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22 research outputs found

Co-extraction of high-quality RNA and DNA from rubber tree (Hevea brasiliensis)

Author: An Zewei
Cheng Han
Hu Yanshi
Huang Huasun
Li Yacao
Wang Qitong
Zhao Yanhong
Publication venue: 'African Journals Online (AJOL)'
Publication date: 14/12/2015
Field of study

High-quality nucleic acids are the basic requirement for performing genomic research. A reliable and efficient method was developed for co-extracting high-quality DNA and RNA from rubber tree (Hevea brasiliensis) in this study. Polyethylene glycol (PEG) and cetyltrimethylammonium bromide (CTAB) extraction buffer with high concentrations of polyvinylpyrrolidone (PVP) and β-mercaptoethanol was used in this study. The results show that 3.2% polyethylene glycol 8000 is the optimal concentration for successful separation of DNA and RNA. Spectrophotometric determination (A260/A280 and A260/A230 ratios), agarose electrophoresis analysis and reverse transcription (RT-PCR) of isolated nucleic acids indicate that high-quality DNA and RNA were extracted by this method. The general applicability of this method was also evaluated, and the results show that it was suitable for a variety of plants.Key words: Hevea brasiliensis, polyethylene glycol (PEG), nucleic acid, co-extraction, higher plants

AJOL - African Journals Online

A rapid method for isolation of low-molecular-weight RNA from Arabidopsis using low salt concentration buffer

Author: Han Cheng
Huasun Huang
Jing Gao
Zewei An
Publication venue: PAGEPress Publications
Publication date: 01/08/2010
Field of study

We have developed a rapid extraction method using low salt concentration buffer for the isolation of low-molecular-weight RNA from Arabidopsis tissues. The method was quick and efficient, and the small scale extraction process took no more than 1 hour, while yield and RNA quality were comparable with those of previously reported. The LMW RNA isolated using this method was high quality, abundant in small RNA and free of high molecular weight RNA. This method can be used to extract low-molecular-weight RNA for the purpose of small RNA cloning and detection, and library construction.</span

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A Rapid and Economical Method for Low Molecular Weight RNA Isolation from a Wide Variety of Plant Species

Author: Huasun HUANG
Lili XIE
Qilin ZHAI
Yachao LI
Zewei AN
Publication venue: 'Japan Society for Bioscience, Biotechnology, and Agrochemistry'
Publication date
Field of study

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Cloning, Expression Analysis, and Functional Characterization of Candidate Oxalate Transporter Genes of HbOT1 and HbOT2 from Rubber Tree (Hevea brasiliensis)

Author: Feng An
Guishui Xie
Pingjuan Zhao
Wentao Peng
Xiaowei Ma
Zewei An
Zifan Liu
Zongming Yang
Publication venue: 'MDPI AG'
Publication date: 01/11/2022
Field of study

Secretion of oxalic acid from roots is an important aluminum detoxification mechanism for many plants such as Hevea brasiliensis (rubber tree). However, the underlying molecular mechanism and oxalate transporter genes in plants have not yet been reported. In this study, the oxalate transporter candidate genes HbOT1 and HbOT2 from the rubber tree were cloned and preliminarily identified. It was found that HbOT1 had a full length of 1163 bp with CDS size of 792 bp, encoding 263 amino acids, and HbOT2 had a full length of 1647 bp with a CDS region length of 840 bp, encoding 279 amino acid residues. HbOT1 and HbOT2 were both stable hydrophobic proteins with transmembrane structure and SNARE_assoc domains, possibly belonging to the SNARE_assoc subfamily proteins of the SNARE superfamily. qRT-PCR assays revealed that HbOT1 and HbOT2 were constitutively expressed in different tissues, with HbOT1 highly expressed in roots, stems, barks, and latex, while HbOT2 was highly expressed in latex. In addition, the expressions of HbOT1 and HbOT2 were up-regulated in response to aluminum stress, and they were inducible by metals, such as copper and manganese. Heterologous expression of HbOT1 and HbOT2 in the yeast mutant AD12345678 enhanced the tolerance to oxalic acid and high concentration aluminum stress, which was closely correlated with the secretion of oxalic acid. This study is the first report on oxalate transporter genes in plants, which provides a theoretical reference for the study on the molecular mechanism of oxalic acid secretion to relieve aluminum toxicity and on aluminum-tolerance genetic engineering breeding

Multidisciplinary Digital Publishing Institute

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PubMed Central

Synthesis of Crystal-Controlled TiO 2

Author: Bing Wang
Fang Y.
Hao An
Hao Cui
Jianping Zhai
Ohno T.
Yang H. G.
Yang Pan
Zewei Yang
Publication venue: 'American Chemical Society (ACS)'
Publication date
Field of study

Crossref

HbCBF1 overexpression in Arabidopsis.

Author: Dianjing Guo (143463)
Haibin Cai (794409)
Haitian Fu (794410)
Han Cheng (475615)
Huasun Huang (794414)
Jialin Fang (794412)
Yanshi Hu (794413)
Zewei An (794411)
Publication venue
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Field of study

(A) Southern blotting of HbCBF1 in transgenic plants. (B) HbCBF1 expression in OE-3, OE-6 and OE-13 lines. The gene expression was detected by Northern blotting analysis using the HbCBF1 probe. (C) Thirty days old seedlings of col-0, OE-3, OE-6 and OE-13 lines showed decreased plant height.</p

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Gel mobility shift assay of recombinant HbCBF1 protein indicated CRT/DRE binding activity.

