Ear shape of maize (Zea mays L.) is related to ear perimeter, kernel row number, and then kernel yield. To study the inheritance of fasciated ear and the relationship between ear flatness and other ear traits is of great importance in improving ear shape and then increasing yield of maize. In this study, the fasciated ear inbred line Yi16 and normal ear inbred line B73 were used to construct F 2 and F 2:3 populations to study the correlations between ear flatness and other ear traits, and identify quantitative trait loci (QTL) of fasciated ear-related traits in two environments. The results showed that ear flatness was positively correlated with cob flatness, ear perimeter, cob perimeter, and kernel row number and negatively correlated with ear length. The broad-sense heritabilities of ear flatness and cob flatness were 59.01 and 67.38%, respectively. A total of 50 QTL were detected. Three QTL (qCF1, qCF4, and qKRN4a) were detected in all generations and environments, and five QTL (qEF4, qCF7, qCP2, qCP4, and qHKW7) were detected in two generations or two environments. The consistent QTL qEF4, qCF4, and qKRN4a were located in the common region of umc1940-umc1631 (bins 4.09) within a distance of 2.4 cM, explaining >10% of phenotypic variation except qKRN4a in F 2. The five pairs of QTL (qEF3-HC × qEF4-HC, qCF1-BB × qCF4-BB, qCF1-BB × qCF7-BB, qKRN1a-BB × qKRN4a-BB, and qHKW3-HC × qHKW7-HC) were detected to have epistatic effects, contributing 7.31-13.95% of phenotypic variance.

Exopolysaccharides (EPSs) Pea is essential for wrinkly colony morphology, pellicle formation, and robust biofilm production in Pseudomonas putida . The second messenger cyclic diguanylate monophosphate (c-di-GMP) induces wrinkly colony morphology in P. putida through unknown mechanism(s). Herein, we found that c-di-GMP modulated wrinkly colony morphology via regulating expression of eppA ( PP_5586 ), a small individually transcribed gene with 177 base pairs, and this gene was adjacent to the upstream of pea cluster. Phenotype observation revealed that eppA was essential for Pea-dependent phenotypes. The deletion of eppA led to smooth colony morphology and impaired biofilm, which was analogous to the phenotypes with the loss of the entire pea operon. EppA expression was positively regulated by c-di-GMP via the transcriptional effector FleQ, and eppA was essential for the c-di-GMP-induced wrinkly colony morphology. Structure prediction results implied that EppA had two transmembrane regions, and Western blot revealed that EppA was located on cell membrane. Transcriptomic analysis indicated that EppA had no significant effect on transcriptomic profile of P. putida . Bacterial two-hybrid (BTH) assay suggested that there was no direct interaction between EppA and the proteins in pea cluster and adjacent operons. Overall, these findings reveal that EppA is essential for Pea-dependent phenotypes, and that c-di-GMP modulates Pea-dependent phenotypes via regulating eppA expression in P. putida . IMPORTANCE Microbe-secreted EPSs are high molecular weight polysaccharides that have the potential to be used as industrially important biomaterials. The EPS Pea in P. putida is essential for wrinkly colony morphology and pellicle formation. Here, we identified a function-unknown protein EppA, which was also essential for Pea-dependent wrinkly colony morphology and pellicle formation, and EppA was probably involved in Pea secretion. Meanwhile, our results indicated that the second messenger c-di-GMP positively regulated the expression of EppA, resulting in Pea-dependent wrinkly colony morphology. Our results reveal the relationship of c-di-GMP, EppA, and Pea-dependent phenotypes, and provide possible pathway to construct genetically engineered strain for high Pea production.

Two-component systems (TCSs) are predominant means by which bacteria sense and respond to environment signals. Genome of Pseudomonas putida contains dozens of putative TCS-encoding genes, but phenotypical-genotypical correlation and transcriptional regulation of these genes are largely unknown. Herein, we characterized function and transcriptional regulation of a conserved P. putida TCS, named TarR-TarS. TarS (PP_0769) encodes a potential histidine kinase, and tarR (PP_0768) encodes a potential response regulator. Protein-protein interaction assay and phosphorylation assay confirmed that TarR-TarS was a functional TCS. Growth assay under antibiotics revealed that TarR-TarS positively regulated bacterial resistance to multiple antibiotics. Pull-down assay revealed that TarR directly interacted with PP_0800 (a hypothetical protein) and GroEL (the chaperonin). GroEL played a positive role in antibiotic resistance, while PP_0800 seemed to have no effect on antibiotic resistance. The regulator FleQ indirectly activated tarR-tarS transcription. However, the second messenger c-di-GMP antagonized FleQ activation to inhibit tarR-tarS transcription. The sigma factor FliA directly activated tarR-tarS transcription via a consensus motif. These findings reveal function and transcriptional regulation of TarR-TarS, and enrich knowledge regarding the relationship between c-di-GMP and antibiotic susceptibility in P. putida.

Zeaxanthin, a natural fat-soluble pigment, not only increases plant resistance, but also has vital significance for human health. However, quantitative trait loci (QTL) and the epistatic effects of zeaxanthin concentration in maize kernel have not been well studied. To identify QTLs and analyse the epistatic effects of zeaxanthin concentration in maize kernel, two sets of segregating generations derived from the cross between HuangC (a high zeaxanthin concentration inbred line) and Rezi1 (a low zeaxanthin concentration inbred line) were evaluated in three different environments. One major-effect QTL, qZea6a, explains 41.4-71.4% of the phenotypic variation and two QTLs, qZea4a and qZea3a, show LOD > 3 for zeaxanthin concentration detected over two generations and three different environments. Four of the ten QTL pairs show epistatic effects, explaining 7.34-14.3% of the phenotypic variance. Furthermore, additivity was the major allelic action at zeaxanthin concentration QTLs located in F 2 and F 2:3 populations and plants with homozygous HuangC alleles have a strong genetic ability in enhancing zeaxanthin concentration in maize kernel. These results will contribute to understanding these complex loci better and provide awareness about zeaxanthin concentration to maize breeders and scientists involved in maize research.