Murugan Theivanai, Awasthi Om Prakash, Singh Bhupinder, Rudra Shalini Gaur
Division of Fruits and Horticultural Technology, ICAR-Indian Agricultural Research Institute, New Delhi, 110012 India.
Division of Environment Science, ICAR-Indian Agricultural Research Institute, New Delhi, 110012 India.
Physiol Mol Biol Plants. 2024 Aug;30(8):1297-1312. doi: 10.1007/s12298-024-01498-7. Epub 2024 Aug 8.
Solid mutant induction using specialized habituation and PBR (Plant bio-regulator) autotrophy-mediated suspension-based ISE system was the prime aim of present investigation. Based on survival of cell clumps after mutagen treatment, the probit analysis was calculated. The result revealed LD at 54.31 Gy in gamma, while for EMS (ethyl methanesulfonate), it was 0.1% for 3 h and 0.5% for 1 h. Based on embryogenesis efficiency, a dose rate of 100 Gy and 0.1% EMS for a 3-h exposure were selected for regeneration. As compared to control, significant decrease in the embryogenesis efficiency was recorded at 100 Gy (85.92%) with similar reduction trends in embryo production (79.49%), germination (13.43%), conversion (2.48%), establishment (15.78%) and acclimatization (60.92%). The growth-related parameters such as root and shoot length and number of leaves/regenerant were also significantly reduced to 67.29%, 30.19% and 5.03%, respectively, in the regenerated plants after gamma irradiation as compared to control. In the EMS treatment, at the dose rate of 0.1% for 3-h, the embryogenesis efficiency was reduced to 43.67% with similar diminution trends in embryo production (59.49%), germination (8.95%), conversion (1.94%), establishment (4.37%) and acclimatization (29.9%). The growth-related parameters in the EMS treatment, decreased to 91.00% (root length), 71.34% (shoot length) and 35.03% (no. of leaves). The molecular marker based varied amplifications confirmed the occurrence of mutations in both gamma and EMS induced M regenerants. The study highlights the alternative high frequency in vitro mutagenesis protocol for induction of solid mutants in Kinnow mandarin and related citrus species.
利用特殊驯化和基于植物生物调节剂（PBR）自养介导的悬浮培养ISE系统诱导固态突变体是本研究的主要目标。根据诱变处理后细胞团的存活率进行了概率分析。结果显示，伽马射线诱变的致死剂量为54.31 Gy，而对于甲基磺酸乙酯（EMS），3小时处理的剂量为0.1%，1小时处理的剂量为0.5%。基于胚胎发生效率，选择100 Gy的剂量率和0.1%的EMS处理3小时用于再生。与对照相比，100 Gy时胚胎发生效率显著降低（85.92%），胚胎产生（79.49%）、发芽（13.43%）、转化（2.48%）、定植（15.78%）和驯化（60.92%）也呈现类似的降低趋势。与对照相比，伽马射线辐照后再生植株中与生长相关的参数，如根长、茎长和每株再生苗的叶片数也分别显著降低至67.29%、30.19%和5.03%。在EMS处理中，剂量率为0.1%处理3小时时，胚胎发生效率降至43.67%，胚胎产生（59.49%）、发芽（8.95%）、转化（1.94%）、定植（4.37%）和驯化（29.9%）也呈现类似的降低趋势。EMS处理中与生长相关的参数，根长降至91.00%，茎长降至71.34%，叶片数降至35.03%。基于分子标记的不同扩增证实了伽马射线和EMS诱导的M再生植株中发生了突变。该研究突出了一种用于诱导金诺橘和相关柑橘品种固态突变体的高频体外诱变替代方案。
