. The sequence was identified as a strain of gentamicin producing Micromonosperma echinospora with 99 % identify. The sequence was submitted to GenBank and obtained an Accession no. MF536417.1. A phylogenetic tree was generated with six strains of same species available in database. The actinomycetes strain (DST-4) was screened for its antagonistic activity against three streptomycin resistant Staphylococcal aureus strains. ‘Triplicate patch inoculums diffusion’ and ‘cell immersed tube dilution’ techniques were used. Streptomycin was taken as reference drug. A quantitative cocultured-incubation method was applied for induction of bioactive compounds from the isolated actinomycete strain. The growth curve analysis of test staphylococcal strains through spectrophotometric as well as subculture experiments had inferred about antibacterial activity of putative bioactive compounds present in cell free extract (CFE) of DST-4 strain. The H¹ NMR study of high performance liquid chromatography (HPLC) chromatogram peak fraction of CFE, was carried out. The nuclear magnetic resonance (NMR) signals had inferred about induction bioactive compounds with of R₂NH and ROH as functional groups.

10 illus, 1 table, 40 ref

028129   NGUYEN T Q, DUONG T H, DANG T N H, LE N G, LE Q G, DO T H, NGUYEN V D, LE T T H, TRUONG N H (Institute of Biotechnology, Ha Noi, Viet Nam, Email: tnhai@ibt.ac.vn) : Enhanced soluble expression and effective purification of recombinant human interleukin-11 by SUMO fusion in Escherichia coli. Indian J Biotechnol 2018, 17(4), 579-85.

Human interleukin-11 is a multifunctional cytokine applied for the clinical treatment of thrombocytopenia. However, IL-11 has been considered a difficult protein in to express in an Escherichia coli expression system. Here, we demonstrate a suitable construction for high production of recombinant human interleukin-11 (rhIL-11) in E. coli. An optimized codon gene encoding human IL-11 was inserted in-frame with the small ubiquitin like modifier (SUMO) protein in the pE-SUMO3 vector. The SUMO IL-11 fusion protein was entirely expressed in soluble form and reached 31.6 % of total soluble protein in E. coli. The rhIL-11 protein with a purity of over 99 % was obtained at high protein yields of 320 mg rhIL-11 per liter of bacterial culture. Bioactivity of rhIL-11, as determined by proliferation of a TF-1 cytokine-dependent cell line, was 4.17 x 10⁵ unit/mg, similar to the activity of the natural protein. Interestingly, the rhIL-11 was purified easily and effectively due to its selective precipitation from the reaction mixture. To the best of our knowledge, this is the first report demonstrating self-aggregating recombinant protein after cleavage from SUMO. Thus, expression of rhIL-11 fused with SUMO yielded greatly increased soluble production and convenient purification, and could offer a potential drug candidate for deployment in clinical trials.

5 illus, 1 table, 25 ref

028140   SHRUTHI N, INDHU M, SHENDE A M, PAWDE A M, SINGH P, BHURE S K (ICAR-Indian Veterinary Research Institute, Izatnagar- 243 122, Email: sdbhure@rediffmail.com) : The anti-peptide relaxin antibodies for monitoring the well-being of the fetus in pregnant bitches. Indian J Biotechnol 2018, 17(4), 569-78.

Pregnancy management is difficult in canines and there is a lack of methodology which would allow monitoring embryonic well-being. Relaxin (Rlx) is being reported for use in pregnancy diagnosis in different species. Here, we evaluated seven anti-Rlx peptide antibodies for detection of well-being of the fetus in bitches. Peptides were synthesized using solid phase peptide synthesis chemistry and the hyper-immune sera raised in chicken against peptides. In sandwich enzyme linked immunsorbent assay (ELISA), the chicken anti-Rlx P4 as a capture and rabbit anti-prorelaxin as detection antibody, gave better results in terms of differentiating the pregnant from non-pregnant bitches. Thirty five canine serum samples (21 non-pregnant, 11 pregnant and 3 males) were screened. Among the 11 pregnant, six delivered normally and the rest of the bitches aborted a few days after the serum collection with one or two dead fetuses. Among the 11 pregnant serum samples, 4 showed the absorbance above the cut-off value set for pregnancy, which delivered healthy puppies and five bitches showed the absorbance below the set cut-off value aborted after a few days of blood collection with one or two dead fetuses. The specificity of the assay was found to be 90.47 % and the sensitivity of the assay for detecting the well-being of the fetus is 82 %. The serum samples collected from those bitches having problems related to normal whelping, had shown absorbance below the set cut-off value. The results of the study indicate that relaxin is a useful marker, specifically for monitoring the pregnancy and tells about the well-being of the fetus.

