In order to find the optimal bacteriostatic agents and selectors and their concentrations in genetic transformation of Lespedeza bicolor with BADH as a marker gene, an resistance experiment was carried out with four c...In order to find the optimal bacteriostatic agents and selectors and their concentrations in genetic transformation of Lespedeza bicolor with BADH as a marker gene, an resistance experiment was carried out with four chemicals, NaCl, betaine aldehyde, PEG and cefotaxime （Cef） added into the media, Different concentrations and combinations of different chemicals were added into the media in the stage of adventitious bud regeneration of cotyledonary nodes, Shoot segment multiplication, and rooting. The results showed that NaCI and betaine aldehyde could inhibit tissue differentiation of L. bicolor, and could thus be used as selectors. Furthermore, 0.8 g/L NaCI could completely inhibit the cotytedonary node differentiation and shoot segment multiplication, and 0.5 g/L NaCI was optimal for inhibition of rooting. It was also found that with the concentration increase of betaine aldehyde in media, adventitious buds regenerated from cotyledonary nodes decreased, and 1.5 g/L was confirmed to be the optimum concentration. Moreover, the mixture of 0.7 g/L NaCI and 0.3 g/L betaine aldehyde significantly inhibited shoot segment multiplication and rooting, so this mixture could be used asa selector. The Cef ex- periment showed that 200-300 mg/L C, ef in media did not inhibit significantly the regeneration of adventitious buds from cotyledonary nodes, and 100 mg/L Cef had little effect on the shoot segment multiplication and rooting.展开更多
基金Supported by The Agricultural Science and Technology Project of Beijing(PXM2016-014207-000053)~~
文摘In order to find the optimal bacteriostatic agents and selectors and their concentrations in genetic transformation of Lespedeza bicolor with BADH as a marker gene, an resistance experiment was carried out with four chemicals, NaCl, betaine aldehyde, PEG and cefotaxime （Cef） added into the media, Different concentrations and combinations of different chemicals were added into the media in the stage of adventitious bud regeneration of cotyledonary nodes, Shoot segment multiplication, and rooting. The results showed that NaCI and betaine aldehyde could inhibit tissue differentiation of L. bicolor, and could thus be used as selectors. Furthermore, 0.8 g/L NaCI could completely inhibit the cotytedonary node differentiation and shoot segment multiplication, and 0.5 g/L NaCI was optimal for inhibition of rooting. It was also found that with the concentration increase of betaine aldehyde in media, adventitious buds regenerated from cotyledonary nodes decreased, and 1.5 g/L was confirmed to be the optimum concentration. Moreover, the mixture of 0.7 g/L NaCI and 0.3 g/L betaine aldehyde significantly inhibited shoot segment multiplication and rooting, so this mixture could be used asa selector. The Cef ex- periment showed that 200-300 mg/L C, ef in media did not inhibit significantly the regeneration of adventitious buds from cotyledonary nodes, and 100 mg/L Cef had little effect on the shoot segment multiplication and rooting.
