Long-distance transport or systemic silencing effects of exogenous biologically active RNA molecules in higher plants have not been reported. Here, we report that cationized bovine serum albumin (cBSA) avidly binds double-stranded beta-glucuronidase RNA (dsGUS RNA) to form nucleic acid–protein nanocomplexes. In our experiments with tobacco and poplar plants, we have successfully demonstrated systemic gene silencing effects of cBSA/dsGUS RNA nanocomplexes when we locally applied the nanocomplexes from the basal ends of leaf petioles or shoots. We have further demonstrated that the cBSA/dsGUS RNA nanocomplexes are highly effective in silencing both the conditionally inducible DR5-GUS gene and the constitutively active 35S-GUS gene in leaf, shoot, and shoot meristem tissues. This cBSA/dsRNA delivery technology may provide a convenient, fast, and inexpensive tool for characterizing gene functions in plants and potentially for in planta gene editing.

Deciduous woody plants like poplar (Populus spp.) have seasonal bud dormancy. It has been challenging to simultaneously delay the onset of bud dormancy in the fall and advance bud break in the spring, as bud dormancy, and bud break were thought to be controlled by different genetic factors. Here, we demonstrate that heterologous expression of the REVEILLE1 gene (named AaRVE1) from Agave (Agave americana) not only delays the onset of bud dormancy but also accelerates bud break in poplar in field trials. AaRVE1 heterologous expression increases poplar biomass yield by 166% in the greenhouse. Furthermore, we reveal that heterologous expression of AaRVE1 increases cytokinin contents, represses multiple dormancy-related genes, and up-regulates bud break-related genes, and that AaRVE1 functions as a transcriptional repressor and regulates the activity of the DORMANCY-ASSOCIATED PROTEIN 1 (DRM1) promoter. Our findings demonstrate that AaRVE1 appears to function as a regulator of bud dormancy and bud break, which has important implications for extending the growing season of deciduous trees in frost-free temperate and subtropical regions to increase crop yield.

Although several protocols for genetic transformation of citrus have been published, it is highly desirable to further improve its efficiency. Here we report treatments of Agrobacterium cells and citrus explants prior to and during co-cultivation process to enhance transformation efficiency using a commercially used rootstock ‘Carrizo’ citrange [Citrus sinensis (L.) Osb. × Poncirius trifoliata (L.) Raf.] as a model plant. We found explants from light-grown seedlings exhibited higher transformation efficiency than those from etiolated seedlings. We pre-cultured Agrobacterium cells in a 1/10 MS, 0.5 g/L 2-(N-morpholino) ethanesulfonic acid (MES) and 100 µM acetosyringone liquid medium for 6 h at 25 °C before used to infect citrus explants. We incubated epicotyl segments in an MS liquid medium containing 13.2 µM 6-BA, 4.5 µM 2,4-D, 0.5 µM NAA for 3 h at 25 °C prior to Agrobacterium infection. In the co-cultivation medium, we added 30 µM paclobutrazol and 10 µM lipoic acid. Each of these treatments significantly increased the efficiencies of transformation up to 30.4% (treating Agrobacterium with acetosyringone), 31.8% (treating explants with cytokinin and auxin), 34.9% (paclobutrazol) and 38.6% (lipoic acid), respectively. When the three treatments were combined, we observed that the transformation efficiency was enhanced from 11.5% to 52.3%. The improvement of genetic transformation efficiency mediated by these three simple treatments may facilitate more efficient applications of transgenic and gene editing technologies for functional characterization of citrus genes and for genetic improvement of citrus cultivars.

