Analysis of transgene(s) (psy+crtI) inheritance and its stability over generations in the genetic background of indica rice cultivar Swarna

  • G. K. Chikkappa
  • N. K. Tyagi
  • K. Venkatesh
  • M. Ashish
  • K. V. Prabhu
  • T. Mohapatra
  • A. K. Singh
Original Article

Abstract

Rice the major staple food crop which feeds more than half of the world’s population but, lacks pathway to synthesize and accumulate provitamin A in endosperm therefore rice eaters particularly children, and pregnant women suffer due to vitamin A deficiency. The pathway for provitamin A synthesis in rice endosperm has been engineered and transgenic rice lines have yellow endosperm, called ‘Golden Rice’. The present study aimed at studying the inheritance of transgene(s) in six transgenic events of ‘Golden Rice’ and transfer of provitamin A trait from transgenic lines to a widely grown mega rice variety Swarna. The events E1, R1 and W1 showed normal Mendelian inheritance in F2, BC1F1 and BC1F2 generations. The event W1 was studied in BC1F3 as well and showed normal Mendelian inheritance of 3:1. The inheritance pattern in L1 event in BC1F1 and BC1F2 showed normal Mendelian inheritance following expected ratio 1:1 and 3:1 respectively. The two events G1 and T1 showed distorted segregation in BC1F2 and BC2F2 respectively in Swarna genetic background. In G1 event, transgene inheritance showed segregation distortion in BC1F2 in favour of transgene negative plants. In T1 event, inheritance followed expected Mendelian segregation in BC1F1, BC2F1 and BC2F2, generations. However, when tested against co-dominant inheritance 1:2:1 pattern in BC2F2, segregation distortion was observed with less than the expected transgene homozygotes. While against 3:1 ratio, it showed the expected segregation pattern in BC2F2 generation. Segregation distortion probably due to differential transmission of transgene positive/negative gametes through either/both parents which needs further study.

Keywords

Provitamin A Transgenic event Phytoene synthase (psy) Phytoene desaturase (crtITransgene inheritance Segregation distortion 

Abbreviations

bp

Base pair

mM

Milli molar

UV

Ultraviolet (light)

