CoChat
Sign Up
Cited by 16Plant tissue culture and regenerationGenetics and Plant BreedingDate Palm Research Studies

The DOI matches its Crossref record; 36 of the 36 references it cites were found in a registry or an index; 4 of the 59 sources its claims cite were checked for support.

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

Open access
Mirzaei, Leila, Yadollahi, Abbas, Kermani, Maryam Jafarkhani, Naderpour, Masoud, Zeinanloo, Ali Asghar, Farsi, Maryam, Davoodi, Dariush·2021·Plant Methods
DOI 10.1186/s13007-021-00724-7PMC PMC8008589

Published version of a preprint

Open this paper in CoChat — free accountOpen PDF

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.

Overview

AI snapshot

Written from the paper’s full text.

TL;DR

In three olive cultivars, micropropagation was optimized by cultivar-specific media/zeatin, culture system, carbon source and light, and genetic stability of callus-induced vs meristem-induced shoots was assessed by FCM and AFLP. True-to-typeness was cultivar-dependent, with ˈArbequinaˈ showing the greatest AFLP dispersal and ˈMeshkatˈ the highest stability.

Key findings

  • Cultivar-specific proliferation: ˈAminˈ in MS + 4 mg/L zeatin produced the highest axillary shoots (1.50), main shoot length (2.23 cm) and growth quality (4.08); ˈArbequinaˈ performed best in OM + 1 mg/L zeatin; ˈMeshkatˈ in OM and MS + 2 mg/L zeatin produced higher axillary shoots (1.48 and 1.45). High zeatin depressed ˈMeshkatˈ but not ˈAminˈ.
  • Periodical mini bioreactor (PMB) improved most growth indices of ˈAminˈ, especially in VS medium, while OM showed no significant PMB vs SSM difference; for ˈArbequinaˈ, only axillary shoots in MS and VS were significantly higher in PMB, and PMB reduced growth quality due to suffocated plantlets.
  • Mannitol and 2000 cd sr m−2 enhanced ˈArbequinaˈ main shoot length and growth quality; mannitol induced 100% callus vs 81.3% for sucrose, and 2000 cd sr m−2 induced 87.5% callus vs 75% at 5000 cd sr m−2.
  • Genetic stability: FCM total DNA ranged 2.87–3.12 pg; the abstract reports 2C DNA discrepancies of 0.01, 0.03 and 0.08 pg for ˈAminˈ, ˈArbequinaˈ and ˈMeshkatˈ, respectively. AFLP clustered samples by cultivar regardless of CIS/MIS origin except for ˈArbequinaˈ; similarity levels were 62–98%, with ˈArbequinaˈ showing the highest dispersal (7–38%), ˈMeshkatˈ the highest stability (5–9%), and ˈAminˈ 2–32%; ˈAminˈ MIS and ˈArbequinaˈ MIS were >97% similar.

Methods

  • Three commercial olive cultivars (ˈAminˈ, ˈMeshkatˈ, ˈArbequinaˈ) were established on OM + 4 mg/L zeatin + 0.5 mg/L 2iP. Proliferation experiments tested media (OM, MS, VS), zeatin (0, 1, 2, 4 mg/L), culture system (PMB vs semi-solid media), carbon source (sucrose vs mannitol) and light intensity (2000 vs 5000 cd sr m−2) under a completely randomized design with at least 25 replications; growth indices were recorded after 6 weeks and analyzed by ANOVA (GLM, SAS) with LSD at P ≤ 0.01.
  • Genetic stability compared callus-induced shoots (CIS) and meristem-induced shoots (MIS) subcultured separately for 6 months. AFLP used CTAB-extracted leaf DNA, 12 EcoRI/MseI primer combinations, binary band scoring, Jaccard/Dice similarity and UPGMA clustering in NTSYS-PC; FCM used parsley as internal standard (2C = 4.46 pg), PI staining, ~10,000 nuclei per sample and accepted peaks with CV < 5%.
  • Callus production percentage and callus volume were recorded; volume was measured by sphere formula or water displacement.

Limitations

  • Stated by the paper: FCM DNA amount discrepancies may be due to cytosolic phenolic compounds interfering with DNA-PI staining, are genotype-dependent, and should be considered cautiously; PVP and careful chopping were used to mitigate this. The paper also notes no correlation between genetic somaclonal variation and phenotypic changes and recommends complementary phenotypic analysis.
  • Evident from the text: only three cultivars and 12 AFLP primer combinations were tested; the paper notes several primers should be tested to increase genome coverage, and no field/agronomic validation beyond in vitro growth indices is reported.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1186/s13007-021-00724-7

Cites

36 of 36 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Verification

Claims the paper makes

4 of 59 cited sources checked · 3 supported · 0 flagged · accuracy not yet measured

  • Findingown data

    In cv Meshkat, OM and MS containing 2 mg/L zeatin produced well-grown plantlets.

