005533   PONNUSAMY P, REETHA T L, SASIKALA M, RONALD B S M, SELVARAJ J, PUVARAJAN B, MANICKAM R (Veterinary Microbiology Dep, Tamil Nadu Veterinary and Animal Sciences Univ, Orathanadu- 614 625, Email: ponsvhl@gmail.com) : Molecular detection and phylogenetic analysis of avian reticuloendotheliosis virus from formalin fixed tissue by PCR in fancy chicken. Indian J Anim Res 2019, 53(9), 1243-5.

The present study was carried out to identify avian neoplastic viruses in the formalin fixed tissues of fancy chicken. The tissue samples collected during necropsy were examined by histopathology. It showed lymphoid and reticular cell infiltration in kidney, liver and lungs. For identification and differentiation of avian neoplastic viruses, PCR was performed using primer sets specific for Mareks disease virus, avian leukosis complex and reticuloendotheliosis virus. It was found that tumors were REV originated. Further confirmation, purified PCR product was subjected to sequencing. It showed 99 % homology with other REV isolates available in the NCBI database. The present communication describes infection of a fancy chicken with REV on the basis of histopathological findings as well as molecular methods.

4 illus, 1 table, 13 ref

005496   DIXIT P, RAO M L V, DIXIT A K, GUPTA R, SHUKLA P C (Veterinary Medicine Dep, Nanaji Deshmukh Veterinary Science Univ, Jabalpur- 482 001, Email: alokdixit7@yahoo.com) : Prevalence and molecular characterisation of Cryptosporidium spp in goat kids. Indian J Anim Res 2019, 53(9), 1234-8.

Prevalence and molecular characterisation of Cryptosporidium species was done in kids belonging to organised and nonorganised goat farms at Jabalpur. The overall prevalence of Cryptosporidium was 14.63 %. The prevalence was nonsignificantly higher in male kids (16.16 %) as compared to that of female kids (13.21 %). Age wise prevalence was higher in kids up to one month age (16.13 %) than that of kids upto 3 months age (13.99 %). No significant difference was found in prevalence among different breeds and in kids kept in farm or field conditions. The prevalence was non-significantly higher in non-diarrhoeic kids than diarrhoeic kids. Most of the infections were of one score (76.6 %). Molecular characterisation by PCR-RFLP of 18S SSU rRNA gene revealed presence of Cryptosporidium parvum species in positive faecal samples.

2 illus, 1 table, 14 ref

005510   JYOTHI J S, PUTTY K, PATIL S R, REDDY Y N (P V N R Telangana Veterinary Univ, Hyderabad- 500 030, Email: kalyaniputty@gmail.com) : In-silico approach to determine the possibility of a recombinant broad-spectrum vaccine for bluetongue disease. Indian J Anim Res 2019, 53(9), 1229-33.

Bluetongue (BT) disease caused by BT virus (BTV) is an acute haemorrhagic disease of domesticated and wild ruminants. 29 BTV serotypes have been identified so far throughout the world with 24 circulating in India conferring no cross protective immunity. Hence, the objective of the current study was to investigate the possibility of developing a recombinant subunit protein based vaccine for BT that is cross protective to all the known serotypes of BTV. VP2 is a major serotype determining protein and vaccination with VP2 was known to induce neutralizing antibody response. Since BTV circulates as 29 immunologically distinct serotypes, the current study aimed to identify conserved region of VP2. The amino acid sequence alignment showed that although VP2 was very variable among serotypes, several regions were relatively more conserved between serotypes; most conserved region (cVP2) of which was evident from 338-383aa. Moreover, selection pressure analysis revealed that all the codons in cVP2 region were under strong negative selection (dN/dS=0.16) suggesting strong structural and functional constraints on cVP2. In addition, two MHCI, MHCII each, and three B cell epitopes were predicted based on the physicochemical properties of the amino acids in cVP2 region. This suggests strong immunogenicity of cVP2 making it a suitable vaccine candidate. Taken together, cVP2 appears to be a promising target for a broad-spectrum subunit vaccine moiety for BT that can confer protection against the available serotypes of BTV. Since all the structural proteins of BTV will not be used in such subunit vaccines, observations from current study can also be extended to develop a marker vaccine for DIVA against BT.

2 illus, 1 table, 27 ref

005542   REVANASIDDU D, RAMESHA K P, KOUR R J, KUMAR A M, DIVYA P, KATAKTALWARE M A, BASAVARAJU M, DAS D N, ANANDKUMAR N, SAPNA N (ICAR - National Dairy Research Institute, Bengaluru- 560 030, Email: rsdeginal@gmail.com) : Genetic variants in male specific region (MSY) of ZNF280BY gene and their association with semen quality traits in Murrah buffalo bulls. Indian J Anim Res 2019, 53(9), 1135-9.

Bull fertility is an important factor for genetic improvement; therefore prediction of fertility an early age of the bulls by using genetic markers is a goal for livestock breeding. Zinc finger protein280BY linked gene is predominantly expressed in testis and has a major role in spermatogenesis. An investigation was carried out to detect the genetic variants in the male specific region (MSY) of ZNF280BY gene and their association with semen quality traits in Murrah buffaloes. Polymerase Chain Reaction-Single Strand Conformation Polymorphism technique (PCR-SSCP) and DNA sequencing methods revealed three different SSCP band patterns (AA, BB, CC); one transition (C10375T) and an indel (insertion of T at 10460-10461). The associations of the genetic variants with semen volume, sperm concentration, Hypo Osmotic Swelling Test (HOST) in fresh and frozen semen were observed. AA and CC genotype bulls had high volume (P$\le$0.05) of semen per ejaculation than BB genotype bulls. BB genotype bulls had high sperm concentration (106 /mL) per ejaculate in comparison to AA and CC genotype bulls (P$\le$0.05). AA and BB genotype bulls had a higher per cent of HOST (P$\le$0.05) reacted sperms of fresh semen and per cent acrosomal integrity of frozen semen in comparison to CC genotype bulls. The observed association between SSCP variants in MSY region of ZNF280BY gene with semen quality parameters indicated the possibilities of using ZNF280BY as a candidate gene for identification of markers for semen quality traits in Murrah buffaloes.

6 illus, 3 tables, 14 ref

005486   BATRA K, NANDA T, KUMAR A, KUMAR V, GOPAL G J, MAAN S (Animal Biotechnology Dep, Lala Lajpat Rai Univ of Veterinary and Animal Sciences, Hisar-125 004) : Molecular cloning and expression kinetics of serum interferon stimulated gene for early pregnancy detection. Indian J Anim Res 2019, 53(9), 1129-34.

Interferon stimulated protein 15 is a well-known ubiquitin cross-reactive protein which is released from the ruminant uterus in response to interferon-tau (IFN-t) during conceptus implantation. This protein increases significantly during the early stages of pregnancy. Therefore, in the present study, Interferon stimulated gene (ISG15) coding region was isolated from buffalo blood at day 18 of pregnancy after artificial insemination. ISG15 was cloned in pET100 vector, overexpressed in E.coli and purified to investigate its properties. The protein was immunoblotted with mouse anti-His antibody to evaluate its characteristics after purification. To conclude, elucidation of protein properties in the prokaryotic system will provide an opportunity for understanding the basic biology of pre and post-implantation and development of a tool for early pregnancy diagnosis.

4 illus, 36 ref

005548   SINHA G, TIWARI S, JADHAV S K (Pt. Ravishankar Shukla Univ, Raipur- 492 010, Email: shubhratiwari238@yahoo.co.in) : Simultaneous sachharification and fermentation of rice residues and its comparative analysis for bioethanol production. Def Life Sci J 2019, 4(3), 158-62.

Energy consumption has inflated steadily over the last century because the world population has fully grown and additional countries became industrialised. Bioethanol is an alcohol produced by fermentation of plant biomass, containing carbohydrate and its production depends upon feedstock availability, variability, and sustainability. The selection  of  feedstock  and  its  pretreatment  is  an  important  part  of  bioethanol  production  process.  In  present  work, the  exploration  of  the  potential  of  agro-waste  rice  residues such as,  rice bran and rice  husk was done,  because it contains sufficient amount of carbohydrate which can be ferment into bioethanol. The aim of the research was also to investigate how different pretreatment methods with moderate conditions differ in hydrolysis and fermentation efficiencies. Pretreatment plays an important role in the hydrolysis of cellulose and lignocellulose. It was found that biological pretreatment was a most effective method in terms of production of bioethanol and it enhances the production as well as fermentation efficiency.

3 illus, 1 table, 30 ref

005541   REDDY A R, KRUPANIDHI S, VENKATESWARULU T C, KUMAR R B, SUDHAKAR P, PRABHAKAR K V (Biotechnology Dep, Vignan’s Foundation for Science, Technology & Research, Valdamudi- 522 213, Email: kodalividyaprabhakar@gmail.com) : Molecular characterization of a biopolymer producing bacterium isolated from sewage sample. Curr Trends Biotechnol Pharm 2019, 13(3), 325-35.

Plastics and polypropylene polymers are synthesized from non renewable resources and persist in environment long after intended use,resulting into problems of global environmental pollution. Hence, the present study focused on production of polyhydroxybutyrate (PHB)microbial polyester by bacterial fermentation. In the present study, 05 PHB producing bacterial species were isolated from sewage waste, Guntur, India. Among all one isolate showed the maximum PHB yield of 4g/L. The high PHB producing bacterium was identified as Acinetobacternosocomialis RR20, based on biochemical and molecular methods. Further, the characterization of PHB produced from this strain was also studied by analytical methods namely, FT-IR, DTA, TGA,¹H NMR, ¹³C NMR and LC-MS.

