Genome-wide identification of RUB activating enzyme and conjugating enzyme gene families and their expression analysis under abiotic stresses in Capsicum annuum.

NEDD8/RUB, as a ubiquitin-like protein, participates in the post-translational modification of protein and requires unique E1, E2, and E3 enzymes to bind to its substrate. The RUB E1 activating enzyme and E2 conjugating enzyme play a significant role in the neddylation. However, it is unknown whether RUB E1 and E2 exist in pepper and what its function is. In this study, a total of three putative RUB E1 and five RUB E2 genes have been identified in the pepper genome. Subsequently, their physical and chemical properties, gene structure, conserved domains and motifs, phylogenetic relationship, and cis-acting elements were analyzed. The structure and conserved domain of RUB E1 and E2 are similar to that of Arabidopsis and tomato. The RUB E1 and E2 genes were randomly distributed on seven chromosomes, and there were two pairs of collinearity between pepper and Arabidopsis and eight pairs of collinearity between pepper and tomato. Phylogenetic analysis reveals that RUB E1 and E2 genes of pepper have a closer relationship with that of tomato, potato, and Nicotiana attenuate. The cis-elements of RUB E1 and E2 genes contained hormone response and stress response. RUB E1 and E2 genes were expressed in at least one tissue and CaRCE1.3 and CaRCE2.1 were exclusively expressed in flowers and anthers. Moreover, the expression of RUB E1 genes (CaECR1, CaAXR1.1, and CaAXR1.2) and RUB E2 genes (CaRCE1.1, CaRCE1.2, and CaRCE2.1) was increased to varying degrees under low-temperature, drought, salt, ABA, and IAA treatments, while CaRCE1.3 and CaRCE2.2 were down-regulated under low-temperature treatment. In addition, these genes were hardly expressed under MeJA treatment. In summary, this study provides a theoretical foundation to explore the role of RUB E1 and E2 in the response of plants to stress.

Identification and bioinformatic analysis of the CaCesA/Csls family members and the expression of the CaCslD1 in the flower buds of CMS/Rf system in pepper.

The cellulose synthase gene superfamily contains cellulose synthase (CesA) and cellulose synthase-like (Csl) gene families, which synthesize cellulose and hemicellulose in plant cell walls and play a crucial role in plant growth and development. However, the CesA/Csl gene family has not been reported in pepper. Therefore, the genome-wide research of the CaCesA/CaCsl gene family was conducted in pepper. In this study, a total of 39 CaCesA/CaCsls genes (10 CesAs genes and 29 Csls genes) were identified in pepper and unevenly distributed on 11 chromosomes. These CaCesA/Csls were divided into seven subfamilies (CesAs, CslAs, CslBs, CslCs, CslDs, CslEs, CslGs), and most of CaCesA/Csls genes are closely related to AtCesA/Csls genes. The cis-acting elements in the promoters of CaCesA/Csls genes are mainly related to hormone response and stress response. There are ten collinear gene pairs between the CesA/Csls gene family of pepper and Arabidopsis, and four fragment duplication gene pairs of the CaCesA/Csls genes were discovered. RNA-seq analysis shows that the majority of CaCesA/Csls are expressed in a variety of plant tissues, indicating that most CaCesA/Csls gene expression patterns are not organ-specific, and CaCslD1/D4 have the highest expression in anthers, followed by petal, ovary, and F9. RNA-seq analysis shows that most CaCesA/Csls are responsive to five hormones (IAA, GA3, ABA, SA, and MeJA). The tissue-specific expression analysis of the CaCslD1 gene shows that the CaCslD1 gene is expressed specifically in flowers. In the flower buds IV of cytoplasmic male sterility (CMS) and its restoration of fertility (Rf) system, CaCslD1 reach the highest expression respectively. However, the relative expression level of CaCslD1 in the fertile accessions is extremely significantly higher than in the sterile accessions. This study shows an overall understanding of the CaCesA/Csls gene family and provides a new insight for understanding the function of CaCslD1 in pollen development and exploring the fertility restoration of CMS in pepper.

Identification and relative expression analysis of CaFRK gene family in pepper.

Fructokinase is the main catalytic enzyme for fructose phosphorylation and can also act as a glucose receptor and signal molecule to regulate the metabolism of plants, which plays an important role in plant growth and development. In this study, the CaFRK gene family and their molecular characteristics are systematically identified and analyzed, and the specific expression of CaFRKs under different tissues, abiotic stresses and hormone treatments were explored. Nine FRK genes were authenticated in pepper genome database, which were dispersedly distributed on eight reference chromosomes and predicted to localize in the cytoplasm. Many cis-acting elements that respond to light, different stresses, hormones and tissue-specific expression were found in the promoters of CaFRKs. FRK proteins of four species including Capsicum annuum, Arabidopsis thaliana, Solanum lycopersicum and Oryza sativa were divided into four groups via phylogenetic analysis. The collinearity analysis showed that there were two collinear gene pairs between CaFRKs and AtFRKs. In addition, it was significantly found that CaFRK9 expressed far higher in flower than other tissues, and the relative expression of CaFRK9 was gradually enhanced with the development of flower buds in fertile accessions, 8B, R1 and F1. Nevertheless, CaFRK9 hardly expressed in all stages of cytoplasmic male sterile lines. Based on the quantitative real-time PCR, most of CaFRK genes showed significant up-regulation under low-temperature, NaCl and PEG6000 treatments. On the contrary, the expression levels of most CaFRKs revealed a various trend in response to hormone treatments (IAA, ABA, GA3, SA and MeJA). This study systematically analyzed CaFRK gene family and studied its expression pattern, which lay the foundation of CaFRK genes cloning and functional verification response to abiotic stresses, and provides new insights into exploring the CaFRK genes on the pollen development in pepper. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03196-1.

Identification and expression analysis of phospholipase C family genes between different male fertility accessions in pepper.

Phospholipase C (PLC) is one of the major lipid-hydrolyzing enzymes, involved in lipid-mediating signal pathway. PLCs have been found to play a significant role in the growth and development of plants. In this study, the genome-wide identification and characteristic analysis of CaPLC family genes in pepper were conducted and the expression of two CaPLC genes were investigated. The results showed that a total of 11 CaPLC family genes were systematically identified, which were distributed on five chromosomes and divided into two groups based on their evolutionary relevance. Some cis-elements responding to different hormones and stresses were screened in the promoters of CaPLC genes. Quantitative real-time PCR indicated that the expression of CaPIPLC1 and CaPIPLC5 in flowers were dozens of times higher than in other tissues. In addition, with the development of flower buds, the relative expressions of CaPIPLC1 and CaPIPLC5 gradually increased in fertile materials R1 and F1. However, no expression of CaPIPLC1 and CaPIPLC5 were detected at all developmental stages of cytoplasmic male sterile lines (CMS) compared with fertile accessions. The study revealed the number and characteristics of the CaPLC family genes, which supplied a basic and systematic understanding of CaPLC family. In addition, these findings provided new insights into the role of CaPLC genes in pollen development and fertility restoration in pepper.