Exploring Genetic Variability among and within Hail Tomato Landraces Based on Sequence-Related Amplified Polymorphism Markers

Landraces are valuable sources of genetic characteristics, which are of plant breeders’ interest to include in breeding programs for crop improvement. We assess the inter- and intra-genetic variability among 96 accessions representing three Hail tomato landrace using DNA-based marker sequence-related amplified polymorphism (SRAP). Seven SRAP primer combinations generated 55 alleles with a polymorphism of 100%, and an average of 7.86 polymorphic alleles per pair of primers. All primers showed an average of 0.68 polymorphic information content (PIC) value and discrimination power (DP) of 14.29. Principal coordinate analysis (PCoA) confirmed the clustering produced via the UPGMA similarity dendrogram allowed for the grouping of the 96 accessions according to its gene bank accession numbers and showed relatively good separation between landraces. A similarity value ranged from 0.04 to 1.0 among accessions of Hail 747, 0.05 to 1.0 in Hail 1072, and from 0.16 to 0.92 in Hail 548. These results showed the landraces harbor a wide range of genetic diversity at both inter and intra-variation levels. AMOVA showed that most of the genetic variation was because of differences within populations (87%). Tomato Hail landraces have well-differentiated genetic populations and admixtures, where Hail 747 formed their separate group, and both Hail 548 and Hail 1072 were admixed, and some accessions showed more diversity pattern. We have to take the SRAP technique’s effectiveness in the study of genetic variability among and within landraces into consideration in the tomato-breeding programs through marker-assessed selection.

Screening for Probiotic Potential of Lactobacillus Rhamnosus Strain CRD4

Lactic acid bacteria are the foremost used probiotic worldwide. Its successful application in manufacturing dairy products and probiotic foods makes it a promising industrial prerequisite. The objective of the present investigation was to isolate, identify, and molecularly characterize Lactic acid bacteria from local dairy samples of Odisha state of India and explores its probiotic traits. One potential strain was isolated using a selective Lactobacillus-MRS agar medium. The biochemical studies illustrated the bacteria were gram-positive, catalyze (-ve), and non-motile. The taxonomical diversity of the bacterium was analyzed by 16s RNA sequencing and classified as Lactobacillus rhamnosus strain CRD4 with gene bank accession no [MG573074]. Further, the selected strain was screened for its probiotic competence of lower acid and bile tolerance. The result confirmed that lactobacillus strain successfully defended the low pH and bile stress and acclaimed 70% cell surface hydrophobicity. Antibiotic studies obtained confirmed the possible resistance of the strain. The maximum zone of inhibition was expressed in diameter 42mm against Ciprofloxacin. In conclusion, based upon the above results, Lactobacillus rhamnosus can be a profound probiotic candidate.

Pectinolytic Potential of Differentbacillus Species Isolated From Lepidopteran Insects and Infected Soil

Background: The microorganisms are preferred over plants and animals for the bulk production of industrially important enzymes like pectinases because of their low production-cost, availability of fermentation raw material and the easily controllable production conditions. The pectinolytic potential of a large pool (130) of Bacillus cereus group and non-Bacillus cereus was evaluated. The effect of different physicochemical parameters on polygalacturonase(PGase) production by three promising strains, B. pumilusS76, B.thuringiensis S140(b) and B.taquilensis S140(c) with gene bank accession no. KU981112, KU981113 and KU981114, were studied and the enzyme was partially characterized. The enzyme activity was assayed using dinitrosalicylic acid method.Results: The polygalacturonases produced by Bp S76, Bt S140(b) and BtqS140(c) were thermostable at temperature above 55ºC with a melting temperature (Tm) of 72ºC,62ºC and 57ºC, respectively and the half-life (T1/2) at 60ºC of 50, 65 and 30 min., respectively. The catabolite repression was noted in presence of glucose while the production increased upon addition of 1% yeast extract to the growth medium. The enzyme activity was also induced in presence of Na+ and Ca+2.Conclusion: The strain Bp S76 appeared a better candidate for future biotechnological application owing to the production of higher titers (6.11 IU/L.hrs) of polygalacturonase with greater stability.

Isolation and identification of Bifidobacterium spp. from infant intestinal tract

As one of probiotic bacteria, Bifidobacterium is supplement in food industry such as in milk and yogurt. The advantageous impact of these bacteria for human health was reported. However, in Vietnam, supplement of these bacteria as probiotic source to the food was not much in food industry. In our experiment, Bifidobacterium are isolated from a feces sample of 7 days-child which only had mother milk. The bacterial isolated from infant intestinal tract was Bifidobacterium infantis (B. infantis, closest to Bifidobacterium longum ssp. infantis strain Bi-26 with gene bank accession CP054425.1). The techniques to identify these bacteria were MALDI Biotyper, PCR and sequencing. This experiment is original for further study the characters of these bacteria to apply them in our daily life.

