Expression Pattern of FAE1s and Erucic Acid Accumulation in Developing Seeds of Crambe abyssinica

doi:10.7668/hbnxb.20194753

Abstract

Abstract:

To explore the expression patterns of the FAE1s controlling the biosynthesis of very-long-chain fatty acids and their effects on the synthesis of erucic acid in the developing seeds of Crambe abyssinica,the expression levels of the five cloned FAE1 homologous genes(CaFAE1-1—CaFAE1-5)were analyzed in the roots,stems,and leaves of seedlings and in the developing seeds(5—25 d after flowering)by using Real-time Quantitative PCR(qRT-PCR)technology,and the fatty acid components in seeds at different developmental stages were detected by gas chromatography-mass spectrometry(GC-MS)method,and finally the correlation between the expression level of individual CaFAE1 and the erucic acid content in seeds was emphasized.The results showed that,compared to seeds,all five CaFAE1s expressed at an extremely low level in root,stem,and leaf tissues.In the 5—25 d after flowering seeds,CaFAE1-3 always exhibited the highest expression level,and it increased gradually with the seed development process;while the expression levels of the other four CaFAE1s showed a trend of first increasing and then decreasing,all reached their highest values in 20 d after flowering seeds,and then CaFAE1-5 and CaFAE1-4 decreased rapidly.The GC-MS analysis results showed that the relative contents of long-chain saturated fatty acids C16:0 and C18∶0,as well as monounsaturated fatty acid C18∶1,were significantly higher than those of very-long-chain fatty acids in 5—15 d after flowering seeds,afterward their contents rapidly decreased,indicating that the carbon chain elongation of fatty acids by CaFAE1 using C18∶0 and C18∶1 as substrates mainly occurred after 15 d after flowering,and the end-products were dominantly C22∶0 and C22∶1.The correlation analysis between the expression levels of CaFAE1s and the content of fatty acid components demonstrated that in 5—25 d after flowering seeds,the expression levels of the five CaFAE1s showed highly significant or significant positive correlation with the content of very-long-chain fatty acids,and CaFAE1-1,CaFAE1-2,and CaFAE1-3 showed highly significant or significant positive correlation with the content of erucic acid.Collectively,it is speculated that the convergence of multiple functional copies of CaFAE1 accompanied by high-level expression of high-active CaFAE1 may be the main cause of high erucic acid content in Crambe abyssinica seeds.

Key words: Crambe abyssinica, Developing seeds, Erucic acid, Fatty acid elongase 1, Expression pattern, qRT-PCR

HU Xinyue, WANG Jin, NIE Wanhong, GONG Chao, XU Benbo, ZHANG Xuekun, ZHAO Fuyong. Expression Pattern of FAE1s and Erucic Acid Accumulation in Developing Seeds of Crambe abyssinica[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(3): 35-42. doi: 10.7668/hbnxb.20194753.

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Figures/Tables 7

Tab.1 Information of primers of CaFAE1 and CaActin2 for qRT-PCR

引物名称 Primer name	引物序列(5'—3') Sequence (5'—3')	扩增子/bp Amplicon	扩增效率/% Amplification efficiency
CaFAE1-1-Fq	AACGCATACCGGAGCTGAC	156	96.60
CaFAE1-1-Rq	GGAAGAATCAACGGACCCAAC
CaFAE1-2-Fq	TTCACACGGTCCGAACACAT	119	98.80
CaFAE1-2-Rq	GGCAACATCGGTTATGTCCTTAG
CaFAE1-3-Fq	TGAGTTGGCATACATAGAGGCA	110	95.30
CaFAE1-3-Rq	TTAAAGCTACCCAAACCGCACT
CaFAE1-4-Fq	AACGCATACCGGAGCTGAC	118	88.20
CaFAE1-4-Rq	AACCGTTCGACCAGCAACC
CaFAE1-5-Fq	GGTGCGCTGGAAAATCTATTCA	137	93.50
CaFAE1-5-Rq	TTGCTTCGGAGCTTGAAAGTATTG
CaActin2-Fq	CAGTGTCTGGATCGGTGGTT	91	96.30
CaActin2-Rq	TGGACCTGCCTCGTCATACT

Fig.1 DNA sequence alignment results of β-Actin2 among different genera in the Cruciferae family (partial) All the DNA sequences of β-Actin2 in the Cruciferae family displayed in the figure were downloaded from the NCBI website (https://www.ncbi.nlm.nih.gov/).The sequence regions indicated by the arrows were the designed qPCR primers for CaActin2.

Fig.2 The expression levels of CaFAE1s in the root, stem, and leaf tissues of Crambe abyssinica Different lowercase letters indicate significant differences between different genes at the P<0.05 level. The same as Fig.3.

Fig.3 Relative expression levels of CaFAE1s in Crambe abyssinica seeds at different developmental stages

Fig.4 Dynamic changes in the expression level of CaFAE1s during seed development process

Fig.5 Fatty acid components and relative abundance in seeds of Crambe abyssinica at different developmental stages

Fig.6 The correlation between the expression levels of CaFAE1 genes and the relative content of fatty acids in the developing seeds of Crambe abyssinica ^*.P<0.05;^**.P<0.01;^***.P<0.001。

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