    “In ˈMeshkatˈ cultivar, OM and MS containing 2 mg/L zeatin produced well-grown plantlets (Table 1).”
    Quote in the paperNot a substantial claim
  • Findingown data

    Calli obtained from different media and genotype had different morphological characteristics like color and texture.

    “Calli obtained from different media and genotype had different morphological characteristics like color and texture.”
    Quote in the paperNot a substantial claim
  • Findingown data

    In cv Arbequina, OM with 1 mg/L zeatin produced significantly higher number of axillary shoots than other media.

    “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.”
    Quote in the paperNot a substantial claim
  • Findingown data

    In cv Amin, most growth indices were higher in PMB compared to SSM.

    “The results showed that most growth indices of ˈAminˈ cultivar were higher in the periodical mini bioreactor (PMB) compared to semi-solid media (SSM) (Fig. 1).”
    Quote in the paperNot a substantial claim
  • Findingown data

    Callus formation at 2000 cd sr m−2 (87.5%) was more than at 5000 cd sr m−2 (75%).

    “Callus formation at 2000 cd s m−2 (87.5%) was more than 5000 cd sr m−2 (75%) (Table 3).”
    Quote in the paperNot a substantial claim
  • Probably not a substantial claim. 36
  • It may state two separate claims as one. 26
  • The quote only partly states this claim. 1

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    Using a bioreactor is advantageous due to lower production cost, less labor, lower zeatin concentration, increased proliferation efficiency, and removal of gelling agent.

    “Using bioreactor is advantageous because of lower production cost, less labor, lower zeatin concentration and increased efficiency of proliferation rate as well as removing gelling agent [11].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [11] Benelli C, De Carlo A. In vitro multiplication and growth improvement of Olea europaea L. cv Canino with temporary immersion system (PlantformTM). 3 Biotech. 2018. https://doi.org/10.1007/s13205-018-1346-4. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    TIS economizes plant production by reducing zeatin amount (50%) in olive tissue culture as reported in Canino.

    “TIS also economizes plant production by reducing the amount of zeatin (50%) in olive tissue culture as reported in ˈCaninoˈ cultivar [11].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [11] Benelli C, De Carlo A. In vitro multiplication and growth improvement of Olea europaea L. cv Canino with temporary immersion system (PlantformTM). 3 Biotech. 2018. https://doi.org/10.1007/s13205-018-1346-4. source

  • Probably not a substantial claim. It may be too vague to check against evidence.

    Premisecitation

    Detection of genetic uniformity of in vitro-raised plants at an early stage is desirable.

    “Therefore, the detection of genetic uniformity of in vitro-raised plants at an early stage is desirable [22].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [22] Krishna H, Alizadeh M, Singh D, Singh U, Chauhan N, Eftekhari M, Sadh RK. Somaclonal variations and their applications in horticultural crops improvement. 3 Biotech. 2016. https://doi.org/10.1007/s13205-016-0389-7. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    Temporary immersion system overcomes gas accumulation, reduces mass production costs, and causes the least hyperhydricity.

    “It overcomes gas accumulation, reduces the mass production costs [11] and causes the least hyperhydricity [12].”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [11] Benelli C, De Carlo A. In vitro multiplication and growth improvement of Olea europaea L. cv Canino with temporary immersion system (PlantformTM). 3 Biotech. 2018. https://doi.org/10.1007/s13205-018-1346-4. source

      The Introduction says Plantform™ ventilation prevents gas accumulation, and the Results and discussion describe lower production costs and prevention of shoot hyperhydricity under the study’s culture conditions. The paper does not establish that temporary immersion causes the *least* hyperhydricity relative to other methods. Page numbers were not available in our copy. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [12] Grigoriadou K, Vasilakakis M, Tzoulis T, Eleftheriou EP. Experimental use of a novel temporary immersion system for liquid culture of olive microshoots. In "Liquid Culture Systems for in vitro Plant Propagation". Springer 2005; doi: https://doi.org/10.1007/1-4020-3200-5-18 source

  • It may state two separate claims as one.

    Findingown data

    In some cases, calli were green and strict solid resulting in indirect organogenesis and their callus-differentiated shoots were morphologically healthy, but some were crystallized, spongy and vitrified which never gave rise to indirect shoot formation.

