Cloning the full-length CDNA of actin gene and analysing alliinase gene expression in tillering onion
Keywords:Actin, Alliinase, RACE, Tillering onion.
Tillering onion is a herbaceous plant belonging to the Liliaceae family. We cloned the cDNAs of the actin gene (AcACT, GenBank: MF919598) of tillering onion using rapid amplification of the cDNA ends. The full-length cDNA of AcACT was 1,357 bp long with an open reading frame of 1,131 bp encoding 376 amino acids. The amino acid sequence of AcACT shared > 96% similarity with the amino acid sequences of other ACTs and was found (by means of phylogenetic tree analysis) to be closely related to those of Ananas comosus and Papaver somniferum. AcACT expressions showed no significant differences (p > 0.01) in two cultivars L-SH and L-SY over three growth periods and under suitable conditions, low temperature, and short-day conditions. In addition, AcACT was used as an internal reference gene to analyse the expression of the alliinase gene (AcALL). AcALL expression trends in the roots, stems and leaves were consistent with those of diallyl disulphide and diallyl trisulphide. Thus, AcACT is highly conserved and can be used as a suitable internal reference gene when analysing gene expression in tillering onion.
BASHLINE, L., et al. Cell wall, cytoskeleton, and cell expansion in higher plants. Molecular plant. 2014, 7(4), 586-600. https://doi.org/10.1093/mp/ssu018
CHENG, F. and CHENG, Z.H. Research progress on the use of plant allelopathy in agriculture and the physiological and ecological mechanisms of allelopathy. Frontiers in plant science. 2016, 7, 1697. https://doi.org/10.3389/fpls.2016.01697
DUAN, Z.R. and TOMINAGA, M. Actin-myosin XI: An intracellular control network in plants. Biochemical and Biophysical Research Communications.2018, 506(2), 403-408. https://doi.org/10.1016/j.bbrc.2017.12.169
FU, X.P., et al. Physiological response and sulfur metabolism of the V. dahliae-infected tomato plants in tomato/potato onion companion cropping. Scientific Reports. 2016, 6, 36445. https://doi.org/10.1038/srep36445
FU, X.P., et al. Transcriptomic comparison of Allium cepa var. agrogatum Don. cultivars with different facilitating potentials on tomato seedlings. Journal of Plant Interactions. 2019, 14(1), 54-60. https://doi.org/10.1080/17429145.2018.1550216
Fu, Y.X., et al. Effects of concomitant tillering onion on microbial community structure in tomato rhizosphere soil. China Vegetables. 2020, 6, 49-57.
HUGGETT, J., et al. Real-time RT-PCR normalization; strategies and considerations. Genes & Immunity. 2005, 6(4), 279-284.
KIM, S.J., et al. Abiotic stress and tissue-specific reference genes for quantitative reverse transcription PCR analysis in Korean native watermelons, Citrullus lanatus ‘Black-King’ and ‘Speed-Plus-Honey’. Applied Biological Chemistry. 2018, 61, 593-598.
KOZERA, B. and RAPACZ, M. Reference genes in real-time PCR. Journal of Applied Genetics. 2013, 54(4), 391-406.
KUMAR, S., et al. Mega X: molecular evolutionary genetics analysis across computing platforms. Molecular Biology & Evolution. 2018, 35(6), 1547-1549. https://doi.org/10.1093/molbev/msy096
LI, N.H., et al. Intercropping with potato-onion enhanced the soil microbial diversity of tomato. Microorganisms. 2020, 8(6), 834. https://doi.org/10.3390/microorganisms8060834
LI, Q.H., et al. Effects of rotation and interplanting on soil bacterial communities and cucumber yield. Acta Agriculturae Scandinavica Section B - Soil and Plant Science. 2009, 59(5), 431-439. https://doi.org/10.1080/09064710802342319
LIU, M., WU, Z. and JIANG, F. L. Selection and validation of garlic reference genes for quantitative real-time PCR normalization. Plant Cell Tissue and Organ Culture. 2015, 122(2), 435-444. https://doi.org/10.1007/s11240-015-0780-9
LIVAK, K.J. and SCHMITTGEN, T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods. 2001, 25(4), 402-408.
MA, Z.M., et al. Formin nanoclustering-mediated actin assembly during plant flagellin and DSF signaling. Cell Reports. 2021,34(13), 108884.