5 illus, 3 tables, 39 ref

028122   KIM D H, KIM K S, RAMAKRISHNA S (Hanyang Univ, Seoul, Korea, Email: ks66kim@hanyang.ac.kr) : Transcriptional activation of LGR5 gene by an engineered CRISPR-Cas9-based system induces hepatic-specific factors. Indian J Biotechnol 2018, 17(4), 561-68.

Several new approaches for reprogramming or direct reprogramming somatic cells have been implemented during the last few years. Endogenous gene activation or repression can be achieved using dead clustered regularly interspaced short palindromic repeats associated protein 9 (dCas9) system fused with a transcriptional activating or repression domain. This study was undertaken to screen and validate efficient sgRNAs targeting reprogramming or direct reprogramming transcription factors by CRISPR-Cas9 based system. In this study, we designed and validated several individual single-guide RNA (sgRNA) targeting LGR5 and Yamanaka factors, such as Oct3/4, Sox2, c-Myc and Klf4, for effective transcriptional activation in mouse cells. Furthermore, we investigated the combination of effective sgRNAs for the synchronized effect of transcriptional activation on LGR5 gene and Yamanaka factors and achieved approximately 8.4-fold and 38-fold higher levels of mouse LGR5 and Oct3/4 upregulation, respectively, compared with the control. Further, we demonstrate that the activation LGR5 gene promoter induces known defined factors responsible for direct conversion of somatic cells to hepatocyte-like cells. We envision that our validation of effective sgRNAs will facilitate the development of mouse induced pluripotent stem cells (iPSCs) and novel findings of LGR5 as a potential candidate for direct conversion of somatic cells to hepatocyte-like cells.

5 illus, 2 tables, 27 ref

028130   PARK S W, DO H J, CHOI W, KIM H J, KANG M J, SEO H G, KIM J H (Biomedical Science Dep, CHA Univ, Gyeonggi-Do 13488, Republic of Korea, Email: jaehwan_k@cha.ac.kr) : CRISPR/Cas9 knock-in of GST-tagged human noggin in the β-casein gene locus of bovine ear fibroblast cells. Indian J Biotechnol 2018, 17(4), 553-60.

We developed knock-in vector system of human Noggin mature sequence with glutathione S-transferase (GST) containing factor Xa protease linker to facilitate the subsequent purification of recombinant protein. To achieve this, bovine ear fibroblast cells were isolated and transfection conditions were optimized by electroporation. To generate knock-in vector, human Noggin lacking its native signal peptide is fused to GST and foot and month disease virus 2A (F2A), and then inserted into bovine β-casein gene exon 3. We also generated enhanced green fluorescent protein (EGFP) expression vector of GST-human Noggin mature fused to β-casein signal peptide and F2A, and successfully detected recombinant human Noggin protein secreted into culture media, followed by factor Xa cleavage. Then, we co-transfected human Noggin knock-in vector with single-guided RNA and Cas9 expression vectors into bovine ear fibroblasts and obtained the stably-integrated colonies by antibiotic selection. PCR screening analysis revealed that 26 out of 35 colonies positively integrated human Noggin knock-in vector into bovine β-casein locus. One of positive clones was subjected to chromosome analysis, presenting normal karyotypes. Our data may provide the additional purification guideline of recombinant proteins by tagging GST with a protease linker sequence in the upstream of target genes and a high efficiency of integration ratio into bovine β-casein locus.

4 illus, 1 table, 29 ref

028106   AN-WEN L, ZHANG H, LIN J, FANG-HAI W (Sun Yat-sen Univ, Guangzhou 510075, China, Email: lsswfh@mail.sysu.edu.cn) : De novo assembly and analysis of the white-backed planthopper (Sogatella furcifera) transcriptome. J Insect Sci 2018, 18(4), 11.

The white-backed planthopper Sogatella furcifera (Horváth) has become an important pest on rice in China and Southeast Asian countries. White-backed planthopper wing bud length is in relation to adult wing length, but little is known about the development and differentiation of wing buds at the molecular level. Using Illumina HiSeq high-throughput sequencing technology, we sequenced four cDNA libraries, two biological replicates of longwinged female fifth-instar nymphs (LW), and two of short-winged nymphs (SW). In total, 62,154 unigenes with an average length of 984 bp and N50 length of 1,878 bp were obtained by de novo transcriptome assembly. A total of 18,416 open reading frames (ORFs) were predicted based on the unigenes. Ninety-three percentage of these ORFs could be annotated by searching for homology in six protein databases. A total of 184 differentially expressed genes (DEGs) with 129 upregulated and 55 downregulated were found in SW compared to LW. Gene Ontology and euKaryotic Orthologous Group classification provided comprehensive information about the function of each gene. Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed five enriched pathways including three metabolic pathways. In addition, we found that some DEGs were relevant to muscle movement and cuticle and likely involved in development and differentiation of wing buds. This study provided transcriptome resource of female fifth-instar nymphs of white-backed planthopper including long-winged and short-winged nymphs, and different molecular features between them lay the foundation for adult wing morph prediction, promoting further studies on planthopper population management.

