Generation and Characterization of Overexpression Lines of a Rice Pyruvate Kinase Gene OsPK1

doi:10.3969/j.issn.1000-7091.2012.02.001

Abstract

Abstract: We have reported that downregulation of OsPK1，a rice pyruvate kinase，causes dwarfism，reduced?seed set and so on． Therefore，the aim in this study is to explore the effect of the overexpression of OsPK1 on rice．?The construct containing a CaMV 35S-driven full-length CDS of OsPK1 was introduced into the genome of wild-type?(WT) Nipponbare via an Agrobacterium-mediated method． Three overexpression lines were selected for further characterization and analysis． Statistics showed that these overexpression plants approach WT plants in plant height，tiller?number，panicle length and thousand seed weight，while present slightly reduced seed set and significantly increased?viable seed number per panicle． Quantitative RT-PCR results exhibited that the expressions of four metabolic enzymes were downregulated or upregulated in the young leaves of overexpression plants． The sugar contents were?measured using GC-MS，and the results indicated that the contents of glucose，fructose，galactose and sucrose in OsPK1 overexpression plants showed few differences compared with those of WT plants． Together，the characterization?of OsPK1 overexpression plants can help us better understand the role of OsPK1，and increased viable seed number?per panicle should be a prospective application．?

Key words: OsPK1, Overexpression, Pyruvate kinase, Rice (Oryza sativa L.)

CLC Number:

ZHANG Yan, PANG Jin-huan, WU Jia-he, HE Chao-zu. Generation and Characterization of Overexpression Lines of a Rice Pyruvate Kinase Gene OsPK1[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(2): 1-5. doi: 10.3969/j.issn.1000-7091.2012.02.001.

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References

[1] Plaxton W C． The organization and regulation of plant glycolysis[J]． Annu Rev Plant Physiol Plant Mol Biol，1996, 47:185－214．
[2] Ruuska S A，Girke T，Benning C，et al． Contrapuntal net- works of gene expression during Arabidopsis seedfilling
[J]． Plant Cell，2002，14:1191－1206．
[3] ?Andre C，Froehlich J E，Moll M R，et al． A heteromeric?plastidic pyruvate kinase complex involved in seed oil biosynthesis in Arabidopsis[J]． Plant Cell，2007，19:2006?－2022．
[4] Baud S，Wuilleme S，Dubreucq B，et al． Function of plastidial pyruvate kinases in seeds of Arabidopsis thaliana
[J]． Plant J，2007，52:405－419．
[5] Grodzinski B，Jiao J，Knowles V L，et al． Photosynthesis?and carbon partitioning in transgenic tobacco plants deficient in leaf cytosolic pyruvate kinase[J]． Plant Physiol, 1999，120:887－895．
[6] Knowles V L，McHugh S G，Hu Z Y，et al． Altered growth?of transgenic tobacco lacking leaf cytosolic pyruvate kinase[J]． Plant Physiol，1998，116:45－51．
[7] Oliver S N，Lunn J E，Urbanczyk-Wochniak E，et al． Decreased expression of cytosolic pyruvate kinase in potato?tubers leads to a decline in pyruvate resulting in an in vivo repression of the alternative oxidase[J]． Plant Physiol, 2008，148:1640－1654．
[8] Zhang Y，Xiao W K，Luo L J，et al． Downregulation of OsPK1，a cytosolic pyruvate kinase，by T-DNA insertion causes dwarfism and panicle enclosure in rice[J]． Planta, 2011，doi:10． 1007/s00425－011－1471－3．
[9] Hiei Y，Ohta S，Komari T，et al． Efficient transformation of?rice (Oryza sativa L． ) mediated by Agrobacterium and?sequence analysis of the boundaries of the T-DNA[J]．?Plant J，1994，6:271－282．
[10] Dai M Q，Zhao Y，Ma Q，et al． The rice YABBY1 gene is?involved in the feedback regulation of gibberellin metabolism[J]． Plant Physiol，2007，144:121－133．
[11] Tan Y P，Li K，Hu L，et al． Fast and simple droplet sampling of sap from plant tissues and capillary microextraction of soluble saccharides for picogram-scale quantitative determination with GC-MS[J]． J Agric Food Chem, 2010，58:9931－9935．
[12] ?Collakova E，Goyer A，Naponelli V，et al． Arabidopsis?10-formyl tetrahydrofolate deformylases are essential for?photorespiration[J]． Plant Cell，2008，20(7):1818－?1832．?

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