Soybean is an important oil and protein crop in the world. Cotton bollworm (Helicoverpa armigera) is one of primary defoliating insects which seriously damage the soybean production. The wild soybean, ED059, was reported to be highly resistant to H. armigera. However, the molecular mechanism of resistance to H. armigera remains unknown. In this study, we conducted the transcript and protein profilings for ED059 and a susceptible soybean cultivar (cv.) Tianlong 2. In summary, 2213 genes and 116 proteins were specifically expressed in response to H. armigera in ED059, while 2179 genes and 9 proteins were specifically expressed in cv. Tianlong 2. Four hundred and four genes were significantly up-regulated in response to H. armigera in ED059 while down-regulated or not changed in cv. Tianlong 2. GO analyses showed the specifically expressed genes in ED059 were mainly enriched in metabolism-related pathways. The function of one potential resistant gene involved in the phenylpropanoid pathway was confirmed by use of transgenic Arabidopsis. Our study provided useful resources for the soybean improvement, which could contribute to the elucidation of defense mechanisms against chewing insects in plants.

SNP markers represent the most extensively distributed and abundant type of polymorphic markers. Although researchers have developed high-throughput detection methods for SNP markers, there remains a lack of simple, flexible, and cost-effective approaches for detecting small numbers of SNPs. To address this need, this study proposes a method for detecting SNPs based on T-C mismatch. Specifically, two upstream primers were designed with a single base difference at the 3’ end. Additionally, a T-C mismatch was introduced within 5 bp upstream of the 3’ end to distinguish SNPs. Detection was achieved through PCR amplification and subsequent gel electrophoresis analysis. In this study, twelve random SNPs from bitter gourd were selected, and the detection efficiency was found to be 83.3%. The proposed SNP detection method is characterized by its simplicity and flexibility, offering an effective tool for molecular marker-assisted selection breeding in applications such as stress resistance improvement and agricultural productivity enhancement.

MAP30, a ribosome-inactivating protein mainly isolated from the fruits and seeds of bitter gourd (Momordica charantia), has many pharmacological properties, such as anti-HIV and anti-tumor activities. Using qPCR analysis, we found that MAP30 transcription was higher in light, but drastically reduced in darkness. However, the molecular mechanism by which MAP30 is regulated remains unclear. Using yeast one-hybrid (Y1H) library screening, we found that McHY5, a homolog of AtHY5, bound to the MAP30 promoter. In addition, McHY5 and its homolog McHY5-X1 are identified as positive regulators binding to the MAP30 promoter by using Y1H, Electrophoretic mobility shift assay (EMSA) and Dual-Luciferase Reporter (DLR) assay. Due to MAP30 is not present in Arabidopsis we transformed pMAP30:MAP30 to Col and obtained transgenic line 18. Compared with pMAP30:MAP30/Col 18#, MAP30 transcription was largely reduced in pMAP30:MAP30 18#/Athy5–215 whether under light or dark conditions. Moreover, approximately 70 % of the tested bitter gourd germplasms showed consistent expression patterns of McHY5/McHY5-X1 and MAP30 under light conditions. In contrast to McHY5, McPIF1-X1 and McPIF3 acted as inactivators to inhibit MAP30 transcription. In this study, we found that MAP30 transcription was activated by McHY5 and McHY5-X1 in light, while McPIF1-X1 and McPIF3 inhibited MAP30 transcription mainly in darkness. Conclusively, we established the antagonistic module of McHY5/McHY5-X1 and McPIF1-X1/McPIF3 in MAP30 transcription regulation. This work expands our understanding of the regulatory mechanism of MAP30 and provides a valuable conceptual guidance for cultivating high pharmacological activity bitter gourd varieties with high MAP30 protein levels.