5 illus, 4 tables, 32 ref

028105   ALLEN S, GINWAL H S, BARTHWAL S (Genetics and True Propagation Div, Forest Research Institute, Dehradun-248195, Email: allenswati@gmail.com) : Identification of allele specific primers in Chir pine (Pinus roxburghii Sarg.) through data mining. Indian J Biotechnol 2018, 17(3), 538-40.

Pinus roxburghii Sarg. commonly known as Chir pine, is a commercially exploited species for resin in India. Resin is a phenotypic trait which is expressed only in mature trees. Due to this a large number of trees are cut every year which pose a serious ecological threat. Recent advancements in bioinformatics and molecular biology have enabled scientists to identify high resin yielding pine genotypes at nursery stage using molecular markers. In the present study, investigation, 337 terpene coding sequences from the database were analyzed using data mining and 45 microsatellites allele specific primers were designed for marker assisted selection. Out of these 45 primers, only 22.2 % primer pairs showed polymorphism within the 53 genotypes of Chir pine examined. A wide range of fragment size  was  observed from 100 - 600 bp. PCR amplification using allele specific markers produced a total of 22 bands, out of which 14 were found to be polymorphic. The total number of bands amplified per marker varied from of 1 to 3 with an average of 1.4. Genetic divergence in terms of percent polymorphism ranged from 50 to 100 % with a mean of 63.3 % per marker.

1 illus, 1 table, 10 ref

028145   SWETHA V P,PARVATHY V A, SHEEJA T E, SASIKUMAR B (Crop Improvement and Biotechnology Div, ICAR-Indian Institute of Spices Research, Kozhikode-673012, Email: bhaskaransasikumar@yahoo.com) : Isolation and amplification of genomic DNA from nutmeg mace. Indian J Biotechnol 2018, 17(3), 533-7.

A reliable and efficient protocol for isolation and amplification of genomic DNA from dried mace of Myristica fragrans, was developed. The yield of genomic DNA was 231.4 µg g^-1 and 306.8 µg g^-1, respectively for the samples procured from the farm and market. The absorbance ratio at A260/A280 was greater than 1.8 indicating the good quality of DNA. Complete restriction digestion and PCR amplification of genomic DNA further confirmed the quality of isolated DNA.

5 illus, 2 tables, 20 ref

028147   VALADEZ-MOCTEZUMA E, LÓPEZ-LÓPEZ A, TEODORO-PARDO C V D (Departamento de Fitotecnia, Chapingo Autónoma Univ, Mexico, Email: evaladez@chapingo.mx) : Usefulness of three DNA-PCR techniques to differentiate Jalapeno pepper varieties. Indian J Biotechnol 2018, 17(3), 527-32.

The cultivation of pepper (Capsicum annuum) is one of the oldest in America. However, out of the 25 species described only 5 have been domesticated. The varieties registered and legally protected must comply with International Union for the Protection of New Varieties of Plants (UPOV) guidelines. Nevertheless, simple sequence repeat the morphological descriptors used for this purpose are sometimes subjective and they are altered by the environment conditions. Molecular genetic markers have been used as alternative to reduce these limitations. The aim of this study was to compare the effectiveness of three techniques based on PCR to discriminate varieties of jalapeño pepper with little phenotypic differences and with very similar shape of the fruit. The techniques used were random amplified polymorphic DNA-DNA amplification fingerprinting (RAPD-DAF), inter simple sequence repeat (ISSR) and simple sequen repeat (SSR) through capillary and conventional electrophoresis. Results indicated that the values of genetic variation in the studied jalapeno varieties are different depending on the technique used, which is due to the capacity of each technique to sample the genome.

5 illus, 2 tables, 29 ref

028127   MAL C, KUNDU S (Biophysics Dep, Calcutta Univ, Kolkata-700009, Email: skbmbg@caluniv.ac.in) : Comparative study on sequence characteristics of mature and precursor miRNAs of monocot and dicot plants. Indian J Biotechnol 2018, 17(3), 520-6.

MicroRNAs (miRNAs) negatively regulate mRNAs at post-transcriptional level and thus can regulate different biological processes. Here we analyze the sequence characteristics and length distribution of miRNA sequences of six monocot and six dicot plants. We observe a species specific nucleotide preference of miRNAs. Although the first position of 5' end of mature miRNAs is U rich, there exists a wide variation. While the GC % of monocot mature miRNAs in general is nearly equal to the AU %, the GC % of stress induced miRNAs is significantly higher than AU %. Thus, higher content of GC % can be used as a signature of stress induced miRNAs in monocot. The length distribution of mature miRNAs shows the highest peak at 21 nucleotides with some other minor peaks at 20, 22 and 24 nucleotides. The synthesis of some miRNAs has positional preference either to 5' or to 3' arm of their precursors, but they are different in monocot and dicot.

