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Phytochemistry, anti-asthmatic and antioxidant activities of Anchomanes difformis(Blume) Engl. leaf extract

Asian Pacific Journal of Tropical Biomedicine 2016年第3期6卷 225-231页

作者： Ovuakporie-Uvo Oghale Mac Donald Idu Department of plant Biology and biotechnology University of Benin

Objective: To study the phytochemistry, anti-asthmatic and antioxidant activities of the aqueous leaf extract of Anchomanes difformis(Blume) Engl.(A. difformis) and to verify claims of use in folk ***: For anti-asthmatic activity, male and female guinea pigs with average body weight of(451.4 ± 118.1) g were divided into six groups of six animals each. Group 1served as control(distilled water); Group 2 was administered with salbutamol(reference drug) only; Group 3 served as ovalbumin sensitized group, Group 4, 5 and 6 were treated with A. difformis extract at doses of 100, 200 and 400 mg/kg, respectively. Described methods were used to test fluid viscosity, fluid volume and quantitative phytochemistry analysis. Absorbance was read using a UV–Vis spectrophotometer and results computed in percentage. Total antioxidant assays [2, 2-diphenyl-1-picrylhydrazyl(DPPH) and lipid peroxidation assay], were carried out using reported ***: The anti-asthmatic evaluation showed that protection from asthma of the animals in Group 6(400 mg/kg, 32.7%) were similar to that in Group 2(salbutamol, 33.0%).Excised trachea was free of mucus secretion in Group 5(200 mg/kg) as was observed in the control group. Fluid volume increase in Groups 3 and 6 indicated mucus *** radical scavenging activity of extract was effective as ascorbic acid which served as standard at 20 mg/m L. But, the extract elicited low lipid peroxidation activity compared with the reference(tocopherol) at concentrations ***: A. difformis aqueous leaf extract is safe and possesses positive antiasthmatic and antioxidant activities as claimed by traditional herbal practitioners in Delta State.

关键词： Anti-asthmatic Anti-oxidant Phytochemistry Anchomanes difformis

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Heterologous expression of Lolium perenne antifreeze protein confers chilling tolerance in tomato

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Journal of Integrative Agriculture 2018年第5期17卷 1128-1136页

作者： Srinivasan Balamurugan Jayan Susan Ann Inchakalody P Varghese Shanmugaraj Bala Murugan Mani ChANDra Harish Sarma Rajeev Kumar Ramalingam Sathishkumar plant Genetic Engineering Laboratory Department of BiotechnologyBharathiar University

Antifreeze proteins（AFP） are produced by certain plants, animals, fungi and bacteria that enable them to survive upon extremely low temperature. Perennial rye grass, Lolium perenne, was reported to possess AFP which protects them from cold environments. In the present investigation, we isolated AFP gene from L. perenne and expressed it in tomato plants to elucidate its role upon chilling stress. The T1 transgenic tomato lines were selected and subjected to molecular, biochemical and physiological analyses. Stable integration and transcription of Lp AFP in transgenic tomato plants was confirmed by Southern blot hybridization and RT-PCR, respectively. Physiological analyses under chilling conditions showed that the chilling stress induced physiological damage in wild type（WT） plants, while the transgenic plants remained healthy. Total sugar content increased gradually in both WT and transgenic plants throughout the chilling treatment. Interestingly, transgenic plants exhibited remarkable alterations in terms of relative water content（RWC） and electrolyte leakage index（ELI） than those of WT. RWC increased significantly by 3-fold and the electrolyte leakage was reduced by 2.6-fold in transgenic plants comparing with WT. Overall, this report proved that Lp AFP gene confers chilling tolerance in transgenic tomato plants and it could be a potential candidate to extrapolate the chilling tolerance on other chilling-sensitive food crops.

关键词： Lolium perenne antifreeze protein chilling tolerance genetic transformation transgenic tomato

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Characterization and functional analysis of β-1,3-galactosyltransferase involved in Cry1Ac resistance from Helicoverpa armigera(Hübner)

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Journal of Integrative Agriculture 2015年第2期14卷 337-346页

作者： ZHANG Li-li LIANG Ge-mei GAO Xi-wu CAO Guang-chun GUO Yu-yuan College of Agriculture and biotechnology China Agricultural University State Key Laboratory for Biology of plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

