Summary of Study ST003567

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002201. The data can be accessed directly via it's Project DOI: 10.21228/M84517 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST003567
Study Title	Enoyl-acyl carrier protein reductase (ZmENR1) expression level is critical for de novo fatty acid biosynthesis and anther cuticle and pollen exine formation in maize.
Study Summary	Lipid metabolism is very important for plant male reproduction. Many lipid metabolism genes, male sterility (GMS) genes play a role in the endoplasmic reticulum of anther tapetum, but little is known about GMS genes involved in the de novo synthesis of fatty acids in anther tapetum. Using gene mapping, we cloned a new GMS gene, enoyl-acyl carrier protein (ACP) reductase (ZmENR1). enr1 (mutation of enoyl-ACP reductase gene) has early tapetum degradation, anther epidermis and pollen exine defects. In order to further study the function of ZmENR1, we compared the changes of lipid content in anthers of wild-type (WT) and enr1 mutants at maturity. The expression of ZmENR1 significantly affected the contents of cutin, wax and total fatty acids (TFA) in anthers. In addition, enr1 mutant reduced the expression of cutin/wax and sporopollen related genes to inhibit the formation of anther epidermis and pollen exine. Therefore, our researches provide a new insight into the synthesis of fatty acids mediated by ZmENR1, and provide an insight into the molecular mechanism of ZmENR1 regulating maize male sterility.
Institute	University of Science and Technology Beijing
Last Name	Zhang
First Name	Shaowei
Address	30 XUEYUAN ROAD, HAIDIAN DISTRICT, BEIJING 100083, CHINA
Email	b20200413@xs.ustb.edu.cn
Phone	0086-13271969211
Submit Date	2024-11-05
Raw Data Available	Yes
Raw Data File Type(s)	cdf
Analysis Type Detail	GC-MS
Release Date	2024-11-12
Release Version	1

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Project:

Project ID:	PR002201
Project DOI:	doi: 10.21228/M84517
Project Title:	Analysis of maize Anther Wax, Cutin, and total fatty acids (TFA) of WT and enr1 mutant
Project Type:	Detection of small molecular compounds in cutin and wax. Detection of free fatty acids and dissociated long-chain fatty acids in TFA.
Project Summary:	Lipid metabolism is very important for plant male reproduction. Many lipid metabolism genes, male sterility (GMS) genes play a role in the endoplasmic reticulum of anther tapetum, but little is known about GMS genes involved in the de novo synthesis of fatty acids in anther tapetum. Using gene mapping, we cloned a new GMS gene, enoyl-acyl carrier protein (ACP) reductase (ZmENR1). enr1 (mutation of enoyl-ACP reductase gene) has early tapetum degradation, anther epidermis and pollen exine defects. In order to further study the function of ZmENR1, we compared the changes of lipid content in anthers of wild-type (WT) and enr1 mutants at maturity. The expression of ZmENR1 significantly affected the contents of cutin, wax and total fatty acids (TFA) in anthers. In addition, enr1 mutant reduced the expression of cutin/wax and sporopollen related genes to inhibit the formation of anther epidermis and pollen exine. Therefore, our researches provide a new insight into the synthesis of fatty acids mediated by ZmENR1, and provide an insight into the molecular mechanism of ZmENR1 regulating maize male sterility.
Institute:	University of Science and Technology Beijing
Last Name:	Zhang
First Name:	Shaowei
Address:	30 XUEYUAN ROAD, HAIDIAN DISTRICT, BEIJING 100083, CHINA
Email:	b20200413@xs.ustb.edu.cn
Phone:	0086-13271969211

Subject:

Subject ID:	SU003696
Subject Type:	Plant
Subject Species:	Zea mays
Taxonomy ID:	4577

Factors:

