Advertisement

Theoretical and Applied Genetics

, Volume 122, Issue 2, pp 421–428 | Cite as

Identification, fine mapping and characterisation of a dwarf mutant (bnaC.dwf) in Brassica napus

  • Xinhua Zeng
  • Lixia Zhu
  • Yanli Chen
  • Liping Qi
  • Yuanyuan Pu
  • Jing Wen
  • Bin Yi
  • Jinxiong Shen
  • Chaozhi Ma
  • Jinxing Tu
  • Tingdong FuEmail author
Original Paper

Abstract

In the present study, we have obtained one dwarf mutant (bnaC.dwf) from the Brassica napus inbred line T6 through chemical mutagen ethyl methanesulfonate (EMS). We have determined the phenotypic effects and genetic characteristics of dwarf mutant (bnaC.dwf). The dwarf mutant was insensitive to exogenous GA3 for plant height, suggesting that it is significantly playing a crucial role in the gibberellins response pathway. Genetic analysis revealed that one recessive gene is responsible for controlling the phenotypic expression of dwarf mutant. Amplified Fragment Length Polymorphism (AFLP) technique was applied for selecting markers linked to the BnaC.DWF gene which assisted in screening of dwarf and normal individuals in the BC4 population. We have screened 1,024 primer combinations and then identified nine AFLP markers linked to the BnaC.DWF gene. Identification and linkage of the markers were carried out by analysing 2,000 individuals from a larger population of the BC4. Two markers EA10MC09 and EA12MC02 were located on the flanking region of the BnaC.DWF gene at a distance of 0.2 and 0.05 cM, respectively. Four AFLP markers EA09MG05, EA02MC07, EA01MC01 and EC04MC07 were successfully converted into Sequence Characterised Amplified Region markers namely SCA9G5, SCA2C7, SCA1C1 and SCC4C7. We further integrated BnaC.DWF linked Simple Sequence Repeat markers into two populations (Piquemal et al. Theor Appl Genet 111:1514–1523, 2005; Cheng et al. Theor Appl Genet 118:1121–1131, 2009). BnaC.DWF was mapped to the linkage region N18. The molecular markers developed from these investigations will greatly accelerate the selection process for developing dwarf varieties in B. napus by Marker Assisted Selection and genetic engineering.

Keywords

Plant Height Amplify Fragment Length Polymorphism Amplify Fragment Length Polymorphism Marker Scar Marker DELLA Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We greatly acknowledge the assistance of Prof. Meng in providing us DNA of DH populations for our experimental studies. I greatly acknowledge the efforts of Gautm Mayank for editing through English version of the manuscript. We are grateful for the financial support of the National key Basic Research Special Foundation of China (No. 2006CB101604 and No. 2007CB109006), the National High-tech Research and Development Program (2009AA101105), the Modern Agriculture Industrialisation System Construction (nycytx-00501) and the National key Technology R&D Program (2008BAD97B04).

