玉米粉质胚乳突变体

1、宋任涛宋任涛上海大学上海大学Walbot and Evans (2003) Nature Reviews Genetics, 4: 369-379Sabelli and Larkins, Plant Physiol 149 (2009) 14-26 淀粉粒淀粉粒(S)和蛋白体和蛋白体(P)是主要的储藏形式是主要的储藏形式SSP胚乳是主要的储藏组织胚乳是主要的储藏组织Herman and Larkins (1999) Plant Cell, 11: 601-613Adapted from Lending and Larkins (1989) Plant Cell First report by

2、Singleton WR and Jones DF in the early of last century, including opaque endosperm1 and opaque endosperm2. Emerson RA, Beadle GW and Fraser AC(1935) A summary of linkage studies in maize. Cornell Univ Agric Exp Stn Memoir 180:1-83, described 40 classic genetic loci. Mertz, ET et al. (1964) Mutant ge

3、ne that changes protein composition and increases lysine content of maize endosperm. Science 145:279-280 (o2). Nelson, OE Jr et al. (1965) Second mutant gene affecting the amino acid pattern of maize endosperm proteins. Science 150:1469-1470 (floury2 (fl2), o1 and o2). Misra, PS et al. (1972) Endosp

4、erm protein synthesis in maize mutants with increased lysine content. Science 176:1425-1426 (bt2, fl2, o2 and o7).nRecessive opaque mutants (o1,o2, o5, o7, o9o11, o13o16),nSemi-dominant floury mutants (fl1, fl2 and fl3)nDominant mutants (Mc and De-B30) Bryan C. Gibbon and Brian A. Larkins (2005) Tre

5、nds Genet 21, 227-233. MutantProductFunctionReferenceo2bZIP transcription factorDown-regulation 22KDa zein, LKR, and etc.Schmidt, et al. (1987) Sciencefl222KDa-zeinWith a defective SP, accumulation on ERColeman, et al. (1995) PNASDe-B3019KDa-zeinWith a S15P mutation in the SP, un-cleavedKim, et al.

6、(2004) Plant Physiol.Mc16-kDa -zeinA frame-shift mutation (38-bp deletion), changed the last 63 amino acidsKim, et al. (2006) Plant J.fl1A novel ER proteinA-to-G point mutation,resulting in a Ser-to-Glysubstitution in the first transmembrane domainHolding, et al. (2007) Plant Cell McWhirter, KS (197

7、1) A floury endosperm, high lysine locus on chromosome 10. MNL 45:184 Misra, PS et al. (1972) Endosperm protein synthesis in maize mutants with increased lysine content. Science 176:1425-1426 Burr, FA and Burr, B (1982) Three mutations in Zea mays affecting zein accumulation: a comparison of zein po

8、lypeptides, in vitro synthesis and processing, mRNA levels, and genomic organization. J Cell Biol 94:201-206 (o2, fl2, o7) zeins33.4%, non-zeins15.1% , total proteins20.7%o7wt2500 x6000 x21 DAP蛋白体变少变小蛋白体变少变小Gene2 appeared to be the candidate gene for o7. 28kb The cloned gene is the O7 gene. Suppress

9、ion of O7 gene function by RNAi reproduced the opaque endosperm phenotype. Acyl-CoA Synthetase (ACS) superfamily Acyl Activating Enzyme (AAE) family Substrate screening for purified GST-O7. 草酸ABCFP-PTS1GFP-O7No significant difference at transcript level between mutant and wild type. The 12-bp deleti

10、on in o7 mutant allele caused significant reduction of o7 protein in developing seeds. CK NTG NTG NTG TG TG TG The content of total protein, zein, starch and fat of wt and mutantWT 21DAPSTPBmu 21DAPSTPBSzabados and Savour. (2009) Trands in Plant sciencexThe subcellular localization of O6 proteinOnio

11、n CellsTobacco CellsThe enzyme activity analysis of recombinant O6 protein The free and total proline content of wt and mutant kernelThe expression analysis of zeins in wt and mutant endospermThe expression analysis of zeins in wt and mutant endosperm15KDa16KDa50KDa27KDa22KDa19KDaTublin18DAP21DAP24D

12、AP27DAPWTpro1WTpro1WTpro1WTpro1The total protein analysis of wt and pro1 mutant endosperm of different alleleTublineIF2-PeIF2-Total27DAP24DAPWTmuWTmuP5CSProlineXUncharged tRNAproTranslational stresseIF2phosphorylationProtein synthesis结构基因 fl2, Mc1, DeB30转录层面 o2（转录因子）翻译层面 o7, o6（氨基酸供给） o1（蛋白体形成） fl1（蛋白体装配）储运层面Maize Genetics Stock CenterBrookhaven National Laboratory Dr. Frances BurrChina Agricultural UniversityDr. Jinsheng LaiInstitute of Genetics and Developmental Biology, CASDr. Lihuang ZhuUniversity of ArizonaDr. Brian LarkinsShanghai UniversityDr. Gang