Carbohydrate chains are the principal antigens by which Bacillus thuringiensis（Bt） identify receptor proteins. The interaction between the antigen and Bt causes a pore in the membrane of midgut epithelial cells of insects. Receptor proteins, such as aminopeptidase N and alkaline phosphatase, are glycoproteins. Cadherin is another cell surface receptor protein which has potential glycosylation sites. Glycosyltransferase is very important for the synthesis and modification of receptor proteins. It can indirectly influence the function of Bt. The 1 950 bp full-length c DNA encoding β-1,3-galactosyltransferase was cloned from the the midgut of Helicoverpa armigera by degenerative PCR combined with RACE techniques（GAL-Harm, Gen Bank accession no.： GQ904195.1） with two potential N-glycosylation sites（^157NNTI^160 and ^272NKTL^275）. Protein sequence alignments revealed that H. armigera β-1,3-galactosyltransferase shared high identity with β-1,3-galactosyltransferase in other insect species. The expression level of the β-1,3-galactosyltransferase gene in Cry1Ac-resistant H. armigera larvae was 9.2-fold higher than that in susceptible strain. The function of β-1,3-galactosyltransferase was investigated using RNAi technique. The result showed Cry1 Ac enhanced the toxicity against the si RNA-treated larvae compared with non-si RNA-treated ones, which indicated β-1,3-galactosyltransferase played an important role for the insecticidal toxicity of Cry1 Ac in H. armigera.

关键词： β-1 3-galactosyltransferase Cry1Ac resistance Helicoverpa armigera RNAi

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Morphology and ontogeny of directly differentiating shoot buds and somatic embryos in Santalum album L.

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Journal of Forestry Research 2019年第4期30卷 1179-1189页

作者： Rakhshanda Akhtar Anwar Shahzad plant biotechnology Section Department of BotanyAligarh Muslim University

Santalum album *** a commercially important tree that yields essential oil of high medicinal *** research through organogenesis and embryogenesis has been documented but no report depicts comparative ontogeny of directly differentiating shoot buds(SB)and somatic embryos(SE).In the present study aseptic seedling derived hypocotyl segments(HC)and hypocotyl+root junction(HC+R)were used to induce SBs and SEs,*** differences between the structures were confirmed using scanning electron microscopy and histological *** medium containing 6-benzyladenine or BA(2.5μM)produced highest number of direct SB,while MS+BA(7.5μM)proved suitable for higher frequency of SE *** differentiating structures attained growth when transferred to MS medium containing a combination of BA and anaphthalene acetic acid or silver nitrate(AgNO3).A combination of indole-3-butyric acid and silver nitrate(AgNO3)in half-strength woody plant medium and lesser osmotic concentration(2%sucrose),induced rhizogenesis.

关键词： Direct somatic embryos Emblings Histology Hypocotyl Ontogenic

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Expression profiling of transgenes(Cry1Ac and Cry2A) in cotton genotypes under different genetic backgrounds

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Journal of Integrative Agriculture 2022年第10期21卷 2818-2832页

作者： Kashif NOOR Hafiza Masooma Naseer CHEEMA Asif Ali KHAN Rao Sohail Ahmad KHAN Department of plant Breeding and Genetics University of AgricultureFaisalabad 38040Pakistan institute of plant Breeding and biotechnology Muhammad Nawaz Shareef University of AgricultureMultan 60000Pakistan Center of Agricultural Biochemistry and biotechnology University of AgricultureFaisalabad 38040Pakistan

Transgenic cotton carrying the CrylAc gene has revolutionized insect pest control since its adoption,although the development of resistance in insect pests has reduced its *** 10 years of cultivating Bacillus thuringiensis(Bt)cotton with a single Cry1 Ac gene,growers are on the verge of adopting Bt cotton that carries the double gene(Cry1 Ac+Cry2 A)due to its better effectiveness against insect ***,the current study was designed to evaluate the role of each gene in the effectiveness of Bt cotton carrying the double *** expression levels of the Cry1 Ac and Cry2 A genes were evaluated in the leaves of 10 genotypes(2 parents and 8 Fhybrids)at 30 days after sowing(DAS),while samples of leaves,bolls and flowers were taken from the upper and lower canopies at 70 and 110 *** Fhybrids were developed through reciprocal crosses between two Bt(CKC-1,CKC-2)and two non-Bt(MNH-786,FH-942)*** differential expression of transgenes was evaluated through Enzyme Linked Immuno-Sorbent Assay(ELISA).The results showed that the MNH786 xCKC-1 hybrid had the highest concentrations of Cry1 Ac gene at30 DAS(3.08μg g^(-1))and 110 DAS(1.01μg g^(-1))in *** contrast,the CKC-2 xMNH-786 hybrid showed the lowest concentrations of Cry1 Ac gene at 30 DAS(2.30μg g^(-1))and 110 DAS(0.86μg g^(-1)).The Fhybrid FH-942×CKC-2 showed the highest concentrations of Cry2 A gene at 30 DAS(8.39μg g^(-1))and 110 DAS(7.74μg g^(-1))in leaves,while the CKC-1 xMNH-786 hybrid expressed the lowest concentrations of Cry2 A gene at 30 DAS(7.10μg g^(-1))and 110 DAS(8.31μg g^(-1)).A comparison between the two stages of plant growth showed that leaves had the highest concentrations at 30 DAS,whereas the lowest concentrations were observed at 110 DAS for both genes in *** the expression pattern was compared between various plant parts in genotype CKC-2,it was found that leaves had higher concentrations of Cry1 Ac(3.12μg g^(-1))and Cry2 A(8.31μg g^(-1))at 70 DAS,followed by bolls

