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Genetic manipulation of lupins

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  1. Tez No: 400578
  2. Yazar: MEHMET BABAOĞLU
  3. Danışmanlar: DR. M. R. DAVEY
  4. Tez Türü: Doktora
  5. Konular: Ziraat, Botanik, Agriculture, Botany
  6. Anahtar Kelimeler: Belirtilmemiş.
  7. Yıl: 1996
  8. Dil: İngilizce
  9. Üniversite: The Unıversıty Of Nottıngham
  10. Enstitü: Yurtdışı Enstitü
  11. Ana Bilim Dalı: Bitki Koruma Ana Bilim Dalı
  12. Bilim Dalı: Belirtilmemiş.
  13. Sayfa Sayısı: Belirtilmemiş.

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Özet (Çeviri)

The available gene pool in the genus Liqrimts, as in other grain legume genera, is restricted by the sexual incompatibility of many interspecific and iiitergeneric crosses. Genetic manipulation and in vitro culture provide substantial advantages in broadening the genetic variability and reducing the time required for the introgression of new traits, compared to conventional breeding. Reproducible protocols were developed for callus induction and maintenance, and for plant regeneration, from pre-existing meristems, for the two important hipimis species, L mutabilis cv. Potosi andL angiistifolius cv. Kubesa. Excision of the shoot apical layer(s) resulted in the induction of multiple buds covering the whole surface of the shoot apical explant (sa2) in L. mutabilis cv. Potosi. Multiple buds produced shoots. Leaf mesophyll was the most suitable tissue as a protoplast source in both cultivars. However, sustained division of protoplasts of lupin was not achieved. Protoplasts remain recalcitrant to culture. Currently, they do not offer any reasonable prospect as a basis for the genetic manipulation of Lupimts species. This study presents the first establishment of hairy root cultures in both Lupimis species, Transgenic hairy roots were induced by inoculation of explants from both L angiistifolius cv. Kubesa wAL. mutabilis cv. Potosi with Agrobacterium rhizogenes R1601. Hairy root cultures exhibited resistance to kanamycin, but still retained their transformed nature after 14 months with or without kanamycin selection. All root cultures produced NFTII protein as detected by the NPTH. Elisa assay. The integration of the nptTL gene into the genomes of both cultures was confirmed by non-radioactive Southern hybridisation. This study describes the first production of transgenic plants of lupin. Transgenic plants were regenerated after inoculation of shoot apical explant of L. mutabilis cv. Potosi with Agrobacterium twnefaciens LBA4404. Such plants were kanamycin resistant, expressed transient GUS activity and sysnthesised the NPTII protein. The integration of the nptW and gus-intron genes were detected by analysis of genomic DNA Double transgenic hairy roots were obtained by the inoculation of transgenic shoots (Kanamycin resistant) with A. rhizogenes R1601 (DTRs). DTRs were more resistant to kanamycin than donor transgenic shoots and contained one extra copy of the nptTL gene in their genomes. These organs also expressed transient GUS activity. Mcroprojectile-mediated gene transfer was unsuccessful compared to Agrobacterhim-me&aUd transformation.

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