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24 research outputs found

In vitro culture method of powdery mildew (Oidium heveae Steinmann) of Hevea brasiliensis

Author: Cai Haibin
Hua Yuwei
Huang Huasun
Sun Aihua
Tu Min
Publication venue: 'African Journals Online (AJOL)'
Publication date: 22/01/2016
Field of study

A method for culturing powdery mildew (Oidium heveae) from isolated leaves of Hevea brasiliensis was evaluated, which included three steps: Leaves and fungi selection, nutrient solution and culture dish preparation, fungi inoculation and culture. The culture time and produced conidia number were considered as decision index. We tested the influence of micro components of nutrient solution including 6-benzylaminopurine (6-BA), salicylic acid (SA) and vitamin C (VC) and evaluated the culture difference of various leaf phenological phases and rubber tree clones. The results show that the longest culture time of isolated leaves emerged on modified Murashige and Skoog (MS) macro elements with 4 mg/L 6-BA, 20 mg/L SA, 1 mg/L VC. The colour phase leaf was the preferable choice for culturing average 15 to 16 days and producing 3.2222 × 106 mL-1 conidia. The culture effects of using various rubber clones were different and higher resistance clones cultured less conidia. The method leading to mass production of powdery mildew was simple using a climate incubator to resolve problems linked to season and space limitation and preservation of powdery mildew. This method could improve rubber resistance breeding process.Key words: Hevea brasiliensis, Oidium heveae, in vitro culture, nutrient solution, phenological phase

AJOL - African Journals Online

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

Overexpression of a Hevea brasiliensis ErbB-3 binding protein 1 Gene Increases Drought Tolerance and Organ Size in Arabidopsis

Author: Han Cheng
Huasun Huang
Jianshun Zhu
Xiang Chen
Publication venue: 'Frontiers Media SA'
Publication date: 01/01/2016
Field of study

Rubber trees are economically important tropical tree species and the major source of natural rubber, which is an essential industrial material. This tropical perennial tree is susceptible to cold stress and other abiotic stresses, especially in the marginal northern tropics. Recent years, the genome sequencing and RNA-seq projects produced huge amount of sequence data, which greatly facilitated the functional genomics study. However, the characterization of individual functional gene is in urgent demands, especially for those involved in stress resistance. Here we identified and characterized the rubber tree gene ErbB-3 binding protein 1, which undergoes changes in expression in response to cold, drought stress and ABA treatment. HbEBP1 overexpression in Arabidopsis increased organ size, facilitated root growth and increased adult leaf number by delaying the vegetative-to-reproductive transition. In addition, HbEBP1 overexpression enhanced the resistance of the Arabidopsis plants to freezing and drought stress, demonstrating that this gene participates in the regulation of abiotic stress resistance. RD29a, RD22 and CYCD3;1 expression was also greatly enhanced by HbEBP1 overexpression, which explains its regulatory roles in organ size and stress resistance. The regulation of drought stress resistance is a novel function identified in plant EBP1 genes, which expands our understanding of the roles of EBP1 gene in response to the environment. Our results provide information that may lead to the use of HbEBP1 in genetically engineered crops to increase both biomass and abiotic stress resistance

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PubMed Central

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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Identification, Functional Study, and Promoter Analysis of HbMFT1, a Homolog of MFT from Rubber Tree (Hevea brasiliensis)

Author: Huasun Huang
Xiang Li
Yuwei Hua
Zhenghong Bi
Publication venue: MDPI AG
Publication date: 01/03/2016
Field of study

A homolog of MOTHER OF FT AND TFL1 (MFT) was isolated from Hevea brasiliensis and its biological function was investigated. Protein multiple sequence alignment and phylogenetic analysis revealed that HbMFT1 conserved critical amino acid residues to distinguish MFT, FLOWERING LOCUS T (FT) and TERMINAL FLOWER1 (TFL1)-like proteins and showed a closer genetic relationship to the MFT-like group. The accumulation of HbMFT1 was generally detected in various tissues except pericarps, with the highest expression in embryos and relatively higher expression in roots and stems of seedlings, flowering inflorescences, and male and female flowers. HbMFT1 putative promoter analysis showed that tissue-specific, environmental change responsive and hormone-signaling responsive elements were generally present. HbMFT1 was strongly induced under a short-day condition at 28 °C, with the highest expression after the onset of a day. Overexpression of HbMFT1 inhibited seed germination, seedling growth, and flowering in transgenic Arabidopsis. The qRT-PCR further confirmed that APETALA1 (AP1) and FRUITFULL (FUL) were drastically down-regulated in 35S::HbMFT1 plants. A histochemical β-glucuronidase (GUS) assay showed that HbMFT1::GUS activity was mainly detected in stamens and mature seeds coinciding with its original expression and notably induced in rosette leaves and seedlings of transgenic Arabidopsis by exogenous abscisic acid (ABA) due to the presence of ABA cis-elements in HbMFT1 promoter. These results suggested that HbMFT1 was mainly involved in maintenance of seed maturation and stamen development, but negatively controlled germination, growth and development of seedlings and flowering. In addition, the HbMFT1 promoter can be utilized in controlling transgene expression in stamens and seeds of rubber tree or other plant species