    “In some cases, calli were green in color and strict solid in texture resulting in indirect organogenesis and their callus-differentiated shoots were morphologically healthy, but some of them were crystallized, spongy and vitrified which never gave rise to indirect shoot formation (Fig. 4).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    Light intensity at 2000 cd sr m−2 enhanced main shoot length and growth quality of Arbequina plantlets, but not number of axillary shoots or main shoot leaves.

    “Light intensity at 2000 cd sr m−2 enhanced main shoot length, and growth quality of plantlets, whereas the number of axillary shoots and the number of main shoot leaves were not significantly different (Table 2).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    Mannitol and sucrose induced 100% and 81.3% callus initiation at the base of explants, respectively.

    “The results indicated that mannitol and sucrose induced 100% and 81.3% callus initiation at the base of explants, respectively (Table 3).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    In cv Arbequina, PMB caused suffocated plantlets and reduced growth quality compared to SSM.

    “Suffocated plantlets resulted in a reduction in growth quality of plantlets in PMB were compared to SSM (Fig. 3).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    In cv Amin, VS medium with PMB showed absolute superiority in main shoot length, number of axillary shoots and growth quality.

    “Regarding VS, it revealed absolute superiority in main shoot length, number of axillary shoots and growth quality using PMB for ˈAminˈ cultivar”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    In cv Amin, MS containing 4 mg/L zeatin produced the highest number of axillary shoots, main shoot length, and growth quality among tested media/zeatin.

    “MS containing 4 mg/L zeatin in ˈAminˈ cultivar produced the highest number of axillary shoots, main shoot length, and growth quality (Table 1).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    In Arbequina and Meshkat, there was a direct correlation between zeatin concentration and callus formation, but in Amin there was no correlation in callus production.

    “In ˈArbequinaˈ and ˈMeshkatˈ cultivars, there was a direct correlation between zeatin concentration and callus formation, but in ˈAminˈ there was no correlation in callus production (Table 3).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    High zeatin concentration (4 mg/L) depressed Meshkat plantlets in both media, increasing callus production as a negative factor.

    “ˈMeshkatˈ plantlets were depressed by high zeatin concentration (4 mg/L) in both media, though the increment of zeatin only increased callus production as a negative factor (Tables 1 and 3).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    Phylogenetic analysis split the samples into 8 distinct groups indicating that callus-regenerated plants had highly monomorphic amplicons similar to those of meristem-regenerated plants.

    “The phylogenetic analysis split the samples into 8 distinct groups indicating that callus-regenerated plants had highly monomorphic amplicons similar to those of the meristem-regenerated plants (Fig. 5).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one. The quote only partly states this claim.

    Findingown data

    The 2C DNA value assessed by FCM had discrepancies between CIS and MIS of 0.01, 0.03 and 0.08 pg in Amin, Arbequina and Meshkat, respectively, with total DNA amounts 2.87-3.12 pg.

    “There was some discrepancy (0.01–0.08 pg) between CIS and MIS in total DNA amount (2.87–3.12 pg) that varied among cultivars (Table 5).”
    • Also stated in: § Results · plain
      “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.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Findingown data

    Each primer pair resulted in 28 to 55 markers with an average of 42 amplified fragments.

    “Each primer pair resulted in 28 to 55 markers with an average of 42 amplified fragments (Table 4).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    Arbequina showed the highest dispersal (7-38%) in CIS and MIS, while Iranian cultivars Amin (2-32%) and Meshkat (5-9%) had more stability.

    “In this study, ˈArbequinaˈ showed the highest dispersal (7–38%) in CIS and MIS, but both Iranian cultivars (ˈAminˈ (2–32%) and ˈMeshkatˈ (5–9%)) had more stability (Fig. 6).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    From 12 primer combinations, 259 bands (50 bp to 1000 bp) were analyzed in all tested cultivars consisting of MIS and CIS, of which 238 bands were polymorphic.

    “From 12 primer combinations tested, 259 bands (50 bp to 1000 bp) were analyzed in all tested cultivars consisting of MIS and CIS of which 238 bands were polymorphic.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    One CIS in Arbequina appeared in a separate branch similar to all groups consisting 3 cultivars with 62% genetic similarity, and one of its MIS samples appeared in a separate category with 79% resemblance with both CIS and MIS of Meshkat.

    “One of the CIS in ˈArbequinaˈ cultivar appeared in a separate branch of phylogenetic tree which was similar to all groups consisting 3 cultivars with 62% genetic similarity and one of its MIS samples appeared in a separate category with 79% resemblance with both CIS and MIS of ˈMeshkatˈ.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Findingreasoning

    In this study, 84% of samples had lower DNA amounts.