8 illus, 3 tables, 46 ref

005551   TUFCHI N, PANT K, WAHEED S M, DEVVRET (Biotechnology Dep, Graphic Era Univ, Dehradun, Uttarakhand, Email: neematufchi@gmail.com) : In-Silico analysis and identification of functional singlenucleotide polymorphism (SNPS) of the DISC1 gene. Curr Trends Biotechnol Pharm 2019, 13(3), 317-24.

SNPs (Single-nucleotide polymorphisms) are essential for understanding the genetic origin of various complex human diseases. The identification of the functional SNPs responsible for the disease is still a challenge so there is an urgent need for identification of functional SNPs. In this work, analysis was made on the genetic variations which can alter both the function and expression of the DISC1 gene using computational approaches. The total of 23 SNPs was found, out of which 12 are missense (non-synonymous ornsSNPs), 8 occurred in 3’UTR region and 3 are synonymous SNPs. The 2 nsSNPs (rs6675281and rs821616) were found to be damaging by PolyPhen and SIFT servers. I-mutant server showed decrease in the stability of rs6675281 and increase in the stability of rs821616 protein upon mutations. Structural analysis of proteins with mutations was done using SPDBV (Swiss PDBviewer), MUSTER (MUlti—Sources Thread ER) and PyMol tools for the detection of molecular dynamics and energy minimization calculations.This study revealed that L607F and S704C variants could indirectly or directly destabilize the amino acid interactions.

2 illus, 5 tables, 26 ref

005523   LUKKANI N J, REDDY E S (Genetics and Genomics Dep, Yogi Vemana Univ, Kadapa, Andhra Pradesh) : Screening of ACC-deaminase and antifungal metabolites producing fluorescent pseudomonads isolated from rhizosphere soil of groundnut. Curr Trends Biotechnol Pharm 2019, 13(3), 309-16.

Plant Growth Promoting Rhizobacteria that can colonize the root systems are an efficient group of beneficial bacteria. Fluorescent pseudomonads belong to this category that can enhance plant growth and disease suppression by different types of mechanisms. Fifty five fluorescent pseudomonads (JS 1....JS 55) identified from soil samples collected from different places of Rayalaseema region located in Andhra Pradesh, screened for ACC- deaminase enzyme activity and antifungal metabolites production. Results showed that it is highly likely that these pseudomonads isolates deaminated endogenous ACC. Furthermore, these isolates remarkably producing antifungal metabolites like siderophores, chitinases and antibiotics. Among these, 5 isolates (JS 7, JS 16, JS 24, JS 31 andJS 44) are efficient producers of ACC-deaminase and antifungal metabolites.

37 ref

005487   BESAN M, GAUTAM M K, SHRIVASTAVA S K (Pharmaceutical Engineering and Technology Dep, Indian Institute of Technology, Varanasi- 221 005, Email: skshrivastava.phe@iitbhu.ac.in) : Development of 1, 4-naphthoquinones as potential epidermal growth factor receptor inhibitors for the treatment of cancer. Curr Trends Biotechnol Pharm 2019, 13(3), 283-308.

In this manuscript, we have designed,synthesized and characterized 1, 4-naphthoquinone derivatives. The synthesized compounds (MB1-MB19) were further subjected for evaluation of their anticancer activity using MCF-7, HeLa and HepG2 cancer cell lines. The compound MB-9 was observed to be most active against these three cancer cell lines i.e. MCF-7(IC₅₀ = 15.63 ± 0.47 μM), HeLa (IC₅₀ =13.45 ± 0.48μM), and HepG2 (IC₅₀ = 23.87 ± 0.59μM). Compound MB-9 has also shown potent tyrosine kinase inhibitory activity with IC₅₀ = 1.80 ± 0.06μM. Moreover, molecular docking investigations revealed that compound MB-9 has strong binding affinity to the active site residues of tyrosine kinase.These outcomes give a promising beginning to assist in the improvement of novel and powerful anticancer agents.

7 illus, 6 tables, 39 ref

005534   POONATI R, MALLEPADDI P C, PUNATI R D, MAITY S N, ALAMURI A, MANCHIKALAPUDI S, ALAPATI K S, KAVI KISHOR P B, POLAVARAPU R (Biotechnology Dep, Acharya Nagarjuna Univ, Guntur- 522 510, Email: pbkavi@yahoo.com) : Development and validation of point of care diagnostics for the rapid detection of multiple species of Leptospira at resource-limited areas. Curr Trends Biotechnol Pharm 2019, 13(3), 270-82.

Leptospirosis is a life threatening,emerging, infectious zoonotic disease of humans and livestock all over the world. The diagnosis of this disease is frequently ineffective. Conventional methods of diagnosis, gold standard bacterial culture and microscopic agglutination test (MAT)are being used. These tests are time-consumingand the test for the disease can be positive more than 2 weeks after the onset of the disease. Present study was taken up to circumvent the problems and disadvantages in the methods/protocols being used currently in leptospirosis diagnosis. This study aimed to develop a point of care lateral flow (LFA) and indirect ELISA assays to detect disease-specific antibodies in the wholeblood/serum/plasma for the rapid diagnosis of acute infection. The Leptospira specific lipopolysaccharide (LPS) was used in combination with recombinant multi-epitope membrane protein as an antigen candidate for the detection of disease specific antibodies. The developed lateral flow (LFA) and indirect ELISA assays were compared with gold standard MAT and commercial ELISA kit. The MAT confirmed standard references era were used for validation of developed assays. A total of 223 positive and 115 non-positive samples for leptospirosis were used in the present validation. Indirect ELISA assay developed in the present study showed higher sensitivity andspecificity of 98.65 % and 100 % respectively. Incontrast, the sensitivity and specificity of the commercial ELISA and LFA were 96.41 %, 97.39 %, 87.44 % and 98.26 % respectively. The results obtained proved that LFA and indirect ELISA assays were more effective than the conventional methods for the diagnosis of acute leptospirosis, especially within the onset and useful for the point of care diagnosis at resource-limited areas.

5 illus, 2 tables, 30 ref

005537   PUNATI R D, MALLEPADDI P C, POONATI R, JAIN M, MAITY S N, SOHAL J S, PODHA S, KAVI KISHOR P B, POLAVARAPU R (Biotechnology Dep, Acharya Nagarjuna Univ, Guntur- 522 510, Email: pbkavi@yahoo.com) : Development and validation of rapid, sensitive and in-expensive protein g-based point of care diagnostic assay for serodiagnosis of paratuberculosis at resource-limited areas. Curr Trends Biotechnol Pharm 2019, 13(3), 232-42.

Paratuberculosis is caused by Myco-bacterium avium subspecies paratuberculosis (MAP). It affects domestic cattle, ruminant livestock species with emerging zoonotic concerns and also causes human Crohn’s disease. It is a chronic granulomatous enteritis and results in huge economic loss to domestic dairy cattle and ruminant industry. Further, there is mounting evidence on the zoonotic role of MAP in human Crohn’s disease. MAP is not killed by pasteurization, therefore, milk and milk products are major sources of infection for humans. The disease affects cattle and small ruminant industries, thus impacting farmer’s economy. Control of the disease is hampered by the lack of rapid and accurate diagnostic tests. Therefore, it is urgently needed to develop affordable as well as high-performing diagnostic tools. So, the present study was aimed to develop a simple,inexpensive, rapid and robust point of care (POC) lateral flow diagnostic test kit for sero diagnosis of paratuberculosis infection in bovine cattle and in small ruminant livestock species. We developed a rapid, pen-side lateral flow antibody (Ab) diagnostic test kit for onsite screening of paratuberculosis. The rapid kit detects infection in 20 minutes and can be used directly by a farmer without the requirement of an expert or equipment. We evaluated 2,502 samples using lateral flow assay (LFA) which included 243 reference sera and 2,259 field samples. The results suggest that the sensitivity and the specificity of LFA were 91.01% and 98.94 % respectively in comparison with the gold standard culture, polymerase chain reaction (PCR) and delayed-type hypersensitivity (DTH) methods. Thus, LFA aids to detect the MAP specific antibodies (Ab) in the collected sample sat POC resource-limited areas and help the bovine and small ruminant livestock healthcare systems.This assay is well suited for the early diagnosis of MAP in less equipped laboratories and in resource-limited POC settings suitable for the Indian environment.

3 illus, 2 tables, 36 ref

005483   ALTAAE H H (Plant Protection Dep, Mosul Univ, Mosul, Iraq, Email: dr.hudataae@yahoo.com) : Using nested PCR to detect the non-defoliating pathotype of Verticillium dahliae on olive orchard. Crop Res 2019, 54(5&6), 139-42.

Verticillium wilt caused by Verticillium dahliae infects olive at any time point of its life cycle. The severity of verticillium wilt on olive trees depends upon the virulence of the pathogen isolates. Isolates of V. dahliae infecting cotton and olive showed cross virulence, which was classified into defoliating (D) and non-defoliating (ND) pathotypes based on their ability to completely defoliate the plant or only cause wilt (i. e. no defoliation). In recent years, several different types of molecular techniques have been used for identifying V. dahliae isolates in a wide range of hosts or virulence. The polymerase chain reaction (PCR) procedure was used in this study for the detection and quantification of verticillium wilt pathogens. The incidences of V. dahliae on olive in different areas of Iraq and Jordan were investigated. The samples from the same trees were used to detect the type of pathogenic strain [defoliating pathotype (D) versus non-defoliating pathotype (ND)] using PCR with specific primer pairs. The isolates showed positive results using nested PCR reactions, proving the existence of V. dahliae pathogen in tested olive trees in Iraq. The results confirm that the pathotype of V. dahliae is from the ND type. This is the first report which showed the existence of ND V. dahliae in Iraq.

