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Effect of cDNA fragments in different length derived from Potato Virus Y coat protein gene on the induction of RNA-mediated virus resistance

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Science China(Life Sciences) 2004年第4期47卷 382-388页

作者： ZHU Junhua ZHU Xiaoping WEN Fujiang BAI Qingrong ZHU Changxiang SONG Yunzhi Laboratory of plant genetic Engineering College of Plant ProtectionShandong Agricultural UniversityTaian 271018China

We have reported that cDNA derived from entire coat protein (CP) gene of potato virus Y (PVY) could induce resistance to PVY infection in transgenic tobacco plants, and the resistance was further demonstrated to be RNA-mediated rather than coat protein-mediated. In this study, we cloned cDNA fragments of 202 bp, 417 bp, and 603 bp in length derived from the 3′ end of the PVY CP gene, and the cDNA fragments were introduced into tobacco (var. NC89) plants via Agrobacterium-mediated transformation system. The results of resistance assay showed that the CP cDNA fragments of 417 bp and 603 bp could confer resistance of the trans-genic plants to PVY infection, but the fragment of 202 bp in length could not. Molecular analysis revealed that the resistance was RNA-mediated, which is believed to be a result of post-transcriptional gene silencing. The results indicate that the length of cDNA fragments needed for resistance induction was located somewhere between 202 bp and 417 bp from the 3′ end of PVY CP gene.

关键词： PVY partial length of CP gene fragments transgenic tobacco plants RNA-mediated resistance post-transcrip-tional gene silencing.

Determination of Essential Fatty Acid Composition among Mutant Lines of Canola (Brassica napus),through High Pressure Liquid Chromatography

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Journal of Integrative plant Biology 2009年第12期51卷 1080-1085页

作者： Ghulam Raza Aquil Siddique Imtiaz Ahmad Khan Muhammed Yasin Ashraf Abdullah Khatri plant genetic division Nuclear Institute of Agriculture (NIA) Tando Jam 70060 Pakistan Nuclear Institute for Agriculture and Biology Faisalabad 38000 Pakistan

The present study aimed to quantify the methyl esters of lenoleic acid （LA）, γ-lenolenic acid （LNA） and oleic acid （OL） in the oil of Brassica napus mutants. Five stable mutants （ROO-75/1, ROO-100/6, ROO-125/12, ROO-125/14, and ROO-125/17） of B. napus cv. ＇Rainbow＇（P） and three mutants （W97-95/16, W97-0.75/11 and W97-.075/13） of B. napus cv. ＇Westar＇（P） at M6 stage, exhibiting better yield and yield components, were analyzed for essential fatty acids. The highest seed yield was observed in the mutant （ROO-100/6） followed by ROO-125/14 of Rainbow, that is, 34% and 32% higher than their parent plants, respectively. Westar mutant W97-75/11 also showed 30% higher seed yield than its parent plant. High performance liquid chromatography analysis of the composition of fatty acids indicated that OL was the most dominant fatty acid, ranging from 39.t to 66.3%; LA was second （15.3-41.6%） and LNA was third （18.1-28.9%）. Mutant ROO-125/14 showed higher OL contents than parent （Rainbow）. These results are expected to support the approval of ROO-125/14 in the National Uniform Varietal Yield Trials （NUVYT） as a new variety based on high oil quality.

关键词： Brassica high performance liquid chromatography lenoleic acid lenolenic acid oleic acid.

Protection of Mice with a Divalent Tuberculosis DNA Vaccine Encoding Antigens Ag85B and MPT64

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Acta Biochimica et Biophysica Sinica 2004年第4期36卷 269-276页

作者： Xia TIAN, Hong CAI, and Yu-Xian ZHU* The National Laboratory of Protein Engineering and plant genetic Engineering, Peking University, Beijing 100871, China The National Laboratory of Protein Engineering and plant genetic Engineering Peking UniversityBeijing 100871 China

