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Zuo L.-H.,Agricultural University of Hebei | Zuo L.-H.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | Shang A.-Q.,Agricultural University of Hebei | Zhang S.,Agricultural University of Hebei | And 9 more authors.
PLoS ONE | Year: 2017

Elm (Ulmus) has a long history of use as a high-quality heavy hardwood famous for its resistance to drought, cold, and salt. It grows in temperate, warm temperate, and subtropical regions. This is the first report of Ulmaceae chloroplast genomes by de novo sequencing. The Ulmus chloroplast genomes exhibited a typical quadripartite structure with two single-copy regions (long single copy [LSC] and short single copy [SSC] sections) separated by a pair of inverted repeats (IRs). The lengths of the chloroplast genomes from five Ulmus ranged from 158,953 to 159,453 bp, with the largest observed in Ulmus davidiana and the smallest in Ulmus laciniata. The genomes contained 137-145 protein-coding genes, of which Ulmus davidiana var. japonica and U. davidiana had the most and U. pumila had the fewest. The five Ulmus species exhibited different evolutionary routes, as some genes had been lost. In total, 18 genes contained introns, 13 of which (trnL-TAA+, trnL-TAA-, rpoC1-, rpl2-, ndhA-, ycf1, rps12-, rps12+, trnA-TGC+, trnA-TGC-, trnV-TAC-, trnI-GAT+, and trnI-GAT) were shared among all five species. The intron of ycf1 was the longest (5,675bp) while that of trnF-AAA was the smallest (53bp). All Ulmus species except U. davidiana exhibited the same degree of amplification in the IR region. To determine the phylogenetic positions of the Ulmus species, we performed phylogenetic analyses using common protein-coding genes in chloroplast sequences of 42 other species published in NCBI. The cluster results showed the closest plants to Ulmaceae were Moraceae and Cannabaceae, followed by Rosaceae. Ulmaceae and Moraceae both belonged to Urticales, and the chloroplast genome clustering results were consistent with their traditional taxonomy. The results strongly supported the position of Ulmaceae as a member of the order Urticales. In addition, we found a potential error in the traditional taxonomies of U. davidiana and U. davidiana var. japonica, which should be confirmed with a further analysis of their nuclear genomes. This study is the first report on Ulmus chloroplast genomes, which has significance for understanding photosynthesis, evolution, and chloroplast transgenic engineering. © 2017 Zuo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Zhang Y.,Agricultural University of Hebei | Zhang Y.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | Zhang J.,Agricultural University of Hebei | Zhang J.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | And 7 more authors.
Journal of Forestry Research | Year: 2016

Extensive planting of Bacillus thuringiensis (Bt)-transgenic plants economically benefits society; however, the potential risk they pose is receiving increasing attention. This study used enzyme-linked immunosorbent assay and fluorescence quantitative PCR (RT-PCR) to monitor the temporal and spatial dynamics of the expression of Bt toxic protein in a forest of 6- to 8-year-old trees of transgenic insect-resistant poplar 741 for three consecutive years. The enrichment, distribution, and degradation of Bt toxic protein and the influence of transgenic poplars on the targeted insect population, Hyphantria cunea, were investigated. The content of Cry1Ac toxic protein dynamically changed in transgenic poplar. During the annual growth cycle, the content initially increased, then decreased in the long and the short branches of the crown and in the root system, peaking in August. During the study, the protein did not accumulate overtime. The mRNA transcription of gene Cry1Ac was almost consistent with the level of the protein, but transcription peaked in July. In the transgenic and control forestland, microscale levels of the Cry1Ac toxic protein were detected from the soil, but increased accumulation was not observed with the planting year of transgenic poplar. Meanwhile, Bt was isolated and detected molecularly from the soil in the experimental forestland. A systematic investigation of the density of H. cunea in the experimental transgenic poplar forest indicated that transgenic Pb29 poplar could resist insects to a certain degree. At peak occurrence of the targeted insects, the density of H. cunea in the experimental forest was significantly lower than in the nontransgenic poplar forest. © 2016 Northeast Forestry University and Springer-Verlag Berlin Heidelberg


Yang R.L.,Agricultural University of Hebei | Yang R.L.,Hebei University | Wang A.X.,Agricultural University of Hebei | Wang A.X.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | And 8 more authors.
Genetics and Molecular Research | Year: 2016

