scholarly journals Mammalian species identification using ISSR-HRM technique

2021 ◽  
Vol 104 (2) ◽  
pp. 003685042110261
Author(s):  
Wannapimol Kriangwanich ◽  
Korakot Nganvongpanit ◽  
Kittisak Buddhachat ◽  
Puntita Siengdee ◽  
Siriwadee Chomdej ◽  
...  

Wildlife trading and the illegal hunting of wildlife are contributing factors to the biodiversity crisis that is presently unfolding across the world. The inability to control the trade of animal body parts or available biological materials is a major challenge for those who investigate wildlife crime. The effective management of this illegal trade is an important facet of wildlife forensic sciences and can be a key factor in the enforcement of effective legislation surrounding the illegal trade of protected and endangered species. However, the science of wildlife forensics is limited by the absence of a comprehensive database for wildlife investigations. Inter-simple sequence repeat markers (ISSR) coupled with high resolution melting analysis (HRM) have been effectively used for species identification of 38 mammalian species. Six primers of the ISSR markers were chosen for species identification analysis. From six ISSR primers resulting in a range of accuracy of 33.3%–100% and 100% in terms of precision in every primer. Furthermore, 161 mammalian samples were 100% distinguished to the correct species using these six ISSR primers. ISSR-HRM analysis was successfully employed in determining mammal identification among varying mammalian species, and thus could serve as an effective alternative tool or technique in the species identification process. This option would offer researchers a heightened level of convenience in terms of its performance and the ease with which researchers or field practice veterinarians would be able to interpret results in effectively identifying animal parts at wildlife investigation crime scenes.

2010 ◽  
Vol 39 (1) ◽  
pp. 119-122 ◽  
Author(s):  
Mahmudul Islam Nazrul ◽  
Fan Xiao Lin ◽  
Bian Yin-Bing

Among ten slow-growing protoclones of Agaricus bisporus (J. Lge) Imbach, all appressed colonies showed slower growth rate and spawn run, and inability to produce fruiting bodies in substrate. Seven of 40 inter-simple sequence repeat (ISSR) primers amplified 78 reproducible fragments, 48.93% were polymorphic, each producing 7 to 16 bands ranging from 0.10 to 2.10 kbp, sufficient to differentiate the protoclones from each other. Appressed protoclones were homoallelic at a number of loci that were heteroallelic in the parent, suggesting that they represented rare homokaryons. Thus, using morphological characters along with ISSR, polymorphisms could be useful for quick, easy, and accurate in distinguishing homo- and heterokaryotic isolates. Key words: Agaricus bisporus (J. Lge) Imbach; Homokaryon; ISSR; Protoclone DOI: 10.3329/bjb.v39i1.5537Bangladesh J. Bot. 39(1): 119-122, 2010 (June)


Author(s):  
S.S. Alghamdi ◽  
H.M. Migdadi ◽  
M.A. Khan ◽  
E.H. El-Harty ◽  
Y.H. Dewir

Background: Somaclonal variation is considered as a source of genetic variation for crop improvement. It has been investigated using cytological, biochemical and molecular techniques.Methods: Genetic stability in the embryo-derived axillary shoots of 4 chickpea genotypes was assessed using eight inter-simple sequence repeat (ISSR) and 19 random amplified polymorphic DNA (RAPD) primers. Result: RAPD primers produced 94 and ISSR primers produced 38 distinct and scorable alleles, with an average of 4.9 alleles for RAPD and 4.75 for ISSR primers. The polymorphic information content (PIC) ranged from 0.36 to 0.90 for RAPD and from 0.50 to 0.87 for ISSR. ISSR recognized a 90%, but RAPD recognized 82% similarity value. No absolute similarity value was between the mother plant and the regenerated shoots for the overall genotypes. At a 90% similarity value, 15 out of the 20 regenerated shoots from ‘Giza 88’ group with their mother plant using ISSR markers; however, 11 regenerated shoots grouped with their mother plant in one central cluster for ‘Giza 4’ using RAPD markers. The observed variations in the total number of polymorphic RAPD and ISSR bands and the number of bands specific to the mother and regenerated shoots, detected intra-clonal variation and genetic instability seem to be genotype-dependent.


CORD ◽  
2005 ◽  
Vol 21 (02) ◽  
pp. 34 ◽  
Author(s):  
R. Manimekalai

Inter Simple Sequence Repeat (ISSR) markers are versatile and used in a number of applications viz. genetic diversity estimation, phylogenetic relationship and gene tagging in different crops. In coconut, the ISSR markers are being reported here for the first time. In the present paper, thirty-five  primers targeting to amplify the inter microsatellite  regions were screened  using  thirty  different  coconut  germplasm  accessions.  The  ISSR  primers  were evaluated based on polymorphism information content and marker index.  Out of 35 primers screened,  19 primers  produced  clear amplification  pattern.  The polymorphism  information content varied between 0.019 and 0.386, whereas, the marker index ranged from 0.019 to 5.673  among  the  primers.    Based  on  the  high  marker  index,  five  and  ten  primers  were selected.  The similarity  matrices  were constructed  separately  for five, ten and 19 primers using NYSYS software and the correlation was tested using Mantel’s test. There was high correlation  between five and ten primers with 19 primers. Hence, the primers with higher marker index (5 and 10 primers) were regarded as informative  primers. These informative primers can be used to develop more polymorphic markers in coconut for rapid screening of germplasm materials.