4 illus, 3 tables, 15 ref

005511   KAI Z, PEI Z, HONGMEI W, YUNLEI Z, WEI C, HAIYAN G, XIAOHUI S, YANLI C (Institute of Cotton Research of Chinese Academy of Agricultural Sci, Anyang- 455 000, Email: aywhm@163.com) : Isolation and characterization of the GbVIP1 gene and response to Verticillium wilt in cotton and tobacco. J Cotton Res 2019, 2, 2.

Verticillium wilt is a serious soil-borne vascular disease that causes major losses to upland cotton (Gossypium hirutum L.) worldwidely every year. The protein VIP1 (VirE2 interaction protein 1), a bZIP transcription factor, is involved in plant response to many stress conditions, especially pathogenic bacteria. However, its roles in cotton response to Verticillium wilt are poorly understood. Results: The GbVIP1 gene was cloned from resistant sea-island cotton (G. barbadense) cv. Hai 7124. Expression of GbVIP1 was up-regulated by inoculation with Verticillium dahliae and exogenous treatment with ethylene. Results of virus-induced gene silencing suggested that silencing of GbVIP1 weakened cotton resistance to Verticillium wilt. The heterologous expression of GbVIP1 in tobacco showed enhanced resistance to Verticillium wilt. The PR1, PR1-like and HSP70 genes were up-regulated in GbVIP1 transgenic tobacco after Verticillium wilt infection. Our results suggested that GbVIP1 increased plant resistance to Verticillium wilt through up-regulating expressions of PR1, PR1-like, and HSP70. These results provide new approaches to improving resistance to Verticillium wilt in upland cotton and also have great potential for disease-resistance breeding of cotton.

6 illus, 44 ref

005498   FAIZA A, GHULAM Q, YONGHUI L, SHUYA M, LILI L, ZUOREN Y, ZHI W, FUGUANG L (Chinese Academy of Agricultural Sciences, Henan- 455 000, Email: wangzhi.12@163.com) : Genome-wide identification of Gossypium indeterminate domain genes and their expression profiles in ovule development and abiotic stress responses. J Cotton Res 2019, 2, 3.

INDETERMINATE DOMAIN (IDD) transcription factors form one of the largest and most conserved gene families in plant kingdom and play important roles in various processes of plant growth and development, such as flower induction in term of flowering control. Till date, systematic and functional analysis of IDD genes remained infancy in cotton. In this study, we identified total of 162 IDD genes from eight different plant species including 65 IDD genes in Gossypium hirsutum. Phylogenetic analysis divided IDDs genes into seven well distinct groups. The gene structures and conserved motifs of GhIDD genes depicted highly conserved exon-intron and protein motif distribution patterns. Gene duplication analysis revealed that among 142 orthologous gene pairs, 54 pairs have been derived by segmental duplication events and four pairs by tandem duplication events. Further, Ka/Ks values of most of orthologous/ paralogous gene pairs were less than one suggested the purifying selection pressure during evolution. Spatiotemporal expression pattern by qRT-PCR revealed that most of the investigated GhIDD genes showed higher transcript levels in ovule of seven days post anthesis, and upregulated response under the treatments of multiple abiotic stresses. Evolutionary analysis revealed that IDD gene family was highly conserved in plant during the rapid phase of evolution. Whole genome duplication, segmental as well as tandem duplication significantly contributed to the expansion of IDD gene family in upland cotton. Some distinct genes evolved into special subfamily and indicated potential role in the allotetraploidy Gossypium hisutum evolution and development. High transcript levels of GhIDD genes in ovules illustrated their potential roles in seed and fiber development. Further, upregulated responses of GhIDD genes under the treatments of various abiotic stresses suggested them as important genetic regulators to improve stress resistance in cotton breeding.

8 illus, 59 ref

005545   SINGH A, KAUR A (Botany Dep, Akal Univ, Talwandi Sabo, Bathinda, Email: arvinder_bot@auts.ac.in) : Comparative studies on seed protein characteristics in eight lines of two Gossypium species. J Cotton Res 2019, 2, 6.

In order to achieve the targets aiming at the improvement of protein quality, knowledge regarding seed protein fractions and polypeptides constituting them in different crops is essential. Besides having high nutritional value as animal feed and human food, the protein isolates from cottonseed meal have also been proven promising as industrial raw materials for a number of applications. As far as Indian work on the characterization of cotton seed proteins is concerned, relatively meagre reports are available. Keeping in mind the importance of cotton seed proteins, lines belonging to Gossypium arboreum L. (Indian cotton) and G. hirsutum L. (American cotton) which are grown in all the major cotton growing states in India were selected for analysing their seed protein characteristics.  Whereas G. arboreum (A-genome) lines revealed a lower range of seed protein content i.e. 19.5~24.3 %, an upper range (21.8~29.5 %) could be observed in lines of G. hirsutum (AD-genome). Globulins represented dominating fraction in both species followed by albumins, glutelins and prolamins. A significant positive correlation between albumins/globulins and seed protein content in G. arboreum /G. hirsutum, respectively, was observed. Intraspecific electrophoretic variation in seed protein extracts was observed in the region of molecular weight 22 kDa - 27 kDa in lines of both the species; however some lines with A-genome showed similarity in banding pattern with AD-genome. Four polypeptides with disulphide-linkages were also reported for the first time. Albumins were observed to reveal more variations in their electrophoretic pattern between the lines of two species followed by globulins. On the basis of present and previous studies, screening the lines with low or high protein content will lead the selection of lines with superior polypeptide fraction important for nutritional and industrial purposes. On comparing the composition and behaviour of four 2-S linked polypeptides with other plant groups, these were suggested to be legumin-like in nature. The similarity in banding patterns between the lines of A-genome and AD-genome species marked towards the close evolutionary relationship between these two. Albumin fractions on the basis of our results could be taken for cultivar differentiation in cotton crop.

3 illus, 9 tables, 50 ref

005549   TAMBEL LEILA I M, MINGYUAN Z, YUAN C, XIANG Z, CHEN YUAN, DEHUA C (Yangzhou Univ, Yangzhou- 225 100, Email: cdh@yzu.edu.cn) : Amino acids application enhances flowers insecticidal protein content in Bt cotton. J Cotton Res 2019, 2, 7.

Low insecticidal protein expression at reproductive organs affect insect resistance in Bt transgenic cotton. In order to enhance flower insecticidal protein expression, the conventional cultivar Sikang1 (S1) and the hybrid cultivar Sikang3 (S3) were used as experimental materials; the applications of selected 5 types of amino acids and 21 types of amino acids were sprayed on the flowers in 2016 and 2017 cotton growing seasons. The flower Bt protein contents increased significantly under the two amino acid treatments in both cultivars, the Bt protein concentration increased by 15.2 to 25.8 % compared with the control. However, no significant differences were detected between the two treatments of amino acid application. Increased amino acid and soluble protein contents, enhanced GPT, GOT, protease,and peptidase activities were observed under the amino acid application at the flowering stage. These results suggest that exterior application of the amino acids treatments could bolster the flower insecticidal protein expression.

1 illus, 4 tables, 30 ref

005558   YI S, PENG Z, ZHAO L, FANGJUN L, XIAOLI T (China Agricultural Univ, Beijing- 100 193, Email: lifangjun@cau.edu.cn) : An isopentyl transferase gene driven by the senescence-inducible SAG12 promoter improves salinity stress tolerance in cotton. J Cotton Res 2019, 2, 15.

Soil salinity seriously affects cotton growth, leading to the reduction of yield and fiber quality. Recently, genetic engineering has become an efficient tool to increase abiotic stress tolerance in crops. In this study, isopentyl transferase (IPT), a key enzyme involved in cytokinin (CTK) biosynthesis from Agrobacterium tumefaciens, was selected to generate transgenic cotton via Agrobacterium-mediated transformation. A senescence-inducible SAG12 promoter from Arabidopsis was fused with the IPT gene. Ectopic-expression of SAG12::IPT significantly promoted seed germination or seedling tolerance to salt stress. Two IPT transgenic lines, OE₃ as a tolerant line during seed germination, and OE₈ as a tolerant line at seedling stage, were selected for further physiological analysis. The data showed that ectopic-expression of SAG12::IPT induced the accumulation of CTKs not only in leaves and roots, but also in germinating seeds. Moreover, ectopic-expressing IPT increased the activity of antioxidant enzymes, which was associated with the less reactive oxygen species (ROS) accumulation compared with control plants. Also, ectopic-expression of IPT produced higher K⁺ /Na⁺ ratio in cotton shoot and root. The senescence-induced CTK accumulation in cotton seeds and seedlings positively regulates salt stress partially by elevating ROS scavenging capability.

6 illus, 45 ref

005555   XIAOMIN Y, XUKE L, XIUGUI C, DELONG W, JUNJUAN W, SHUAI W, LIXUE G, CHAO C, XIAOGE W, XINLEI W, WUWEI Y (Institute of Cotton Research of Chinese Academy of Agricultural Sci, Anyang- 455 000, Email: yew158@163.com) : Genome-wide identification and expression analysis of DNA demethylase family in cotton. J Cotton Res 2019, 2, 16.

