Assessing genetic stability of a range of terrestrial microalgae after cryopreservation using amplified fragment length polymorphism (AFLP)

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.

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

Plant Methods ◽

10.1186/s13007-021-00724-7 ◽

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.

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