关键词： transgenic cotton breeding transgene expression double gene insect resistance

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Highly Sensitive and Specific Monoclonal Antibody-Based Serological Methods for Rice Ragged Stunt Virus Detection in Rice plants and Rice Brown planthopper Vectors

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Journal of Integrative Agriculture 2014年第9期13卷 1943-1951页

作者： LIU Huan SONG Xi-jiao NI Yue-qun LU Li-na ZHOU Xue-ping WU Jian-xiang institute of biotechnology College of Agriculture & Biotechnology Zhejiang University

Rice ragged stunt virus（RRSV） is a serious rice disease in Asia, causing serious yield losses on rice. The capsid protein（CP） gene of the major outer capsid protein of RRSV was expressed in Escherichia coli BL21（DE3） using the pMAL-C2 X expression vector. The recombinant protein was used as the immunogen to immunize BALB/c mice. A hybridoma cell line 8A12 secreting monoclonal antibody（MAb） against RRSV was obtained by fusing mouse myeloma cells（Sp 2/0） with spleen cells from the immunized BALB/c mice. Western blot analysis showed that the MAb 8A12 can specifically react with RRSV CP. Using the MAb, an antigen-coated-plate enzyme-linked immunosorbent assay（ACP-ELISA）, a dot enzyme-linked immunosorbent assay（dot-ELISA）, and immunocapture-RT-PCR（IC-RT-PCR） assay were developed to detect RRSV. The established ACP-ELISA, dot-blot ELISA and IC-RT-PCR methods could detect RRSV in infected rice tissue crude extracts with dilutions of 1：40 960, 1：1 280 and 1：655 360（w/v, g mL-1）, respectively. The ACP-ELISA and dot-blot ELISA methods could detect RRSV in infected insect vector crude extracts with dilutions of 1：12 800 and 1：1 600（an individual planthopper μL-1）, respectively. The field survey revealed that Rice ragged stunt disease occurs on rice in Hainan, Yunnan, Guangxi, Sichuan, Guizhou, Fujian, Hunan, Jiangxi and Zhejiang in China.

关键词： Rice ragged stunt virus rice brown planthopper monoclonal antibody antigen-coated-plate enzyme-linked immunosorbent assay(ACP-ELISA) dot-blot ELISA immunocapture RT-PCR

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The methyl jasmonate-responsive transcription factor SmMYB1 promotes phenolic acid biosynthesis in Salvia miltiorrhiza

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Horticulture Research 2021年第1期8卷 147-159页

作者： Wei Zhou Min Shi Changping Deng Sunjie Lu Fenfen Huang Yao Wang Guoyin Kai Laboratory of Medicinal plant biotechnology College of PharmacyZhejiang Chinese Medical University310053 HangzhouZhejiangChina institute of plant biotechnology School of Life SciencesShanghai Normal University200234 ShanghaiChina