We characterized new transgenic varieties of poplar with multiple insect-resistant and salt stress tolerant genes. Two insect-resistant Bacillus thuringiensis (Bt) genes, Cry1Ac and Cry3A, and a salt-tolerant gene, Betaine aldehyde dehydrogenase (BADH) were inserted into a vector, p209-Cry1Ac-Cry3A-BADH. The clone of Populus × euramericana was transformed by the vector using the Agrobacterium-mediated method. Three transgenic lines were assessed using genetic detection and resistance expression analysis. PCR revealed that exogenous genes Cry1Ac, Cry3A, BADH and selective marker gene NPTII were present in three transgenic lines. Quantitative real-time PCR (qPCR) showed significant differences in the transcriptional abundance of three exogenous genes in different lines. Results of assays for Bt toxic proteins showed that the Cry1Ac and Cry3A toxic protein content of each line was 12.83-26.32 and 2108.91-2724.79 ng/g, respectively. The Cry1Ac toxic protein content of different lines was significantly different; the Cry3A toxic protein content was about 100 times higher than that of the Cry1Ac toxic protein. The insect-resistance test revealed the mortality rate of transgenic lines to Hyphantria cunea L1 larvae varied by 42.2-66.7%, which was significantly higher than non-transgenic lines. The mortality rate of L1 and L2 Plagiodera versicolora larvae was 100%. The insecticidal effect of transgenic lines to P. versicolora larvae was higher than that to H. cunea larvae. NaCl stress tolerance of three transgenic lines under 3-6% NaCl concentration was significantly higher than that of non-transgenic lines. © FUNPEC-RP.


Zuo L.,Agricultural University of Hebei | Zuo L.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | Yang M.,Agricultural University of Hebei | Yang M.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | And 4 more authors.
Journal of Animal and Plant Sciences | Year: 2015

Thirty pairs of polymorphism SSR primers were used for cluster analysis on 18 Xinjiang wild apple clones, 23 edible apple varieties, and 7 apple rootstock varieties. Forty eight varieties were classified into three groups: edible apple, Xinjiang wild apple, and rootstock apple, which do not interlace with each other. Varieties with genetic relationships were closely clustered inside the groups, consistent with the traditional family tree. This paper classified the primers nto three types. The relevant analysis of the comprehensive cluster results of all primers indicated the second primer group had the best correlation, with the coefficient of 0.957, whereas the first premier group had the lowest correlation, with the coefficient of 0.871. The second primer group had better distinguishing ability for edible apple and rootstock apple; the third primer group had poor distinguishing ability for rootstock apple; the first primer group can be applied to distinguish Xinjiang wild apple. The cluster results of the combination of 6 highly-effective primers on 48 apple samples had the correlation coefficient of 0.940 with the comprehensive cluster results of 30 pairs of primers. © 2015, Pakistan Agricultural Scientists Forum. All rights reserved.


Ren Y.,Agricultural University of Hebei | Ren Y.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | Dong Y.,Agricultural University of Hebei | Dong Y.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | And 6 more authors.
Journal of Animal and Plant Sciences | Year: 2015

Tobacco was transformed by employing the plant transformation vector p209-Cry1Ac-Cry3A-BADH by using Agrobacterium-mediated method to obtain completely regenerated plants screened by kanamycin sulfate. PCR detection indicated out of the nine lines in which NPT II gene was detected, Cry1Ac, Cry3A, and BADH were detected in seven lines; Cry1Ac and Cry3A were detected in one line; and Cry1Ac was detected in one line. Fluorescence quantitative PCR detection indicated in all target gene lines, three target genes were differentially expressed at the transcriptional level in four lines. BADH expression was absent in three lines. ELISA analysis revealed Cry1Ac and Cry3A toxin expression were detected in all target gene lines. The content of Cry3A toxin (up to 13,749.30 ng·g-1) was significantly higher than that of Cry1Ac toxin (up to 290.70 ng·g-1). The indoor insect-resistance test showed each transgenic line with insectresistant gene inhibited the survival, growth, and development of Prodenia litura (Fabricius) lar vae to varying degrees. Average corrected mortality of five lines was significantly higher than that of control, reaching up to 70.6%. Two lines were selected for further salt-tolerance research, and results showed transgenic lines had an individual salt tolerance compared with control. © 2015, Pakistan Agricultural Scientists Forum. All rights reserved.


Dong Y.,Agricultural University of Hebei | Dong Y.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | Du S.S.,Agricultural University of Hebei | Du S.S.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | And 6 more authors.
Canadian Journal of Forest Research | Year: 2014

To increase the expression efficiency of the Bt gene in transgenic poplar and expand the insect resistance spectrum, Cry1Ac and Cry3A were simultaneously constructed in two different plant transformation vectors. Two Bt genes on vector p71A68Y71 were separately driven by promoters CAMV35S and CoYMV, and a matrix attachment region (MAR) sequence was added to both sides; in contrast, two Bt genes on vector p05A68A71 were driven by the promoter CAMV35S without a MAR sequence structure. With the agrobacterium mediation method, two vectors transformed the high-quality hybrid clone poplar Juba of Populus deltoides (W. Bartram ex Marshall) and thus produced four transgenic resistant lines. Polymerase chain reaction (PCR) showed that the targeted genes were integrated into the poplar Juba genome. Fluorescence quantitative PCR and enzymelinked immunosorbent assay techniques were used to detect the transcription abundance and toxic protein expression of the Bt gene in the leaves of eight transgenic leaves. Results show the significant difference in transcription abundance of the two Bt genes between different lines. The transcription abundance of the gene Cry1Ac of the K series lines from vector p71A68Y71 was significantly higher than that of the H series lines from the other vector; however, no significant difference in the transcription abundance of gene Cry3A between the two vectors was observed. The transcription abundance of gene Cry3A was higher than that of gene Cry1Ac. Significant differences in the toxic expression of the two Bt genes existed among the lines but not in the H and K series lines. The Cry3A toxic protein content was much higher than the Cry1Ac toxic protein content in all lines, and the expression of the latter at 0.13 ng·g−1to 0.69 ng·g−1 was extremely low. The insect-resistant effect of the transgenic lines to lepidoptera pests such as the Hyphantria cunea (Drury) larva was unremarkable, and no significant difference was found in different lines and vectors. The lines strongly resisted coleoptera pests, e.g., Plagiodera versicolora (Laicharting), and their insectresistant effect was significantly higher than that of the highly resistant trans-Cry3A poplar line CC84. Significant differences were found between different lines, but no significant difference was found between the K and H series lines, which were from different vectors. Trangenic lines H4 and K1 had a higher transcription abundance, toxic protein expression, and lethal effect on P. versicolora than the other lines. © 2015 National Research Council of Canada. All rights reserved.