DNA vaccine may be a promising tool for controlling tuberculosis development. However, vaccines encoding single antigens of mycobacterium did not produce protective effect as BCG did. In the present study, we evaluated the immunogenicity and protective efficacy of a divalent DNA vaccine encoding two immunodominant antigens Ag85B and MPT64 of Mycobacterium tuberculosis. We found that both humoral and Th1-type (high IFN-γ, low IL-4) cellular responses obtained from the divalent DNA vaccine group were significantly higher than that conferred by BCG. RT-PCR results showed that antigens were expressed differentially in various organs in divalent DNA vaccine group. The survival rate for mice treated with the divalent DNA vaccine after challenging with high doses of virulent M. tuberculosis H37Rv was significantly higher than that of the BCG group or any of the single DNA vaccine group. Significant differ- ences were also found between the single and divalent DNA vaccinated mice in terms of body, spleen and lung weight. Bacterial loading decreased about 2000-fold in lungs and about 100-fold in spleens of divalent DNA vaccinated mice when compared with that of the control group. We conclude that our divalent DNA vaccine may be a better choice for controlling tuberculosis disease in animals.

关键词： tuberculosis divalent DNA vaccine immunogenicity protection

Chikungunya infection:A potential re-emerging global threat

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Asian Pacific Journal of Tropical Medicine 2016年第10期9卷 910-915页

作者： Shanmugaraj Bala Murugan Ramalingam Sathishkumar plant genetic Engineering Laboratory Department of BiotechnologyBharathiar University

Infectious diseases are indeed a lifelong threat to everyone irrespective of age, sex, lifestyle and socio-economic status. The infectious diseases have persisted among the prominent causes of death globally. Recently, re-emergence of Chikungunya viral infection harmed many in Asian and African countries. Chikungunya was considered as a major threat in developing and underdeveloped countries; the recent epidemiological outbreak of Chikungunya in La Reunion urges the global researchers to develop effective vaccine against this viral disease. In this review, Chikungunya, pathogenesis and epidemiology were briefly described.

关键词： Aedes mosquitoes Alphavirus Arthropod-borne disease Chikungunya Non-Structural Proteins Structural Proteins

Studies on gene structure, enzymatic activity and regulatory mechanism of acetohydroxy acid isomeroreductase from G2 pea

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Science China(Life Sciences) 2003年第4期46卷 389-398页

作者：徐云剑顾雪松李珺李晴 PeterJ.Davies 朱玉贤 The National Laboratory of Protein Engineering and plant genetic Engineering Peking University Department of plant Biology Cornell University Ithaca

The AAIR genomic DNA of G2 pea (Pisum sativum L.) was amplified by PCR method. Sequence analysis showed that it was composed of 8 introns and 9 exons with three of the introns containing specific A/T-rich endogenous promoter regions. Molecular hybridization experiments revealed that the expression of AAIR remained at a high level before and after flowering if grown in short day growth chambers. However, when grown under long day conditions, the level of AAIR expression declined very rapidly after flowering. This variation of AAIR expression is consistent with the change of enzymatic activity of acetohydroxy acid isomeroreductase. Functional complementation experiments carried out using an acetohydroxy acid isomeroreductase deficient E. coli strain showed that these cells could not grow on M9 medium without addition of branched-chain amino acids unless they were transformed with the AAIR expression vector. Further study revealed that overexpression of the pea AAIR cDNA in acetohydroxy acid isomeroreductase deficient E. coli strain enhanced significantly its branched-chain amino acid biosynthetic capacity. Results from gel shift experiments showed that fractions of pea nuclear protein extracts could bind specifically to some A/T rich regions present in introns of the AAIR gene. The A/T-rich-region-binding proteins remained at a steady level in the non-senescing apical buds of short-day grown G2 pea. In the rapid-senescing apical buds of long-day grown G2 pea, the levels of these proteins declined rapidly after flower initiation. Therefore, the nuclear protein binding capacities to endogenous promoter regions may constitute an important mechanism to regulate AAIR gene expression.

