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MB Sample ID: SA189393

Local Sample ID:	I.16
Subject ID:	SU002103
Subject Type:	Plant
Subject Species:	Alkanna tinctoria

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Subject:

Subject ID:	SU002103
Subject Type:	Plant
Subject Species:	Alkanna tinctoria

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
I.16	SA189393	FL023249	growth	developmental stage
I.16	SA189393	FL023249	Austrian	soil type

Collection:

Collection ID:	CO002096
Collection Summary:	Alkanna tinctoria plants were provided as rooted acclimatized individuals, originally collected and identified from natural populations
Sample Type:	Plant

Treatment:

Treatment ID:	TR002115
Treatment Summary:	Plants were produced by micropropagation from several mother plants by the Hellenic Agricultural Organization (HAO, Thessaloniki, Greece). The plants were transferred to 5 L pots containing 4.5 L of sterilized (121° C for 15 min) peat moss and perlite (volume ratio 2:1), mixed with 200 g field soil collected either in Austria or in Greece. Thus, all plants were grown in a substrate with highly similar chemical and physical characteristics, but hosting those microbial communities prevailing in these three distinct soils. Plants were grown in the greenhouse at 16 h light / 8 h dark photoperiod, 25°C with 50% relative humidity (RH) and a photosynthetic photon flux density (PPFD) of 96 μmolm^-2 s^-1. Plants were watered twice per week with deionized water and moved randomly once per week. Plants were harvested at four different defined developmental stages, the first stage (“vegetative growth”) was defined when more than 50% of the individuals started to produce new leaves, “blooming” was the stage when more than 50% of the individuals had flowers, “fruiting” when more than 50% of the individual plants began to produce fruits.

Treatment ID:

TR002115

Treatment Summary:

Plants were produced by micropropagation from several mother plants by the Hellenic Agricultural Organization (HAO, Thessaloniki, Greece). The plants were transferred to 5 L pots containing 4.5 L of sterilized (121° C for 15 min) peat moss and perlite (volume ratio 2:1), mixed with 200 g field soil collected either in Austria or in Greece. Thus, all plants were grown in a substrate with highly similar chemical and physical characteristics, but hosting those microbial communities prevailing in these three distinct soils. Plants were grown in the greenhouse at 16 h light / 8 h dark photoperiod, 25°C with 50% relative humidity (RH) and a photosynthetic photon flux density (PPFD) of 96 μmolm^-2 s^-1. Plants were watered twice per week with deionized water and moved randomly once per week. Plants were harvested at four different defined developmental stages, the first stage (“vegetative growth”) was defined when more than 50% of the individuals started to produce new leaves, “blooming” was the stage when more than 50% of the individuals had flowers, “fruiting” when more than 50% of the individual plants began to produce fruits.

Sample Preparation:

Sampleprep ID:	SP002109
Sampleprep Summary:	Plant roots were ground to a fine powder using a ball mill (Fritsch Pulverisette 0, Germany). Each powdered sample was weighed (70 mg) into microcentrifuge tubes, followed by extraction with 3 mL of methanol by ultrasound at 10% power for 3 h (Bandelin Sonorex Digital 10P, Berlin, Germany) and centrifugation for 10 minutes at 12.500 rpm (Hermle Z 216 MK, Wehingen, Germany). The supernatants were collected and subjected to UHPLC-HRMS analysis after filtering with 0.22 μm syringe filters.

Sampleprep ID:

SP002109

Sampleprep Summary:

Plant roots were ground to a fine powder using a ball mill (Fritsch Pulverisette 0, Germany). Each powdered sample was weighed (70 mg) into microcentrifuge tubes, followed by extraction with 3 mL of methanol by ultrasound at 10% power for 3 h (Bandelin Sonorex Digital 10P, Berlin, Germany) and centrifugation for 10 minutes at 12.500 rpm (Hermle Z 216 MK, Wehingen, Germany). The supernatants were collected and subjected to UHPLC-HRMS analysis after filtering with 0.22 μm syringe filters.

Combined analysis:

Analysis ID	AN003291
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Thermo Accela 1250
Column	Waters Acquity UPLC HSS C18 SB (100 x 2.1mm, 1.8um)
MS Type	ESI
MS instrument type	Orbitrap
MS instrument name	Thermo LTQ Discovery Orbitrap
Ion Mode	POSITIVE
Units	intensity units

Chromatography:

Chromatography ID:	CH002431
Instrument Name:	Thermo Accela 1250
Column Name:	Waters Acquity UPLC HSS C18 SB (100 x 2.1mm, 1.8um)
Column Temperature:	50
Flow Rate:	0.3mL/min
Chromatography Type:	Reversed phase

MS:

MS ID:	MS003062
Analysis ID:	AN003291
Instrument Name:	Thermo LTQ Discovery Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	The MS/MS data were obtained for the six most intense m/z peaks in each full scan, with the normalized collision energy set to 35 eV. The acquisition and initial processing of the data were by means of XcaliburTM (Thermo Scientific, USA) software, while data alignment and feature extraction were performed utilizing the XCMS Online platform (The Scripps Research Institute, USA). The batch error correction was done with the help of MetaboAnalyst 5.0.
Ion Mode:	POSITIVE