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mutants, Overexpressors, and Interactors of Arabidopsis Plastocyanin Isoforms： Revised Roles of Plastocyanin in Photosynthetic Electron Flow and Thylakoid Redox State

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Molecular Plant 2009年第2期2卷 236-248页

作者： Paolo Pesaresi Michael Scharfenberg Martin Weigel Irene Granlund Wolfgang R Schroder Giovanni Finazzi Fabrice Rappaport Simona Masiero Antonella Furini Peter Jahns Dario Leister Dipartimento di Produzione Vegetale Universita degli studi di Milano c/o Parco Tecnologico Padano Via Einstein Loc Cascina Codazza 1-26900 Lodi Italy Lehrstuhl fur Botanik Department Biologie I Ludwig-Maximilians-Universitat Munchen GroBhaderner Str. 2 D-82152 Planegg-Martinsried Germany Abteilung fhr Pflanzenzuchtung und Genetik Max-Planck-lnstitut fur Zhchtungsforschung Carl-von-Linne-Weg 10 D-50829 Koln Germany Institute of Chemistry and Umea Plant Science Centre (UPSC) Umea University SE-901 87 Umea Sweden Institut de Biologie Physico-Chimique UMR 7141 CNRS-Universite R et M. Curie 13 rue Pierre et Marie Curie F-75005 Paris France Dipatimento di Biologia Universita degli studi di Milano Via Celoria 26 L-20133 Milano Italy Dipartimento Scientifico e Tecnologico Universita degli studi di Verona Strade le Grazie 15 1-37134 Verona Italy Institut fur Biochemie der Pflanzen Heinrich-Heine-Universitat Dusseldorf Universitatsstr. 1 D-40225 Dusseldorf Germany

Two homologous plastocyanin isoforms are encoded by the genes PETE1 and PETE2 in the nuclear genome of Arabidopsis thaliana. The PETE2 transcript is expressed at considerably higher levels and the PETE2 protein is the more abundant isoform. Null mutations in the PETE genes resulted in plants, designated pete1 and pete2, with decreased plastocyanin contents. However, despite reducing plastocyanin levels by over -90%, a pete2 null mutation on its own affects rates of photosynthesis and growth only slightly, whereas pete1 knockout plants, with about 60-80% of the wild-type plastocyanin level, did not show any alteration. Hence, plastocyanin concentration is not limiting for photosynthetic elec- tron flow under optimal growth conditions, perhaps implying other possible physiological roles for the protein. Indeed, plastocyanin has been proposed previously to cooperate with cytochrome C6A （Cyt C6A） in thylakoid redox reactions, but we find no evidence for a physical interaction between the two proteins, using interaction assays in yeast. We observed homodimerization of Cyt C6A in yeast interaction assays, but also Cyt C6A homodimers failed to interact with plastocyanin. Moreover, phenotypic analysis of atc6-1 pete1 and atc6-1 pete2 double mutants, each lacking Cyt C6A and one of the two piastocyanin-encoding genes, failed to reveal any genetic interaction. Overexpression of either PETE1 or PETE2 in the pete1 pete2 double knockout mutant background results in essentially wild-type photosynthetic performance, excluding the possibility that the two plastocyanin isoforms could have distinct functions in thylakoid electron flow.

关键词： mutants Overexpressors nteractors of Arabidopsis Plastocyanin Isoforms

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PEX16 contributions to peroxisome import and metabolism revealed by viable Arabidopsis pex16 mutants

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Journal of Integrative Plant Biology 2019年第7期61卷 853-870页

作者： Sarah E.Burkhart Roxanna J.Llinas Bonnie Bartel Department of BioSciences Rice University