关键词： AAIR G2 pea A/T rich region enzymatic activity senescence. D

Agro-Morphological, Physico-Chemical and Molecular Characterization of Rice Germplasm with Similar Names of Bangladesh

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Rice science 2016年第4期23卷 211-218页

作者： Mir Sharf Uddin AHMED Mohammad KHALEQUZZAMAN Md.Khairul BASHAR Abul Kalam Mohammad SHAMSUDDIN genetic Resources and Seed division Bangladesh Rice Research Institute HarvestP lus International Centre for Tropical Agriculture Department of genetics and plant Breeding Bangladesh Agricultural University

Thirty-one duplicate and similar named rice germplasms of Bangladesh were studied to assess the genetic variation for the agro-morphological and physico-chemical traits and simple sequence repeat banding patterns during 2009–2012 at Bangladesh Rice Research Institute. The range of variations within the cultivar groups showed higher degree. The principal component analysis showed that the first five components with vector values > 1 contributed 82.90% of the total variations. The cluster analysis grouped the genotypes into four clusters, where no duplicate germplasm was found. The highest number(11) of genotypes was constellated in cluster I and the lowest(3) in cluster II. The intra- and inter-cluster distances were the maximum in cluster I(0.93) and between clusters I and IV(24.61), respectively, and the minimum in cluster IV(0.62) and between clusters I and III(5.07), respectively. The cluster mean revealed that the crosses between the genotypes of cluster I with those of clusters II and IV would exhibit high heterosis for maximum good characters. A total of 350 alleles varied from 3(RM277) to 14(RM21) with an average of 7.8 per locus were detected at 45 microsatellite loci across the 31 rice accessions. The gene diversity ranged from 0.48 to 0.90 with an average of 0.77, and the polymorphism information content values from 0.44(RM133) to 0.89(RM206) with an average of 0.74. RM206, RM21, RM55, RM258 and RM433 were considered as the best markers on the basis of their higher polymorphism information content values. The dendrogram from unweighted pair-group method with arithmetic average clustering also classified the genotypes into four groups, where group IV comprised of 20 genotypes and group III of one genotype, but no duplicate was found. Finally, similar and duplicate named rice germplasms need to be conserved in gene bank as are distinct from each other.

关键词： diversity simple sequence repeat similar name landrace rice agro-morphological and physico-chemical trait

Characterization of chickpea germplasm conserved in the Indian National Genebank and development of a core set using qualitative and quantitative trait data

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The Crop Journal 2016年第5期4卷 417-424页

作者： Sunil Archak Rishi K.Tyagi P.N.Harer L.B.Mahase Neeta Singh Om P.Dahiya M.Abdul Nizar Mohar Singh Vrushali Tilekar Vikas Kumar Manoranjan Dutta Narendra P.Singh Kailash C.Bansal ICAR-National Bureau of plant genetic Resources Pulses Improvement Project Mahatma Phule Krishi Vidyapeeth ICAR-National Bureau of plant genetic Resources Regional Station ICAR-Indian Institute of Pulse Research

Chickpea is the third most important pulse crop as a source of dietary protein. Ever-increasing demand in Asian countries calls for breeding superior desi-type varieties, in turn necessitating the availability of characterized germplasm to breeders. The Indian National Genebank,located at the National Bureau of plant genetic Resources, New Delhi, conserves 14,651 accessions of chickpea. The entire set was characterized in a single large-scale *** variation was observed for eight quantitative and 12 qualitative agro-morphological traits. Allelic richness procedure was employed to assemble a core set comprising 1103 accessions, 70.0% of which were of Indian origin. Comparable values of total variation explained by the first three principal components in the entire collection(51.1%) and the core(52.4%)together with conservation of nine pairwise r values among quantitative traits in the core collection and a coincidence rate around 99.7% indicated that the chickpea core was indeed an excellent representation of the entire chickpea collection in the National Genebank. The chickpea core exhibited greater diversity than the entire collection in agro-morphological traits, as assessed by higher variance and Shannon–Weaver diversity indices, indicating that the chickpea core maximized the phenotypic diversity available in the Indian chickpea germplasm. The chickpea core, comprising mainly indigenous desi genotypes, is expected to be an excellent resource for chickpea breeders. Information on the chickpea core can be accessed at http://***/pgrportal.

