Populus wulianensis mainly grows in hilly and sloped areas and has strong resistance to adversity. Previous transcriptome studies have shown that a WRKY gene PwuWRKY48 is expression-induced under drought stress. In this study, we aim to characterize the gene’s structure and investigate its role in plant drought resistance. The results show that PwuWRKY48 (1113 bp) belongs to a class IIc WRKY subfamily and it was determined as a nuclear localization protein. The gene promoter region contains a variety of cis-elements in relation to stress resistance. Under drought stress, PwuWRKY48 was expression-induced in leaves and stems, 29.7 and 16.6 times those before treatment, respectively. Overexpressing PwuWRKY48 lines were associated with increased activities of peroxidase (POD) and superoxide dismutase (SOD), 2.5 and 1.6 times higher than those of the wild type. While malondialdehyde content (MDA), superoxide anion radical (O₂^·−), and relative conductivity were decreased by 20%, 30%, and 21.3%, proline and chlorophyll contents increased by 37.5% and 11.2%, respectively. This indicates that PwuWRKY48 efficiently improved the drought tolerance of transgenic plants. PwuWRKY48 can be used as a gene resource for molecular breeding of plant drought resistance.

The expansin genes are commonly expressed in plant cells, and the encoded proteins influence plant growth and stress resistance by loosening the structure and increasing the flexibility of the cell wall. The objective of this study was to characterize expansin gene promoters in Populus trichocarpa to clarify the regulatory mechanisms underlying gene expression and evolution. Sequence alignments revealed that the similarity among 36 poplar expansin genes was greater for the coding sequences than for the promoter sequences, which suggested these promoter sequences evolved asynchronously. The bases flanking the start codon exhibited a usage bias, with sites +3, +4, and +5 biased toward GC, whereas the other sites were biased toward AT. The flanking sites were significantly correlated with gene expression, especially sites −10 and −17, in which C and G are the bases positively associated with gene expression. A total of 435 regulatory elements (61 types) were identified on the promoters of the poplar expansin genes; Skn-1 was the most common element in 23 promoters. Some expansin genes had more regulatory elements on their promoters (e.g., PtrEXPA4, PtrEXPA3, PtrEXPB3, and PtrEXPB1), whereas some others had less (e.g., PtrEXLA2, PtrEXLB1, and PtrEXPA23). Furthermore, 26 types of elements were involved in expansin gene expression, 25 of which positively affected expression in all analyzed samples. The exception was the endosperm expression-related element Skn-1, which negatively regulated expression in four tissues or treatments. Expression analysis showed that the expansin genes in Populus trichocarpa performed much differently under regular and abiotic stress conditions, which well matched the diversity of their promoter sequences. The results show that expansin genes play an important role in plant growth and development and stress resistance through expression adjustment.

Dehydrins exist widely in plants and play an important role in abiotic stress resistance. Two low-temperature-induced dehydrin-like genes, PmCAP and LpCAP, from the pine species Pseudotsuga menziesii and Larix principis-rupprechtii Mayr were cloned and found to contain 576 bp and 687 bp, encoding 191 and 228 amino acids, respectively. Both genes were individually assembled into prokaryotic expression vectors and transferred into E. coli cells. When transgenic stains were cultured at −5 °C, the lethal time 50% (LT50) was 50 h and 54 h for PmCAP and LpCAP, respectively, compared with 32 h for the control. When cultured at −20 °C, the LT50 was 38 h, 41 h, and 25 h for PmCAP, LpCAP, and the control. Thermal hysteresis (TH) testing of PmCAP and LpCAP proteins revealed TH values of 0.27 °C and 0.72 °C, respectively, relative to 0.05 °C for the BSA control. These results indicate that the two pine dehydrin proteins have antifreeze characteristics and that their antifreeze levels were well in relation to the environmental conditions of pine growth (Larix principis-rupprechtii Mayr mostly grows in cold and high-altitude zones, while Pseudotsuga menziesii grows in temperate and low-altitude zones). LpCAP, especially, could be a better gene resource for the molecular breeding of plant cold resistance.

Bursaphelenchus xylophilus is a pine wood nematode capable of destroying pine forests. Exploring the genes providing resistance to this pathogen and understanding their resistance mechanisms is thus necessary and constitutes an effective way to tackle this problem. We used Pinus tabuliformis Carriere to dissect its response to B. xylophilus strain BxFC. The 30 d inoculation results showed that the P. tabuliformis germplasms exhibited a wide resistance spectrum. Some lines were sensitive with the needles fully wilted and the MDA content and the relative conductivity of needles greatly increased, while some lines demonstrated strong resistance with good needle vigor and better physiological conditions. Moreover, the transcriptome analysis revealed 7928 differentially expressed genes (DEGs) between the resistant and sensitive germplasm pools, including 3754 upregulated and 4174 downregulated genes in the resistant lines. These DEGs were specially enriched in the pathways of plant–pathogen interaction (318 genes), phenylpropanoid biosynthesis (108 genes), ubiquitin-mediated proteolysis (47 genes), carotenoid biosynthesis (18 genes), and monoterpenoid biosynthesis (9 genes). Accordingly, P. tabuliformis utilized multiple ways to control the proliferation and activity of B. xylophilus, such as immune response, ubiquitination, thickening plant cell walls, and increasing its terpenoid and antioxidant contents. Our results could thus help in better understanding the resistance process of P. tabuliformis against B. xylophilus and offer some new strategies and gene resources for a molecular breeding program of resistant P. tabuliformis.