Macleaya cordata, (Willd.) R. Br. (Willd.) R. Br.

Chen, Jinjun, Liu, Zihao, Liu, Yisong, Zhang, Xuewen & Zeng, Jianguo, 2021, Preliminary investigations on the pathogenesis-related protein expression profile of the medicinal herb Macleaya cordata and anti-bacterial properties of recombinant proteins, Phytochemistry (112667) 184, pp. 1-9 : 2

2.1. An overview of the analysis of M. cordata (willd.) R. Br. leaf ESTs

The cDNA library from M. cordata leaf was non-normalized, and the titer was 1.0 × 106 cfu/mL with recombinant clones. After the poorquality sequences were discarded, 511 high-quality ESTs were used to investigate the gene expression profile of M. cordata leaf. The initial sequences were grouped into 364 non-redundancy sequences, of which 78 clusters exhibited more than one EST and 286 singlets (Supplementary Table 1). The average length of the non-redundant sequence was 513 nucleotides (ranging from 105 to 1013 bp) (Supplementary Figure 1 ). Additionally, the newly discovered EST sequences of the cDNA library were submitted to Genbank with the accession number from JK752513 to JK752907 .

With the development of high-throughput next-generation sequencing technologies, sequencing the transcriptomes of medicinal plants with immense pace has become a hot pursuit, generating more generalized insights. However, de novo assembly is a big challenge, especially for the assembly of cysteine-rich defensins, because these peptides are hypervariable sequences using conserved scaffolds. The cDNA library with ESTs Sanger sequencing approach has been verified to be a rapid and reliable method for uncovering undescribed genes, and the interesting candidates provide the gene templates for subsequent recombinant expression.

Figures

Fig. 1. The alignment analysis of McDef1-5. The underlined amino acids represent the signal sequence predicted with SignalP. The cysteines are marked in yellow background and the putative disulfide bonds are also shown. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)