Yeasts from Chinese strong flavour Daqu samples: isolation and evaluation of their potential for fortified Daqu production

In total, 16 yeast were isolated from Chinese strong flavour Daqu samples and underwent RAPD analysis and identification. Totally, 11 different species were identified among these isolates including Saccharomyces cerevisiae, Hanseniaspora vineae, Pichia kluyveri, Trichosporon asahii, Wickerhamomyces anomalus, Kluyveromyces lactis, Yarrowia lipolytica, Wickerhamomyces mori, Galactomyces geotrichum, Dabaryomyces hansenii, and Saccharomyces kudriavzevii. To understand the impact of these yeast strains on the quality and flavour of Daqu, we then assessed volatile compounds associated with Daqu samples fermented with corresponding strains. These analyses revealed strain YE006 exhibited the most robust ability to produce ethanol via fermentation but yielded relatively low quantities of volatile compounds, whereas strain YE010 exhibited relatively poor fermentation efficiency but produced the greatest quantity of volatile compounds. These two yeast strains were then utilized in a mixed culture to produce fortified Daqu, with the optimal inoculum size being assessed experimentally. These analyses revealed that maximal fermentation, saccharifying, liquefying, and esterifying power as well as high levels of volatile compounds were achieved when using a 2% inoculum composed of YE006/YE010 at a 1:2 (v/v) ratio. When the liquor prepared using this optimized fortified Daqu was compared to unfortified control Daqu, the former was found to exhibit significantly higher levels of flavour compounds and better sensory scores. Overall, our findings may provide a reliable approach to ensuring Daqu quality and improving the consistency and flavour of Chinese strong-flavour liquor through bioaugmentation.

The genetic potential of Taxus sumatrana medicinal plants in Kerinci Regency

Taxus -a taxol-producing medicinal plant that mostly found in highland area- is a species in genus Taxus and family Taxaceae. This study was aimed to determine the genetic diversity within and between population of T. sumatrana in Kerinci Regency, i.e. Mount Kerinci and Mount Tujuh, based on altitude. The genetic diversity was analyzed with RAPD analysis. The altitude was categorized as low (<2000 m asl) and high (>2000 m asl). The cambium extraction was carried out based on CTAB method. DNA amplification was conducted in RAPD method on machine of PCR System 9700 Applied Biosystems. Nine RAPD primers were used in this study. The results revealed that the average of polymorphic locus was 53.89%. Genetic diversity within population was fairly high with value of 0.1799 and Shannon index of 0.2746. Among the four populations, the population of High Tujuh showed the highest level of variability (He=0.2044). The Nei genetic distance between populations was ranging from 0.0567 to 0.1302. The potential of High Tujuh population is still large enough so that it can still be explored for genetic conservation and cultivated as a taxol-producing material which is useful for medicine.

Strain Typing of Trichosporon asahii Clinical Isolates by Random Amplification of Polymorphic DNA (RAPD) Analysis

Objective The aim of this study was to identify and isolate Trichosporon asahii (T. asahii) from clinical samples and to assess the genetic relatedness of the most frequently isolated strains of T. asahii using random amplification of polymorphic DNA (RAPD) primers GAC-1 and M13. Methods All the clinical samples that grew Trichosporon species, identified and confirmed by polymerase chain reaction (PCR) using Trichosporon genus-specific primers, were considered for the study. Confirmation of the species T. asahii was carried out by T. asahii-specific PCR. Fingerprinting of the most frequently isolated T. asahii isolates was carried out by RAPD using random primers GAC-1 and M13. Results Among the 72 clinical isolates of Trichosporon sp. confirmed by Trichosporon-specific PCR, 65 were found to be T. asahii as identified by T. asahii-specific PCR. Fingerprinting of the 65 isolates confirmed as T. asahii using GAC-1 RAPD primer yielded 11 different patterns, whereas that of M13 primer produced only 5 patterns. The pattern I was found to be the most predominant type (29.2%) followed by pattern III (16.9%) by GAC-1 primer. Conclusions This study being the first of its kind in India on strain typing of T. asahii isolates by adopting RAPD analysis throws light on genetic diversity among the T. asahii isolates from clinical samples. Fingerprinting by RAPD primer GAC-1 identified more heterogeneity among the T. asahii isolates than M13.