Isolation and characterization of novel and efficient protease producing bacteria from drinking water resources

An attempt has been made to explore the stability of protease enzyme (isolated from Bacillus sp.) by statistical method. More than 100 isolates were screened for extracellular protease activity derived from various potable water samples of Mahabubnagar district, Telangana State, India. The activity of protease is found to be varying from sample to sample, the highest being reported by the isolate from water sample of Kalwakurthy mandal, Mahabubnagar district and therefore was selected for further studies. The 16S rRNA (ribosomal ribonucleic acid) gene sequence of the isolate showed closest similarity with Bacillus sp. and the sequence was submitted to National Center for Biotechnology Information (NCBI) gene bank (accession number GU566359) and the culture was deposited in three international culture deposition centers (KCTC-13725: MTCC-10465: JCM-16713). The present study revealed that, this Bacillus sp. showed a greater amount of protease production with the inherent characters of thermo, alkali and oxidant stability which makes it a potential alternative protease producing strain in various industrial applications.

Molecular Identification of Talaromyces islandicus Isolated from Clinical Sample (First Record in Iraq)

The analysis of ITS4 gene sequences has been the technique generally used to study and confirm the identification and taxonomy of fungi However, fungal species cannot always be distinguished from each other using cultural methods. Thus, clinical samples were collected from cases with aspergillosis infections, were applied for microbiological and molecular identification. DNA was extracted from Talaromyces islandicus and the ITS4 gene were amplified by using specific primer, then sequencing of nucleic acid of genes was performed by machine is AB13730XL, Applied Biosystem, Macro gen company, the DNA sequencing results of flank sense of ITS4 gene from Talaromyces islandicus was confirm the identification into species level: Talaromyces islandicus. analysis of the sequences appeared that there two substitution (Transversion, Transition) in the Talaromyces islandicus strain with Sequence ID LC540.1 location at Range of nucleotide from 9 to 77, 100% compatibility with NCBI while no substitution appeared in the Talaromyces islandicus Gene Bank accession number: KY30.1.

Isolation and Molecular Characterization of a Model Antagonistic Pseudomonas aeruginosa Divulging In Vitro Plant Growth Promoting Characteristics

The use of microbial technologies in agriculture is currently expanding quite rapidly with the identification of new bacterial strains, which are more effective in promoting plant growth. In the present study 18 strains of Pseudomonas were isolated from soil sample of Balochistan coastline. Among isolated Pseudomonas strains four designated as SP19, SP22, PS24, and SP25 exhibited biocontrol activities against phytopathogenic fungi, that is, Rhizopus microsporus, Fusarium oxysporum, Aspergillus niger, Alternaria alternata, and Penicillium digitatum; PS24 identified as Pseudomonas aeruginosa by 16srRNA gene bank accession number EU081518 was selected on the basis of its antifungal activity to explore its potential as plant growth promotion. PS24 showed multiple plant growth promoting attributes such as phosphate solubilization activity, indole acetic acid (IAA), siderophore, and HCN production. In order to determine the basis for antifungal properties, antibiotics were extracted from King B broth of PS24 and analyzed by TLC. Pyrrolnitrin antibiotic was detected in the culture of strain PS24. PS24 exhibited antifungal activities found to be positive for hydrogen cyanide synthase Hcn BC gene. Sequencing of gene of Hcn BC gene of strain PS24 revealed 99% homology with the Pseudomonas aeruginosa strain PA01. The sequence of PS24 had been submitted in gene bank accession number KR605499. Ps. aeruginosa PS24 with its multifunctional biocontrol possessions can be used to bioprotect the crop plants from phytopathogens.

158. EFFECT OF A59V LOCUS IN THE LEPTIN GENE ON LENGTH OF PREGNANCY IN IRANIAN HOLSTEIN COWS

We investigated effect of A59V polymorphism in the leptin gene on length of pregnancy. Blood was collected from 255 Holstein cattles belonging to four different herd managements in Isfahan province. Genomic DNA extracted from whole blood. Genotypes of A59V locus were identified PCR-RFLP technique. Amplified region is located in exon three of leptin gene. The genomic bovine leptin sequences, which consist of three exons, were obtained from Gene Bank (Accession number U50365). The polymerase chain reaction was used to amplify the 331 bp DNA fragments from genomic DNA. The PCR reaction contained 100 ng of genomic DNA, 0.3 µM of each primer, 1.5 mM MgCl2, 200 µM dNTP, 10mM Tris HCl, 50 mM KCl and 1 U Taq-polymerase in total volume of 20 µL. Sequences of primers that were used in PCR were reported previously by Haegeman et al. (2000). Conditions for PCR were 94°C for 2 min, followed by 35 cycles of 94°C for 30 s, 55°C for 1 min, and 72°C for 30 s. Followed by final extension for at for 15 min 72°C. Digestion of PCR product of 331 bp with 5 U of HphI (Fermentas) in 20 µL of reaction volume at 37°c for 8 h and analyzed on 8% no denature polyacrylamyde gel. Allele A in the A59V locus was the allele not digested by restriction enzyme, allele B was the restriction enzyme-digested PCR product. Digestion revealed 3 genotypes, AA (331 bp), AB (331, 311, and 20 bp), and BB (311 and 20 bp). Significances of the genotype effects were estimated by GLM procedure of SAS. This study showed that genotype effect on length of pregnancy were significant (P<0.01). Animals homozygous for allele A had higher length of pregnancy ((P<0.01, AA=279.17±0.47, AB=276.96±0.57, BB=274.8±2.2).