DNA methylation is an important epigenetic factor that maintains and regulates gene expression. The mode and level of DNA methylation depend on the roles of DNA methyltransferase and demethylase, while DNA demethylase plays a key role in the process of DNA demethylation. The results showed that the plant’s DNA demethylase all contained conserved DNA glycosidase domain. This study identified the cotton DNA demethylase gene family and analyzed it using bioinformatics methods to lay the foundation for further study of cotton demethylase gene function. This study used genomic information from diploid Gossypium raimondii JGI (D), Gossypium arboreum L. CRI (A), Gossypium hirsutum L. JGI (AD1) and Gossypium barbadebse L. NAU (AD2) to Arabidopsis thaliana. Using DNA demethylase genes sequence of Arabidopsis as reference, 25 DNA demethylase genes were identified in cotton by BLAST analysis. There are 4 genes in the genome D, 5 genes in the genome A, 10 genes in the genome AD1, and 6 genes in the genome AD2. The gene structure and evolution were analyzed by bioinformatics, and the expression patterns of DNA demethylase gene family in Gossypium hirsutum L. were analyzed. From the phylogenetic tree analysis, the DNA demethylase gene family of cotton can be divided into four subfamilies: REPRESSOR of SILENCING 1 (ROS1), DEMETER (DME), DEMETER-LIKE 2 (DML2), and DEMETER-LIKE3 (DML3). The sequence similarity of DNA demethylase genes in the same species was higher, and the genetic relationship was also relatively close. Analysis of the gene structure revealed that the DNA demethylase gene family members of the four subfamilies varied greatly. Among them, the number of introns of ROS1 and DME subfamily was larger, and the gene structure was more complex. For the analysis of the conserved domain, it was known that the DNA demethylase family gene member has an endonuclease III (ENDO3c) domain. The genes of the DNA demethylase family are distributed differently in different cotton species, and the gene structure is very different. High expression of ROS1 genes in cotton were under abiotic stress. The expression levels of ROS1 genes were higher during the formation of cotton ovule. The transcription levels of ROS1 family genes were higher during cotton fiber development.

4 illus, 1 table, 37 ref

005509   JICHAO J, SHUAI Z, JUNYU L, LI W, XIANGZHEN Z, KAIXIN Z, LIJUAN Z, JINJIE C (Chinese Academy of Agricultural Sciences, Anyang- 455 000, Email: aycuijinjie@163.com) : Comparative transcriptional analysis provides insights of possible molecular mechanisms of wing polyphenism induced by postnatal crowding in Aphis gossypii. J Cotton Res 2019, 2, 17.

Aphis gossypii is a worldwide sap-sucking pest with a variety of hosts and a vector of more than 50 plant viruses. The strategy of wing polyphenism, mostly resulting from population density increasing, contributes to the evolutionary success of this pest. However, the related molecular basis remains unclear. Here, we identified the effects of postnatal crowding on wing morph determination in cotton aphid, and examined the transcriptomic differences between wingless and wing morphs. Effect of postnatal crowding on wing determination in A. gossypii was evaluated firstly. Under the density of 5 nymphs·cm^{− 2} , no wing aphids appeared. Proportion of wing morphs rised with the increase of density in a certain extent, and peaked to 56.1 % at the density of 20 nymphs·cm^{− 2} , and reduced afterwards. Then, transcriptomes of wingless and wing morphs were assembled and annotated separately to identify potentially exclusively or differentially expressed transcripts between these two morphs, in which 3 126 and 3 392 unigenes annotated in Nr (Non-redundant protein sequence) database were found in wingless or wing morphs exclusively. Moreover, 3 187 up- and 1 880 down-regulated genes were identified in wing versus wingless aphid. Pathways analysis suggested the involvement of differentially expressed genes in multiple cellular signaling pathways involved in wing morphs determination, including lipid catabolic and metabolism, insulin, ecdysone and juvenile hormone biosynthesis. The expression levels of related genes were validated by the reverse transcription quantitative real time polymerase chain reaction (RT-qPCR) soon afterwards. The present study identified the effects of postnatal crowding on wing morphs induction and demonstrated that the critical population density for wing morphs formation in A. gossypii was 20 nymphs·cm^{− 2} . Comparative transcriptome analysis provides transcripts potentially expressed exclusively in wingless or wing morph, respectively. Differentially expressed genes between wingless and wing morphs were identified and several signaling pathways potentially involved in cotton aphid wing differentiation were obtained.

6 illus, 1 table, 54 ref

005559   YUHUAN M, LONGFU Z, XIANLONG Z (Huazhong Agricultural Univ, Wuhan- 430 070, Email: xlzhang@mail.hzau.edu.cn) : Down regulation of cotton GbTRP1 leads to accumulation of anthranilates and confers resistance to Verticillium dahliae. J Cotton Res 2019, 2, 19.

Verticillium wilt, caused by Verticillium dahliae, is called a “cancer” disease of cotton. The discovery and identification of defense-related genes is essential for the breeding of Verticillium wilt-resistant varieties. In previous research we identified some possible broad-spectrum resistance genes. Here, we report a tryptophan synthesis-related gene GbTRP1 and its functional analysis in relation to the resistance of cotton to V. dahliae. Expression analysis shows that GbTRP1 is suppressed at 1 h and 6 h post V. dahliae infection, but activated at 12 h and 24 h, and the expression of GbTRP1 is highly induced by treatment with salicylic acid and jasmonic acid. Sub-cellular localization studies show that GbTRP1 is localized in the chloroplast. Suppression of GbTRP1 expression leads to lesion-mimic phenotypes and activates the immune response in cotton by showing enhanced resistance to V. dahliae and B. cinerea. Metabolomic analysis shows that anthranilic compounds significantly accumulated in GbTRP1-silenced plants, and these metabolites can inhibit the growth of V. dahliae and B. cinerea in vitro. Our results show that suppression of GbTRP1 expression dramatically activates the immune response and increases resistance of cotton to V. dahliae and B. cinerea, possibly due to the accumulation of anthranilate compounds. This study not only provides genetic resources for disease resistance breeding, but also may provide a basis for new chemical control methods for combatting of fungal disease in cotton.

8 illus, 28 ref

005560   ZHENYU L, ABIDALLHA E H M A, HUIMIN W, MINGYUAN Z, XIANG Z, YUAN C, DEHUA C (Yangzhou Univ, Yangzhou- 225 009, Email: cdh@yzu.edu.cn) : Bt insecticidal efficacy variation and agronomic regulation in Bt cotton. J Cotton Res 2019, 2, 23.

The bollworm can be controlled effectively with Bacillus thuringiensis transgenic cotton (Bt cotton) which is being applied worldwide. However, the insecticidal efficacy is not stable. Here we give a summary of research progress for the mechanism of the altered insecticidal efficacy, factors affecting the expression of insect resistance, agronomic practices on regulation of insecticidal efficacy in Bt cotton. To realize the transgenic potential of Bt cotton cultivars, future research may be conducted by increasing synthesis and reducing degradation of Bt protein to maintain high insecticidal ability in the transgenic cotton by agronomic management.

64 ref

005519   KUMARI A, AMERI S, RAVIKRISHNA , DHAYALAN A, SELVANKUMAR T (Biotechnology Dep, Mahendra Arts and Science Coll, Namakkal- 637 501, Email: anjalisoni458@gmail.com) : Isoenzyme and anticancer screening of conotoxin proteins extracted from sea snails. Int J Pharm Biol Sci 2019, 9(3), 32-8.

The present investigation has been reported with the cone snail Conus inscriptus venom proteins. MTT assay with normal and SK-BR-3 cancer cell lines evidenced anticancer properties in Conus inscriptus venom protein. Hemolytic, Phospholipase (isoenzyme) and hyaluronidase assays were also performed for biomedical applications.

4 illus, 33 ref

005482   ABULJADAYEL D, ATEF A, AL-MATARY M, EDRIS S, AL-GHAMDI K M, HAJRAH N H, SABIR J S M, HALL N, BAHIELDIN A (Biological Sciences Dep, King Abdulaziz Univ, Jeddah- 215 89, Email: abmahmed@kau.edu.sa) : A new PCR-based species genotyping differentiation approach in entamoeaba. Biosci Biotech Res Asia 2019, 16(3), 491-508.

The most commonly used approach for Entamoeba species differentiation up to date is the tRNA-linked STR regions of the parasite’s genome. In the present study, a new reliable, fast and easy molecular tool for species differentiation was developed. DNA was isolated from fecal samples collected from infected subjects with either Entamoeba histolytica (EH) or Entamoeba disper (ED) in Saudi Arabia. Two types of primer sets were compared in which the first targeted tRNA-linked STR regions, while the second was designed after multiple contig alignment of the two genomes using NUCmer program in aligned areas with high similarity (~90 %) and difference between of ~90 bp. The selection criteria secures that designed primers should pair with both EH and ED contig sequences at homologous regions of 200-500 bp of both species except for the presence of indels that result in the recovery of amplicons of two species with different sizes. Banding patterns in the tRNA-linked STR region resulted in the occurrence of several common amplicons. We speculate that primers mismatch with regions other than the specified STR arrays of Entamoeba histolytica or Entamoeba disper with organisms other than Entamoeba existed in the fecal sample. However, the STR-based approach looked very useful in studying strain differentiation and parasite diversity. The results for the new approach complemented those of the STR-based approach, except that the latter failed to detect coinfected subjects. The new approach proved to be useful at the species level, while the tRNA-linked STR approach can still be a good choice for strain differentiation.

6 illus, 4 tables, 20 ref

005501   GIZAW T M, VEMURI P K (Biotechnology Dep, Koneru Lakshmaiah Education Foundation, Vaddeswaram, Andhra Pradesh, Email: vemuripraveen@kluniversity.in) : Effect of plasmid presence and stability on growth of Escherichia coli DH5α with use of drugs, chemicals, and radiation. Asian J Pharm 2019, 13(3), 209-16.

The purpose of the study delineates the growth and plasmid stability of E. coli DH5α host system.  Different concentrations of drugs, chemicals, and various frequency of radiations were subjected to the host system to verify the colony-forming units along with plasmid concentration and stability. Among chemicals, acridine orange showed the highest effect on growth of DH5α, while among the drugs, danthrone showed maximum effect on the growth of the organism. Radiofrequency of 2 GHz and lowintensity microwave radiation were recorded as the highest inhibitory effects. However, there is no significant effect in growth observed in exposure to UV rays. The present work discussed that drugs, chemicals, radiofrequency, and microwave radiation have a huge effect on the growth of organism and also on the concentration and stability of plasmid.