Genetic Characterization of Swamp Eel of Bangladesh Through Dna Barcoding and Rapd Techniques

The freshwater air-breathing swamp eel Monopterus spp. are native to the freshwater of Bangladesh and throughout the Indian subcontinent. To identify the different swamp eel species and to check the genetic diversity among them, a total of twelve swamp eel specimens were collected from four districts (Tangail, Bogura, Bagerhat and Sylhet) representing the four division of Bangladesh. The extracted DNA from twelve fish samples was amplified by the PCR technique for DNA barcoding and RAPD analysis. Among 12 specimens, 8 specimens showed a 95-100% similarity with M. cuchia species published in the NCBI GenBank database and BOLD system. The studied mct3 (collected from Tangail region), mcs1, mcs2 and mcs3 (collected from Sylhet region) specimens showed about 83% homology with Ophisternon sp. MFIV306-10 as per BLAST search; whereas BOLD private database showed 99% similarity with Ophisternon bengalense (Bengal eel). From the phylogenetic tree analysis, 8 samples were clustered with M. cuchia and 4 samples showed similarity with Ophisternon sp. MFIV306-10 and Ophisternon bengalense _ANGBF45828-19. In RAPD-PCR based analysis, it was found that the maximum genetic distance (1.6094) was observed between mcba2 and mcs3, while between mct1 and mct2, the minimum genetic distance was 0.000. A total of 192 bands, of which 35 were polymorphic with 17.88% polymorphisms among swamp eel species and the size of the amplified DNA fragments ranged from 250 to 1800 bp. The information on DNA barcoding and RAPD analysis help measure the genetic diversity among swamp eel species, ensure the reliability of the published taxonomic information, and initiate proper management programs to conserve these vulnerable species to meet future export demand. Asiat. Soc. Bangladesh, Sci. 46(2): 117-131, December 2020

Assessment of polymorphism using RAPD in the generalized hemiparasitic plant Helicanthes elasticus (Desv.)Danser collected from six different hosts

RAPD analysis was carried out to determine genetic diversity that occurred among the selected accessions of H.elasticus obtained from six different hosts. All together 10 random primers were used for the RAPD assay, of which primer S11 produced 12 amplified bands in which 10 are polymorphic. Primer S9 produced the least number of amplified fragments. 10 primers together resulted in 88 amplified fragments, of which 26 are found to be polymorphic. Considerable polymorphism was shown by 3 primers that can be represented in a descending order S11>S79>S10. Accessions of H.elasticus obtained from Anacardium occidentale and Citrus maxima produced more polymorphic bands with S11. The overall polymorphism obtained in the selected samples of this hemiparasite was 29.54 indicating that genetic similarity was more among these samples rather than genetic diversity.

Molecular characterization of Canna indica L. based on random amplified polymorphic DNA markers

Random amplified polymorphic DNA (RAPD) markers were employed for characterization, assessment of genetic variation and inferring relationships among six variants of Canna indica L. A total of 198 RAPD bands ranging from 200 bp to 3 kbp were generated by all the six variants. Among them, most of the bands were found to be polymorphic, four band were unique of which two bands (OPA022000 and OPA043000) were observed in the variant 2 (small red) and the other two (OPA013000 and OPA053000) were noticed in the variant 4 (orange), and the remaining bands were found to be monomorphic. The pair-wise genetic distance was determined among the six variants that ranged from 0.1446 to 0.6554. A dendogram was constructed based on the RAPD profiling to infer the relationship among the six variants of C. indica that resulted in two major clusters: the first one contained two variants, viz. variant 1 (local red) and variant 2 (small red), while the second cluster composed of the remaining four variants. The results as revealed from the RAPD analysis were found congruent with those of morphological and anatomical investigation of the species. Bangladesh J. Plant Taxon. 28(1): 75-81, 2021 (June)

Comparative Analysis Between Paramecium Strains with Different Syngens Using the RAPD Method

Paramecium spp. is types of free-living protists that live in freshwater environments. They are ciliates with high motility and phagocytosis and have been used to analyze cell motility and as a host model for pathogens. Besides such biological characteristics, apart from the usual morphological and genetic classification of species, the existence of taxonomies (such as syngens) and mating types related to Paramecium’s unique reproduction is known. In this study, we attempted to develop a simple method to identify Paramecium strains, which are difficult to distinguish morphologically, using random amplified polymorphic DNA (RAPD) analysis. Consequently, we can observe strain-specific band patterns. We also confirm that the presence of endosymbiotic Chlorella cells affects the band pattern of P. bursaria. Furthermore, the results of the RAPD analysis using several P. caudatum strains with different syngens show that it is possible to detect a band specific to a certain syngen. By improving the reaction conditions and random primers, based on the results of this study, RAPD analysis can be applied to the identification of Paramecium strains and their syngen confirmation tests.