Author: Dianjing Guo (143463)
Haibin Cai (794409)
Haitian Fu (794410)
Han Cheng (475615)
Huasun Huang (794414)
Jialin Fang (794412)
Yanshi Hu (794413)
Zewei An (794411)
Publication venue
Publication date
Field of study

(A) Probes sequences used in this analysis. Bolded nucleotides are CRT/DRE motif, while underlined nucleotides are the mutation sites. (B) HbCBF1 bound with CRT/DRE elements specifically in vitro. Four microgram total proteins of GST (lane 1 from the left) or GST::HbCBF1 (lane 2–9 from the left) were used for the binding with labeled hot COR15a probe. One hundred and twenty-five folds molar unlabelled cold competitor probes were added as indicated (labe 3–9 from the left) in each reactions. The retarded binding bands and free non-binding probe were indicated as "B" and "F" respectively.</p

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HbCBF1 overexpression conferred cold resistance and relatively slow dehydration in Arabidopsis.

Author: Dianjing Guo (143463)
Haibin Cai (794409)
Haitian Fu (794410)
Han Cheng (475615)
Huasun Huang (794414)
Jialin Fang (794412)
Yanshi Hu (794413)
Zewei An (794411)
Publication venue
Publication date
Field of study

(A) The electrolyte leakage rates of col-0, OE-3, OE-6 and OE-13 lines under nonacclimated (warm) or 5-days cold—acclimated conditions (cold). Electrolyte leakage was measured using leaves from two-week-old seedlings frozen to -7°C at a refrigerating rate of 1°C per h from -1°C. The star symbol (*) indicates significant difference between each individual OE line and wild type plants (WT). (B) Representative seedlings of col-0, OE-3, OE-6 and OE-13 lines recovered from 18 hours frozen at -6°C. The survival rates of each line were listed. (C) The dehydration rates of the detached leaves from col-0, OE-3, OE-6 and OE-13 seedlings. The star (*) symbol indicates significant difference between WT and each individual OE line, while plus symbols (+) shows significant difference between WT and individual OE-6 and OE-13 line. Data in A and C were presented as mean ± standard error of mean of five biological replicates. Significance was determined by Student’s t test at the probability levels of P < 0.05. Comparisons were made between the wild type plant and each individual overexpressing line. For B, totally thirty to fifty plants were tested for each line and about ten seedlings were grown in each pot.</p

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Functional Characterization of Hevea brasiliensis CRT/DRE Binding Factor 1 Gene Revealed Regulation Potential in the CBF Pathway of Tropical Perennial Tree

Author: Dianjing Guo (143463)
Haibin Cai (794409)
Haitian Fu (794410)
Han Cheng (475615)
Huasun Huang (794414)
Jialin Fang (794412)
Yanshi Hu (794413)
Zewei An (794411)
Publication venue
Publication date
Field of study

<div>Rubber trees (Hevea brasiliensis) are susceptible to low temperature and therefore are only planted in the tropical regions. In the past few decades, although rubber trees have been successfully planted in the northern margin of tropical area in China, they suffered from cold injury during the winter. To understand the physiological response under cold stress, we isolated a C-repeat binding factor 1 (CBF1) gene from the rubber tree. This gene (HbCBF1) was found to respond to cold stress but not drought or ABA stress. The corresponding HbCBF1 protein showed CRT/DRE binding activity in gel shift experiment. To further characterize its molecular function, the HbCBF1 gene was overexpressed in Arabidopsis. The HbCBF1 over expression (OE) line showed enhanced cold resistance and relatively slow dehydration, and the expression of Arabidopsis CBF pathway downstream target genes, e.g. AtCOR15a and AtRD29a, were significantly activated under non-acclimation condition. These data suggest HbCBF1 gene is a functional member of the CBF gene family, and may play important regulation function in rubber tree.</div

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FigShare

Co-extraction of high-quality RNA and DNA from rubber tree (Hevea brasiliensis)

A rapid method for isolation of low-molecular-weight RNA from Arabidopsis using low salt concentration buffer

A Rapid and Economical Method for Low Molecular Weight RNA Isolation from a Wide Variety of Plant Species

Cloning, Expression Analysis, and Functional Characterization of Candidate Oxalate Transporter Genes of HbOT1 and HbOT2 from Rubber Tree (Hevea brasiliensis)

Synthesis of Crystal-Controlled TiO 2

<i>HbCBF1</i> overexpression in <i>Arabidopsis</i>.

Gel mobility shift assay of recombinant HbCBF1 protein indicated CRT/DRE binding activity.

<i>HbCBF1</i> overexpression conferred cold resistance and relatively slow dehydration in <i>Arabidopsis</i>.

Functional Characterization of <i>Hevea brasiliensis CRT/DRE Binding Factor 1</i> Gene Revealed Regulation Potential in the CBF Pathway of Tropical Perennial Tree