    “In this study, 84% of samples had lower DNA amounts which might be due to PI staining.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Mannitol promoted bud sprouting and secondary shoot uniformity rather than sucrose regardless of its concentration.

    “Mannitol promoted bud sprouting and secondary shoot uniformity rather than sucrose regardless of its concentration [19].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [19] Leva A, Sadeghi H, Petruccelli R. Carbohydrates modulate the in vitro growth of olive microshoots. I. The analysis of shoot growth and Branching patterns. J Plant Growth Regul. 2013. https://doi.org/10.1007/s00344-012-9275-7. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    Shoot development pattern, plantlet survival, growth quality and secondary shoot formation in olive are affected by carbon sources.

    “It is proved that shoot development pattern, plantlet survival, growth quality and secondary shoot formation in olive are affected by carbon sources [19].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [19] Leva A, Sadeghi H, Petruccelli R. Carbohydrates modulate the in vitro growth of olive microshoots. I. The analysis of shoot growth and Branching patterns. J Plant Growth Regul. 2013. https://doi.org/10.1007/s00344-012-9275-7. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    AFLP is the most efficient marker for olive in comparison with SSR and RAPD.

    “For revealing numerous bands per reaction and valuable number of indices, AFLP is considered the most efficient marker for olive in comparison with SSR and RAPD [24].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [24] De la Rosa R, Angiolillo A, Guerrero C, Pellegrini M, Rallo L, Besnard G, Bervillé A, Martin A, Baldoni L. A first linkage map of olive (Olea europaea L) cultivars using RAPD, AFLP, RFLP and SSR markers. Theor Appl Genet. 2003. https://doi.org/10.1007/s00122-002-1189-5. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    Mannitol is involved in activating osmotic effect, and plants with high mannitol or acclimatized in mannitol-enriched media have negligible osmotic stresses.

    “As mannitol is involved in activating osmotic effect, plants with high amount of mannitol or well-acclimatized in mannitol enriched media have negligible osmotic stresses [15].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [15] Moradnezhad M, Hosseini R, Zarrabi M M, Golmohammadi FG. A New Approach for Olive (Arbequina cv.) Micropropagation: Effect of Dikegulac, Light and Carbon Source. Int J Horticultural Sci Technol 2017; https://ijhst.ut.ac.ir/article_65937 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    Zeatin is the most effective PGR in olive shoot regeneration and plays an important role in salvaging the endangered diploid and triploid Laperrinei olive.

    “zeatin, which is the most effective PGR in shoot regeneration and plays an important role in salvaging the endangered diploid and triploid ˈLaperrineiˈ olive [16]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [16] Haddad B, Carra A, Saadi A, Haddad N, Mercati F, Gristina A, Boukhalfa S, Djillali A, Carimi F. In vitro propagation of the relict laperinne’s olive (Olea europaea L. subsp. Laperrinei). Plant Biosystems . 2018. https://doi.org/10.1080/11263504.2017.1306002. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    Haddad et al. reported mannitol enrichment resulted in greener foliage and morphologically healthy plantlets similar to mother plants.

    “Haddad et al. [16] reported mannitol enrichment resulted in greener foliage and morphologically healthy plantlets similar to their mother plants.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [16] Haddad B, Carra A, Saadi A, Haddad N, Mercati F, Gristina A, Boukhalfa S, Djillali A, Carimi F. In vitro propagation of the relict laperinne’s olive (Olea europaea L. subsp. Laperrinei). Plant Biosystems . 2018. https://doi.org/10.1080/11263504.2017.1306002. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    In Laperrinei olive, regeneration is highly related to media and PGRs, and microshoot formation does not develop without PGRs.

    “In other subspecies of olive (Laperrinei olive), regeneration was highly related to media and PGRs, and only in case of not applying PGRs, microshoot formation did not develop [16].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [16] Haddad B, Carra A, Saadi A, Haddad N, Mercati F, Gristina A, Boukhalfa S, Djillali A, Carimi F. In vitro propagation of the relict laperinne’s olive (Olea europaea L. subsp. Laperrinei). Plant Biosystems . 2018. https://doi.org/10.1080/11263504.2017.1306002. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Sadder reported suffocation of olive cultivar Nabali in liquid OM.

    “However, Sadder [9] reported the suffocation of olive cultivar ˈNabaliˈ in liquid OM.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [9] Sadder MT. In vitro establishment of olive (Olea europaea L.) ˈNabaliˈ in still liquid medium and callus culture (Research Note). Dirasat, Agricultural Sci. 2002;29:59–64.