Peroxisomes rely on peroxins(PEX proteins) for biogenesis, importing membrane and matrix proteins, and fission. PEX16, which is implicated in peroxisomal membrane protein targeting and forming nascent peroxisomes from the endoplasmic reticulum(ER), is unusual among peroxins because it is inserted co-translationally into the ER and localizes to both ER and peroxisomal membranes. PEX16 mutations in humans, yeast, and plants confer some common peroxisomal defects; however, apparent functional differences have impeded the development of a unified model for PEX16 action. The only reported pex16 mutant in plants, the Arabidopsis shrunken seed1 mutant, is inviable,complicating analysis of PEX16 function after embryogenesis. Here, we characterized two viable Arabidopsis pex16 alleles that accumulate negligible PEX16 protein levels. Both mutants displayed impaired peroxisome function-slowed consumption of stored oil bodies, decreased import of matrix proteins, and increased peroxisome size. Moreover,one pex16 allele exhibited reduced growth that could be alleviated by an external fixed carbon source, decreased responsiveness to peroxisomally processed hormone precursors, and worsened or improved peroxisome function in combination with other pex mutants. Because the mutations impact different regions of the PEX16 gene, these viable pex16 alleles allow assessment of the importance of Arabidopsis PEX16 and its functional domains.

关键词： pex16 mutants peroxisome metabolism

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Chemical mutagenesis and soybean mutants potential for identification of novel genes conferring resistance to soybean cyst nematode

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Journal of Integrative Agriculture 2018年第12期17卷 2734-2744页

作者： GE Feng-yong ZHENG Na ZHANG Liu-ping HUANG Wen-kun PENG De-liang LIU Shi-ming Institute of Plant Protection Chinese Academy of Agricultural Sciences College of Plant Protection Hunan Agricultural University

The resistance of soybean(Glycine max(L.) Merr.) to soybean cyst nematode(SCN, Heterodera glycines Ichinohe), which is a devastating pathogen in soybean production and causes a large quantity of annual yield loss worldwide, can shift during the long-term interaction and domestication. It is vital to identify more new resistance genetic sources for identification of novel genes underlying resistance to SCN for management of this pathogen. In the present study, first, two ethane methylsulfonate-mutagenesis soybean M2 populations of PI 437654, which shows a broad resistance to almost all of SCN races, and Zhonghuang 13, which is a soybean cultivar in China conferring strong resistance to lodging, were developed. Many types of morphological phenotypes such as four-and five-leaflet leaves were observed from these two soybean M2 populations. Second, 13 mutants were identified and confirmed to exhibit alteration of resistance to SCN race 4 through the forward genetic screening of 400 mutants of the PI 437654 M2 population, the rate of mutants with alteration of SCNinfection phenotype is 3.25%. Third, these identified mutants were further verified not to show any changes in the genomic sequences of the three known SCN-resistant genes, GmSHMT08, GmSNAP18 and GmSANP11, compared to the wildtype soybean; and all of them were still resistant to SCN race 3 similar to the wild-type soybean. Taken together, we can conclude that the 13 mutants identified in the present study carry the mutations of the new gene(s) which contribute(s) to the resistance to SCN race 4 in PI 437654 and can be potentially used as the genetic soybean sources to further identify the novel SCN-resistant gene(s).

关键词： soybean ethane methylsulfonate-mutagenesis populations mutants soybean cyst nematode race 4 resistance

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Genetic Analysis of Two Weak Dormancy mutants Derived from Strong Seed Dormancy Wild Type Rice N22 (Oryza sativa)

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Journal of Integrative Plant Biology 2011年第5期53卷 338-346页

作者： Bingyue Lu KunXie Chunyan Yang Long Zhang Tao Wu Xi Liu Ling Jiang Jianmin Wan National Key Laboratory for Crop Genetics and Germplasm Enhancement Jiangsu Plant Gene Engineering Research Center NanjingAgricultural University Nanjing 210095 China National Key Facility for Crop Gene Resources and Genetic Improvement Institute of Crop Science Chinese Academy of AgriculturalSciences Beijing 100081 China

Two weak dormancy mutants, designated Q4359 and Q4646, were obtained from the rice cultivar N22 after treatment with 400 Gy 6Co gamma-radiation. Compared to the N22 cultivar, the dormancy of the mutant seeds was more readily broken when exposed to a period of room temperature storage. The mutants also showed a reduced level of sensitivity to abscisic acid compared to the N22 cultivar, although Q4359 was more insensitive than Q4646. A genetic analysis indicated that in both mutants, the reduced dormancy trait was caused by a single recessive allele of a nuclear gene, but that the mutated locus was different in each case. The results of quantitative trait locus （QTL） mapping, based on the F2 population from Q4359 x Nanjing35, suggested that Q4359 lacks the QTL qSdn-1 and carries a novel allele at QTL qSdn-9, while a similar analysis of the Q4646 x Nanjing35 F2 population suggested that Q4646 lacks QTL qSdn-5, both qSdn-1 and qSdn-5 are major effect seed dormancy QTL in N22. Therefore, these two mutants were helpful to understand the mechanism of seed dormancy in N22.

