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Jiang R.-B.,Chinese Academy of Forestry | Jiang J.-M.,Chinese Academy of Forestry | Liu J.,Chinese Academy of Forestry | Chen Y.-T.,Chinese Academy of Forestry | And 2 more authors.
Forest Research | Year: 2011

In order to establish stable ISSR-PCR reaction system of Machilus thunbergii, single factor and orthogonal experiment design were used to optimize the reaction system after exploring M. thunbergii DNA extraction. For finding optimal concentration of factors of ISSR-PCR, the different levels of concentration of Mg2+, primer, template DNA, dNTPs were trailed by single factor experiment. Meanwhile, in order to improve the reliability of the results of the single-factor test, the authors also adopted orthogonal design [L9(34)] by four factors, three levels for further optimizing and screening the best conditions of Mg2+, primer, template DNA, and dNTPs. The best ISSR-PCR reaction system of M. thunbergii was eventually established by comparing the two method's results, which included 20 μL reaction system, Taq enzyme, 0.05 U · μL-1, Mg2+ 2.0 mmol · L-1 template DNA 1 ng · L-1, dNTPs 0.3 mmol · L-1, primers (835) 0.5 μmol · L-1, and 1 × PCR buffer. The establishment of the better repeatability and stability ISSR-PCR reaction system could provide technical support for further research of genetic structure and genetic variety of M. thunbergii group.

Liu X.,CAS South China Botanical Garden | Meng W.,Paulownia Research and Development Center | Liang G.,South China Agricultural University | Li K.,CAS South China Botanical Garden | And 3 more authors.
PLoS ONE | Year: 2014

This paper aims to establish evidence for available phosphorous (AP) binding with total nitrogen (N) in subtropical forest soils. Soil organic carbon (SOC), total N, total phosphorous (P) and AP concentration were measured for three contrasting forest types in southern China: Masson pine forest (MPF), coniferous and broadleaved mixed forest (CBMF) and monsoon evergreen broadleaved forest (MEBF). A pot experiment with N addition was conducted to confirm the dominant factor to affect on soil AP concentration. The results showed that mean soil total N concentration in 0-10 cm soil layer was 440±50 for MPF, 840±80 for CBMF and 1020±50 mg kg-1 for MEBF, respectively. The mean soil AP concentration in 0-10 cm soil layer was 2.67±0.87 for MPF, 2.65±0.58 for CBMF, 4.10±0.29 mg kg -1 for MEBF, respectively. The soil total N concentration could explain about 70% of the variations in soil AP concentration in the top 20 cm soil layers in the three forest types. A pot experiment with N addition also showed an increase of AP concentration from 2.56 to 5.63 mg kg-1, when N addition increased from 5 g to 17 g NH4NO3. Our results therefore suggested that N addition significantly increased soil AP concentration, which might be beneficial for stabilizing the net primary production of subtropical forests that were limited by soil AP. This finding may provide a theory basis for tropical and subtropical forests management. © 2014 Liu et al.

Feng Y.,Paulownia Research and Development Center | Feng Y.,Central South University of forestry and Technology | Zhang L.,Central South University of forestry and Technology | Fu J.,Paulownia Research and Development Center | And 5 more authors.
Journal of Agricultural and Food Chemistry | Year: 2016

Eucommia ulmoides Oliver, the only member of the Eucommiaceae family, is a rare and valuable tree used to produce a highly valued traditional Chinese medicine and contains α-linolenic acid (ALA) up to 60% of the total fatty acids in the kernels (embryos). Glycolysis provides both cellular energy and the intermediates for other biosynthetic processes. However, nothing was known about the molecular basis of the glycolytic pathway in E. ulmoides kernels. The purposes of this study were to identify novel genes of E. ulmoides related to glycolytic metabolism and to analyze the expression patterns of selected genes in the kernels. Transcriptome sequencing based on the Illumina platform generated 96,469 unigenes in four cDNA libraries constructed using RNAs from 70 and 160 days after flowering kernels of both low- and high-ALA varieties. We identified and characterized the digital expression of 120 unigenes coding for 24 protein families involved in kernel glycolytic pathway. The expression levels of glycolytic genes were generally higher in younger kernels than in more mature kernels. Importantly, several unigenes from kernels of the high-ALA variety were expressed more than those from the low-ALA variety. The expression of 10 unigenes encoding key enzymes in the glycolytic pathway was validated by qPCR using RNAs from six kernel stages of each variety. The qPCR data were well consistent with their digital expression in transcriptomic analyses. This study identified a comprehensive set of genes for glycolytic metabolism and suggests that several glycolytic genes may play key roles in ALA accumulation in the kernels of E. ulmoides. © 2016 American Chemical Society.

Yue H.-F.,Chinese Academy of Forestry | Yue H.-F.,Paulownia Research and Development Center | Shao W.-H.,Chinese Academy of Forestry | Jing Z.-H.,Chinese Academy of Forestry | And 3 more authors.
Forest Research | Year: 2010

The Castanopsis sclerophylla seed length, seed diameter, seed length x seed diameter, seed length/seed diameter and hundred grain weight from nine areas were studied. The results showed that extremely significant difference were found in seed characters of C. sclerophylla from different areas. The most coefficient of variation is hundred grain weight and the least is seed length/seed diameter. Correlation analysis showed diat the seed length and seed diameter had the significant positive correlation widi hundred grain weight ; seed became bigger and heavier as latitude became lower, the annual average temperature and frost free period become longer; the hundred grain weight had significant positive correlation with rainfall, seed length/seed diameter had significant negative correlation with the rainfall, And clustering analysis showed that phenotypic variation was basicaly continuous variation with the geography, which was consistent with the result of relevant analysis.

Feng Y.-Z.,Paulownia Research and Development Center | Feng Y.-Z.,Non Timber Forestry Research and Development Center | Wang L.,Paulownia Research and Development Center | Wang L.,Non Timber Forestry Research and Development Center | And 10 more authors.
Genes and Genomics | Year: 2016

Eucommia ulmoides is a focus of considerable attention because it is used to produce gutta-percha latex and raw materials for traditional Chinese medicine. The species has also been widely planted for vegetable oil production due to the high content of alpha-linolenic acid (ALA) in the seeds. However, little was known about fatty acid biosynthesis in the seeds. Here, we present transcriptome analyses that identified genes related to the fatty acid biosynthesis pathways in the seeds collected 70 and 160 days after flowering (DAF) from two varieties: ‘Huazhong No. 6’ (ALA content of 38.7 %) and ‘Huazhong No. 10’ (ALA content of 67.6 %). De novo assembly generated 96,469 unigenes with an average length of 690 bp. Among them, 11,260 unigenes identified by functional annotation against the Kyoto Encyclopedia of Genes and Genomes database were mapped to 117 metabolic pathways. 65 unigenes involved in fatty acid biosynthesis include 5 coding for 3-oxoacyl-ACP reductase, 2 coding for β-hydroxyacyl-ACP dehydratase, 10 coding for β-ketoacyl-ACP synthase II, and 2 coding for enoyl-ACP reductase I. Expression of the nine differentially expressed unigenes coding for major enzymes in fatty acid biosynthesis was validated by real-time fluorescent quantitative PCR (qPCR) using RNA from six seed stages (70, 88, 106, 124, 142, and 160 DAF) in ‘Huazhong No. 10’. The qPCR analysis showed that expression of these 9 unigenes all changed significantly at 88 DAF. This study provides a large-scale transcriptome annotation of E. ulmoides, which is fundamental information for further characterizing gene functionals involved in the process of seed development. © 2016, The Genetics Society of Korea and Springer-Science and Media.

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