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    High calcium in OM plays an important role in cytokinin signal transduction leading to better cell division.

    “High calcium (CaCl2 and Ca(NO3)2) in OM plays an important role in cytokinin signal transduction in cells leading to better cell division [13].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [13] Ali A, Ahmad T, Abbasi NA, Hafiz IA. Effect of different media and growth regulators on in vitro shoot proliferation of olive cultivar 'moraiolo'. Pakistan Journal of Botany 2009; http://www.pakbs.org//contents.html

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Leva et al. showed more undifferentiated parenchyma-like callus growth in medium containing sucrose compared with mannitol.

    “Leva et al. [19] showed more undifferentiated parenchyma-like callus growth in medium containing sucrose compared with mannitol.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [19] Leva A, Sadeghi H, Petruccelli R. Carbohydrates modulate the in vitro growth of olive microshoots. I. The analysis of shoot growth and Branching patterns. J Plant Growth Regul. 2013. https://doi.org/10.1007/s00344-012-9275-7. source

  • It may state two separate claims as one.

    Premisecitation

    High zeatin concentration reduces shoot proliferation and elongation, and this effect is genotype-dependent.

    “High zeatin concentration reduced shoot proliferation and elongation which are genotype-dependent [16].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [16] Haddad B, Carra A, Saadi A, Haddad N, Mercati F, Gristina A, Boukhalfa S, Djillali A, Carimi F. In vitro propagation of the relict laperinne’s olive (Olea europaea L. subsp. Laperrinei). Plant Biosystems . 2018. https://doi.org/10.1080/11263504.2017.1306002. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Ploidy level stability assessment in olive cultivar sylvestris multiplied on OM with zeatin after a PGR shock showed DNA content reduced 2.5-9% in micropropagated plants.

    “Ploidy level stability assessment in olive cultivar ˈsylvestrisˈ multiplied on OM with zeatin after a PGR shock followed by transferring to the same medium without PGRs showed the DNA content reduced 2.5–9% in the micropropagated plants [20].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [20] Brito G, Lopes T, Loureiro J, Rodriguez E, Santos C. Assessment of genetic stability of two micropropagated wild olive species using flow cytometry and microsatellite markers. Trees. 2010. https://doi.org/10.1007/s00468-010-0442-9. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Mannitol in combination with OM significantly increases shoots and node numbers per shoot.

    “and in combination with OM significantly increases shoots and node numbers per shoot [15]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [15] Moradnezhad M, Hosseini R, Zarrabi M M, Golmohammadi FG. A New Approach for Olive (Arbequina cv.) Micropropagation: Effect of Dikegulac, Light and Carbon Source. Int J Horticultural Sci Technol 2017; https://ijhst.ut.ac.ir/article_65937 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Mannitol is the main photosynthesis product in olive.

    “Mannitol, a sugar alcohol consisting of 30% of total carbohydrates in leaves and branches, is the main photosynthesis product in olive [18]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [18] Bustan A, Avni A, Lavee S, Zipori I, Yeselson Y, Schaffer AA, Riov J, Dag A. Role of carbohydrate reserves in yield production of intensively cultivated oil olive (Olea europaea L.) trees. Tree Physiol. 2011. https://doi.org/10.1093/treephys/tpr036. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Light type is a determinative environmental factor in olive growth.

    “It is also reported that light type is a determinative environmental factor in olive growth [15].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [15] Moradnezhad M, Hosseini R, Zarrabi M M, Golmohammadi FG. A New Approach for Olive (Arbequina cv.) Micropropagation: Effect of Dikegulac, Light and Carbon Source. Int J Horticultural Sci Technol 2017; https://ijhst.ut.ac.ir/article_65937 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    OM is the best medium for olive micropropagation, high in Ca, Mg, S, Cu and Zn.

    “Analysis of olive shoot tip (4–5 cm) by Rugini [10] resulted in introducing the best medium for olive micropropagation called OM which is high in Ca, Mg, S, Cu and Zn elements.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [10] Rugini E. In vitro propagation of some olive (Olea europaea sativa L.) cultivars with different root-ability, and medium development using analytical data from developing shoots and embryos. Sci Hortic. 1984. https://doi.org/10.1016/0304-4238(84)90143-2. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Callus formation is mainly dependent on cultivars, media and PGRs.

