An efficient transformation protocol is a primary requisite to study and utilize the genetic potential of any plant species. A quick transformation system is also crucial for the functional analysis of genes along with the study of proteins and their interactions in vivo. Presently, however, quick and effective transformation systems are still lacking for many plant species including pineapple. This has limited the full exploration of the genetic repository of pineapple as well as the study of its genes, protein localization and protein interactions. To address the above limitations, we have developed an efficient system for protoplast isolation and subcellular localization of desired proteins using pineapple plants derived from tissue culture. A cocktail of 1.5% (W/V) Cellulase R-10 and 0.5% (W/V) Macerozyme R-10 resulted in 51% viable protoplasts with 3 h digestion. Compared to previously reported protocols, our protoplast isolation method is markedly faster (saving 4.5 h), requires only a small quantity of tissue sample (1 g of leaves) and has high yield (6.5 × 105). The quality of the isolated protoplasts was verified using organelle localization in protoplasts with different organelle markers. Additionally, colocalization analysis of two pineapple Mg2+ transporter genes in pineapple protoplasts was consistent with the results in a tobacco transient expression system, confirming that the protoplast isolation method can be used to study subcellular localization. Further findings showed that the system is also suitable for protein–protein interaction studies. Based on our findings, the presently described method is an efficient and effective strategy for pineapple protoplast isolation and transformation; it is convenient and time saving and provides a greater platform for transformation studies.

The tea plant (Camellia sinensis) is an economically important crop that plays an important role not only in the beverage industry but also in the pharmaceutical industry. The environment has a great influence on the quality of the tea plant. Heat shock factors (Hsfs) are transcriptional regulators that control the plant response to adversity. However, only a limited number of studies have reported the Hsf gene in Camellia sinensis, and most of these reports involve high-temperature, drought, and salt stress. Research on light, dark, and cold stress is limited. In this study, 22 CsHsf genes were obtained by whole genome sequencing and found to be located on 11 chromosomes. In addition, the gene structure, protein motif, and phylogeny were studied. We classified the genes into three major subfamilies: CsHsfA, CsHsfB, and CsHsfC. Interestingly, we found that there was more alignment between CsHsf and Hsf genes in dicotyledons, including Arabidopsis thaliana and Solanum lycopersicum, than in the monocotyledon Oryza sativa. The expression of many CsHsf genes was affected by low-temperature, light, and dark abiotic stresses. Notably, CsHsf15 and CsHsf16 showed high induction rates under both light and cold stress, and both genes carried cis-acting elements associated with light and low-temperature responses. These results lay a solid groundwork for further investigations into the involvement of CsHsf genes in the response of Camellia sinensis to abiotic stresses.

In the original version of this article [1], the spelling of the tenth author name, Moakai Yan, was incorrect. The corrected name is given in this correction article.

Lead and zinc pollution is a prevalent issue in agricultural soils surrounding lead and zinc mines, posing a serious risk to crop growth and soil health. Heavy metal-resistant, plant growth-promoting bacteria (PGPB) capable of supporting plant development under high metal exposure have significant potential for mitigating these deleterious effects. Here we isolated and identified the Pb- and Zn-resistant and plant growth-promoting bacterial strain Serratia ureilytica T6 based on 16S rRNA and average nucleotide identity (ANI) analysis. Furthermore, 14 strains (T1–T14) from a rice paddy soil irrigated by Pb-Zn mine effluent were isolated and identified, and their phytopromoting characteristics were determined. Genome analysis of S. ureilytica T6 showed a genome size of 5,102,941 bp, with G + C content of 59.74%. A total of 4822 genes were annotated by RAST, among which 15 genes were putatively associated with Pb-Zn resistance. The genome of S. ureilytica T6 was found to possess multiple genes associated with probiotic properties by a comparative analysis of KEGG, GO, and COG databases. Several taxonomic identifications of S. ureilytica T6 revealed that strain T6 is Gram-negative, facultative anaerobic and motile. The pH growth range of S. ureilytica T6 was between 4.00 and 9.50; temperature growth range was 4–37 °C; NaCl tolerance was 0–9%. S. ureilytica T6 displayed a high tolerance to a variety of heavy metals, with minimum inhibitory concentrations of 1.5 and 9 mmol·L⁻¹ for Pb and Zn. S. ureilytica T6 can utilize a variety of carbon sources and nitrogen sources. T6 has the ability to produce indole-3-acetic acid (IAA), siderophore, and phosphorus and potassium solubilization, and it was initially judged that strain T6 has the potential for plant growth-promoting ability. Different plant growth-promoting effects of T6 inoculations were observed in improving rice biomass, plant height, etc. We observed that with increasing Pb and Zn stress, SOD activity first increased and then decreased, while POD and CAT activities gradually decreased. The addition of S. ureilytica T6 significantly enhanced the activities of SOD, POD, and CAT in rice seedlings under low to moderate Pb and Zn stress but had no significant effect under high concentrations (150 mg·L⁻¹) of Pb or Zn. In addition, S. ureilytica T6 has the potential to be used as a phytoremediation tool.

The TCP transcription factor (TF) family is a vital set of plant-specific regulators involved in plant growth, development, and responses to environmental stresses. Despite the extensive research on TCP transcription factors in numerous plant species, the functions they fulfill in Betula platyphylla are still not well understood. In this study, 21 BpTCP genes were identified via genome-wide analysis. Bioinformatics analysis was used to examine the physicochemical properties of these transcription factors, including molecular weight, isoelectric point, chromosomal distribution, and predicted subcellular localization. We expected that most BpTCP transcription factors would be located in the nucleus. Collinearity analysis revealed that gene fragment duplication events played a major role in the evolutionary expansion and diversification of the BpTCP gene family. Promoter analysis identified diverse cis-acting elements in BpTCP, suggesting that they play a role in stress responses, hormonal regulation, and plant growth and development. qRT-PCR analysis showed that BpTCP genes displayed tissue-specific expression patterns in the roots, stems, and leaves, displaying remarkable differences in expression levels when subjected to abiotic stresses, including drought and high- and low-temperature conditions. Notably, BpTCP17 and BpTCP18 showed markedly higher expression levels under multiple stress conditions. Subcellular localization experiments confirmed that both BpTCP17 and BpTCP18 localize in the nucleus, consistent with bioinformatic predictions. These findings emphasize the potential roles of BpTCP17 and BpTCP18 in mediating abiotic stress responses, highlighting their potential as candidate genes for improving stress tolerance in B. platyphylla.