Altered sucrose metabolism and plant growth in transgenic Populus tomentosa with altered sucrose synthase PtSS3.

Improvement of wood quality is an important focus of forest genetics and breeding research. Sucrose synthase (SS) catalyzes the reaction of sucrose and uridine diphosphate into uridine diphosphate glucose and fructose. It is a key enzyme involved in cell wall formation during secondary growth by providing the UDP-Glucose substrate for cellulose biosynthesis. In this study, we isolated the single-copy gene PtSS3 from the SS gene family of Populus tomentosa and analyzed its structure. To identify its function in secondary growth, we generated 19 transgenic lines of P. tomentosa using PtSS3 overexpression (OE) and artificial microRNA (amiRNA) constructs. We also performed comprehensive analyses of the transgenic P. tomentosa plants, including phenotypic analyses, quantitative real-time PCR, enzyme activity assays and sugar metabolism. We found significantly higher PtSS3 enzyme activity, fructose, and glucose levels and significantly lower sucrose levels in the stems and leaves of OE-PtSS3 plants. The opposite trend was observed in the amiRNA-PtSS3 lines. Gene expression analyses showed that PtSS3 transcript levels in stems and leaves were up-regulated in the OE-PtSS3 lines and down-regulated in the amiRNA-PtSS3 lines, and the OE-PtSS3 plants grew taller than the wild-type and amiRNA-PtSS3 plants. These findings indicate that PtSS3 plays an important role in sucrose metabolism and growth of trees.

Overexpression of AtAP1M3 regulates flowering time and floral development in Arabidopsis and effects key flowering-related genes in poplar.

APETALA1 plays a crucial role in the transition from vegetative to reproductive phase and in floral development. In this study, to determine the effect of AP1 expression on flowering time and floral organ development, transgenic Arabidopsis and poplar overexpressing of AtAP1M3 (Arabidopsis AP1 mutant by dominant negative mutation) were generated. Transgenic Arabidopsis with e35Spro::AtAP1M3 displayed phenotypes with delayed-flowering compared to wild-type and flowers with abnormal sepals, petals and stamens. In addition, transgenic Arabidopsis plants exhibited reduced growth vigor compared to the wild-type plants. Ectopic expression of AtAP1M3 in poplar resulted in up- or down-regulation of some endogenous key flowering-related genes, including floral meristems identity gene LFY, B-class floral organ identity genes AP3 and PI, flowering pathway integrator FT1 and flower repressors TFL1 and SVP. These results suggest that AtAP1M3 regulates flowering time and floral development in plants.

High quality haplotype-resolved genome assemblies of Populus tomentosa Carr., a stabilized interspecific hybrid species widespread in Asia.

Populus has a wide ecogeographical range spanning the Northern Hemisphere, and interspecific hybrids are common. Populus tomentosa Carr. is widely distributed and cultivated in the eastern region of Asia, where it plays multiple important roles in forestry, agriculture, conservation, and urban horticulture. Reference genomes are available for several Populus species, however, our goals were to produce a very high quality de novo chromosome-level genome assembly in P. tomentosa genome that could serve as a reference for evolutionary and ecological studies of hybrid speciation throughout the genus. Here, combining long-read sequencing and Hi-C scaffolding, we present a high-quality, haplotype-resolved genome assembly. The genome size was 740.2 Mb, with a contig N50 size of 5.47 Mb and a scaffold N50 size of 46.68 Mb, consisting of 38 chromosomes, as expected with the known diploid chromosome number (2n = 2x = 38). A total of 59,124 protein-coding genes were identified. Phylogenomic analyses revealed that P. tomentosa is comprised of two distinct subgenomes, which we deomonstrate is likely to have resulted from hybridization between Populus adenopoda as the female parent and Populus alba var. pyramidalis as the male parent, with an origin of approximately 3.93 Ma. Although highly colinear, significant structural variation was found between the two subgenomes. Our study provides a valuable resource for ecological genetics and forest biotechnology.