Genetic diversity, population structure, and DNA fingerprinting of Ailanthus altissima var. erythrocarpa based on EST-SSR markers.

Ailanthus altissima var. erythrocarpa is an A. altissima variety with high economic, ecological and ornamental value, but there have been no reports on the development of SSR primers for it. According to the SSR primer information provided by the transcriptome of A. altissima var. erythrocarpa, 120 individuals with different redness levels were used to screen polymorphic primers. Transcriptomic analysis revealed 10,681 SSR loci, of which mononucleotide repeats were dominant (58.3%), followed by dinucleotide and trinucleotide repeats (16.6%, 15.1%) and pentanucleotide repeats (0.2%). Among 140 pairs of randomly selected primers, nineteen pairs of core primers with high polymorphism were obtained. The average number of alleles (Na), average number of effective alleles (Ne), average Shannon's diversity index (I), average observed heterozygosity (Ho), average expected heterozygosity (He), fixation index (F) and polymorphic information content (PIC) were 11.623, 4.098, 1.626, 0.516, 0.696, 0.232 and 0.671, respectively. Nineteen EST-SSR markers were used to study the genetic diversity and population structure of A. altissima var. erythrocarpa. The phylogenetic tree, PCoA, and structure analysis all divided the tested resources into two categories, clearly showing the genetic variation between individuals. The population showed high genetic diversity, mainly derived from intraspecific variation. Among nineteen pairs of primers, 4 pairs (p33, p15, p46, p92) could effectively distinguish and be used for fingerprinting of the tested materials. This study is of great significance for genetic diversity analysis and molecular-assisted breeding of A. altissima var. erythrocarpa.

Effect of T-DNA Integration on Growth of Transgenic Populus × euramericana cv. Neva Underlying Field Stands.

Multigene cotransformation has been widely used in the study of genetic improvement in crops and trees. However, little is known about the unintended effects and causes of multigene cotransformation in poplars. To gain insight into the unintended effects of T-DNA integration during multigene cotransformation in field stands, here, three lines (A1-A3) of Populus × euramericana cv. Neva (PEN) carrying Cry1Ac-Cry3A-BADH genes and three lines (B1-B3) of PEN carrying Cry1Ac-Cry3A-NTHK1 genes were used as research objects, with non-transgenic PEN as the control. Experimental stands were established at three common gardens in three locations and next generation sequencing (NGS) was used to identify the insertion sites of exogenous genes in six transgenic lines. We compared the growth data of the transgenic and control lines for four consecutive years. The results demonstrated that the tree height and diameter at breast height (DBH) of transgenic lines were significantly lower than those of the control, and the adaptability of transgenic lines in different locations varied significantly. The genotype and the experimental environment showed an interaction effect. A total of seven insertion sites were detected in the six transgenic lines, with B3 having a double-site insertion and the other lines having single copies. There are four insertion sites in the gene region and three insertion sites in the intergenic region. Analysis of the bases near the insertion sites showed that AT content was higher than the average chromosome content in four of the seven insertion sites within 1000 bp. Transcriptome analysis suggested that the differential expression of genes related to plant hormone transduction and lignin synthesis might be responsible for the slow development of plant height and DBH in transgenic lines. This study provides an integrated analysis of the unintended effects of transgenic poplar, which will benefit the safety assessment and reasonable application of genetically modified trees.

Tolerance of phyllospheric Wickerhamomyces anomalus to BDE-3 and heavy metals.

Few research have focused on the potential microorganism and gene resources for plant resistance to polybrominated diphenyl ether (PBDE) and heavy metal (HM) co-contamination. The purpose of this study was to investigate the impact of phyllospheric Wickerhamomyces anomalus bioremediation ability on PBDE and HM co-contamination. The results showed that the toleration capability of W. anomalus to cadmium (Cd2+) was higher than that to chromium (Cr) or 4-bromodiphenyl ether (BDE-3) contamination. The threshold levels of W. anomalus tolerance to BDE-3, Cd2+, and Cr were 30 mg/L, 500 mg/L, 30 mg/L, respectively. The use of the higher concentration of BDE-3 (30 mg/L) as a carbon source may improve tolerance to Cd2+ and Cr (10 mg/L Cd2+ and 10 mg/L Cr). Overexpression of Wapdr15 gene of ABCG subfamily from W. anomalus improved the tolerance to BDE-3 (10 mg/mL) and Cd2+ (0.5 mg/mL) significantly in transgenic tobacco lines. The synergism effect of BDE-3 and Cd2+ stress existed similarly in W. anomalus and transgenic lines. The findings suggest that W. anomalus should be taken into account for providing an efficient method in improving crops' tolerance during PBDE and HM co-contamination in soil.

