scholarly journals Evaluation of Genetic Stability in Olive Callus-induced and Meristem-induced Shoots using Flow Cytometry and Amplified Fragment Length Polymorphism Techniques

2020 ◽  
Author(s):  
Leila Mirzaei ◽  
Abbas Yadollahi ◽  
Maryam Jafarkhani Kermani ◽  
Masoud Naderpour ◽  
Ali Asghar Zeinanloo ◽  
...  
Keyword(s):  
Length Polymorphism ◽  
Fragment Length ◽  
Genetic Stability ◽  
Fragment Length Polymorphism ◽  
Culture Media ◽  
Carbon Sources ◽  
Amplified Fragment Length Polymorphism ◽  
Dna Amount ◽  
Indirect Organogenesis

Abstract BackgroundIn vitro culture of olive, as an economically valuable tree, has fundamentally genotype-dependant low micropropagation rate which need to be improved in already established cultivars, while needing to be struggled in firstly considered cultivars. Various plant tissue culture media, planting systems and growth factors were evaluated in two promissing Iranian olive cultivars ˈAminˈ and ˈMeshkatˈ plus a commercial Spanish cultivar ˈArbequinaˈ. ResultsThe results showed that in ˈAminˈ cultivar MS enriched with zeatin (4 mg/L), in ˈArbequinaˈ cultivar OM enriched with zeatin (1 mg/L) and in ˈMeshkatˈ cultivar OM and MS enriched with zeatin (2 mg/L) produced reasonably healthier in vitro plantlets. The results indicated that using Periodical Mini Bioreactor (PMB) system in the presence of VS medium in ˈAminˈ cultivar resulted in growth indicies amelioration compared to semi-solid media. Regarding carbon sources and light intensities, mannitol and 2000 cd·sr·m-² greatly enhancedˈArbequinaˈ growth indices. The genetic stability of callus induced shoots (CIS) was compared with meristem induced shoots (MIS) using Amplified Fragment Length Polymorphism (AFLP) marker and flow cytometery (FCM). The results revealed that 2C DNA value assessed by partec FCM was mostly depended on the cultivars rather than the origin of generation -callus or meristem-. The AFLP results also showed that the cultivars were classified regardless of the micropropagation origin (CIS or MIS), except for ˈArbequinaˈ. However, ˈArbequinaˈ CIS and MIS still had 62- 80% of genetic similarity with three other categories and the highest resemblance (97%) was seen with MIS of ˈAminˈ cultivar. ConclusionsBriefly,olive in vitro propagation optimization co-produced large volume and percent of calli resulting in more undesired indirect organogenesis. True to typeness of CIS was assessed showing that no changes in DNA amount occurred. Nonetheless, in phylogenetic analysis cultivars were clustered separately even some discrepancies were seen in ˈArbequinaˈ MIS and CIS, however, ˈAminˈ and ˈMeshkatˈ cultivars were more stable in clustering.

scholarly journals Evaluation of genetic stability in olive callus-induced and meristem-induced shoots using flow cytometry and amplified fragment length polymorphism techniques

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Leila Mirzaei ◽  
Abbas Yadollahi ◽  
Maryam Jafarkhani Kermani ◽  
Masoud Naderpour ◽  
Ali Asghar Zeinanloo ◽  
...  
Keyword(s):  
Length Polymorphism ◽  
Fragment Length ◽  
Genetic Stability ◽  
Fragment Length Polymorphism ◽  
Amplified Fragment Length Polymorphism ◽  
Aflp Analysis ◽  
Growth Indices ◽  
Axillary Shoots ◽  
Olive Cultivars

Abstract Background In vitro culture of olive, as an economically valuable tree, has fundamentally a genotype-dependant low micropropagation rate which needs to be improved in already established and newly released cultivars. Various plant tissue culture media, planting systems and growth factors were evaluated in two promissing Iranian olive cultivars ˈAminˈ and ˈMeshkatˈ and the commercial Spanish cultivar ˈArbequinaˈ. Results The results showed that cultivars have their specific optimal media, i.e. ˈAminˈ in the MS with 4 mg/L zeatin, ˈArbequinaˈ in the OM with 1 mg/L zeatin, and ˈMeshkatˈ in the OM and MS with 2 mg/L zeatin, which produced significantly a higher number of axillary shoots than other media. The results also indicated a significant improvement in the growth indices of ˈAminˈ (number of axillary shoots) when cultured using periodical mini bioreactor (PMB) in the VS medium. In comparison with VS, OM did not reveal any significant differences when both culturing systems (PMB and semi-solid media (SSM)) were used. Regarding the effect of carbon source and light intensity, mannitol and 2000 cd sr m−2 greatly enhanced ˈArbequinaˈ growth indices (main shoot length and growth quality). The results of genetic stability of callus induced shoots (CIS) and meristem induced shoots (MIS) revealed that 2C DNA value assessed by partec flow cytometery (FCM) had 0.01, 0.03 and 0.08 pg discrepencies in ˈAminˈ, ˈArbequinaˈ and ˈMeshkatˈ, repectively. The Amplified Fragment Length Polymorphism (AFLP) results also indicated that the cultivars were classified regardless of the micropropagation origin (CIS or MIS), except for ˈArbequinaˈ. The AFLP findings showed that ˈArbequinaˈ had the highest dispersal (7–38%) in CIS and MIS, while the Iranian cultivar of ˈMeshkatˈ (5–9%) had the highest stability. Conclusions This study indicated the importance of in vitro growth parameters for improving the micropropagation indices of olive cultivars. It showed that optimized protocols (OM, PMB, zeatin, mannitol and 2000 cd sr m−2) co-produced larger calli resulting in indirect organogenesis. Based on FCM and AFLP analysis, it can be concluded that true-to-typeness of micropropagated olive was cultivar-dependent.