14 illus, 26 ref

005525   NAWFA R, PURNOMO A S, PUTRO H S (Chemistry Dep, Institut Teknologi Sepuluh Nopember, Surabaya- 60111, Email: refnawfa@chem.its.ac.id) : Synthesis of antibiotic Penicillin-G enzymatically by Penicillium chrysogenum. Asian J Chem 2019, 31(10), 2367-9.

Penicillin-G antibiotic was used as the basic ingredient of making antibiotic type β-lactam such as tetracycline, amoxicillin, ampicillin and other antibiotics. Penicillin-G was splited into 6-amino penicillanic acid as the source of β-lactam. The biosynthetic pathway for the formation of penicillin-G in Penicillium chrysogenum cell through the formation of intermediates was carried out in the form of amino acids such as α-aminoadipate, L-cysteine, L-valine which are formed from glucose (food ingredients).The formation of 6-amino penicillanic acid is an amino acid combination of L-cysteine and L-valine, a step part of the formation of antibiotic penicillin-G in P. chrysogenum cells, thus, it is obvious that there are enzymes involved in its formation. The objective of this study was to examine the use of enzymes present in P. chrysogenum cells to produce penicillin-G and 6-amino penicillanic acid using the intermediate compounds α-aminoadipate, L-cysteine, L-valine and phenylacetic acid assisted by NAFA^® coenzymes in P. chrysogenum cells which is more permeable. The research method started from producing biomass of P. chrysogenum cells that demonstrated penicillin-producing antibiotic capability, as the source of the enzyme, followed by addition of permeability treatment of P. chrysogenum cell membrane to get immobile of enzyme by its own cell therefore it can be used more than once. After that the enzyme activity was proven by adding α-aminoadipate, L-cysteine, Lvaline, phenylacetic acid and NAFA® coenzyme for the formation of penicillin-G, whereas the addition of L-cystein, L-valine and NAFA^® coenzyme were aimed to form 6-amino penicillanic acid. The results showed that P. chrysogenum is able to produce antibiotics with stationary early phase on day 6. The best increased permeability of P. chrysogenum cell membranes was obtained using a 1:4 of toluene:ethanol ratio mixture with the highest antibiotic concentration (130.06 mg/L) after testing for the enzymatic formation of antibacterial penicillin-G.

4 tables, 7 ref

005505   IWUALA E, ODJEGBA V, UMEBESE C, SHARMA V, ALAM A (Bioscience and Biotechnology Dep, Banasthali Vidyapith, Rajasthan- 304 022, Email: afrozalamsafvi@gmail.com) : Physiological and gene expression studies of selected Zea mays L. and Pennisetum glaucum (L.) R. Br. genotypes to simulated drought stress condition. Vegetos 2019, 32(3), 397-406.

Drought is a major environmental stress that significantly obstructs productivity of various crops worldwide. Among several remedial methods to conquer this abiotic stress, use of resistant cultivars is most preferred. This study compares the response in selected genotypes of pearl millet and maize under a progressive drought stress conditions at specific intervals. Three genotypes (IP14599, IP14787 and LRNO3) of pearl millet recorded relatively higher water content (RWC) than the three genotypes (DTSYN11, LRN03 and LRIO1) of maize. Leaf water potential (Ψ_L), leaf osmotic potential (Ψ_π), leaf turgor potential (Ψ_p) and PSII were measured in harvests to attain comparative observations. Furthermore, to correlate these results expression of three genes were measured. It was observed that Ψ_π decreased over time and Ψ_p recorded a decrease with Ψ_Lat a higher rate in maize compared to pearl millet. A more declining trend in maximum fluorescence (∆F/Fm′) and electron transport rate (ETR) in LRIOI, LRNO1 and DTSYN11 was recorded compared to IP14599, IP14787 and LRNO3. The study of gene CBF in leaves and roots revealed it’s responsiveness to drought in genotypes of pearl millet IP14599, LRNO3 and IP14787 while it was absent in maize genotypes LRNO1 and LRIO1, with the exception of DTSYN11. Expression of RubSc gene showed a noteworthy decline in reactive oxygen species in the genotypes IP14599, LRNO3, DTSYN11 and IP14787, while a marked increase was observed in LRNO1 and LRIO1. Likewise gene PIP2;3 were highly responsive to drought in pearl millet but not in maize, where they might support greater water transport. Overall, the results indicate remarkable activation of mRNA expression of these genes under drought stress which provides the resistance against drought.

5 illus, 2 tables, 38 ref

005513   KHAN M S, DAS K, RAJASEKHARAN P E (Pharmacognosy and Phytochemistry Dep, Krupanidhi Coll of Pharmacy, Bangalore- 560 035, Karnataka, Email: drkkdsd@gmail.com) : Chromatographic quantification of polyphenol in relation to potential antioxidant activity and isolation of DNA from in vitro cultivated Decalepis nervosa Wight & Arn. leaf explant. Ann Phytomed 2019, 8(2), 141-9.

The objective of the present study was to perform in vitro micropropagation as well as the growth of callus from the leaf explant of Decalepis nervosa Wight & Arn. (DN) (Family: Apocynaceae), a climber woody medicinal plant. Addition of silicon (Si) as sodium and potassium silicate in the MS media helped in fast micropropagation, callus development and organogenesis of the explant which was modified technique in plant tissue culture of the said plant. Silicon ion also helped in avoiding browning of callus. Therefore, an alternate in vitro tissue culture method was established using sodium silicate and potassium silicate using varying concentrations of 0.5, 1.0, 2.0, 2.5, 3.0 and 3.5 mg/l to overcome the challenges when compared with the normal MS media. Half strength MS medium supplemented with BAP at 0.2 mg/l and NAA at 2.5 mg/l gave callus growth in 21 days and resulted in more accumulation of gallic acid. Thereafter, direct rooting and shoots of meristem occurred with IBA and Si as sodium silicate (IBA at 2 mg/l and Na₂SiO₃ at 2.5 mg/l), supplemented in half-strength MS media along with coconut water, within 18 days. Further, gallic acid in the methanol callus extract was estimated by the HPLC method and analyzed for antioxidant activity, using DPPH assay method. Finally, leaf DNA extraction was carried out (starting of genotype analysis) by CTAB (cetyl trimethylammonium bromide) method for purification of DNA and nanodrop method for quantification of the same. Finally, the results concluded that potassium silicate enhanced the production of phenolic content when analyzed by HPLC method, proved antioxidant activity as well as improved root formation in a very short time. Thereafter, single DNA pure band was identified and quantified which is essential for future research on new drug discovery.

9 illus, 4 tables, 42 ref

005540   RANI A, AZMI W (Biotechnology Dep, Himachal Pradesh Univ, Shimla- 171 005, Himachal Pradesh, Email: wamikazmi@rediffmail.com) : An overview on biosynthesis and applications of extracellular pyocyanin pigment and its role in Pseudomonas aeruginosa pathogenesis. Ann Phytomed 2019, 8(2), 28-42.

Microbial pigments are chemical moieties capable of absorbing light in the visible range. The demand for natural colorants is increasing in every day as the present trend throughout the world is shifting towards the use of ecofriendly and biodegradable commodities. Among the natural sources, the pigment producing microorganisms hold a promising potential to meet present day challenges. Pigments are compounds that come in a wide variety and extensively used in industries. Industrial production of natural pigments by microbial fermentation has several advantages over extraction of pigments of plant origin, such as easy availability of inexpensive raw materials for their production, independence from crop seasons and their seasonal variations, etc. The Pseudomonas aeruginosa has ability to produce a number of redox-active phenazine compounds including pyocyanin. The pyocyanin pigment possesses antimicrobial, anticancerous, antioxidant, anti-inflammatory and immunosupperessive properties. This review covers the challenges and new insights into pyocyanin from P. aeruginosa with emphasis on the role of pyocyanin in P. aeruginosa infection with special attention to applications of pyocyanin pigment.

6 illus, 4 table, 95 ref

005494   DAS K (Pharmacognosy and Phytochemistry Dep, Pharmacy Coll, Bangalore-560035, Karnataka, Email: drkkdsd@gmail.com) : Authentic identification and new drug discovery from natural plant based constituents through DNA bar-coding: A challenging task to the researchers. Ann Phytomed 2019, 8(2), 19-27.

Education provides thumb impression to signature whereas technology provides signature to thumb impression. Signature is copied in many cases but thumb impression never be copied. Such an important technology is DNA bar coding in plant species which a new biological tool for organismal biologists to increase their understanding of the environment especially authentication of all individual plants and phylogenetic construction. DNA bar code helps to determine the correct identification of a plant sample in a rapid, repeatable, and reliable fashion to conserve world biodiversity. Not only that, it is a powerful tool in systematics, ecology, evolutionary biology, including community assembly, species interaction networks, taxonomic discovery, and assessing priority areas for ecological and environmental protection. Furthermore, plant DNA bar codes are useful in the regulatory areas where endangered species and commercial products (viz., Foods and plant based supplements) are observed by the forensic investigators. Even though, application of genetic markers in the field of biological and commercial products by adopted genomic sequencing technologies are more efficient and cost effective workflow. Therefore, plant based DNA barcode is necessary and essential to preserve in the form of a library (through DNA amplification) which is the major challenges ahead in future, i.e., on building the global plant DNA barcode library to contribute toward the discovery of overlooked plant species around the globe.

7 illus, 1 table, 41 ref

005515   KHARAYAT B, SINGH P (Bioscience and Biotechnology Dep, Banasthali Vidyapith, Tonk, Rajasthan) : Study of media optimization and kinetic modeling of L-methioninase from Pseudomonas stutzeri. Vegetos 2019, 32(3), 370-80.