Water-soluble phenolic acids are major bioactive compounds in the medicinal plant species Salvia *** acid biosynthesis is induced by methyl jasmonate(MeJA)in this important Chinese ***,we investigated the mechanism underlying this induction by analyzing a transcriptome library of *** in response to *** transcriptome analysis identified the MeJA-responsive R2R3-MYB transcription factor-encoding gene *** SmMYB1 significantly promoted phenolic acid accumulation and upregulated the expression of genes encoding key enzymes in the phenolic acid biosynthesis pathway,including cytochrome P450-dependent monooxygenase(CYP98A14).Dual-luciferase(dual-LUC)assays and/or an electrophoretic mobility shift assays(EMSAs)indicated that SmMYB1 activated the expression of CYP98A14,as well as the expression of genes encoding anthocyanin biosynthesis pathway enzymes,including chalcone isomerase(CHI)and anthocyanidin synthase(ANS).In addition,SmMYB1 was shown to interact with SmMYC2 to additively promote CYP98A14 expression compared to the action of SmMYB1 *** together,these results demonstrate that SmMYB1 is an activator that improves the accumulation of phenolic acids and anthocyanins in *** findings lay the foundation for in-depth studies of the molecular mechanism underlying MeJA-mediated phenolic acid biosynthesis and for the metabolic engineering of bioactive ingredients in ***.

关键词： phenolic additive promoted

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Getting lost in the weeds:How VVQX9 has shed light on gene functional divergence

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Molecular plant 2021年第5期14卷 717-718页

作者： Maida Romera-Branchat institute of plant Biology and biotechnology University of MunsterSchlossplatz 748149 MunsterGermany

Studying functional divergence between different species is key to understanding gene evolution,but it is challenging when comparison of distant phyla or when allelic diversity is limiting.

关键词： functional divergence light

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Expression of. Arabidopsis tryptophan biosynthetic pathway genes: effect of the 5' coding region of phosphoribosylanthranilate isomerase gene

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Science China(Life Sciences) 1999年第3期42卷 274-280,337-338页

作者：何奕昆刘新仿李家洋 plant biotechnology Laboratory Institute of Genetics Chinese Academy of Sciences China Chinese Center De Montfort University Norman Borlaug Institute for Plant Science Reseach Beijing China

There are three non-allelic isogenes encoding phosphoribosylanthranilate isomerase (PAI) in Arabidopsis thaliana. The expression plasmids were constructed by fusion of the GUS reporter gene to the three PAI promoters with or without the 5' region encoding PAI N-terminal polypeptides and transferred into Arabidopsis plants by Agrobacterium tumefaciens. Analysis of GUS activity revealed that the PAI 5' coding region was necessary for high expression of GUS activity. GUS activity in transgenic plants transformed with the expression plasmids containing the 5' coding region of PAH or PAD was 60-100-fold higher than that without the corresponding 5' region. However, the effect of 5' coding region of PAI2 gene on the GUS activity was very small (only about 1 time difference) . The GUS histochemical staining showed a similar result as revealed by GUS activity assay. It was expressed in the mesophyll cells and guard cells, but not in the epidermic cells, indicating that the N-terminal polypeptides encoded by the 5' region of PAI genes have the function of PTP.

关键词： phosphoribosylanthranilate isomerase (PAI) tryptophan biosynthetic pathway gene expression and regulation plastid targeting peptide (PTP) Arabidopsis thaliana.

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The new effector AbSCP1 of foliar nematode(Aphelenchoides besseyi) is required for parasitism rice

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Journal of Integrative Agriculture 2022年第4期21卷 1084-1093页

作者： HUANG Xin CHI Yuan-kai Addisie Abate BIRHAN ZHAO Wei QI Ren-de PENG De-liang institute of plant Protection and Agro-products Safety Anhui Academy of Agricultural SciencesHefei 230001P.R.China State Key Laboratory for Biology of plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193P.R.China Agricultural biotechnology Directorate Ethiopian Biotechnology InstituteAddis Ababa 5954Ethiopia

plant parasitic nematodes secrete effector proteins to parasitize hosts successfully. Of these proteins, serine carboxypeptidases have critical roles in pathogenicity. This study investigated the role of new effector AbSCP1 in Aphelenchoides besseyi pathogenicity. In situ hybridization and qRT-PCR analyses indicated that AbSCP1 was exclusively expressed in the esophageal glands and upregulated in juveniles. Subcellular localization assays indicated that the protein was expressed in the nucleus. The ability to hydrolyze C-terminal amino acid residues was proven for AbSCP1. Moreover, RNAi significantly reduced the expression of AbSCP1 and RNAi-treated nematodes’ reproductive potential. Pathogenicity assays on rice showed that RNAi-treated nematodes were less pathogenic than the untreated control groups. These results suggest the important role of AbSCP1 in the A. besseyi infection process.

关键词： Aphelenchoides besseyi serine carboxypeptidases effector nematode pathogenicity

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