Xu L.N.,Agricultural University of Hebei | Xu L.N.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | Dong Y.,Agricultural University of Hebei | Dong Y.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | And 7 more authors.
Genetics and Molecular Research | Year: 2016

This study aimed to determine the influence of vector structure on dual Bt gene expression and establish an efficient expression vector using Cry1Ac and Cry3A genes. Four vectors (N4, N5, N10, and S23) were developed and used for genetic transformation of tobacco to obtain insect-resistant transgenic lines. The vectors were constructed using the MAR structure, applying different promoter and enhancer sequences, and changing the transgene open-reading frame sequence. The average Cry1Ac toxalbumin expression quantity was 67 times higher in N5 than in N4 transgenic lines (8.77 and 0.13 μg/g, respectively). In contrast, the average Cry3A toxalbumin expression quantity was 1.5 times higher in N4 than in N5 lines (12.70 and 8.21 μg/g, respectively). The sequences of both Bt genes significantly influenced toxalbumin expression, although upstream Bt genes presented lower expression levels. The average Cry1Ac toxalbumin content was 13 times higher in the transgenic lines of AtADH 5'-nontranslated sequence N5 (8.77 mg/g) than in the omega N10 lines (0.67 mg/g). Furthermore, the average Cry1Ac toxalbumin content was 5 times higher in MAR N5 than in non-MAR S23 lines (8.77 and 1.63 mg/g, respectively). The average Cry3A toxalbumin content was 1.3 times higher in N5 than in S23 lines (8.21 and 6.48 mg/g, respectively). Moreover, toxalbumin expression levels differed significantly among the S23-transformed lines. The MAR structure applied on both ends of the genes increased both the level and stability of exogenous gene expression. In conclusion, N5 was the most optimal of the four tested vectors. © 2016 The Authors.


Dong Y.,Agricultural University of Hebei | Dong Y.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | Ma Y.,Agricultural University of Hebei | Wang H.,Hebei Provincial Academy of Forestry | And 5 more authors.
Forest Science and Practice | Year: 2013

One-year-old potted clone plants of four willow species (Salix matsudana × alba, S. babylonica, S. psammophila and S. cheilophila) were cultivated and irrigated with saline solutions of different concentrations, while their electrical impedance spectroscopy (EIS) parameters and other physical parameters were monitored. The results indicate i) that under salt stress, height and basal diameter of all species are inhibited, and ii) that relative conductivity of cellular exudates increases while intracellular resistance (r i) and extracelluar resistance (r e) drop. Both r i and r e were positively correlated with height growth and basal diameter while they were significantly and negatively correlated with electric conductivity. The concentration of Na+ in the shoots of willows was negatively correlated with both r i and r e, whereas the concentration of K+ in the shoots was positively correlated with both r i and r e. Hence, electrical impedance spectroscopy is a reliable tool for evaluating the capacity of willow species for tolerance to saline soils, with r i as the most accurate parameter. © 2013 Beijing Forestry University and Springer-Verlag Berlin Heidelberg.


Xu L.,Agricultural University of Hebei | Xu L.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | Yang M.,Agricultural University of Hebei | Yang M.,Hebei Key Laboratory for Tree Genetic Resources and Forest Protection | And 2 more authors.
Research Journal of Biotechnology | Year: 2013

Endophytic bacteria can promote the rooting and growing of plants. In this paper, Paenibacillus (P22) and Stenotrophomonas (S16) isolates from hybrid poplar were used to inoculate shoots of Liquidambar formosana in order to investigate the effects of P22 and S16 on the rooting and growth of the plants. The result showed that P22 is stronger than S16 in enhancing the rooting and growth of tissue-cultured Liquidambar formosana and both bacteria equally proved to be plant-growth-promoting bacteria that have capability to grow on a medium with low nitrogen content. Thus, the result demonstrated the possibility of isolating plant-growth-promoting bacteria from its natural host to promote the growth of another plant.

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