scholarly journals Genome-Sequence-Based Fluorescent Amplified-Fragment Length Polymorphism Analysis ofMycobacterium tuberculosis

2000 ◽  
Vol 38 (3) ◽  
pp. 1121-1126 ◽  
Author(s):  
Jonathan N. Goulding ◽  
John Stanley ◽  
Nick Saunders ◽  
Catherine Arnold
Keyword(s):  
Genome Sequence ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Amplified Fragment Length Polymorphism ◽  
Clinical Isolates ◽  
Resolving Power ◽  
Base Substitution ◽  
Polymorphism Analysis

The whole-genome fingerprinting technique, fluorescent amplified-fragment length polymorphism (FAFLP) analysis, was applied toMycobacterium tuberculosis. Sixty-five clinical isolates were analyzed to determine the value of FAFLP as a stand-alone genotyping technique and to compare it with the well-established IS6110 typing system. The genome sequence of M. tuberculosis strain H37Rv (S. T. Cole et al., Nature 393:537–544, 1998) was used to model computer-generated informative primer combination(s), and the precision and reproducibility of FAFLP were evaluated by comparing the results of in vitro and computer-generated experiments. Multiplex FAFLP was used to increase resolving power in a predictable and systematic fashion. FAFLP analysis was broadly congruent with IS6110 typing for those strains with multiple IS6110 copies. It was also able to resolve an epidemiologically unlinked group of strains with only one copy of IS6110; up to 10% of clinical isolates may fall into this category. For certain epidemiological investigations, it was concluded that a combination of FAFLP and IS6110 typing would give higher resolution than would either alone. FAFLP data were digital, precise, reproducible, and suitable for rapid electronic dissemination, manipulation, interlaboratory comparison, and storage in national or international epidemiological databases. Because FAFLP samples and analyzes base substitution across the genome as a whole, FAFLP could generate new information about the microevolution of the M. tuberculosis complex.

scholarly journals In Vitro Antifungal Susceptibilities and Amplified Fragment Length Polymorphism Genotyping of a Worldwide Collection of 350 Clinical, Veterinary, and Environmental Cryptococcus gattii Isolates

2010 ◽  
Vol 54 (12) ◽  
pp. 5139-5145 ◽  
Author(s):  
Ferry Hagen ◽  
Maria-Teresa Illnait-Zaragozi ◽  
Karen H. Bartlett ◽  
Daniëlle Swinne ◽  
Erik Geertsen ◽  
...  
Keyword(s):  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Geometric Mean ◽  
Amplified Fragment Length Polymorphism ◽  
Cryptococcus Gattii ◽  
Antifungal Drugs ◽  
Dilution Step ◽  
Interspecies Hybrids

ABSTRACT The in vitro susceptibilities of a worldwide collection of 350 Cryptococcus gattii isolates to seven antifungal drugs, including the new triazole isavuconazole, were tested. With amplified fragment length polymorphism (AFLP) fingerprinting, human, veterinary, and environmental C. gattii isolates were subdivided into seven AFLP genotypes, including the interspecies hybrids AFLP8 and AFLP9. The majority of clinical isolates (n = 215) comprised genotypes AFLP4 (n = 76) and AFLP6 (n = 103). The clinical AFLP6 isolates had significantly higher geometric mean MICs for flucytosine and fluconazole than the clinical AFLP4 isolates. Of the seven antifungal compounds examined in this study, isavuconazole had the lowest MIC90 (0.125 μg/ml) for all C. gattii isolates, followed by a 1 log2 dilution step increase (MIC90, 0.25 μg/ml) for itraconazole, voriconazole, and posaconazole. Amphotericin B had an acceptable MIC90 of 0.5 μg/ml, but fluconazole and flucytosine had relatively high MIC90s of 8 μg/ml.

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