Impatiens L., the largest genus in the family Balsaminaceae with approximately 1000 species, is a controversial and complex genus that includes many economically important species well known for medicinal and ornamental values. However, there is limited knowledge of molecular phylogeny and chloroplast genomics, and uncertainties still exist at a taxonomic level. In this study, we have assembled four chloroplast genomics specimens of Impatiens cyanantha and Impatiens monticola, which are found at the different altitudes of Guizhou and Yunnan in China, and compared them with previously published three wild Balsaminaceae species (Impatiens piufanensis, Impatiens glandlifera, and Hydrocera triflora). The complete chloroplast genome sequences ranged from 152,236 bp (I. piufanensis) to 154,189 bp (H. triflora) and encoded 115 total distinct genes, of which 81 were protein-coding, 30 were distinct transfer RNA genes(tRNA), and 4 were ribosomal RNA genes (rRNA). A comparative analysis of I. cyanantha (Guizhou) vs. I. cyanantha (Yunnan) and I. monticola (Guizhou) vs. I. monticola (Yunnan) revealed minor changes in lengths; however, similar gene contents, gene orders, and GC contents existed among them. Interestingly, highly coding and non-coding genes, and regions matK, psbK, atpH-atpI, trnC-trnT, petN, psbM, atpE, rbcL, accD, psaL, rps3-rps19, ndhG-ndhA,rpl16, rpoB, ndhB, ndhF, ycf1, and ndhH were found, which could be suitable for identification of species and phylogenetic studies. During the comparison between I. cyanantha (Guizhou) and I. cyanantha (Yunnan), we observed that the rps4, ycf2, ndhF, ycf1, and rpoC2 genes underwent positive selection. Meanwhile, in the comparative study of I. monticola (Guizhou) vs. I. monticola (Yunnan), The accD and ycf1 genes were positively selected. Additionally, phylogenetic relationships based on maximum likelihood (ML) and Bayesian inference (BI) among whole chloroplast genomes showed that a sister relationship with I. monticola (Guizhou) and I. monticola (Yunnan) formed a clade with I.piufanensis proving their close connection. Besides, I.cyanantha (Guizhou) and I. cyanantha (Yunnan) formed a clade with I. glandlifera. Along with the findings and the results, the current study might provide valuable significant genomic resources for systematics and evolution of the genus impatiens in different altitudes of regions.

Transplanted rice cultivation has caused groundwater depletion in several regions globally. Direct-seeded rice under aerobic conditions is a water-saving alternative. However, under aerobic conditions, iron in the soil is oxidized from the ferrous to ferric forms, which are not easily available to rice crops, resulting in iron-deficiency-induced chlorosis (IDIC) and causing significant reductions in yield. Cultivated rice accessions have limited variations in IDIC tolerance, while the wild Oryza germplasm could be a potential source of IDIC tolerance. In this study, 313 Oryza accessions were evaluated for IDIC tolerance at the tillering stage under aerobic conditions and 20 IDIC-tolerant lines were identified. The twenty lines showed no signs of chlorosis and had high levels of iron content and SPAD values, while the eight cultivated controls exhibited varying degrees of chlorosis symptoms and low levels of SPAD and iron content. To confirm their tolerance, the selected lines were evaluated again in a subsequent year, and they showed comparable levels of tolerance, indicating that these lines were efficient in iron uptake and utilization, resulting in maintained high chlorophyll and leaf area index. These accessions may be useful for developing IDIC-tolerant cultivars for aerobic rice cultivation and future studies of the molecular basis of IDIC tolerance.

Tillering and shade tolerance are important traits in turfgrass, influenced by environmental factors, nutrients, and hormones. Shade stress negatively affects tillering. In this study, two dwarf mutants, shadow-1 and shadow-2 , developed via Gamma-ray and fast-neutron mutagenesis, respectively, showed significantly higher tillering than the wild-type under greenhouse conditions. Both mutants demonstrated shade tolerance in plant height, grass quality, and color under 85% and 95% shade conditions, while shade-induced inhibition of tillering was observed in both the mutants and the wild-type. In comparison to wild-type plants under 95% shade conditions, we observed that the cytokinin biosynthetic gene IPT8 is upregulated, while the cytokinin inactivating gene CKX2 is downregulated in shadow-1 . Similarly, the GA biosynthetic genes CPS1, GA2ox3, and GA20ox1 are upregulated, while the GA inactivating gene GA20ox8 is downregulated in the shadow-1 mutant. Furthermore, the ethylene biosynthetic genes ACS and ACO are also downregulated in the shadow-1 mutant. Consistently, we observed that wild-type plants exhibit increased GA and reduced CK levels, while shadow-1 mutant plants have reduced GA but increased CK levels. This explains the shadow-1 mutant’s shade tolerance in terms of plant height, grass quality, and color. Conversely, the tillering inhibitor genes CRY1, MAX2, and SnRK1 are upregulated in both wild-type and shadow-1 mutant plants. Our results provide novel insights into the mechanisms behind tillering and shade tolerance in turfgrasses under shade conditions.