2015 ◽  
Vol 38 (5) ◽  
Author(s):  
Padmavati G. Gore ◽  
M. K. Rana ◽  
Kuldeep Tripathi ◽  
Mohar Singh ◽  
I. S. Bisht ◽  
...  

Genetic diversity was assessed in 50 accessions of seven <italic>Lens</italic> species using ISSR markers. The collection included accessions of the cultivated species <italic>L. culinaris</italic> and six wild species, <italic>viz</italic>., <italic>L. culinaris</italic> ssp. <italic>odemensis, L. culinaris</italic> ssp. <italic>orientalis</italic>, <italic>L.</italic> <italic>orientalis, L. nigricans, L. lamottei</italic> and <italic>L. ervoides.</italic> The 23 ISSR primers amplified a total of 368 bands with an average of 16 bands per primer. Maximum number of 20 bands was amplified using each of the primers ISSR-34 and ISSR-835. All the primers were found to be polymorphic. PIC values ranged from 0.02 to 0.80. The primers ISSR-807, ISSR- 809, ISSR- 827, ISSR- 847, ISSR-28 and ISSR- 37 were found to be very useful for analyzing the molecular diversity of the genus <italic>Lens</italic>. Cluster Analysis and Principal Coordinates Analyses placed the 50 accessions into two groups and complemented each other.


Biologia ◽  
2014 ◽  
Vol 69 (12) ◽  
Author(s):  
Samaneh Abedi ◽  
Reza Darvishzadeh ◽  
Iraj Bernousi ◽  
Babak Abdollahi Mandoulakani ◽  
Hamid Hatami Maleki ◽  
...  

AbstractBroomrapes (Orobanche L.) are holoparasitic plants, parasitizing roots of a wide range of host plants. In this study, genetic polymorphism among 44 Orobanche aegyptiaca Pers. individuals collected from different regions of northwest Iran was investigated using inter-simple sequence repeat (ISSR) markers. Two hundred-sixty one discernible bands were amplified using 20 ISSR primers which 245 (94%) was polymorphic, indicating considerable genetic variation among the examined individuals. The number of polymorphic bands per primer ranged from 4 to 17, averaging 12.25. UPGMA clustering using Jaccard’s similarity coefficients revealed six main groups. Genetic similarity coefficients varied from 0.71 (between individuals 23 and 27) to 0.34 (between 13 and 30). A model-based Bayesian approach subdivided 38 out of 44 broomrape genotypes into 2 genetic groups and the remaining ones were categorized as mixed genotypes based on Q values. According to an analysis of molecular variance, 99% of the total variation was partitioned within genetic groups. The results demonstrated the potential usefulness of ISSR markers for determination of genetic variation in O. aegyptiaca. Understanding the variability in broomrape is important when attempting to develop resistant host crops against this parasite.


Agronomy ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 402
Author(s):  
Noha A. El-Tayeh ◽  
Hanaa K. Galal ◽  
Magda I. Soliman ◽  
Hoida Zaki

Aerva javanica is one of Egypt’s most important traditional medicinal plants used as antidiarrheal and anthelmintic medicine and recently as an anticancer agent. In this study, variations among ten populations of Aerva javanica in different sites in the Eastern Desert of Egypt were analyzed based on morphological and ecological attributes and molecular variation expressed by Inter-Simple Sequence Repeat (ISSR) markers. Morphological diversity was higher for populations in the Wadi El-Markh and Bir Abbady regions than others. The polymorphism revealed by ten ISSR primers was 79.4% among populations. Distance trees created using the results obtained from soil variables, morphological characterizations, and molecular data showed that the highest similarity was 0.974 between Populations 8 and 9, while the lowest similarity was 0.715 between Population 1 and Population 3 regions. In conclusion, the obtained data are important to design a plan for sustainable conservation of Aerva javanica as an important medicinal plant having a wide interspecific genetic variability within various populations.