关键词： mutants Dormancy Sowing Seed Seeds Varieties Loci Political systems Genes Quantitative trait loci

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Study on the Gas Phase Stability of Heme-binding Pocket in Cytochrome Tb_5 and Its mutants by Electrospray Mass Spectrometry

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Chinese Journal of Chemistry 2002年第12期20卷 1540-1545,1462页

作者：余翀天郭寅龙吕龙王韵华姚萍黄仲贤 Shanghai Institute of Organic Chemistry Chinese Academy of Sciences Shanghai 200032 Department of Chemistry Fudan University Shanghai 200433

To elucidate the effect of various amino acid residues on the heme-binding pocket in cytochrome Tb 5, several residues were chosen for replacement by means of site-directed mutagenesis. Comparison of the mass spectrum between the F35Y mutant and the wild type shows that the relative abundance of holo-protein ion of F35Y is lower than that of the wild type in gas phase. It is concluded that mutation from Phe35 residue to tyrosine decreases the hydrophobic character of cytochrome Tb 5 heme pocket, which decreases the stability of heme-binding pocket. ESI-MS spectra of the mutants V61E, V61K, V61H and V61Y show various contribution of amino acid to the stability of heme-binding pocket. The small and non-polar residue Val61 was replaced with large or polar residues, resulting in enhancing the trend of heme leaving from the pocket. In addition, comparison of the mass relative abundance of holo-proteins among all the Val61-mutants, shows that their stability in gas phase appropriately submit the following order: wild type>V61H>V61E>V61K≈V61Y. The extra great stability of quadruple sites mutant E44/48/56A/D60A shows that reduction of electrostatic or hydrogen bond interactions among the residues locating in the outside region of the heme edge remarkably affect the stability of heme. The results of analyzing the oxidation states of heme iron in Tb 5 and its mutants by in-source-CAD experiment suggest that the charge states of heme iron maintain inflexible in mutation process

关键词： electrospray cytochrome Tb 5 mutants gas phase stability

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Rice Brittleness mutants: A Way to Open the ‘Black Box’ of Monocot Cell Wall Biosynthesis

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Journal of Integrative Plant Biology 2011年第2期53卷 136-142页

作者： Baocai Zhang Yihua Zhou State Key Laboratory of Plant Genomics and National Center for Plant Gene Research Institute of Genetics and Developmental Biologythe Chinese Academy of Sciences Beijing 100101 China

Rice is a model organism for studying the mechanism of cell wall biosynthesis and remolding in Gramineae. Mechanical strength is an important agronomy trait of rice （Oryza sativa L.） plants that affects crop lodging and grain yield. As a prominent physical property of cell walls, mechanical strength reflects upon the structure of different wall polymers and how they interact. Studies on the mechanisms that regulate the mechanical strength therefore consequently results in uncovering the genes functioning in cell wall biosynthesis and remodeling. Our group focuses on the study of isolation of brittle culm （bc） mutants and characterization of their corresponding genes. To date, several bc mutants have been reported. The identified genes have covered several pathways of cell wall biosynthesis, revealing many secrets of monocot cell wall biosynthesis. Here, we review the progress achieved in this research field and also highlight the perspectives in expectancy. All of those lend new insights into mechanisms of cell wall formation and are helpful for harnessing the waste rice straws for biofuel production.

关键词： Rice Cell walls Mechanization Mechanics Cells Walls Lymphocytes Strength Biosynthesis mutants

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Field identification of morphological and physiological traits in two special mutants with strong tolerance and high sensitivity to drought stress in upland rice(Oryza sativa L.)