7 illus, 1 table, 19 ref

028117   GOWDA R H, ABDEL-RAZEK A S, MARAPPA P B, SHANKARANARAYANA S K, HOOLAGERI H C, PUTTEGOWDA P H (Life Science Dep, Bangalore Univ, Bangalore-560056, Email: ravikumarh79@gmail.com) : Molecular phylogenetic affiliation of Wolbachia and phage WO in storage pests from India. Indian J Biotechnol 2018, 17(3), 514-9.

The rich distribution and diversity of Wolbachia has been well investigated. The array of reproductive alterations induced by this cytoplasmically inherited α- proteobacteria has made it a successful ‘reproductive parasite’. Despite their obligate intracellular lifestyle which usually protects bacteria from phage infection, Wolbachia harbors a widespread temperate phage called WO. Recent studies implicate the rapid phase of evolutionary changes in Wolbachia genome due to the horizontal phage transfer. These changes directly reflect the varied reproductive phenotypes induced by Wolbachia in their hosts. The distribution and diversity of Wolbachia in same host species across different geographical locations in itself is interesting. Previous studies have demonstrated the absence of Wolbachia in storage pests, Tribolium castaneum and Rhyzopertha dominica in laboratory stocks of Japan. Contrary to their studies, natural infections of Wolbachia and Phage WO in Tribolium castaneum, Rhyzopertha dominica and Sitophilus zeamais have been reported in the current study. Of the above screened three storage pests, the first two harbours A super group and remaining one with B super group. Further, a phage WO infection was common across all the three storage pests. The analysis of the endosymbiont and its phage provides basic information which could be further exploited for biocontrol programs.

2 illus, 1 table, 30 ref

028146   THE C Y, MAZIAH M, SHAHARUDDIN N A, HO C (Biochemistry Dep, Putra Malaysia Univ, Malaysia, Email: maziahm@upm.edu.my) : Efficient in vitro multiple shoots regeneration system from rice shoot apex in recalcitrant Malaysian indica rice cultivars (Oryza sativa L.). Indian J Biotechnol 2018, 17(3), 504-13.

Development of in vitro rice shoot regeneration system can contribute to new advances for the selection of stress tolerant cultivars as well as production of elite cultivar through genetic transformation approach. Multiple shoot induction was carried out in two widely cultivated Malaysian indica rice MR 220 and MR 253 using 10 mm rice shoot apex along with coleoptile isolated from 4 days old seedlings. The explants were cultured on a Murashige and Skoog (MS) media supplemented with various concentrations of 6-benzylaminopurine (BAP) and Kinetin (KIN) at (0 – 10 mg/L) for two culture periods 2 and 4 weeks. The results showed that the optimal multiple shoots growth were observed in media supplemented with 6 mg/L KIN, which induced an average (10.30 ± 0.95) shoots for MR 220 and (6.67 ± 0.57) shoots for MR 253. Results from biochemical changes studies indicated that a relatively higher total chlorophyll and soluble protein content were obtained in rice shoots treated with KIN as compared to BAP suggesting KIN is more suitable in the in vitro multiple shoots regeneration system of Malaysian indica rice.

3 illus, 5 tables, 48 ref

028142   SINGH A, SHARMA J K (Seed Science and Technology Dep, CSK Himachal Pradesh Krishi Vishwavidyalya, Palampur -176062, Email: singhadr@yahoo.com) : Molecular characterization of parental lines and hybrids of maize cultivated in North-Western Himalayan region using microsatellite markers. Indian J Biotechnol 2018, 17(3), 496-503.

Maize is an important cereal crop of North-Western Himalayan region of India with wide ranging and diversified uses which is resulting in the release and recommendation of new hybrids of maize for the region. The use of parental lines with  a narrow genetic base by various private and public sector seed agencies may cause a risk of genetic diversity loss. Molecular characterization of hybrids and parental lines by DNA fingerprinting provides knowledge of the genetic relationship among them thus preventing the risk of increasing uniformity. The technique is helpful for molecular identification and hybrid purity testing. The present study was undertaken to carry out microsatellite marker based DNA fingerprinting of ten maize hybrids and their parental lines for molecular identification and assessment of genetic diversity. Twenty three microsatellite markers were found to be polymorphic. Number of alleles per locus ranged from 2 to 6 with a mean of 3.30. Mean expected heterozygosity and polymorphic information content was 0.58 and 0.52, respectively. Average Jaccard’s similarity coefficient observed was 0.46 revealing an average genetic diversity of 54 among various genotypes studied. Cluster analysis delineated the maize hybrids and parental lines into four main clusters A, B, C and D at 0.32 similarity coefficient.

2 illus, 3 tables, 20 ref