2021 ◽  
Author(s):  
Un-Hyang Ho ◽  
Jung Sam Kye ◽  
Song Im Chae ◽  
Jong Ho Kim ◽  
Myong Ho Kim

Abstract Mulberry (Morus spp.) is a cross-pollinating and highly hybridized plant of which productivity are greatly varied in different varieties. We analysed molecular distinction among four mulberry species and varieties cultivated in DPR Korea by using nuclear ribosomal internal transcribed spacer (ITS) sequences and inter simple sequence repeat (ISSR) markers. ITS sequences didn’t represent a remarkable interspecific distinction among four mulberry species used in our study, suggesting that it could not be employed to identify them. ISSR analysis using 16 random primers generated 158 different markers ranging from 100 to 4000 bp in size. The results showed the inter-specific genetic variation (55.34%) was slightly higher than intra-specific genetic variation(44.66%), with comparatively low average number of migrants per generation (Nm) among populations (0.3886). Using ISSR primers selected in this study, in the future, the suitable breeding strategy might be established in raising of elite mulberry varieties on the basis of interspecific hybridization.


Author(s):  
Jan Vitamvas ◽  
Iva Viehmannova ◽  
Petra Hlasna Cepkova ◽  
Hana Mrhalova ◽  
Katerina Eliasova

Abstract: The objective of this work was to induce and detect somaclonal variation in arracacha (Arracacia xanthorrhiza) plants regenerated via indirect morphogenesis, in order to evaluate the potential of this technique to produce new genotypes for breeding purposes of this crop. Calli were induced from petiole segments on Murashige & Skoog (MS) medium supplied with 0.1 mg L-1 2,4-dichlorophenoxyacetic acid. The regeneration of plants via indirect morphogenesis was carried out on half-strength MS medium without plant growth regulators. Fifteen randomly chosen plants were subjected to flow cytometry and “inter-simple sequence repeat” (ISSR) analysis. Ploidy level remained stable in all tested regenerants (2n=4x=44), with no changes in the genome. Eighteen ISSR primers produced a total of 1,584 fragments in all samples. Two ISSR primers produced four polymorphic fragments in 26.7% of the tested samples. Somaclonal variation in arracacha is a result of plant regeneration via indirect morphogenesis and can be detected by ISSR markers.


2014 ◽  
Vol 6 (3) ◽  
pp. 292-299
Author(s):  
Marcelina KRUPA-MAŁKIEWICZ ◽  
Miłosz Smolik ◽  
Anna BARNIAK ◽  
Beata SMOLIK

Randomly amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers were used to investigate genetic variability within thirteen Viburnum species (Viburnum × hillieri; V. dilatatum; Viburnum × carlcephalum; V. opulus; V. hupehense; Viburnum× bodnantense; Viburnum × burkwoodii; V. sieboldii; Viburnum × globosum ‘Jermyns Globe’; V. alnifolium (lantanoides); V. plicatum ‘Sterile’; V. plicatum f. tomentosum and V. plicatum ‘Watanabe’) of wide geographical distribution, collected in the Dendrological Garden in Przelewice (the north-west part of Poland). Twenty-three RAPD and fourteen ISSR primers generated a total of 690 and 418 reproducible bands, respectively, and 39% (RAPD) and 55.5% (ISSR) of them were polymorphic for the two marker systems, which suggest high genetic variability within Viburnum genus. However, high numbers of genotype-specific bands, i.e. 60.9% (RAPD) and 44.5% (ISSR), were seen in Viburnum. Genetic similarity assessed within Viburnum species with the RAPD and ISSR analyses ranged from 6 to 42% and from 6 to 31%, respectively. Both RAPD and ISSR-based dendrograms clustered in five main groups. The Mantel test between two Nei’s similarity matrices gave correlation coefficient r=0.305*, showing low correlation between RAPD- and ISSR- based matrices. Thus, both marker systems were equally important for the genetic diversity analysis in Viburnum genus.


2018 ◽  
Author(s):  
Shipeng Li ◽  
Mingxin Guo ◽  
Pengcheng Fu ◽  
Hongxia Liu ◽  
Xusheng Zhao

The Chinese jujube (Ziziphus jujuba Mill.) originates from sour jujube (Ziziphus acidojujuba Mill.) and is an economically important genus in the Rhamnaceae family. However, little is known about the genetic relationship between jujube cultivars and wild species. In this study, we estimated the genetic variation and relationships between 85 jujube cultivars and 55 sour jujube individuals by ISSR markers. Of 216 ISSR primers, 110 were able produce amplified product(s) and 28 showed polymorphisms, accounting for 50.9% and 25.5% of total primers respectively. A total of 89 loci were amplified with 28 primers, of which 42 loci (47.2%) were polymorphic, and most of primers exhibited highly PIC values. Cluster analysis and population structure analysis roughly divided the 140 accessions into two major groups. One group included all jujube cultivars and some sour jujube individuals, and the other group included remaining sour jujube individuals. Most jujube cultivars have a certain correlation with their origin, and there are obvious gene exchanges between sour jujube and jujube cultivars. The results provide a useful basis for jujube germplasm conservation, genetic improvement and evolution research.


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