Multidisciplinary Digital Publishing Institute

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PubMed Central

Optimization of the Transformation Protocol for Increased Efficiency of Genetic Transformation in Hevea brasiliensis

Author: Huasun Huang
Jing Cheng
Jinu Udayabhanu
Shichao Xin
Tiandai Huang
Yuwei Hua
Publication venue: MDPI AG
Publication date: 01/04/2022
Field of study

The recurring growth of bacterium in newly developed resistant cells and a minimal level of bacterial infection rate are the main limiting factors of Agrobacterium-mediated transformation experiments in Hevea brasiliensis. The current study aimed to optimize crucial factors of the transformation protocol in order to obtain an efficient transformation experimental model for Hevea using cotyledonary somatic embryos as explants. Transformation conditions such as antibiotic concentration, preculture duration, Agrobacterium concentration, sonication and cocultivation conditions were analyzed using the binary vector pCAMBIA2301. Transient transformation was confirmed by GUS histochemical staining. The best transformation efficiency was observed when the explants were not cultured on a preculture medium that contained acetosyringone at a level of 100 μM. The best results were obtained using a bacterial density of 0.45 at OD 600 nm, 50 s of sonication of explants in a bacterial liquid culture and a total incubation time of 18 min in the same bacterial suspension. Transmission electron microscopical analysis confirmed the impacts of sonication on bacterial infection efficiency. Cocultivation conditions of 22 °C and 84 h of darkness were optimal for the transfer of T-DNA. Agrobacterium was eliminated with 500 mg/L of timentin, and the selection of transformants was performed using 100 mg/L of kanamycin in the selection medium. The presence of transgene was confirmed in the resistant embryos by polymerase chain reaction (PCR). The improved method of genetic transformation established in the present study will be useful for the introduction of foreign genes of interest into the Hevea genome for the breeding of this economically important plant species in the future

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Identification and Functional Evaluation of Three Polyubiquitin Promoters from Hevea brasiliensis

Author: Huasun Huang
Jinu Udayabhanu
Shichao Xin
Tiandai Huang
Xuemei Dai
Yueting Fan
Yuwei Hua
Publication venue: 'MDPI AG'
Publication date: 17/06/2022
Field of study

Hevea brasiliensis is an economically important tree species that provides the only commercial source of natural rubber. The replacement of the CaMV35S promoter by endogenous polyubiquitin promoters may be a viable way to improve the genetic transformation of this species. However, no endogenous polyubiquitin promoters in Hevea have been reported yet. Here, we identified three Hevea polyubiquitin genes HbUBI10.1, HbUBI10.2 and HbUBI10.3, which encode ubiquitin monomers having nearly identical amino acid sequences to that of AtUBQ10. The genomic fragments upstream of these HbUBI genes, including the signature leading introns, were amplified as putative HbUBI promoters. In silico analysis showed that a number of cis-acting elements which are conserved within strong constitutive polyubiquitin promoters were presented in these HbUBI promoters. Transcriptomic data revealed that HbUBI10.1 and HbUBI10.2 had a constitutive expression in Hevea plants. Semi-quantitative RT-PCR showed that these three HbUBI genes were expressed higher than the GUS gene driven by CaMV35S in transgenic Hevea leaves. All three HbUBI promoters exhibited the capability to direct GFP expression in both transient and stable transformation assays, although they produced lower protoplast transformation efficiencies than the CaMV35S promoter. These HbUBI promoters will expand the availability of promoters for driving the transgene expression in Hevea genetic engineering

Multidisciplinary Digital Publishing Institute

An Analysis of the Patents concerning Hevea brasiliensis

Author: CHEN Qing
HUANG Huasun
LI Qiong
LI Xijuan
REN Xinxin
XU Bingjiang
Publication venue
Publication date: 01/04/2017
Field of study

This paper mainly analyzes the application data of patents concerning Hevea brasiliensis in the world and China from the perspectives of time, research field, and the geographical distribution of applicants. It also interprets the patents for invention in China from time, research field, and types of applicants. Based on the study, it shows that although the patent applications fluctuate, the general trend is rising. And the patents are mostly concentrated in plant protection, and agricultural biotechnology. The Chinese applicants of patents for invention largely work in the research institutes, and most of these patents focus on the field of plant protection, agricultural biotechnology, and agro-processing. However, the foreign applicants mainly work in companies, and most of these patents only focus on plant protection. These findings will provide a reference for formulating development policies about natural rubber industry in China

AgEcon Search - Research in Agricultural & Applied Economics

AgEcon Search: Research in Agricultural and Applied Economics