Niches and Seasonal Changes, Rather Than Transgenic Events, Affect the Microbial Community of Populus × euramericana 'Neva'.

Exploring the complex spatiotemporal changes and colonization mechanism of microbial communities will enable microbial communities to be better used to serve agricultural and ecological operations. In addition, evaluating the impact of transgenic plants on endogenous microbial communities is necessary for their commercial application. In this study, microbial communities of Populus × euramericana 'Neva' carrying Cry1Ac-Cry3A-BADH genes (ECAA1 line), Populus × euramericana 'Neva' carrying Cry1Ac-Cry3A-NTHK1 genes (ECAB1 line), and non-transgenic Populus × euramericana 'Neva' from rhizosphere soil, roots, and phloem collected in different seasons were compared and analyzed. Our analyses indicate that the richness and diversity of bacterial communities were higher in the three Populus × euramericana 'Neva' habitats than in those of fungi. Bacterial and fungal genetic-distance-clustering results were similar; rhizosphere soil clustered in one category, with roots and phloem in another. The diversity and evenness values of the microbial community were: rhizosphere soil > phloem > root system. The bacterial communities in the three habitats were dominated by the Proteobacteria, and fungal communities were dominated by the Ascomycota. The community composition and abundance of each part were quite different; those of Populus × euramericana 'Neva' were similar among seasons, but community abundance fluctuated. Seasonal fluctuation in the bacterial community was greatest in rhizosphere soil, while that of the fungal community was greatest in phloem. The transgenic lines ECAA1 and ECAB1 had a bacterial and fungal community composition similar to that of the control samples, with no significant differences in community structure or diversity among the lines. The abundances of operational taxonomic units (OTUs) were low, and differed significantly among the lines. These differences did not affect the functioning of the whole specific community. Sampling time and location were the main driving factors of changes in the Populus × euramericana 'Neva' microbial community. Transgenic events did not affect the Populus × euramericana 'Neva' rhizosphere or endophytic microbial communities. This study provides a reference for the safety evaluation of transgenic plants and the internal colonization mechanism of microorganisms in plants.

Physiological and biochemical responses and microscopic structure changes of Populus tomentosa Carr seedlings to 4-BDE exposure.

Populus species are very effective in remediation of contaminants. Polybrominated diphenyl ethers (PBDEs) are commonly used as flame retardants and are known to be persistent environmental pollutants. Numerous studies have shown that PBDEs are rising in human tissues and biota. 4-Monobrominated diphenyl ether (4-BDE), one of the less brominated PBDEs, was served as a model compound for biodegradation of lower brominated congeners. The present study was designed to clarify the effects of 4-BDE stress on morphological, physiological, and biochemical impacts of Populus tomentosa Carr in a tissue culture condition. Different concentrations of 4-BDE (3 and 30 mg L(-1)) were supplied alone or together with 0.5 mg L(-1) IBA in tissue culture media. With the concentration increased, 4-BDE caused negative effects on the microscopic structure of roots, stem, and leaves. The leaf color became shallow in low concentration of 4-BDE treatments and appeared albinism with 4-BDE concentration increased. The chlorophyll content and the leaf mass per area of albino leaves reduced significantly. 4-BDE also caused positive effects on the adventitious root differentiation and the biomass below 30 mg L(-1). With the 4-BDE treatment time increased (23, 47, and 58 days), the peroxidase (POD) activity displayed the decreasing trend. The proline content decreased first and then increased. Exposure to 4-BDE induced the malondialdehyde (MDA) to increase in leaves. Application of 4-BDE affected the endogenous hormone levels of cuttings in their adventitious roots inducing media. Below 0.3 mg L(-1), 4-BDE caused the faint expression of auxin-sensitive DR5::GUS reporter gene in Arabidopsis thaliana. Additionally, P. tomentosa Carr exhibited the better tolerance against 4-BDE in the range of less than 30 mg L(-1).