关键词： Cicer arietinum Chickpea germplasm Indian National Genebank Agro-morphological variation Chickpea core

Phosphorus Starvation Tolerance in Rice Through Combined Physiological,Biochemical,and Proteome Analyses

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Rice science 2023年第6期30卷 613-631页

作者： Prathap V Suresh KUMAR Nand Lal MEENA Chirag MAHESHWARI Monika DALAL Aruna TYAGI division of Biochemistry Indian Council of Agricultural Research-Indian Agricultural Research InstituteNew Delhi 110012India division of Germplasm Evaluation National Bureau of Plant Genetic ResourcesNew Delhi 110012India National Institute for plant Biotechnology New Delhi 110012India

Phosphorus(P) deficiency limits the growth,development,and productivity of *** better understand the underlying mechanisms in P-deficiency tolerance and the role of Pup1 QTL in enhancing P use efficiency(PUE) for the development of P-efficient rice cultivars,a pair of contrasting rice genotypes(Pusa-44 and NIL-23) was applied to investigate the morpho-physio-biochemical and proteomic variation under P-starvation *** rice genotypes were grown hydroponically in a PusaRich medium with adequate P(16 mg/kg,+P) or without P(0 mg/kg,-P) for 30 d.P-starvation manifested a significant reductions in root and shoot biomass,shoot length,leaf area,total chlorophyll,and P,nitrogen and starch contents,as well as protein kinase *** stress increased root-to-shoot biomass ratio,root length,sucrose content,and acid phosphatase activity,particularly in the P-tolerant genotype(NIL-23).Comparative proteome analysis revealed several P metabolism-associated proteins(including OsCDPKs,OsMAPKs,OsCPKs,OsLecRK2,and OsSAPks) to be expressed in the shoot of NIL-23,indicating that multiple protein kinases were involved in P-starvation/deficiency ***,the up-regulated expression of OsrbcL,OsABCG32,OsSUS5,OsPoll-like B,and ClpC2 proteins in the shoot,and OsACA9,OsACA8,OsSPS2F,OsPP2C15,and OsBiP3 in the root of NIL-23,indicated their role in P-starvation stress control through the Pup1 QTL. Thus,our findings indicated that-P stress-responsive proteins,in conjunction with morpho-physio-biochemical modulations,improved PUE and made NIL-23 a P-deficiency tolerant genotype due to the introgression of the Pup1 QTL in the Pusa-44 background.

关键词： phosphorus deficiency Pup1 QTL protein kinase acid phosphatase proteome analysis rice

Characterization of the Tomato Prosystemin Promoter:Organ-specific Expression,Hormone Specificity and Methyl Jasmonate Responsiveness by Deletion Analysis in Transgenic Tobacco plants

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Journal of Integrative plant Biology 2012年第1期54卷 15-32页

作者： Hamlet Avilés-Arnaut John Paul Délano-Frier Center of Research and Advanced Studies (Cinvestav) at Irapuato:Unit for plant Biotechnology and genetic Engineering

Tomato systemin is a bioactive peptide that regulates the systemic activation of wound-responsive genes. It is released from its 200 amino acid precursor called prosystemin. Initial tissue-localization and hormone-induced expression assays indicated that the tomato prosystemin gene （SIPS） accumulates mainly in floral tissues and in response to exogenous abscisic acid and methyl jasmonate （MeJA） treatments, respectively. Later, the promoter regions of the PS gene in tomato （Solanum lycopersicum L. cv. Castlemart）, pepper （Capsicum annuum） and potato （Solanum tuberosum） were isolated and an in silico analysis of the SIPS promoter revealed an over-representation of stress- and MeJA-responsive motifs. A subsequent 5＇ deletion analysis of the SIPS promoter fused to the/^-glucuronidase reporter （GUS） gene showed that the -221 to ＋40 bp proximal SIPS promoter region was sufficient to direct the stigma, vascular bundle-specific and MeJA-responsive expression of GUS in transgenic tobacco plants. Important ***-specific, light- and MeJA-responsive cis-elements were also present in this region. These findings provide relevant information regarding the transcriptional regulation mechanisms of the SIPS promoter operating in transgenic tobacco plants. They also suggest that its Ussue-specificity and inducible nature could have wide applicability in plant biotechnology.

关键词： cis-regulatory elements flower specific jasmonic acid methyl jasmonate promoter analysis prosystemin wound-responsive genes.