VAD

Vitamin A Deficiency

GGPP

Geranyl geranyl Pyrophosphate

GR

Golden Rice

PCR

Polymerase chain reaction

DNA

Deoxyribonucleic acid

NPF

National Phytoron Facility

IARI

Indian Agricultural Research Institute

NBPGR

National Bureau of Plant Genetic Resources

References

  1. Afolabi AS, Worland B, Snape JW, Vain P (2004) A large-scale study of rice plants transformed with different T-DNAs provides new insights into locus composition and T-DNA linkage configurations. Theor Appl Genet 109:815–826PubMedCrossRefGoogle Scholar
  2. Al-Babili S, Beyer P (2005) Golden rice—five years on the road—five years to go? Trends Plant Sci 10:565–573PubMedCrossRefGoogle Scholar
  3. Bhat SR, Srinivasan S (2002) Molecular and genetic analysis of transgenic plants: considerations and approaches. Plant Sci 163:673–681CrossRefGoogle Scholar
  4. Bregitzer P, Tonks D (2003) Inheritance and expression of transgenes in barley. Crop Sci 43:4–12CrossRefGoogle Scholar
  5. Budar F, Thia-toong I, Van Montagu M, Hernalsteens JP (1986) Agrobacterium mediated gene transfer results mainly in transgenic plants transmitting T-DNA as a single mendelian factor. Genetics 114:303–313PubMedGoogle Scholar
  6. Burkhardt PK, Beyer P, Wunn J, Kloti A, Armstrong GA, Schledz M, von Lintig V, Potrykus I (1997) Transgenic rice (Oyza stiva) endosperm expressing daffodil (Nacissus psecudonarcissus) phytoene synthase accumulates phytoene, a key intermediate of provitamin A biosynthesis. Plant J 11:1071–1078PubMedCrossRefGoogle Scholar
  7. Campbell BT, Baenziger PS, Mitra A, Sato S, Clemente T (2000) Crop Sci 40:1133–1141CrossRefGoogle Scholar
  8. Chareonpornwattana S, Thara KV, Wang L, Datta SK, Panbangred W, Muthukrishanan S (1999) Inheritance, expression, and silencing of a chitinase transgene in rice. Theor Appl Genet 98:371–378CrossRefGoogle Scholar
  9. Cho MJ, Choi HQ, Buchanan BB, Lemaux PG (1999) Inheritance of tissue specific expression of barley hordein promoter uid A fusions in transgenic barley plants. Theor Appl Genet 98:1253–1262CrossRefGoogle Scholar
  10. Christian P, West KP Jr, Khatry SK, Katz JK, Shrestha SR, Pradhan EK, LeClerq SC, Pokhrel RP (1998) Night blindness of pregnancy in rural Nepal—nutritional and health risks. Int J Epidemiol 27:231–237PubMedCrossRefGoogle Scholar
  11. Christian P, West KP Jr, Khatry SK, Kimbrough-Pradhan E, LeClerq SC, Katz J, Shrestha SR, Dali SM, Sommer A (2000) Night blindness during pregnancy and subsequent mortality among women in Nepal: effects of vitamin A and β-carotene supplementation. Am J Epidemiol 152:542–547PubMedCrossRefGoogle Scholar
  12. Cunningham FX Jr, Gantt E (1998) Genes and enzymes of carotenoid biosynthesis in plants. Annu Rev Plant Physiol Mol Biol 49:557–583CrossRefGoogle Scholar
  13. Datta SK, Datta K, Parkhi V, Rai M, Baisakh N, Sahoo G, Rehana S, Bandyopadhyay A, Alamgir Md Ali MS, Abrigo E, Oliva N, Torrizo L (2007) Golden Rice: introgression, breeding, and field evaluation. Euphytica 154:271–278CrossRefGoogle Scholar
  14. Demeke T, Hucl P, Baga M, Casweil K, Leung N, Chibbar RN (1999) Transgene inheritance and silencing in hexaploid spring wheat. Theor Appl Genet 99:947–953CrossRefGoogle Scholar
  15. Ghakawa D, Maqbool SB, Sudhakar D, Christon P, Kohli A (2000) Transgenic rice as a system to study the stability of transgene expression: Multiple heterologous transgene show similar behaviour in diverse genetic backgrounds. Theor Appl Genet 101:388–399CrossRefGoogle Scholar
  16. Jones G, Steketee RW, Black RE, Bhutta ZA, Morris SS, Bellagio Child Survival Study Group (2003) How many child deaths can we prevent this year? Lancet 362:65–71PubMedCrossRefGoogle Scholar
  17. Kim J, Duan X, Wu R, Seok SJ, Boston RS, Jang I, Eun M, Nahm BH (1999) Molecular and genetic analysis of transgenic rice plants expressing the maize ribosome-inactvating protein b-32 gene and the herbicide resistance bar gene. Mol Breed 5:85–94CrossRefGoogle Scholar
  18. Nandakumar N, Tyagi NK, Balram M, Chikkappa GK, Aprajita G, Mohapatra T, Prabhu KV, Singh AK (2008) Molecular analysis of transgene (psy) inheritance in a golden rice line developed in the genetic background of IR64. J Plant Biochem Biotech 17(2):127–132Google Scholar
  19. Paine JA, Shipton CA, Chaggar S, Howells RM, Kennedy MJ, Vernon G, Wright SY, Hinchliffe E, Adams JL, Silverstone AL, Drake R (2005) A new version of Golden Rice with increased pro-vitamin A content. Nat Biotechnol 23:482–487PubMedCrossRefGoogle Scholar
  20. Pawlowski WP, Torbert KA, Rines HW, Somers DA (1998) Irregular patterns of transgene silencing in allohexaploid oat. Plant Mol Biol 38:597–607PubMedCrossRefGoogle Scholar
  21. Peng J, Wen F, Lister RL, Hodges TK (1995) Inheritance of gusA and neo genes in transgenic rice. Plant Mol Biol 27:91–104PubMedCrossRefGoogle Scholar
  22. Perret SJ, Valentine J, Leggett JM, Morris P (2003) Integration, expression and inheritance of transgenes in hexaploid oat (Avena sativa L.). J Plant Physiol 160:931–943PubMedCrossRefGoogle Scholar
  23. Rai M, Datta K, Parkhi V, Tan J, Oliva N, Chawla HS, Datta SK (2007) Variable T-DNA linkage configuration affects inheritance of carotenogenic transgenes and carotenoid accumulation in transgenic indica rice. Plant Cell Rep 26:1221–1231PubMedCrossRefGoogle Scholar
  24. Satoto Sulistyowati Y, Hartanac A, Slamet-Loedin IH (2008) The segregation pattern of insect resistance genes in the progenies and crosses of transgenenic Rojolele Rice. Indonesian Journal of Agricultural Science 9(2):35–43Google Scholar
  25. Spencer TM, O’Brien JV, Start WG, Adams TR, Willian J, Gordon-Kamm WJ, Lemaux PG (1992) Segregation of transgenes in maize. Plant Mol Biol 13:201–210CrossRefGoogle Scholar
  26. Srivastava A, Vasil V, Vasil IK (1996) Molecular characterization of the fate of transgenes in transformed wheat (Triticum aestivum L.). Theor Appl Genet 92:1031–1037CrossRefGoogle Scholar
  27. Vain P, James VA, Worland B, Snape JQ (2002) Transgene behaviour across two generations in a large random plants produced by particle bombardment. Theor Appl Genet 105:878–879PubMedCrossRefGoogle Scholar
  28. Wu L, Nandi S, Chen L, Rodriguez RL, Huang N (2002) Expression and inheritance of nine transgenes in rice. Transgeic Res 11:533–541CrossRefGoogle Scholar
  29. Yin Z, Plader W, Malepszy S (2004) Transgene inheritance in plants. J Appl Genet 45(2):127–144PubMedGoogle Scholar
  30. Zhang Y, Yin X, Yang A, Li G, Zhang J (2005) Stability of inheritance of transgenes in maize (Zea mays L.) lines produced using different transformation methods. Euphytica 144:11–22CrossRefGoogle Scholar

Copyright information

© Optical Society of India 2011

Authors and Affiliations

  • G. K. Chikkappa
    • 1
    • 4
  • N. K. Tyagi
    • 1
    • 3
  • K. Venkatesh
    • 1
  • M. Ashish
    • 1
  • K. V. Prabhu
    • 1
  • T. Mohapatra
    • 2
  • A. K. Singh
    • 1
  1. 1.Division of GeneticsIndian Agricultural Research InstituteNew DelhiIndia
  2. 2.National Research Center on Plant BiotechnologyIndian Agricultural Research InstituteNew DelhiIndia
  3. 3.Basmati Export Development FoundationMeerutIndia
  4. 4.Directorate of Maize ResearchNew DelhiIndia

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