    “Callus formation is mainly dependent on cultivars, media and PGRs [9, 10, 21].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [9] Sadder MT. In vitro establishment of olive (Olea europaea L.) ˈNabaliˈ in still liquid medium and callus culture (Research Note). Dirasat, Agricultural Sci. 2002;29:59–64.

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [10] Rugini E. In vitro propagation of some olive (Olea europaea sativa L.) cultivars with different root-ability, and medium development using analytical data from developing shoots and embryos. Sci Hortic. 1984. https://doi.org/10.1016/0304-4238(84)90143-2. source
    • cites [21] Oulbi S, Toufik I, Belkoura I. Somatic embryogenesis from cultured zygotic explants of some olive cultivars (Olea europaea L.). New Biotechnol. 2014. https://doi.org/10.1016/j.nbt.2014.05.1999. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Nuclear DNA content in olive species was significantly different between donor plants and acclimatized plants in highly reproducible histograms.

    “Moreover, nuclear DNA content in olive species was significantly different between donor plants and acclimatized plants in highly reproducible histograms [20].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [20] Brito G, Lopes T, Loureiro J, Rodriguez E, Santos C. Assessment of genetic stability of two micropropagated wild olive species using flow cytometry and microsatellite markers. Trees. 2010. https://doi.org/10.1007/s00468-010-0442-9. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductnone given

    PVP was added to nuclei isolation buffer to reduce interaction between secondary compounds.

    “To reduce the interaction between secondary compounds, 10 g/L polyvinyl pyrrolidone (PVP) was added to the nuclei isolation buffer.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper may give no reason why it holds.

    Premiseown data

    ˈAminˈ is a highly resistant native cultivar to salinity stress.

    “Because ˈAminˈ is highly resistance to salinity stress and other abiotic and biotic stresses (under submission data), it was suspected to be a polyploid.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites under submission data

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Study design and conductnone given

    The micropropagation experiments were conducted under a completely randomized design with at least 25 replications.

    “The micropropagation experiments were conducted under a completely randomized design with at least 25 replications.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    For wild olive, 2C DNA content was 3.19 ± 0.047 pg/2C.

    “Loureiro et al. [7] reported that 2C DNA content of O. europaea ssp. europaea var. europaea ranged between 2.90 ± 0.020 pg/2C and 3.07 ± 0.018 pg/2C and for wild olive, it was 3.19 ± 0.047 pg/2C.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [7] Loureiro J, Rodriguez E, Costa A, Santos C. Nuclear DNA content estimations in wild olive (Olea europaea L. ssp. europaea var. sylvestris Brot.) and portuguese cultivars of O europaea using flow cytometry. Genet Resour Crop Evol. 2007. https://doi.org/10.1007/s10722-006-9115-3. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Haddad et al. [16] reported all amplicons in ISSR marker in regenerated diploid and triploid endangered olive compared with donor plants were monomorph.

    “Haddad et al. [16] reported all amplicons in ISSR marker in regenerated diploid and triploid endangered olive compared with donor plants were monomorph, and so all the regenerants in high PGRs and their mother plants were grouped in the same category.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [16] Haddad B, Carra A, Saadi A, Haddad N, Mercati F, Gristina A, Boukhalfa S, Djillali A, Carimi F. In vitro propagation of the relict laperinne’s olive (Olea europaea L. subsp. Laperrinei). Plant Biosystems . 2018. https://doi.org/10.1080/11263504.2017.1306002. source

  • It may state two separate claims as one.

    Premisecitation

    In marker investigations, DNA is usually extracted from leaves and so, mutation level in diploids might be underestimated.

    “In marker investigations, DNA is usually extracted from leaves and so, the mutation level in diploids might be underestimated [34].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [34] Besnard G, Baali-Cherif D. Coexistence of diploids and triploids in a Saharan relict olive: Evidence from nuclear microsatellite and flow cytometry analyses. CR Biol. 2009. https://doi.org/10.1016/j.crvi.2009.09.014. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    2C DNA content of O. europaea ssp. europaea var. europaea ranged between 2.90 ± 0.020 pg/2C and 3.07 ± 0.018 pg/2C.

    “Loureiro et al. [7] reported that 2C DNA content of O. europaea ssp. europaea var. europaea ranged between 2.90 ± 0.020 pg/2C and 3.07 ± 0.018 pg/2C and for wild olive, it was 3.19 ± 0.047 pg/2C.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [7] Loureiro J, Rodriguez E, Costa A, Santos C. Nuclear DNA content estimations in wild olive (Olea europaea L. ssp. europaea var. sylvestris Brot.) and portuguese cultivars of O europaea using flow cytometry. Genet Resour Crop Evol. 2007. https://doi.org/10.1007/s10722-006-9115-3. source

  • It may state two separate claims as one.

