Plastome evolution of Engelhardia facilitates phylogeny of Juglandaceae.

BACKGROUND: Engelhardia (Juglandaceae) is a genus of significant ecological and economic importance, prevalent in the tropics and subtropics of East Asia. Although previous efforts based on multiple molecular markers providing profound insights into species delimitation and phylogeography of Engelhardia, the maternal genome evolution and phylogeny of Engelhardia in Juglandaceae still need to be comprehensively evaluated. In this study, we sequenced plastomes from 14 samples of eight Engelhardia species and the outgroup Rhoiptelea chiliantha, and incorporated published data from 36 Juglandaceae and six outgroup species to test phylogenetic resolution. Moreover, comparative analyses of the plastomes were conducted to investigate the plastomes evolution of Engelhardia and the whole Juglandaceae family. RESULTS: The 13 Engelhardia plastomes were highly similar in genome size, gene content, and order. They exhibited a typical quadripartite structure, with lengths from 161,069 bp to 162,336 bp. Three mutation hotspot regions (TrnK-rps16, ndhF-rpl32, and ycf1) could be used as effective molecular markers for further phylogenetic analyses and species identification. Insertion and deletion (InDels) may be an important driving factor for the evolution of plastomes in Juglandoideae and Engelhardioideae. A total of ten codons were identified as the optimal codons in Juglandaceae. The mutation pressure mostly contributed to shaping codon usage. Seventy-eight protein-coding genes in Juglandaceae experienced relaxed purifying selection, only rpl22 and psaI genes showed positive selection (Ka/Ks > 1). Phylogenetic results fully supported Engelhardia as a monophyletic group including two sects and the division of Juglandaceae into three subfamilies. The Engelhardia originated in the Late Cretaceous and diversified in the Late Eocene, and Juglandaceae originated in the Early Cretaceous and differentiated in Middle Cretaceous. The phylogeny and divergence times didn't support rapid radiation occurred in the evolution history of Engelhardia. CONCLUSION: Our study fully supported the taxonomic treatment of at the section for Engelhardia species and three subfamilies for Juglandaceae and confirmed the power of phylogenetic resolution using plastome sequences. Moreover, our results also laid the foundation for further studying the course, tempo and mode of plastome evolution of Engelhardia and the whole Juglandaceae family.

A novel Biochar-PGPB strategy for simultaneous soil remediation and safe vegetable production.

There is currently increasing pressure on agriculture to simultaneously remediate soil and ensure safe agricultural production. In this study, we investigate the potential of a novel combination of biochar and plant growth-promoting bacteria (PGPB) as a promising approach. Two types of biochar, corn stover and rice husk-derived, were used in combination with a PGPB strain, Bacillus sp. PGP5, to remediate Cd and Pb co-contaminated soil and enhance lettuce performance. The contaminated soil was pre-incubated with biochar prior to PGP5 inoculation. The combined application of biochar and PGPB reduced the diethylenetriaminepentaacetic acid (DTPA) -extractable Cd and Pb concentrations in the soil by 46.45%-55.96% and 42.08%-44.83%, respectively. Additionally, this combined application increased lettuce yield by 23.37%-65.39% and decreased Cd and Pb concentrations in the edible parts of the lettuce by 57.39%-68.04% and 13.57%-32.50%. The combined application showed a better promotion on lettuce growth by facilitating chlorophyll synthesis and reducing oxidative stress. These demonstrated a synergistic effect between biochar and PGPB. Furthermore, our study elucidated the specific role of the biochar-PGPB combination in soil microbial communities. Biochar application promoted the survival of PGP5 in the soil. The impact of biochar or PGPB on microbial communities was found to be most significant in the early stage, while the development of plants had a greater influence on rhizosphere microbial communities in later stage. Plants showed a tendency to recruit plant-associated microbes, such as Cyanobacteria, to facilitate growth processes. Notably, the combined application of biochar and PGPB expedited the assembly of microbial communities, enabling them more closely with the rhizosphere microbial communities in late stage of plant development and thus enhancing their effects on promoting plant growth. This study highlights the "accelerating" advantage of the biochar-PGPB combination in the assembly of rhizosphere microbiomes and offers a new strategy for simultaneous soil remediation and safe agricultural production.

Comprehensive Analysis of the Complete Mitochondrial Genome of Rehmannia chingii: An Autotrophic Species in the Orobanchaceae Family.

Rehmannia chingii is an important medicinal plant with immense value in scientific research. However, its mitochondrial genome (mitogenome) has not yet been characterized. Herein, based on whole-genome Illumina short reads and PacBio HiFi reads, we obtained the complete mitogenome of R. chingii through a de novo assembly strategy. We carried out comparative genomic analyses and found that, in comparison with the plastid genome (plastome) showing a high degree of structural conservation, the R. chingii mitogenome structure is relatively complex, showing an intricate ring structure with 16 connections, owing to five repetitive sequences. The R. chingii mitogenome was 783,161 bp with a GC content of 44.8% and contained 77 genes, comprising 47 protein-coding genes (CDS), 27 tRNA genes, and 3 rRNA genes. We counted 579 RNA editing events in 47 CDS and 12,828 codons in all CDSs of the R. chingii mitogenome. Furthermore, 24 unique sequence transfer fragments were found between the mitogenome and plastome, comprising 8 mitogenome CDS genes and 16 plastome CDS genes, corresponding to 2.39% of the R. chingii mitogenome. Mitogenomes had shorter but more collinear regions, evidenced by a comparison of the organelles of non-parasitic R. chingii, hemiparasitic Pedicularis chinensis, and holoparasitic Aeginetia indica in the Orobanchaceae family. Moreover, from non-parasitic to holoparasitic species, the genome size in the mitogenomes of Orobanchaceae species did not decrease gradually. Instead, the smallest mitogenome was found in the hemiparasitic species P. chinensis, with a size of 225,612 bp. The findings fill the gap in the mitogenome research of the medicinal plant R. chingii, promote the progress of the organelle genome research of the Orobanchaceae family, and provide clues for molecular breeding.