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Journal of Integrative Agriculture 2019年第5期18卷 970-981页

作者： HUANG Min XU Yu-hui WANG Hua-qi College of Agronomy and Biotechnology China Agricultural University

The two mutants idr1-1 and 297-28, which were obtained from the radiation mutation of HD297 and IAPAR9, were used as experimental materials in this study for a 2-year(2012 and 2013) experiment about field drought resistance identification in Beijing, China. Key agronomic traits and water-related physiological indexes were observed and measured, including the leaf anti-dead level(LADL), days to heading, plant height, setting percentage, aboveground biomass, leaf water potential(LWP), net photosynthetic rate(Pn) and transpiration rate. The results showed that the mutant idr1-1 that was under drought stress(DS) conditions for 2 years had the highest LADL grades(1.3 and 2.0) among all the materials, and they were 2–3 grades stronger than the wild-type IAPAR9 with an average that was 21.4% higher for the setting percentage than the wild type. Compared with the IAPAR9 for the 2-year average delay in the days to heading and the reduction rates in the plant height, setting percentage, and aboveground biomass under DS compared with the well-watered(WW) treatment, idr1-1 showed 3.2% less delay and 19.1, 16.4, and 6.1% less reduction, respectively. The idr1-1 in the LWP always exhibited the highest performance among all the materials. The Pn of idr1-1 under severe and mild DS comparing with that under WW was slightly decreased and even slightly increased, respectively, leading to an average reduction rate of only 0.92%, which was 26.93% less than that of IAPAR9. Under the severe DS, idr1-1 still showed the highest value of 16.88 μmol CO2 m–2 s–1 among all the materials and was significantly higher than that of IAPAR9(11.66 μmol CO2 m–2 s–1). Furthermore, only idr1-1 had the increased and the highest transpiration rate values(7.6 and 6.04 mmol H2 O m–2 s–1) under both mild and severe DS compared with the values under WW, when the transpiration rate of all the other materials significantly decreased. By contrast, the 297-28 in terms of the LADL grade under DS was the lowest(7.0)

关键词： rice mutants morphological and physiological traits drought resistance field identification

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ERp44 C160S/C212S mutants regulate IP_(3)R_(1) channel activity

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Protein & Cell 2011年第12期2卷 990-996页

作者： Congyan Pan Ji Zheng Yanyun Wu Yingxiao Chen Likun Wang Zhansong Zhou Wenxuan Yin Guangju Ji National Laboratory of Biomacromolecules Institute of Biophysics of Chinese Academy of SciencesBeijing 100101China Urological Surgery Research Institute Southwest HospitalThird Military Medical UniversityChongqing 400038China

Previous studies have indicated that ERp44 inhibits inositol 1,4,5-trisphosphate(IP3)-induced Ca2+release(IICR)via IP3R1,but the mechanism remains largely *** extracellular ATP to induce intracellular calcium transient as an IICR model,Ca2+image,pull down assay,and Western blotting experiments were carried out in the present *** found that extracellular ATP induced calcium transient via IP3Rs(IICR)and the IICR were markedly decreased in ERp44 overexpressed Hela *** inhibitory effect of C160S/C212S but not C29S/T396A/ΔT(331–377)mutants of ERp44 on IICR were significantly decreased compared with ***,the binding capacity of ERp44 to L3V domain of IP3R1(1L3V)was enhanced by ERp44 C160S/C212S *** together,these results suggest that the mutants of ERp44,C160/C212,can more tightly bind to IP3R1 but exhibit a weak inhibition of IP3R1 channel activity in Hela cells.