    Premisecitation

    Interfering phenolic compounds are genotype-dependent and normally micropropagated plants accumulate higher amounts of cytosolic compounds which do not allow proper PI staining resulting in lower DNA amount.

    “These interfering phenolic compounds are genotype-dependent and normally micropropagated plants accumulate higher amounts of cytosolic compounds which do not allow proper PI staining resulting in lower DNA amount [7].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [7] Loureiro J, Rodriguez E, Costa A, Santos C. Nuclear DNA content estimations in wild olive (Olea europaea L. ssp. europaea var. sylvestris Brot.) and portuguese cultivars of O europaea using flow cytometry. Genet Resour Crop Evol. 2007. https://doi.org/10.1007/s10722-006-9115-3. source
    • Rests on an assumption: Olive is extremely high in phenolic compounds.

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. The paper may give no reason why it holds.

    Interpretationnone given

    The lower DNA amounts in 84% of samples might be due to PI staining.

    “In this study, 84% of samples had lower DNA amounts which might be due to PI staining.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim.

    Study design and conductcitation

    The peaks with a coefficient of variation of less than 5% were accepted.

    “The peaks with a coefficient of variation of less than 5% were accepted and the nuclear genome size of each type of plant material was estimated according to Yokoya et al. (2000).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Yokoya et al. (2000) source

  • Probably not a substantial claim.

    Study design and conductnone given

    Only reproducible bands were used to construct the original binary data matrix.

    “Only reproducible bands were used to construct the original binary data matrix.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    All regenerants in high PGRs and their mother plants were grouped in the same category.

    “Haddad et al. [16] reported all amplicons in ISSR marker in regenerated diploid and triploid endangered olive compared with donor plants were monomorph, and so all the regenerants in high PGRs and their mother plants were grouped in the same category.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [16] Haddad B, Carra A, Saadi A, Haddad N, Mercati F, Gristina A, Boukhalfa S, Djillali A, Carimi F. In vitro propagation of the relict laperinne’s olive (Olea europaea L. subsp. Laperrinei). Plant Biosystems . 2018. https://doi.org/10.1080/11263504.2017.1306002. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Meristem mutation position is the main important factor in chimerism maintenance.

    “In addition, Guillaume et al. [34] reported that meristem mutation position is the main important factor in chimerism maintenance and mutation in apical dome normally results in permanent chimera [34].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [34] Besnard G, Baali-Cherif D. Coexistence of diploids and triploids in a Saharan relict olive: Evidence from nuclear microsatellite and flow cytometry analyses. CR Biol. 2009. https://doi.org/10.1016/j.crvi.2009.09.014. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Phenotypical characteristics are controlled by several genes.

    “Similarly, no correlation between genetically somaclonal varied plants with phenotypic changes was found in this study as phenotypical characteristics are controlled by several genes [33].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [33] Bradaï F, Sánchez-Romero C, Martín C. Somaclonal variation in olive (Olea europaea L.) plants regenerated via somatic embryogenesis: Influence of genotype and culture age on genetic stability. Sci Hortic. 2019. https://doi.org/10.1016/j.scienta.2019.03.010. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Mutation in apical dome normally results in permanent chimera.

    “In addition, Guillaume et al. [34] reported that meristem mutation position is the main important factor in chimerism maintenance and mutation in apical dome normally results in permanent chimera [34].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [34] Besnard G, Baali-Cherif D. Coexistence of diploids and triploids in a Saharan relict olive: Evidence from nuclear microsatellite and flow cytometry analyses. CR Biol. 2009. https://doi.org/10.1016/j.crvi.2009.09.014. source

  • It may state two separate claims as one.

    Interpretationown data

    Simultaneous FCM analysis of nuclei from parsley internal standard and olive cultivars in CIS and MIS resulted in highly reproducible histograms with low CV values indicating the priority of cultivar rather than origin of propagation in genetic similarity.

    “Simultaneous analysis of nuclei isolated from parsley (Petroselinum crispum Mill.) (2C DNA amount = 4.46 pg, as internal standard) and olive cv. ˈArbequinaˈ, ˈMeshkatˈ, ˈAminˈ in CIS and MIS resulted in highly reproducible histograms with low CV values indicating the priority of cultivar rather than the origin of propagation in genetic similarity (Table 5).”
    Quote in the paperSubstantial claim
53 of 53

  • The paper relies on this without support.