In this study, Pseudomonas stutzeri has been explored as new bacterial strain for producing intracellular L-methioninase. Kinetic modeling and optimization of fermentation conditions are crucial parameters for improving production of L-methioninase. Response surface methodology has been employed as fast efficient technique for investigating the interactive effect of culture condition parameters for L-methioninase. Plackett–Burman design was first employed for screening significant variables for production media components including glucose, L-methionine, NaCl, malt extract and casein enzymic hydrolysate. The optimal level of these significant media components was further optimized using Central composite design. The high value of determination of coefficient (R² = 0.95) showed the good fitness of these statistical models for explaining the relationship between variables and response. The optimum values for the investigated variables were obtained as 5% glucose, 0.25% L-methionine, 0.2% malt extract, 1% casein enzymic hydrolysate and 1% NaCl. The maximum production of L-methioninase was obtained as 240.97 U/l after performing the experiments at these optimum data variables, which was increased by 1.61-fold in compare to classical method. The kinetic study was further performed at these optimum media components and the production of L-methioninase was found as non-growth associated behavior. Logistic equation and modified Luedeking–Piret equation were further employed to develop mathematical model for growth kinetic and production kinetic. These results would be significant for large scale production of these enzymes using bioreactors in industry.

29 ref

005491   CHIDAMBARAM P, JEYPRAKASH A, CHINNATHAMBI P (Botany Dep, Government Arts Coll, Melur, Tamil Nadu) : Characterisation of carrageenan extracted from fresh and defatted red algae along the Pamban coast, Tamil Nadu, India. Vegetos 2019, 32(3), 281-7.

Studies carried out to breakdown the yield and Fourier-Transform Infrared (FTIR) spectral properties of polysaccharide from Corallina elongata (J. Ellis & Solander), Liagora mannarensis (V. Krishnamurthy & Sundararajan) and Portieria hornemannii (Lyngbye). Carrageenan yield in the range of 16.4 to 24.2% in fresh algae than 14.2 to 23.8% defatted algae was recorded. Spent biomass in the range of 62.0 to 72.0 % in fresh algae than 61.2–70.4 % defatted algae was recorded. Total sugar, reducing sugar and sulphate were maximum in P. hornemannii (Lyngbye) and minimum in C. elongata (J. Ellis & Solander) having fresh algae carrageenan compared to the defatted algae carrageenan. FTIR spectroscopy study shows that the carrageenan extracted from fresh and defatted C. elongata (J. Ellis & Solander) have both kappa and iota type carrageenan. Fresh and defatted carrageenan from L. mannarensis (V. Krishnamurthy & Sundararajan) and P. hornemanni (Lyngbye) have kappa, lamda and iota type carrageenan.

27 ref

005547   SINGH M, BHATTI S, VERMA S K (Biotechnology Dep, Chhatrapati Shahu Ji Maharaj Univ, Uttar Pradesh- 208 024) : Improved plant regeneration method of Artocarpus lakoocha Roxb. from immature seeds. Vegetos 2019, 32(3), 269-74.

An efficient in vitro propagation method is developed for Artocarpus lakoocha Roxb. through direct shoot regeneration from cotyledonary node region of seedlings developed from immature seeds. The immature seeds were germinated on Murashige and Skoog (MS) medium having different concentrations of either 6-benzyl aminopurine (BA) or kinetin or Thidiazuron and subsequently, the cotyledonary node with primary shoot was transferred to MS medium without any growth regulator for shoot multiplication. Maximum (7.23 ± 0.46) shoots regenerated when immature seeds were cultured for 21 days on MS medium having 4.44 µM BA for germination. For rooting, shoots were pulse treated for 48 h with different concentrations of Indole-3-acetic acid (IAA) or Indole-3-butyric acid (IBA) followed by transfer on agar gelled MS basal medium. About 94.45% shoots rooted on pulse treatment with 5.0 µM IBA. Plantlets obtained after rooting were hardened and acclimatized into soil with 85–90% survival. The plantlets established into soil had similar vegetative morphology to the mother plant.

22 ref

005497   ELHADDAD N S, BELKASEM E M, KHATAB H A (Botany Dep, Omer Almukhtar Univ, Al-Baida, Libya, Email: enjesaad@yahoo.com) : Assessing of mutagenicity of monosodium glutamate by using HGPRT gene mutation assay of Coprinopsis cinerea. Ann Biol Res 2019, 10(2), 7-13.

The HPRT gene mutation assay is a remarkable tool for testing genotoxic chemicals, allows for the isolation and screening mutation in different living cells. Here, we report the applicability of HGPRT gene to test the mutagenic activity of different concentrations of monosodium glutamate, MSG, (1, 3, 5 and 7 g/l). The mutagenic effect of monosodium glutamate was assessed by decreasing the viability and increasing the HGPRT gene mutation rate of Coprinopsis cinerea. The alterations were proportional to the concentration of MSG up to reach the optimum concentration (7 g/l) when the maximum rate of mutation was 1.7 and the viability was about 27 %. The highest viability was 37.4 % when oidia treated with 2 g/l of MSG. To determine the optimal mutagenic time of the optimal mutagenic concentration, the period of incubation was ranging from 1 to 4 hours of the cells treated with 7 g/l MSG. Two hours was the optimal mutagenic time that reaches 1.02 mutagenic rate of HGPRT gene mutation and 37.7 % of the viability. The highest viability was gained is 43.6 % when cells treated with 7 g/l MSG and incubated for one hour. In vitro results indicates that the MSG is mutagenic and subsequently may cause DNA damage. These data do provide an indication of potential genotoxic of MSG to human health. Thus its use as a food additive should be completely avoided and look for a safer alternative.

4 illus, 31 ref

005499   FEOKTISTOVA NATALYA A, VASILYEV DMITRY A, MASTILENKO ANDREY V, SULDINA EKATERINA V, MALLYAMOVA ELVIRA S , NAFEEV ALEXANDER A, TOIGILDON A L, TOIGILDINA I A, OBUKHOV IGOR L, SHMORGUN B I (Ulyanovsk State Agrarian Univ, Russia, Email: atoigildin@yandex.ru) : Development of PCR detection system of bacteriophages Pr₄ UGSHA, E₇ ULSAU and Ye₅ ULSAU. Ambient Sci 2019, 6(2), 26-30.

Development of PCR detection system of bacteriophages Proteus phage (Pr₄-UGSHA ), Enterobacter phage (E₇-ULSAU) and Yersinia phage (Ye₅-ULSAU) are presented. Phylogenetic tree of concordance of their genetic organization against each other was built which was between 24 to 31%. It was determined that specific fragment for Proteus phage (Pr₄-UGSHA) is in the genome region of 3700-4500 bps. Highly specific fragments typical only for Enterobacter phage (E₇-ULSAU and Yersinia phage Ye₅-ULSAU in studied genomes were not found, however, we didn’t find regions during sharing of PCR that will allow detecting the only genome of a specific group. Two regions were established, their parallel determination was specific for Enterobacter phage and for Yersinia phage. In the BLAST system for parallel determination of genome fragments specific for groups of studied bacteriophages were determined. Finally, primer systems for PCR typing of bacterio phages were developed. They allowed to carry out identification of bacterio phages proteous, Enterobacter and Yersinia, referring to specific groups in material obtained out of environment samples and pathological material without detachment of pure growth in screenings of specified groups during detection of genome fragment in 125 bps in size over the range of 3700-4500 bps of DNA Proteus phage (Pr₄-UGSHA ); sizes of 294 and 431 bps in size over the range of 17500-18000 and 26500-27500 bps of DNA Enterobacter phage (E₇-ULSAU) and 226 and 85 bps in size over the range of 2000-2500 and 24100-24300 bps of Yersinia phage (Ye₅-ULSAU).

7 illus, 2 tables, 22 ref

005493   CHUNDURI J R, SINGHI N V (Biotechnology Dep, Chauhan Institute of Science, Mumbai- 400 056, Email: jayapradachunduri@gmail.com) : Immobilised aquaprobiotics of marine microbial origin and their efficacy. Ambient Sci 2019, 6(2), Online.

Marine waters contain millions of disease-causing and friendly bacteria which have the capacity to solubilize the insoluble compounds. Such live bacteria are of use to improve the health of wastewaters and their quality. Phosphate solubilizing bacteria solubilize the insoluble phosphate by digesting the compounds which can be considered as aquaprobiotics. An attempt has been made to isolate and characterize the phosphate solubilizing bacteria from the Indian coastal waters. Their capability to remediate the waste waters rich with phosphates. Assessed. Microbiological analysis and molecular identification confirmed that two isolates among the seven variants were Gram-positive and belonged to Klebsiella spp. The immobilized PSBs with sodium alginate on different inert substrata were tested for their phosphate solubilizing activity in the waste waters. The study indicated that both variants have good capability, increased longevity, and reusable conditions. PSB based aquaprobiotics can be of great use in the treatment of waste waters and plants.

3 illus, 1 table, 24 ref

005506   JAHROMI R R, MORADI F, ERFANIAN S, FARAJI S Z, ZARGAR M F, HAGHIGHI B R, SHAKERI M, SHAKERI H, JAHROMI A S (Jahrom Univ of Medical Sciences, Jahrom, Iran, Email: sotodehj2002@yahoo.com) : Molecular analyses of the prevalence of campylobacter detected from the poultry meat and its by products. Ambient Sci 2019, 6(2), 7-10.

Campylobacter is one of the most common bacteria that is usually transmitted between humans and animals. Among the animal source foods (ASF), the presence of the Campylobacter spp. in poultry meat has been extensively reported, while the consumption of under cooked meat is the most important means of transmission of this bacterium to humans. Hence, determining the prevalence of C.jejuni and C.coli in poultry meat and by products through the bacteriological and molecular methods was the pivot of this research. This descriptive cross-sectional study was conducted on poultry carcasses collected from the slaughterhouses of Jahrom-Iran. The presence of Campylobacter spp. in the samples was checked through bacteriological and molecular methods. Obtained data were analyzed by SPSS-16 on the descriptive statistics level. In the bacteriological examination, 0.91 % and 3.98 % of the samples were found to be infected with C.jejuni and other Campylobacter spp., respectively. In the molecular examination, 9.8 %, 1.2 %, and 0.9 % of the samples were found to be infected with C. coli, C.jejuni and other Campylobacter spp., respectively. Thus, it was concluded that the prevalence of the Campylobacter species in poultry meat, Jahrom is lower than their prevalence in the poultry meat in other cities of Iran and other countries.