关键词： ERp44 mutants ATP inositol1 4 5-trisphosphate(IP3)receptors calciumtransient

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Hepatitis B virus,HBx mutants and their role in hepatocellular carcinoma

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World Journal of Gastroenterology 2014年第30期20卷 10238-10248页

作者： Ashraf Ali Hany Abdel-Hafiz Mohd Suhail Amany Al-Mars Mohammad Khalid Zakaria Kaneez Fatima Sultan Ahmad Esam Azhar Adeel Chaudhary Ishtiaq Qadri Department of Medical Biotechnology King Fahad Medical Research CenterKing Abdul Aziz UniversityPO Box 80216Jeddah 21589Saudi Arabia Division of Endocrinology Department of Medicine and MetabolismAnschutz Medical CampusUniversity of Colorado DenverAuroraCO 80045United States Department of Biochemistry All India Institute of Medical SciencesNew Delhi 110029India IQ Institute of Infection and Immunity Lahore 54000Pakistan Special Infectious Agents Unit-Biosafety Level 3 King Fahd Medical Research CenterKing Abdul Aziz UniversityJeddah 21589Saudi Arabia Medical Laboratory Technology Department Faculty of Applied Medical SciencesKing Abdul Aziz UniversityJeddah 21589Saudi Arabia

Hepatocellular carcinoma(HCC)is one of the leading causes of death induced by cancer in the modern world and majority of the cases are related to chronic hepatitis B virus(HBV)***-encoded X protein(HBx)is known to play a pivotal role in the pathogenesis of viral induced *** is a multifunctional protein of17 kDa which modulates several cellular processes by direct or indirect interaction with a repertoire of host factors resulting in *** might interfere with several cellular processes such as oxidative stress,DNA repair,signal transduction,transcription,protein degradation,cell cycle progression and apoptosis.A number of reports have indicated that HBx is one of the most common viral ORFs that is often integrated into the host genome and its sequence variants play a crucial role in *** mutational or deletion analysis it was shown that carboxy terminal of HBx has a likely role in protein-protein interactions,transcriptional transactivation,DNA repair,cell,signaling and pathogenesis of *** accumulated evidence thus far suggests that it is difficult to understand the mechanistic nature of HBx associated HCC,and HBx mediated transcriptional transactivation and signaling pathways may be a major *** article addresses the role of HBx in the development of HCC with particular emphasis on HBx mutants and their putative targets.

关键词： Hepatitis B virus Hepatocellular carcinoma Transcription factors Apoptosis Epigenetics mutants Tumor necrosis factor Activating protein Transforming growth factor Mitogen activated protein kinase

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Mutagenesis and selection of high efficiency hydrogen-producing mutants by ultraviolet radiation

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Journal of Harbin Institute of Technology(New Series) 2006年第6期13卷 635-639页

作者：任南琪郑国香李永峰林海龙李建政 School of Municipal and Environmental Engineering Harbin Institute of Technology

Hydrogen is an ideal, clean and sustainable energy source for the future because of its high conversion and nonpolluting nature. Biohydrogen production by dark-fermentation appears to have a great potential to be developed for practical application. However, one limiting factor affecting the development of hydrogen-production industrialization is that the hydrogen-producing capacity of bacteria is lower, so how to increase bacteria＇ s hydrogen-producing ability will be an urgent issue. In this experiment, 2 mutants, namely UV3 and UV7, were obtained by ultra-violet radiation. They grew and produced hydrogen efficiently on iron-containing medium. The hydrogen evolution of UV3 and UV7 were 2 356. 68 ml/L and 2 219. 62 ml/L at a glucose concentration of 10 g/L, respectively. With wild parent strain Ethanoligenens sp. ZGX4, the hydrogen evolution was 1 806. 02 ml/L under the same conditions. mutants＇ hydrogen-producing capacities were about 29. 71% and 22.22% higher than that of wild parent strain ZGX4. The maximum H2 production rate by mutants UV3 and UV7 were estimated to be 32.57 mmol H2/g cell h and 31.19 mmol H2/g cell h, respectively, which were 38. 18% and 34. 78% higher than the control （23.57 mmol H：/g cell h）. The abundant products of UV3 and UV7 were ethanol and acetic, which accounted for 95% - 98% of total soluble microbial products. In each case, mutant strains UV3 and UV7 evolved hydrogen at a higher rate than the wild type, showing a possible potential for commercial hydrogen production. Another mutant named UV20＇ was also gained whose main end metabolites were butyric acid and acetic acid. This would provide researched material for a discussion of metabolic pathways of hydrogen-producing bacteria.

关键词： biohydrogen UV-mutagenesis mutants

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