    The three olive cultivars Amin, Meshkat and Arbequina are commercially important and their mass propagation optimization ensuring genetic trueness is a serious demand.

    “ˈAminˈ and ˈMeshkatˈ (63% and 66% oil, 14 and 21 kg/tree yields, repectively) are two commercial Iranian cultivars with both table and oil consumption which have similar specifications to the commercial ˈArbequinaˈ (65% oil and 25 kg/tree yield in Iran). Therefore, their mass propagation optimization ensuring ascertained genetic trueness is a serious demand.”

  • The paper relies on this without support.

    FCM-measured 2C DNA amount and AFLP banding patterns are valid indicators of genetic stability/true-to-typeness in micropropagated olive.

    “Based on FCM and AFLP analysis, it can be concluded that true-to-typeness of micropropagated olive was cultivar-dependent.”

  • The paper relies on this without support.

    Callus phase is a prerequisite of variations.

    “Callus phase is a prerequisite of variations and the problem of undesired callus induction at the basal end and petiole end in olive propagation is frequently reported.”

  • The paper relies on this without support.

    Stability of in vitro-raised olives in OM and zeatin enriched media is proved.

    “Since the stability of in vitro-raised olives in OM and zeatin enriched media is proved, MIS was considered as donor plants and CIS was compared to MIS in this study.”

  • The paper relies on this without support.

    Olive is extremely high in phenolic compounds.

    “Olive is extremely high in the above-mentioned compounds and so, its discrepancy in 2C DNA value should be considered cautiously.”

  • The paper relies on this without support.

    Mass propagation of olive cultivars has several difficulties leading to genotype-dependent low micropropagation rate.

    “Mass propagation of olive cultivars has several difficulties leading to genotype-dependent low micropropagation rate.”

  • The paper relies on this without support.

    The genetic stability of olive during in vitro culture is fundamental for elite olive genotypes selection.

    “The genetic stability of olive during in vitro culture is fundamental for the elite olive genotypes selection.”
7 of 7

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Six subcultures conflict with the stated culture schedule

    Major

    Six-week subculturing over six months allows about four subcultures, while the discussion says sampling occurred after six.

  • Stated maximum markers per primer pair excludes a table entry

    Minor

    The stated range ends at 55 markers, but Table 4 lists 71 total bands for MCAG-EAT.

  • AFLP band totals disagree with Table 4

    Minor

    The molecular-assessment text reports 259 total and 238 polymorphic bands, whereas Table 4 totals 499 and 379, respectively.

3 of 3

Citations

Cites

Flagged first, then by citations

Research works printed but not linked: 3

  1. Zohary D, Hopf M. Olive: Olea europaea. Oxford: Domestication of plants in the Old World. Clarendon Press; 1994.
  2. Sadder MT. In vitro establishment of olive (Olea europaea L.) ˈNabaliˈ in still liquid medium and callus culture (Research Note). Dirasat, Agricultural Sci. 2002;29:59–64.
  3. Doyle JJ, Doyle JL. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin. 1987;19:1.

Cited by

Sorted by citations

Related

Metadata

Authors
Mirzaei, L., Yadollahi, A., Kermani, M. J., Naderpour, M., Zeinanloo, A. A., Farsi, M., & Davoodi, D.
Affiliations
  • Tarbiat Modares University (IR)
  • Agricultural Research & Education Organization (IR)
Corresponding author
Yadollahi, Abbas
ORCID
Venue
Plant Methods
Publisher
Springer Science and Business Media LLC
Published
2021-03-29
Volume
17
Issue
1
Pages
31-31
ISSN
1746-4811 (ISSN-L)
DOI
10.1186/s13007-021-00724-7
PMC
PMC8008589
OpenAlex
W3118124287
Cited by
16
References
36
Citations by year
2026: 1 · 2025: 4 · 2024: 6 · 2023: 2 · 2022: 2 · 2021: 1
FWCI
0.86
Type
journal article
Language
en
Open access
Licence
Locations
  • Plant Methods (publishedVersion, cc-by)
  • Research Square (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
Tarbiat Modares University (IG-39806806)
Topics
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant tissue culture and regeneration
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Genetics and Plant Breeding
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Date Palm Research Studies
Keywords
olive, micropropagation, genetic stability, indirect organogenesis, zeatin, mannitol, light intensity, 2C DNA content, AFLP, Arbequina, plant tissue culture media optimization, carbon source

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

A language model drew the claims from the paper on 2026-10-28, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).

A second model labelled each claim on 2026-10-28 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).

Cited sources are checked with a language model’s help (gpt-6-sol).

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review