The tree species Pinus tabulaeformis Carr. (P. tabulaeformis) is commonly planted in China due to its economic and ecological value. In order to identify one or more ectomycorrhizal (ECM) fungal species for future P. tabulaeformis afforestation, we investigated the effects of five ECM fungal species: Laccaria laccata, Boletus edulis, Gomphidius viscidus, Suillus grevillei, and Suillus luteus on the growth of P. tabulaeformis seedlings under greenhouse conditions. The growth parameters of P. tabulaeformis seedlings were evaluated 90 days following fungal colonisation. The majority of seedlings were significantly affected by ECM inoculation. Mycorrhizal inoculated seedlings were taller, had more lateral roots, and a greater biomass compared with the non-mycorrhizal (CK) seedlings. With the exception of G. viscidus, inoculated seedlings exhibited higher phosphorus, potassium, and nitrogen content compared with the CK seedlings. In addition, ECM colonisation increased the enzymatic activity of catalase, acidic phosphatase, protease, and the urease content in the rhizosphere soil. Our study showed that Laccaria laccata, Suillus grevillei, and Suillus luteus may be useful for improving the growth and cultivation of P. tabulaeformis seedlings. Furthermore, we observed that S. luteus inoculation increased the gas exchange parameters of P. tabulaeformis seedlings under field conditions.

Etiolation (a process of growing plants in partial or complete absence of light) promotes adventitious root formation in tetraploid black locust (Robinia pseudoacacia L.) cuttings. We investigated the mechanism underlying how etiolation treatment promotes adventitious root formation in tetraploid black locust and assessed global transcriptional changes after etiolation treatment. Solexa paired-end sequencing of complementary DNAs (cDNAs) from control (non-etiolated, NE) and etiolated (E) samples resulted in 107,564 unigenes. In total, 52,590 transcripts were annotated and 474 transcripts (211 upregulated and 263 downregulated) potentially involved in etiolation were differentially regulated. These genes were associated with hormone metabolism and response, photosynthesis, signaling pathways, and starch and sucrose metabolism. In addition, we also found significant differences of phytohormone contents, activity of following enzymes i.e., peroxidase, polyphenol oxidase and indole acetic acid oxidase between NE and E tissues during some cottage periods. The genes responsive to etiolation stimulus identified in this study will provide the base for further understanding how etiolation triggers adventitious roots formation in tetraploid black locus.

Mulberry (Morus alba L.), a moderately salt-tolerant tree species, is considered to be economically important. In this study, 1-year-old mulberry seedlings cultivated in soil under greenhouse conditions were treated with five concentrations of sodium chloride (NaCl; 0%, 0.1%, 0.2%, 0.3%, and 0.5%) for 3 and 21 days. Plant growth parameters were not affected by 0.1% NaCl, but significant reductions were observed after treatment with 0.2%, 0.3%, and 0.5% NaCl. The malondialdehyde content and cell membrane stability of mulberry seedlings exposed to 0.1% NaCl did not change, indicating that mulberry is not significantly affected by low-salinity conditions. The Na contents of various organs did not increase significantly in response to 0.1% NaCl, but the K:Na, Mg:Na, and Ca:Na ratios of various organs were affected by NaCl. Marked changes in the levels of major compatible solutes (proline, soluble sugars, and soluble proteins) occurred in both the leaves and roots of NaCl-treated seedlings relative to control seedlings. Under severe saline conditions (0.5% NaCl), the ability of mulberry to synthesize enzymatic antioxidants may be impaired.

The DREB transcription factors regulate multiple stress response genes, and are therefore useful for molecular plant breeding. AhDREB, a stress-inducible gene, was isolated from Atriplex hortensis L. and introduced into Populus tomentosa Carrière under the control of the CaMV35S promoter. Under salt stress, the chlorophyll content and net photosynthetic rate were higher in transgenic lines than in the wild type (WT). Moreover, the rate of electrolyte penetration (REC) was lower in the transgenic lines. Additional analyses revealed that the AhDREB transgenic plants generally displayed lower malondialdehyde (MDA) activity but higher superoxide dismutase (SOD) and peroxidase (POD) activities and proline content than the WT under salt stress. RNA sequencing indicated that AhDREB could enhance tolerance to salt by activating various downstream genes in the transgenic plants. Furthermore, no growth inhibition was detected in transgenic plants expressing AhDREB driven by the constitutive CaMV35S promoter. The transcriptome showed 165 and 52 differentially expressed genes in transgenic plants under stress and non-stress conditions, respectively, among which no significant metabolic pathway was enriched and no unintended effects have yet been identified. Together, these results suggest that AhDREB may be a good candidate gene for increasing salt tolerance in transgenic poplar breeding.

Populus tomentosa grow rapidly, but are salt susceptible. To quickly and efficiently gain new poplar breeds with better salt resistance, a DREB transcription factor derived from Atriplex hortensis was transformed into triploid Populus tomentosa by our lab, which significantly improved the salt tolerance of host plants. However, environmental impacts of transgenic plants must be assessed before large-scale cultivation in China. Here, we conducted a field trial of AhDREB1 transgenic and non-transgenic triploid Populus tomentosa to assess the impact of transgenic trees on rhizospheric soil microbial communities and allelopathic activity of leaves. No significant differences in the number of soil microbes present were detected between the transgenic lines and the non-transgenic controls. The allelopathic activity of leaves from both the transgenic and non-transgenic lines varied with sampling time, but did not differ significantly between the transgenic and non-transgenic lines. These results indicate that the impact on the environment of AhDREB1 transgenic P. tomentosa did not differ significantly from that of the non-transformed controls for the variables observed in this field trial. We also investigated the persistence of AhDREB1 genes in decomposing transgenic poplar leaf on the soil under natural conditions for five months, and our data indicated that fragments of the genetically modified DNA were not detectable in the field after more than two months. We used a triphenyl tetrazolium chloride test (TTC) (or pollen germination method) and hybridization to test the pollen viability and fertility, respectively, of the transgenic and non-transgenic trees and the results showed that the pollen viability of both the transgenic and non-transgenic trees was extremely low in 2016; the receptor plant may have been sterile.