Abstract
Endophytes have been defined as microorganisms living inside plant tissues without causing negative effects on their hosts. Endophytic microbes have been extensively studied for their plant growth-promoting traits. However, analyses of endophytes require complete removal of epiphytic microorganisms. We found that the established tests to evaluate surface sterility, polymerase chain reaction, and leaf imprints, are unreliable. Therefore, we used scanning electron microscopy (SEM) as an additional assessment of epiphyte removal. We used a diverse suite of sterilization protocols to remove epiphytic microorganisms from the leaves of a gymnosperm and an angiosperm tree to test the influence of leaf morphology on the efficacy of these methods. Additionally, leaf tissue damage was also evaluated by SEM, as damaging the leaves might have an impact on endophytes and could lead to inaccurate assessment of endophytic communities. Our study indicates, that complete removal of the leaf cuticle by the sterilization technique assures loss of epiphytic microbes, and that leaves of different tree species may require different sterilization protocols. Furthermore, our study demonstrates the importance of choosing the appropriate sterilization protocol to prevent erroneous interpretation of host-endophyte interactions. Moreover, it shows the utility of SEM for evaluating the effectiveness of surface sterilization methods and their impact on leaf tissue integrity.
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Claims 5
Claim · model-written summary
PCR tests for leaf surface sterility were unreliable.
Not reportedComparatorValue
Verbatim
e0“the established tests to evaluate surface sterility, polymerase chain reaction, and leaf imprints, are unreliable”
e1“leaves of a gymnosperm and an angiosperm tree”
Claim · model-written summary
Leaf imprint tests for surface sterility were unreliable.
Not reportedComparatorValue
Verbatim
e0“the established tests to evaluate surface sterility, polymerase chain reaction, and leaf imprints, are unreliable”
e1“leaves of a gymnosperm and an angiosperm tree”
Claim · model-written summary
Complete cuticle removal by sterilization assured loss of epiphytic microbes.
- Taxon
- a gymnosperm and an angiosperm treee1
- Treatment
- complete removal of the leaf cuticle by the sterilization techniquee0
- Interactor
- epiphytic microbese0other
- Outcome
- loss of epiphytic microbese0
- Direction
- losse0
Not reportedComparator
Verbatim
e0“complete removal of the leaf cuticle by the sterilization technique assures loss of epiphytic microbes”
e1“leaves of a gymnosperm and an angiosperm tree”
Claim · model-written summary
Different tree species may require different leaf sterilization protocols.
Verbatim
e0“leaves of different tree species may require different sterilization protocols”
Claim · model-written summary
SEM was useful for evaluating surface sterilization effectiveness and leaf tissue integrity.
- Taxon
- a gymnosperm and an angiosperm treee1
- Outcome
- effectiveness of surface sterilization methods and their impact on leaf tissue integritye0
Not reportedComparatorValue
Verbatim
e0“utility of SEM for evaluating the effectiveness of surface sterilization methods and their impact on leaf tissue integrity”
e1“leaves of a gymnosperm and an angiosperm tree”