PHÂN TÍCH ĐA DẠNG DI TRUYỀN GIỐNG ỚT XIÊM ĐỊA PHƯƠNG Ở QUẢNG NGÃI BẰNG CHỈ THỊ RAPD

Sự đa dạng di truyền của 15 mẫu giống ớt Xiêm địa phương của Quảng Ngãi và mối quan hệ giữa chúng với 5 giống ớt A Riêu và 5 giống ớt Bay đã được đánh giá dựa trên kết quả của 10 mồi RAPD. Hệ số tương đồng di truyền giữa 25 mẫu ớt dao động từ 0,56 đến 1,0. Hệ số tương đồng di truyền và sơ đồ về cây phát sinh loài đã phân ớt A Riêu của Quảng Nam và ớt Bay của Gia Lai thành các nhóm khác nhau. Ớt Xiêm của Quảng Nam thuộc vào cả 2 nhóm và việc phân chia ớt Xiêm vào các nhóm này có mối liên hệ mật thiết với hình dạng quả, Ớt Xiêm với dạng quả lớn thuộc cùng phân nhóm với ớt A Riêu, ớt Xiêm dạng quả nhỏ đến trung bình thuộc cùng phân nhóm với ớt Bay. ABSTRACT Genetic diversity of 15 Xiem chili accessions of Quang Ngai province and its relationship with 5 A Rieu chili accessions of Quang Nam and 5 Bay chili accessions of Gia Lai was assessed by using 10 PCR - RAPD primer. The genetic dissimilarity of the total of 25 accessions was ranged from 0,56 to 1,0. RAPD analysis combined with the construction of phylogenetic tree revealed that big Xiem accessions had the same cluster to A Rieu, while small to medium Xiem accessions had the same cluster to Bay.

Molecular Fingerprinting for Detecting Genetic Relationships among different Accessions of Pterocarpus marsupium

Pterocarpus marsupium Roxb. is a valuable multipurpose forest tree in India. Generally, it is valued greatly for its excellent wood qualities. Due to its significant multipurpose properties, this tree has been overexploited, which ultimately has led to its inclusion in the list of threatened species. In this regard, studying the genetic diversity in P. marsupium is not only significant for the protection of this species, but also necessary for the development and utilization of germplasm resources for its improvement. Before developing any tree improvement program, information on actual genetic diversity and the cryptic number of the differentiated genetic resource are important aids for its conservation and effective utilization. Thus, in the present study, analysis of phylogenetic relationship among P. marsupium species plays an important role in the identification and selection of elite genotype among the wildly distributed accessions. The phylogenetic relationship among 18 genotypes obtained from various forest regions of central India was studied using DNA based molecular markers. In RAPD analysis, out of 40 scorable amplified bands, 29 were polymorphic resulting in expression of polymorphism percentage (73.2%) with an average of 2.90 amplicons per primer. Based on RAPD analysis, the lowest (37%) similarities among accessions were recorded in Anuppur (MAA), Mandla (MMK) and Jabalpur (MJH) and the highest similarity (100%) were observed among Mandla (MMK), Jablapur (MJH); Jashpur (CJM), Surguja (CSA), Bilaspur (CBP) and Durg (CDB) and Raigarh (CRK) accessions. While the ISSR analysis found 66 amplified bands, 45 were polymorphous and average 68.3 percent polymorphic with an average 4.5 bands per oligo. The lowest (36%) similarity was observed among Anuppur (MAA) and Jabalpur (MJH) accessions and the highest similarity (88%) was recorded among Jashpur (CJM), Chhindwara (MCD) and Bilaspur (CBP) accessions. The combined analysis data of RAPD and ISSR showed that Chandrapur (RCC) and Anuppur's (MAA) acessions had the lowest (35%) similarity, with Jabalpur's (MJHs) and Mandla's (MMKs) accession being the highest similarities (100%) reported. As a result, the study of genetic diversity by means of RAPD and ISSR markers alone or in combination, i.e. the MAA, CKB and CRK accessions, was found to be more diverse among 18 accessions of Central India and given greater space for the collection of elite/superior trees to be used in conservation and forest development programs. © 2021 Friends Science Publishers

DETERMINATION OF GENETIC DIVERSITY IN PORTUNUS PELAGICUS (LINNAEUS, 1758) COLLECTED FROM GULF OF MANNAR

In the present study, the marine crab Portunuspelagicus was screened with 17 primers and made to score polymorphism. The main emphasis of the present study was to assess the genetic diversity at intra specific level among the 3 accessions of P. pelagicus species of Gulf of Mannarusing RAPD markers. RAPD analysis shows that there is a high level of polymorphism among different accessions. From this study, it was understood that each location varied with respect to environmental factors and genetic parameters. The OPB-18, OPB-19, OPC-07 and OPN-06 primers produced distinct, highly reproducible amplification profile for all the screened samples. In the present study, amplification bands ranged between 250 and 663 bp. Maximum numbers of bands were produced by OPN-06 and least by OPB-19. Moderate to high genetic diversity was observed in all geographic samples of P. pelagicus from RAPD analysis. RAPD analysis from three different geographical regions shows clear polymorphic patterns. Thoothukudi and Rameshwaram populations appear in one cluster, while the Kanyakumari populations formed the other cluster indicating a genetic variability and diversity in samples collected from different places.