1 illus, 2 tables, 25 ref

005524   NANDHINI S, MANGALA G A, BARANIDHARAN G R, MEENAMBIGAI T V (Animal Biotechnology Dep, Madras Veterinary Coll, Chennai-7, Email: gowrivalavan1@gmail.com) : Real-time expression of stemness and self-renewal genes in canine hematopoietic progenitor cells. Agric Sci Dig 2019, 39(3), 254-6.

Blood cells are responsible for constant maintenance and immune protection of every cell type of the body and this relentless and brutal work requires cells that have the greatest powers of self-renewal and are designated as Hematopoietic progenitor cells (HPCs). Peripheral blood stem cells in circulation have become the most common source of hematopoietic stem cells intended for transplantation after minimal manipulation. Homeo box (Hox), sonic hedgehog (SHH), and Wingless-type MMTV integration site family (Wnt) are known to modulate the self-renewal and expansion of hematopoietic progenitor/stem cells in humans and mice. Unlike cytokines, Hox, SHH, and Wnt are highly conserved among species from flies to humans but studies regarding the self-renewal and expansion of the HSC are extremely limited in dogs.

17 ref

005536   PRIYADHARSHINI B, VIGNESH M, PRAKASH M, ANANDAN R (Genetics and Plant Breeding Dep, Annamalai Univ, Annamalai nagar, Tamil Nadu, Email: geeth_prakash@yahoo.co.in) : Comparison of methods for genomic deoxyribonucleic acid (DNA) extraction suitable for whole-genome genotyping in traditional varieties of rice. Agric Sci Dig 2019, 39(3), 228-31.

The utilization and conservation traditional rice genotypes have attracted global attention. Optimization of DNA isolation protocol for genetic characterization of plants is a necessary and primary step. Over the recent years, next generation sequencing and microarray technologies have revolutionized scientific research, with their applications to high-throughput analysis of biological systems. Isolation of high quantities of pure, intact, double stranded, highly concentrated DNA is prerequisite for successful and reliable large-scale genotyping analysis. Therefore, standardization of DNA isolation is a basic requirement. Here we employed three methods of DNA isolation namely, Dellaporta, Hi-purA and modified CTAB techniques for isolation of genomic DNA from 25 indigenous rice genotypes. From the results, it was found that genomic DNA isolated by modified CTAB method to be the most appropriate for extracting high quality and maximum quantity DNA suitable for genotyping.  The quality and quantity of the differentially extracted DNA was subsequently assessed by spectrophotometric measurements and gel electrophoresis.

15 ref

005512   KAUR R P, DEVI S (Central Potato Resaerch Station, Jalandhar, Punjab) : In planta transformation in plants: A review. Agric Rev 2019, 40(3), 159-74.

In planta transformation has been established as an innovative and simple technique involving direct transformation of plant parts without involving the tedious tissue culture step. Methodologies of in planta transformation involve different plant parts and strategies. Agrobacterium strain carrying the gene of interest is targeted to the specific plant part either directly or in the induction medium. Vacuum infiltration is sometimes used to facilitate foreign gene integration. Initial studies of this novel technique involved treatment of whole plants with the inoculum, and later shifted to treatments of shoot tips and floral parts and finally the female reproductive parts have been targeted. Zygotes, embryos and seeds have recently been used extensively yielding successful transformation. The method is simple, convenient and overcomes the problem of tissue culture induced genetic variability in the transformants. The review traces the origin and development in the in planta methodologies used over the past and the various parameters considered by the workers for increased effectiveness viz. developmental stages, Agrobacterium stain, surfactant, induction medium etc., in the various crops compiled. Based on the review it may be inferred that there is immense potential in planta transformation, and the ease of regeneration and selection of transformants in the methods described, can be utilized for crop improvement.

1 table, 116 ref

005003   POWALE K, KAMBLE B, CHAUHAN N (Biotechnology and Natural Products Dep, Karnavati Univ, Gandhinagar- 382 422, Email: drneelam@ksd.ac.in) : Biocomputational approaches towards deciphering anti dengue viral properties of synthetic and natural moieties. Adv Hum Biol 2019, 9(3), 198-202.

Dengue, an arthropod-borne disease caused due to dengue virus belonging to Flaviviridae, is a serious health problem globally. Currently, there is no licensed vaccine for prophylaxis of the infection or an effective drug regimen for treatment. The virus genome codes for three structural and seven non-structural proteins. Envelope protein is required for the attachment and binding of the virus to the host cells, viral replication and hence, it can act as a good antiviral target.  We intend to evaluate the antiviral activity of compounds from both natural and synthetic sources by using tools of bioinformatics and computational biology. The favourable sites for drug binding, ligand interaction were analysed by various modules of Schrodinger software (2016‑1). Results indicated the amino acids – cysteine 3, arginine 2, threonine 155, tyrosine 132 and asparagine 194 show major interactions such as van der Waals and hydrophobic interaction with the different functional groups of the drug molecules. We observed the natural compounds such as rutin, gallic acid and ellagic acid showed better binding affinity in comparison to the synthetic antiviral drugs such as acyclovir, tenofovir and oseltamivir on different sites of the envelope protein suggesting the plausible anti-dengue viral property.

3 illus, 5 tables, 30 ref

004272   BUNUSHREE B, RAVICHANDRA B, RANGABHASHIYAM S, JAYABALAN R, BALASUBRAMANIAN P (Biotechnology & Medical Dep, National Institute of Technology, Rourkela- 769 008, Email: biobala@nitrkl.ac.in) : Qualitative analysis of biodiesel produced by alkali catalyzed transesterification of waste cooking oil using different alcohols. Indian J Chem Technol 2019, 26(4), 330-6.

The present study evaluates the nature of fatty acid methyl esters (FAMEs) formed through alkali-catalyzed transesterification of waste cooking oil (WCO) using methanol, ethanol as well as in combination, where the sequential addition of ethanol followed by methanol is done keeping the molar ratio of alcohol to oil constant (5:1), with sodium hydroxide as catalyst. A substantial reduction in reaction time from 8 h to 20 min is seen in the latter case. Further, the gas chromatography/mass spectrometry (GC-MS) analysis of the transesterified oil show a significant presence of FAMEs. Transesterified oil obtained from a combination of both the solvents show substantial quantities of unsaturated FAMEs [linoleic acids (41.89%), palmitelaidic acid (7.97%)], saturated FAMEs [stearic acids (4.62%), arachidic acids (2.54%)]and minor fraction of other acids. Hence, the utilization of WCO with the use of combined solvent system for transesterification, appear to have a great potential for replacing the conventional substrates that are being used for biodiesel production without much compromising on engine modifications.

4 illus, 3 tables, 38 ref

004268   AJAMANI A, KUMAR R, BHARGAVA P, VATS S (Kurukshetra Univ, Kurukshetra- 136 119, Email: vatssidd@gmail.com) : Mathematically optimized production, purification and characterization of penicillin G acylase from soil bacterial isolates AA17A and AA17B. Indian J Biotechnol 2019, 18(3), 260-8.

This research article deals with production of industrial enzyme penicillin G acylase from soil bacterial isolates namely AA17A and AA17B, which are selected from 80 soil samples. The strains were selected based on qualitative (turbidity) and quantitative (HPLC) test for 6-aminopenicillanic acid (6APA) production. The enzyme was assayed for its activity and optimized for production of enzyme using design of experiments software (DOE) “Design Expert 8.0.7.1”. Optimization of enzyme production of four carbon sources (glucose, glycerol, sucrose and starch), four nitrogen sources (beef extract, tryptone, peptone and yeast extract), for temperature (25°C, 30°C, 35°C and 40°C), four pH (6, 7, 8, 9), four inoculum volumes (2.5 ml, 5.0 ml, 7.5 ml, 10.0 ml) and the phenyl acetic acid (PAA) level (0.15%, 0.17%, 0.185%, 0.2%). The penicillin acylase activity was enhanced to 1.2 fold under following optimized culture conditions: carbon source - glucose (8%), nitrogen source - beef extract (2%), pH 9.0, temperature 30ºC, phenyl acetic acid 0.185%, inoculum volume 5 ml. Approximately 1.22-fold purification from the initial culture broth was achieved during ammonium sulphate precipitation (70-80%) with a yield of 4.6% enzyme. The specific activity of the final partially purified enzyme was 13.73 IU/mg protein.

2 illus, 6 tables, 37 ref

004302   SAXENA D, SINGH A, SIROHI R (Post Harvest Process and Food Engineering Dep, Govind Ballabh Pant Univ of Agriculture and Technology, Pantnagar- 263 145, Uttarakhand, Email: ranjanabce@gmail.com) : Bioethanol production from pea hull aqueous extract as a novel substrate using Saccharomyces cerevisiae. Indian J Biotechnol 2019, 18(3), 253-9.

Use of agricultural wastes as substrate for the production of bioethanol can help to solve environmental problems caused by inadequate discharge of waste. The production of bioethanol by Saccharomyces cerevisiae using pea hull aqueous extract as a substrate was evaluated in this work. Response surface methodology was employed to study the effect of different parameters such as inoculum concentration (3-7%), fermentation time (24-71 h) and pH (4.4, 4.8 and 5.2) on the production of bioethanol. Results revealed that highest bioethanol yield of 1.65% occurred at 6.8% inoculum concentration; 71 h of fermentation time in the medium with initial pH 5.2. The study highlights that pea hull aqueous extract can be utilized for bioethanol production with extended fermentation times.

