Tissue-specific expression of the sPLA2 and PLA2-like members have been searched in various vegetative (leaf, stem, root, and seeds) and reproductive (flower, anther, pollen, pollen tube, carpels, pistil, ovary, ovule, and egg cells) tissues of Arabidopsis, Amborella, tomato, grape, rice, and maize using the CoNekT database (https://conekt.sbs.ntu.edu.sg/ ) (Proost and Mutwil, 2018 (link)). Gene expression was represented in transcripts per kilobase million (TPM)-based normalization because it can be used for both gene count comparisons within a sample or between samples of the same sample group (Abrams et al., 2019 (link)). The expression values were analyzed in the CIMminer one matrix server (discover.nci.nih.gov/cimminer ).
Total RNA was isolated from tobacco leaves, roots, buds, flowers, imbibed pollen, germinating pollen grains and growing pollen tubes using Qiagen RNAeasy Kit, and Turbo DNA-free Kit (Applied Biosystems, Waltham, MA, USA) was used for DNA removal. cDNA synthesis was carried out using Transcriptor High Fidelity cDNA Synthesis Kit (Roche, Penzberg, Germany) with anchored-oligo (DT)18 primer according to manufacturer’s instructions. Semi-quantitative RT-PCR was performed with NtPLA2 gene-specific oligonucleotides 1-6 (Supplementary Table 2
Total RNA was isolated from tobacco leaves, roots, buds, flowers, imbibed pollen, germinating pollen grains and growing pollen tubes using Qiagen RNAeasy Kit, and Turbo DNA-free Kit (Applied Biosystems, Waltham, MA, USA) was used for DNA removal. cDNA synthesis was carried out using Transcriptor High Fidelity cDNA Synthesis Kit (Roche, Penzberg, Germany) with anchored-oligo (DT)18 primer according to manufacturer’s instructions. Semi-quantitative RT-PCR was performed with NtPLA2 gene-specific oligonucleotides 1-6 (
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