Doubled-haploid production in chickpea (Cicer arietinum L.): role of stress treatments.
This is the first report on the production of double-haploid chickpea embryos and regenerated plants through anther culture using Canadian cultivar CDC Xena (kabuli) and Australian cultivar Sonali (desi). Maximum anther induction rates were 69% for Sonali and 63% for CDC Xena. Under optimal conditions, embryo formation occurred within 15-20 days of culture initiation with 2.3 embryos produced per anther for CDC Xena and 2.0 embryos per anther for Sonali. For anther induction, the following stress treatments were used: (1) flower clusters were treated at 4 degrees C for 4 days, (2) anthers were subjected to electric shock treatment of three exponentially decaying pulses of 50-400 V with 25 microF capacitance and 25 Omega resistance, (3) anthers were centrifuged at 168-1,509g for 2-15 min, and finally (4) anthers were cultured for 4 days in high-osmotic pressure (563 mmol) liquid medium. Anthers were then transferred to a solid embryo development medium and, 15-20 days later, embryo development was observed concomitant with a small amount of callus growth of 0.1-3 mm. Anther-derived embryos were regenerated on plant regeneration medium. Electroporation treatment of anthers enhanced root formation, which is often a major hurdle in legume regeneration protocols. Cytological studies using DAPI staining showed a wide range of ploidy levels from haploid to tetraploid in 10-30-day-old calli. Flow cytometric analysis of calli, embryos and regenerated plants showed haploid profiles and/or spontaneous doubling of the chromosomes during early regeneration stages.
Grewal
Ravinder Kaur
RK
Crop Development Centre, University of Saskatchewan, 51 Campus Drive, Saskatoon, SK, S7N 5A8, Canada. ravinder.grewal@usask.ca
Lulsdorf
Monika
M
Croser
Janine
J
Ochatt
Sergio
S
Vandenberg
Albert
A
Warkentin
Thomas D
TD
D018929:Cell Culture Techniques; D002478:Cells, Cultured; D002498:Centrifugation; D032461:Chromosomes, Plant; D029863:Cicer / Q000196:embryology / Q000235:genetics*; D003080:Cold Temperature; D003470:Culture Media; D018274:Electroporation; D035264:Flowers / Q000196:embryology / Q000235:genetics; D006238:Haploidy; D009997:Osmotic Pressure; D012038:Regeneration; D013312:Stress, Physiological
D003470:Culture Media