Subject type: Plant; Subject species: Zea mays (Factor headings shown in green)

mb_sample_id	local_sample_id	Sample source	Genotype
SA389582	cutin A223-3	Anther	enr1 mutant
SA389583	Wax A223-2	Anther	enr1 mutant
SA389584	Wax A223-3	Anther	enr1 mutant
SA389585	cutin A223-2	Anther	enr1 mutant
SA389586	cutin A223-1	Anther	enr1 mutant
SA389587	TFA A223-1	Anther	enr1 mutant
SA389588	TFA A223-2	Anther	enr1 mutant
SA389589	TFA A223-3	Anther	enr1 mutant
SA389590	Wax A223-1	Anther	enr1 mutant
SA389570	Wax WT-3	Anther	Wild-type
SA389571	TFA WT-4	Anther	Wild-type
SA389572	TFA WT-3	Anther	Wild-type
SA389573	TFA WT-2	Anther	Wild-type
SA389574	TFA WT-1	Anther	Wild-type
SA389575	cutin WT-2	Anther	Wild-type
SA389576	Wax WT-4	Anther	Wild-type
SA389577	cutin WT-1	Anther	Wild-type
SA389578	Wax WT-2	Anther	Wild-type
SA389579	Wax WT-1	Anther	Wild-type
SA389580	cutin WT-4	Anther	Wild-type
SA389581	cutin WT-3	Anther	Wild-type

Showing results 1 to 21 of 21

Showing results 1 to 21 of 21

Collection:

Collection ID:	CO003689
Collection Summary:	Maize mutants of enoyl- ACP reductase gene (enr1) was obtained by ethyl methane sulfonate (EMS) of the maize inbred line Zheng58 in our laboratory. All wild-type (Zheng58) and mutant (enr1) materials are planted in the experimental field of University of Science and Technology Beijing (Beijing, China）. Maize WT and enr1 mutant materials were planted under natural conditions. The anthers of WT and enr1 mutants in pollen mature stage were continuously sampled and freeze-dried for later use.
Sample Type:	Anther
Collection Method:	Use tweezers to peel off the anthers manually.

Treatment:

Treatment ID:	TR003705
Treatment Summary:	No treatment

Sample Preparation:

Sampleprep ID:	SP003703
Sampleprep Summary:	To extract waxes, anthers of WT and enr1 mutant were submersed in 3 mL of chloroform for 1 min. The resulting chloroform extracts were spiked with 30 mg of docosanol and 10 mg of heptadecanoic acid as internal standards and transferred to a new vial. The solvents were evaporated under a gentle stream of nitrogen gas. The residue was derivatized with 100 mLofN-methyl-N-(trimethylsilyl) trifluoroacetamide and incubated for 1 h at 50°C. These derivatized samples were then analyzed by GC-MS (Agilent gas chromatograph coupled to an Agilent 5975C quadrupole mass selective detector). To extract anther cutin, anthers of WT and enr1 mutant were submersed in 3 mL of chloroform: methanol (1:1, v/v). They were first incubated at 50°C for 30 min and then 72 h with constant shaking at room temperature. The anthers were freeze dried and submersed in 1 mL of 1 N methanolic HCl for 2 h at 80°C with 10 mg of heptadecanoic acid as an internal standard. The hydrophobic monomers were subsequently extracted three times with 1 mL of hexane after the addition of 2 mL of saturated NaCl solution. The solvent evaporated, and the remaining samples were derivatized as described above. These derivatized samples were then analyzed by GC-MS (Agilent gas chromatograph coupled to an Agilent 5975C quadrupole mass selective detector). To extract total fatty acids (TFA), anthers WT and enr1 mutant were transesterified in 1mL of 1 N methanolic HCl (containing 50 mg of heptadecanoic acid as an internal standard) for 4 h at 50°C.After the addition of 1.5 mLof 0.9% (w/v) NaCl, the hydrophobic monomers were subsequently extracted with 1.5 mL of hexane. The organic phases were evaporated under a gentle stream of nitrogen gas. The residue was dissolved in 100 mL of hexane and then analyzed by GC-MS (Agilent gas chromatograph coupled to an Agilent 5975C quadrupole mass selective detector).