3 illus, 3 tables, 25 ref

004280   JAYAMUTHUNAGAI J, BHARATHIRAJA B, SELVAKUMARI I A E, VARJANI S (Anna Univ, Chennai- 600 025, Tamil Nadu, Email: btrbio@gmail.com) : Polymerase chain reaction and real-time PCR parameters for amplification of hydrolytic microorganisms and hydrogenotrophic methanogens in anaerobic bioreactor. Indian J Biotechnol 2019, 18(3), 246-52.

The objective of the present research is to envisage microbial assorted variety in the reactor through systems of general polymerase chain reaction (PCR) and real-time PCR, which can be further applied for production of methane using anaerobic reactor by employing vegetable waste as a feed. The outcomes proposed the names of species identified with the hydrolytic microorganisms, for example, Bacillus subtilis and hydrogenotrophic methanogenic archaea like i.e. Methano culleus and i.e. Methano corpusculum of the methanogenic microbes. Results demonstrated that, presence of methanogenic archaea in the reactor was confirmed using general PCR.

5 illus, 2 tables, 25 ref

004285   MOHANTY C S, SINGH V, GUPTA P, SRIVSATAVA A, PATTANAYAK R, JAISWAL V, DIKSHIT N (Plant Molecular Biology and Genetic Engineering Div, CSIR-National Botanical Research Institute, Lucknow-226 001, Uttar Pradesh, Email: cs.mohanti@nbri.res.in) : Estimation of genetic diversity, structure and trait association of winged bean (Psophocarpus tetragonolobus (L.) DC.), genotypes through AFLP and ITS markers. Indian J Biotechnol 2019, 18(3), 235-45.

The genetic diversity among ninety five accessions of winged bean (Psophocarpus tetragonolobus (L.) DC.) belonging to six countries of African and Asiatic origin was detected by employing amplified fragment length polymorphism (AFLP) and internal transcribed spacer (ITS) of nuclear ribosomal DNA markers. A medium to low-level of genetic diversity exists among the accessions of P. tetragonolobus. Association between the AFLP markers and flower, pod and seed traits: days to 50% flowering (DFW), pod length (PDL), pod width (PDW), green pod length (GPL), number of pods per plant (PDSP), number of seeds per pod (SDPD), 100 seed weight (SWT) and seed-oil content (SOC) was carried out by employing the mixed linear model (MLM) based kinship matrix (K-model). Traits like SOC and PDW were found to be strongly associated with AFLP markers. Population structure analysis among the genotypes identified five discrete sub-populations among the studied African and Asiatic lines of P. tetragonolobus.

3 illus, 4 tables, 31 ref

004279   JAISWAL S K, MUTHUSAMY V, HOSSAIN F, ZUNJARE R U, GOSWAMI R, CHHABRA R, CHAND G, DOSAD S, BHOWMICK R, GULERIA S K, et al. (ICAR-Indian Agricultural Research Institute, New Delhi, Email: fh_gpb@yahoo.com) : Characterization of maize genotypes using microsatellite markers associated with QTLs for kernel iron and zinc. Indian J Biotechnol 2019, 18(3), 224-34.

Crop genetic resources rich in Fe and Zn provide sustainable and cost-effective solution to alleviate micronutrient malnutrition. Maize being the leading staple crop assumes great significance as a target crop for biofortification. We report here wide genetic variation for kernel Fe and Zn among 20 diverse maize inbreds lines, majority of which were bred for quality protein maize (QPM) and provitamin-A. Kernel Fe ranged from 30.0 - 46.13 mg/kg, while kernel Zn ranged from 18.68-39.56 mg/kg. Moderate but positive correlation was observed between the micronutrients. Characterization using 25 Single sequence repeats (SSRs) linked to QTLs for kernel Fe produced 58 alleles. Similarly, 86 alleles were identified from 35 SSRs linked to QTLs for kernel Zn. One unique allele for kernel Fe and three unique alleles for kernel Zn were identified. The mean polymorphic information content (PIC) was 0.40 for both kernel Fe and Zn. Jaccard’s dissimilarity coefficients varied from 0.25 - 0.91 with a mean of 0.58 for kernel-Fe while 0.27- 0.88 with a mean of 0.57 for kernel Zn. Principal coordinate analysis depicted diversity of inbreds. Cluster analysis grouped the inbreds into three major clusters for both kernel Fe and Zn. Potential cross combinations have been proposed to develop micronutrient rich hybrids and novel inbreds with higher Fe and Zn. The information generated here would help the maize biofortification programme to develop nutritionally enriched hybrids.

5 illus, 3 tables, 40 ref

004306   SINGH J P, KUMAR R R, GOSWAMI S, RAI G K, SAKHARE A, KUMAR S, SINGH S D, BAKSHI S, JAMHULKAR S J, SAREEN S, ET AL. (Biochemistry Dep, Indian Agricultural Research Institute, New Delhi- 110 012, Email: ranjeetranjaniari@gmail.com) : A putative heat-responsive transcription factor (TaHD97) and its targets in wheat (Triticum aestivum) providing thermotolerance. Indian J Biotechnol 2019, 18(3), 214-23.

Transcription factors (TFs) are protein, which perform their role at transcriptional level by affecting the expression of various genes associated with metabolic pathways, growth and stress-associated genes (SAGs) at different developmental stages. Here, we identified 38 novel heat-responsive transcription factor genes from wheat cv. HD2985 by mining the de novo transcriptome data derived from heat shock (HS) treated wheat. Based on digital gene expression (DGE), a putative transcript (TaHD97) of ~1.1 kb was amplified and cloned from wheat cv. HD2985. The presence of heat stress transcription factor (HSF) DNA binding domain was observed in the amino acid sequence. Differential expression of TaHD97 was observed in HD2985 (thermotolerant) and HD2329 (thermosensitive) under heat stress. Tissue specific expression analysis showed up-regulation of TaHD97 in leaves, stem and endospermic tissues and down-regulation in root under HS. A positive correlation was established between the expression of TaHD97 and its target gene (HSP17 and HSP90) in wheat under heat stress. HSP17 transcripts were observed more in leaves of HD2985, as compared to HD2329. Thermotolerance related biochemical enzymes (SOD, CAT, GPX and TBARS) were observed higher in wheat cv. HD2985 showing maximum expression of TaHD97 under heat stress. There is a need for the functional validation of the gene TaHD97 in order to use it for the regulation of sHSP (catalytic chaperone) - a novel approach towards augmenting thermotolerance in wheat under heat stress.

5 illus, 2 tables, 23 ref

004311   VASANTRAO J M, PANKAJ Y K, KUMAR R (Agricultural Biotechnology and Molecular Biology Dep, Dr. Rajendra Prasad Central Agricultural Univ, Samastipur- 848 125, Bihar, Email: mvjagadale@gmail.com) : Characterization of wheat (Triticum aestivum L.) genotypes unraveled by molecular markers considering heat stress. Indian J Biotechnol 2019, 18(3), 204-13.

The current study focuses and emphasis on the potential of heat stress to negatively affect crop physiology. Here, we have screened 19 wheat (Triticum aestivum L.) genotypes for their tolerance of heat stress. Significant differences were observed among the genotypes for all the traits under consideration. Exploitable extent of genetic variability amongst the entries was present as revealed by considerably higher estimates of mean percentage. On the basis of heat susceptibility index genotypes, Halna, Mon’s Ald’s, Cuo/79/Prulla and K 307 were identified as heat-tolerant whereas SAWSN 3041, SAWSN 3101 and K 0583 were identified as heat-susceptible. Seventeen wheat microsatellite markers were capable of detecting 89 alleles with an average of 4.6 alleles per locus. Polymorphism information content (PIC) value ranged from 0.16 for the primer XGWM 516 to 0.83 for DUPW 117 with an average of 0.60. A perusal of similarity coefficients clearly reflected that a very high degree of similarity exists between wheat variety Mon’s Ald’s and SAWSN 3101 (0.70). On the other hand, the two most distantly related cultivars were found to be AKAW 4008 and PBW 343 (0.034). Marker BARC 4, BARC 170, BARC 311, PSP 3058, WHE014. H04 and GWM 458 were strongly associated with the heat tolerance for traits TGW and BARC 311 were strongly associated with terminal heat tolerance for number of grains/plant, respectively. Considering all the parameters it is adjudged that relatively stable genotypes may be evaluated at various agro-climatic regions for grain yield and heat tolerance along with other contributing characters and ideal plant type.

1 illus, 6 tables, 24 ref

004303   SELOKAR N L, SAINI M, KUMAR D, SHARMA R K, YADAV P S (Animal Physiology and Reproduction Dep, ICAR-Central Institute for Research on Buffaloes, Hisar, Haryana, Email: psycirb@gmail.com) : 10 years after the birth of India’s first cloned farm animal, where is buffalo cloning heading. Indian J Biotechnol 2019, 18(3), 191-2.

India owns the best buffalo breeds, particularly Murrah which is famous all over the world for high milk production. India’s white and pink revolution cannot be imagined without the contribution of buffalo and to achieve these, the best productive animals need to be produced through scientific interventions. Animal cloning is a technique used to produce multiple copies of the best animals without normal reproduction. In India, buffalo cloning has already happened and India’s first cloned buffalo was produced in 2009. Later, several buffalo clones were produced and attempts are ongoing to produce stock of more elite animals. Buffalo cloning has made its way from scientific curiosity to farmer’s farm. In this viewpoint article, we provide an overview of the progress of buffalo cloning and we discuss some of the public perceptions of animal cloning such as aging and food safety of cloned animal products.

1 illus, 8 ref