References

  1. Boss PK, Thomas MR (2002) Association of dwarfism and floral induction with a grape ‘green revolution’ mutation. Nature 416:847–850CrossRefPubMedGoogle Scholar
  2. Brian PW (1959) Effects of gibberellins on plant growth and development. Biol Rev 34:37–77CrossRefGoogle Scholar
  3. Chandler PM, Marion-Poll A, Ellis M, Gubler F (2002) Mutants at the Slender1 locus of barley cv Himalaya. Molecular and physiological characterization. Plant Physiol 129:181–190CrossRefPubMedGoogle Scholar
  4. Cheng X, Xu J, Xia S, Gu J, Yang Y, Fu J, Qian X, Zhang S, Wu J, Liu K (2009) Development and genetic mapping of microsatellite markers from genome survey sequences in Brassica napus. Theor Appl Genet 118:1121–1131CrossRefPubMedGoogle Scholar
  5. Elliott MC (2008) Foreword. In: Slater A (ed) Plant biotechnology: the genetic manipulation of plants, 2nd edn. Oxford University Press, New YorkGoogle Scholar
  6. Evans LT (1998) Feeding the ten billion: plant and population growth. Cambridge University Press, CambridgeGoogle Scholar
  7. Foisset N, Delourme R, Barret P, Renard M (1995) Molecular tagging of the dwarf BREIZH (Bzh) gene in Brassica napus. Theor Appl Genet 91:756–761CrossRefGoogle Scholar
  8. Foisset N, Delourme R, Barret P, Hubert N, Landry BS, Renard M (1996) Molecular-mapping analysis in Brassica napus using isozyme, RAPD and RFLP markers on a doubled-haploid progeny. Theor Appl Genet 93:1017–1025CrossRefGoogle Scholar
  9. Fridborg I, Kuusk S, Moritz T, Sundberg E (1999) The Arabidopsis dwarf mutant shi exhibits reduced gibbrellin responses conferred by over expression of a new putative zinc finger protein. Plant Cell 11:1019–1031CrossRefPubMedGoogle Scholar
  10. Hedden P (2003) The genes of the Green Revolution. Trends Genet 19:5–9CrossRefPubMedGoogle Scholar
  11. Ikeda A, Ueguchi-Tanaka M, Sonoda Y, Kitano H, Koshioka M, Futsuhara Y, Matsuoka M, Yamaguchi J (2001) Slender rice, a constitutive gibberellin response mutant, is caused by a null mutation of the SLR1 gene, an ortholog of the height-regulating gene GAI/RGA/RHT/D8. Plant Cell 13:999–1010CrossRefPubMedGoogle Scholar
  12. Islam N, Evans EJ (1994) Influence of lodging and nitrogen rate on the yield and yield attributes of oilseed rape (Brassica napus L.). Theor Appl Genet 88:530–534CrossRefGoogle Scholar
  13. Ke LP, Sun YQ, Liu PW, Yang GS (2004) Identification of AFLP fragments linked to one recessive genic male sterility (RGMS) in rapeseed (Brassica napus L.) and conversion to SCAR markers for marker-aided selection. Euphytica 138:163–168CrossRefGoogle Scholar
  14. Khush GS (1999) Green revolution: preparing for the 21st century. Genome 42:646–655CrossRefPubMedGoogle Scholar
  15. Khush GS (2001) Green Revolution: the way forward. Nat Rev Genet 2:815–822CrossRefPubMedGoogle Scholar
  16. Lagercrantz U (1998) Comparative mapping between Arabidopsis thaliana and Brassica nigra indicates that Brassica genomes have evolved through extensive genome replication accompanied by chromosome fusions and frequent rearrangements. Genetics 150:1217–1228PubMedGoogle Scholar
  17. Lander ES, Green P, Abrahamson J, Barlow A, Daly MJ, Lincoln S, Newburg L (1987) MAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174–181CrossRefPubMedGoogle Scholar
  18. Li H, Wang Y, Li X, Gao Y, Wang Z, Zhao Y, Wang M (2010) A GA-insensitive dwarf mutant of Brassica napus L. correlated with mutation in pyrimidine box in the promoter of GID1. Mol Biol Rep doi: 10.1007/s11033-010-0094-2
  19. Lincoln S, Daly M, Lander E (1992) Constructing genetic maps with MAPMAKER/EXP 3.0. Whitehead institute technical report, 3rd edn. Whitehead Technical Institute, Cambridge, MAGoogle Scholar
  20. Liu HL, Tang J (1990) A review on the plant ideotype research in oilseed rape. J Crops 4:2–5 (in Chinese)Google Scholar
  21. Liu C, Wang J, Huang T, Wang F, Yuan F, Cheng X, Zhang Y, Shi S, Wu J, Liu K (2010) A missense mutation in the VHYNP motif of a DELLA protein causes a semi-dwarf mutant phenotype in Brassica napus. Theor Appl Genet 121:249–258CrossRefPubMedGoogle Scholar
  22. Lu GY, Yang GS, Fu TD (2001) Silver stained AFLP—a novel assay for DNA fingerprinting in Brassica napus. J Huazhong Agric Univ 20:413–415 (in Chinese)Google Scholar
  23. Lu GY, Yang GS, Fu TD (2004) Molecular mapping of a dominant genic male sterility gene (Ms) in rapeseed (Brassica napus L.). Plant Breed 123:262–265CrossRefGoogle Scholar
  24. Mei DS, Wang HZ, Li YC, Hu Q, Li YD, Xu YS (2006) The discovery and genetic analysis of dwarf mutation 99CDAM in Brassica napus L. Yi Chuan 28(7):851–857 (in Chinese)PubMedGoogle Scholar