Sampleprep ID:

SP003703

Sampleprep Summary:

To extract waxes, anthers of WT and enr1 mutant were submersed in 3 mL of chloroform for 1 min. The resulting chloroform extracts were spiked with 30 mg of docosanol and 10 mg of heptadecanoic acid as internal standards and transferred to a new vial. The solvents were evaporated under a gentle stream of nitrogen gas. The residue was derivatized with 100 mLofN-methyl-N-(trimethylsilyl) trifluoroacetamide and incubated for 1 h at 50°C. These derivatized samples were then analyzed by GC-MS (Agilent gas chromatograph coupled to an Agilent 5975C quadrupole mass selective detector). To extract anther cutin, anthers of WT and enr1 mutant were submersed in 3 mL of chloroform: methanol (1:1, v/v). They were first incubated at 50°C for 30 min and then 72 h with constant shaking at room temperature. The anthers were freeze dried and submersed in 1 mL of 1 N methanolic HCl for 2 h at 80°C with 10 mg of heptadecanoic acid as an internal standard. The hydrophobic monomers were subsequently extracted three times with 1 mL of hexane after the addition of 2 mL of saturated NaCl solution. The solvent evaporated, and the remaining samples were derivatized as described above. These derivatized samples were then analyzed by GC-MS (Agilent gas chromatograph coupled to an Agilent 5975C quadrupole mass selective detector). To extract total fatty acids (TFA), anthers WT and enr1 mutant were transesterified in 1mL of 1 N methanolic HCl (containing 50 mg of heptadecanoic acid as an internal standard) for 4 h at 50°C.After the addition of 1.5 mLof 0.9% (w/v) NaCl, the hydrophobic monomers were subsequently extracted with 1.5 mL of hexane. The organic phases were evaporated under a gentle stream of nitrogen gas. The residue was dissolved in 100 mL of hexane and then analyzed by GC-MS (Agilent gas chromatograph coupled to an Agilent 5975C quadrupole mass selective detector).

Combined analysis:

Analysis ID	AN005860
Analysis type	MS
Chromatography type	GC
Chromatography system	Agilent 7890A
Column	Agilent DB5-MS (30m x 0.25mm, 0.25um)
MS Type	EI
MS instrument type	Single quadrupole
MS instrument name	Agilent 5975C
Ion Mode	POSITIVE
Units	Counts

Chromatography:

Chromatography ID:	CH004451
Chromatography Summary:	These derivatized samples and dissolved fatty acids in hexane were analyzed by GC-MS (Agilent gas chromatograph coupled to an Agilent 5975C quadrupole mass selective detector), with the following parameters: chromatographic column DB5-MS (Agilent, 30mx0.25mx0.25μm). The temperature of injection port was 270℃, the temperature of auxiliary heating device was 290℃, the initial temperature of chromatographic column was 150℃, and it was kept for 5min, and then it was increased by 5C to 325 C per minute for 7 min. The pressure is constantly changing during the heating process. The carrier gas was helium. We used stable flow mode (Agilent gas chromatograph) and the flow rate was 1mL/min. The split ratio was 10:1 and the ion source temperature was 230℃.
Instrument Name:	Agilent 7890A
Column Name:	Agilent DB5-MS (30m x 0.25mm, 0.25um)
Column Temperature:	The initial temperature of chromatographic column was 150℃, and it was kept for 5min, and then it was increased by 5℃ to 325 ℃ per minute for 7 min.
Flow Gradient:	NA
Flow Rate:	1.0mL/min
Solvent A:	NA
Solvent B:	NA
Chromatography Type:	GC

MS:

MS ID:	MS005580
Analysis ID:	AN005860
Instrument Name:	Agilent 5975C
Instrument Type:	Single quadrupole
MS Type:	EI
MS Comments:	The quadrupole temperature is 150℃, and the ion scanning range is m/z50-650. All substances are determined in the NIST 08 and Fiehn databases. All data were collected by ChemStation and analyzed by METIDEA (2.04).
Ion Mode:	POSITIVE