MALANDRAKI, I., et al. Simultaneous detection of three pome fruit tree viruses by one-step multiplex quantitative RT-PCR. PLoS One. 2017,12(7), e0180877. https://doi.org/10.1371/journal.pone.0180877
MITROVÁ, K., et al. Alliinase and cysteine synthase transcription in developing garlic (Allium sativum L.) over time. Food Chemistry. 2018, 251, 103-109. https://doi.org/10.1016/j.foodchem.2017.12.090
NAKAYAMA, T.J., et al. Reference genes for quantitative real-time polymerase chain reaction studies in soybean plants under hypoxic conditions. Genetics and Molecular Research. 2014, 13(1), 860-871. https://doi.org/10.4238/2014.February.13.4
RUSINKO, J. and MCPARTLON, M. Species tree estimation using neighbor joining. Journal of Theoretical Biology. 2017, 414, 5-7. https://doi.org/10.1016/j.jtbi.2016.11.005
SCHMITTGEN, T.D. and ZAKRAJSEK, B.A. Effect of experimental treatment on housekeeping gene expression: validation by real-time, quantitative RT-PCR. Journal of Biochemical Biophysical Methods. 2000, 46(1-2), 69-81. https://doi.org/10.1016/S0165-022X(00)00129-9
SHANG, A., et al. Bioactive compounds and biological functions of garlic (Allium sativum L.). Foods. 2019, 8(7), 246. https://doi.org/10.3390/foods8070246
SOORNI, A., et al. Transcriptome and phytochemical analyses provide insights into the organic sulfur pathway in Allium hirtifolium. Scientific Reports. 2021, 11(1), 768. https://doi.org/10.1038/s41598-020-80837-6
STEPHAN, O.H.S. Actin fringes of polar cell growth. Journal of Experimental Botany. 2017, 68(13), 3303-3320. https://doi.org/10.1093/jxb/erx195
SZYMANSKI, D. and STAIGER, C.J. The actin cytoskeleton: functional arrays for cytoplasmic organization and cell shape control. Plant Physiology. 2018, 176(1), 106-118. https://doi.org/10.1104/pp.17.01519
WANG, G.L., et al. Comparison of nine reference genes for real-time quantitative PCR in roots and leaves during five developmental stages in carrot (Daucus carota L.). The Journal of Horticultural Science and Biotechnology. 2016, 91(3), 264-270. https://doi.org/10.1080/14620316.2016.1148372
WANG, G.L, et al. Selection of reliable reference genes for quantitative RT-PCR in garlic under salt stress. PeerJ. 2019, 7, e7319. https://doi.org/10.7717/peerj.7319
WANG, J.X., et al. Evaluation and selection of suitable qRT-PCR reference genes for light responses in tea plant (Camellia sinensis). Scientia Horticulturae. 2021, 289, 110488. https://doi.org/10.1016/j.scienta.2021.110488
WEN, S.S., et al. Cloning and sequence analysis of an actin gene in aloe. Genetics and Molecular Research. 2014, 13(3), 4949-4955. https://doi.org/10.4238/2014.July.4.9
XU, K.K., et al. Rapid detection of allicin content in Garlic by HPLC. China Condiment. 2016, 41(1), 124-127. http://doi.org/10.3969/j.issn.1000-9973.2016.01.027
YANG, Y., WU, F.Z. and ZHOU, X.G. Protein expression in accessions of Chinese onion with different allelopathic potentials under monocropping and intercropping systems. Acta Physiologiae Plantarum. 2013, 35(7), 2241-2250. https://doi.org/10.1007/s11738-013-1260-7
YANG, Y., XIANG, X.P. and WANG, Q. Cloning and functional analysis of flavanone 3-hydroxylasegene related to allelopathy in tillered onion. Allelopathy Journal. 2018, 45(1), 113-128. http://doi.org/10.26651/allelo. j/2018-45-1-1180
YOSHIMOTO, N. and SAITO, K. S-alk(en)ylcysteine sulfoxides in the genus Allium: proposed biosynthesis, chemical conversion, and bioactivities. Journal of Experimental Botany. 2019, 70(16), 4123-4137. https://doi.org/10.1093/jxb/erz243
ZHANG, X.B., et al. HPLC method for determination of allitridi in garlic oil soft capsule. Chinese Journal of Health Laboratory Technology. 2013, 23(9), 2062-2063, 2066. http://DOI:CNKI:SUN:ZWJZ.0.2013-09-013
ZHAO, X.X., et al. Recent advances in bioactive compounds, health functions, and safety concerns of onion (Allium cepa L.). Frontiers in Nutrition. 2021, 8, 669805. https://doi.org/10.3389/fnut.2021.669805
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