  25. Michelmore RW, Paran I, Kesseli RV (1991) Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl Acad Sci USA 88:9828–9832CrossRefPubMedGoogle Scholar
  26. Monna L, Kitazawa N, Yoshino R, Suzuki J, Masuda H, Maehara Y, Tanji M, Sato M, Nasu S, Minobe Y (2002) Positional cloning of rice semidwarfing gene, sd-1: rice “green revolution gene” encodes a mutant enzyme involved in gibberellin synthesis. DNA Res 9:11–17CrossRefPubMedGoogle Scholar
  27. Muangprom A, Osborn TC (2004) Characterization of a dwarf gene in Brassica rapa, including the identification of a candidate gene. Theor Appl Genet 108:1378–1384CrossRefPubMedGoogle Scholar
  28. Muangprom A, Stephen GT, Sun T, Thomas CO (2005) A novel dwarfing mutation in a Green Revolution gene from Brassica rapa. Plant Physiol 137:931–938CrossRefPubMedGoogle Scholar
  29. Muangprom A, Mauriera I, Thomas CO (2006) Transfer of a dwarf gene from Brassica rapa to oilseed B. napus, effects on agronomic traits, and development of a ‘perfect’ marker for selection. Mol Breed 17:101–110CrossRefGoogle Scholar
  30. Negi MS, Devic M, Delseny M, Lakshmikumaran M (2000) Identification of AFLP fragments linked to seed coat colour in Brassica juncea and conversion to a SCAR marker for rapid selection. Theor Appl Genet 101:146–152CrossRefGoogle Scholar
  31. Peng J, Carol P, Richards DE, Ling KE, Cowling RJ, Murphy GP, Harberd NP (1997) The Arabidopsis GAI gene defines a signaling pathway that negatively regulates gibberellin responses. Gene Dev 11:3194–3205CrossRefPubMedGoogle Scholar
  32. Peng J, Richards DE, Hartley NM, Murphy GP, Devos KM, Flintham JE, Beales J, Fish LJ, Worland AJ, Pelica F, Sudhakar D, Christou P, Snape JW, Gale MD, Harberd NP (1999) Green revolution genes encode mutant gibberellin response modulators. Nature 400:256–261CrossRefPubMedGoogle Scholar
  33. Piquemal J, Cinquin E, Couton F, Rondeau C, Seignoret E, doucet I, Perret D, Villeger MJ, Vincourt P, Blanchard P (2005) Construction of an oilseed rape (Brassica napus L.) genetic map with SSR markers. Theor Appl Genet 111:1514–1523CrossRefPubMedGoogle Scholar
  34. Qiu D, Morgan C, Shi J, Long Y, Liu J, Li R, Zhuang X, Wang Y, Tan X, Dietrich E, Weihmann T, Everett C, Vanstraelen S, Beckett P, Fraser F, Trick M, Barnes S, Wilmer J, Schmidt R, Li J, Li D, Meng J, Bancroft I (2006) A comparative linkage map of oilseed rape and its use for QTL analysis of seed oil and erucic acid content. Theor Appl Genet 114(1):67–80CrossRefPubMedGoogle Scholar
  35. Rozen S, Skaletsky H (1999) Primer 3. Code available at http://wwwgenome.wi.mit.edu/genome_software/other/primer3.html
  36. Sasaki A, Ashikari M, Ueguchi-Tanaka M, Itoh H, Nishimura A, Swapan D, Ishiyama K, Saito T, Kobayashi M, Khush GS, Kitano H, Matsuoka M (2002) Green revolution: a mutant gibberellin-synthesis gene in rice. Nature 416:701–702CrossRefPubMedGoogle Scholar
  37. Silverstone AL, Ciampaglio CN, Sun T (1998) The Arabidopsis RGA gene encodes a transcriptional regulator repressing the gibberellin signal transduction pathway. Plant Cell 10:155–169CrossRefPubMedGoogle Scholar
  38. Spielmeyer W, Ellis MH, Chandler PM (2002) Semidwarf (sd-1), “green revolution” rice, contains a defective gibberellin 20-oxidase gene. Proc Natl Acad Sci USA 99:9043–9048CrossRefPubMedGoogle Scholar
  39. Sun T (2000) Gibberellin signal transduction. Curr Opin Plant Biol 3:374–380CrossRefPubMedGoogle Scholar
  40. Sun T, Gubler F (2004) Molecular mechanism of gibberellin signaling in plants. Annu Rev Plant Biol 55:197–223CrossRefPubMedGoogle Scholar
  41. Wang ML, Zhao Y, Chen F, Yin XC (2004) Inheritance and potentials of a mutated dwarfing gene ndf1 in Brassica napus. Plant Breed 123:449–453CrossRefGoogle Scholar
  42. Winkler RG, Freeling M (1994) Physiological genetics of the dominant gibberellins-nonresponsive maize dwarfs, Dwarf8 and Dwarf9. Planta 193:341–348CrossRefGoogle Scholar
  43. Wu SR, Chen WF, Zhou X (1988) Enzyme linked immunosorbent assay for endogenous plant hormones. Plant Physiol Commun 5:53–57 (in Chinese)Google Scholar
  44. Yi B, Chen YL, Lei SL, Tu JX, Fu TD (2006) Fine mapping of the recessive genic male-sterile gene (Bnms1) in Brassica napus L. Theor Appl Genet 113(4):643–650CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Xinhua Zeng
    • 1
  • Lixia Zhu
    • 1
  • Yanli Chen
    • 1
  • Liping Qi
    • 1
  • Yuanyuan Pu
    • 1
  • Jing Wen
    • 1
  • Bin Yi
    • 1
  • Jinxiong Shen
    • 1
  • Chaozhi Ma
    • 1
  • Jinxing Tu
    • 1
  • Tingdong Fu
    • 1
    Email author
  1. 1.National Key Laboratory of Crop Genetic Improvement, National Center of Rapeseed Improvement in WuhanHuazhong Agricultural UniversityWuhanPeople’s Republic of China

Personalised recommendations