甘蓝根系再生植株技术研究
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摘要
【目的】研究甘蓝根系再生植株技术,为小孢子单胚再生双单倍体(DH)植株当年快速扩繁提供参考。【方法】以甘蓝DH15-1A的根段为外植体,设置预培养后再进行共培养和直接共培养2种培养方式,共培养的培养基为MS+4.5mg/L 6-BA+6mg/L AgNO_3,再向其中添加不同质量浓度(0.045,0.03,0.025,0.018和0.015mg/L)的NAA,筛选适宜的培养方式和NAA质量浓度;选择根龄分别为15,20和25d的DH15-1A的根段在适宜培养基上培养,比较根龄的诱导效果;以DH15-1A、DH15-2B和DH15-3C无菌植株苗的须根作为外植体,分析基因型对植株再生的影响。【结果】采用直接共培养并选用MS+0.030mg/L NAA+4.5mg/L 6-BA+6mg/L AgNO_3培养基可获得甘蓝根段诱导培养的最佳效果,其中愈伤诱导率、外植体诱导率和不定芽诱导率均最高,分别达100.0%,90.0%和430.0%,培养3周后分化芽生长健壮,叶片翠绿;根龄20d外植体的愈伤诱导率最高,达96.0%,并且愈伤分化芽点多,芽点周围褐化少,外植体诱导率和不定芽诱导率均最高,分别为84.0%和420.0%。DH植株基因型是决定根系再生植株效果的一个主要因素,3个供试基因型中,以DH15-2B根段的再生效果最好,愈伤诱导率、外植体诱导率和不定芽诱导率分别为83.3%,76.7%和430.0%。【结论】选用DH15-2B基因型甘蓝20d的根段在MS+0.03mg/LNAA+4.5mg/L 6-BA+6mg/L AgNO_3培养基上培养,最有利于根段再生植株形成,植株再生率高达100%。
【Objective】This study investigated the technology of root regeneration plants in cabbage to create conditions for rapid propagation of DH plants.【Method】The root of Brassica variety DH15-1 Awas used as explant.Two culture methods including pre-culture combined with co-culture and direct co-culture.The co-culture medium was MS+4.5 mg/L 6-BA+6 mg/L AgNO_3,and then NAA was added with different mass concentrations(0.045,0.03,0.025,0.018 and 0.015 mg/L)to screen suitable culture mode and NAA mass concentration.The root segments at the age of 15,20 and 25 days were selected for culture in suitable medium,and the induction effect of root age was compared.The fibrous roots of aseptic seedlings of DH15-1 A,DH15-2 Band DH15-3 Cwere used as explants,and the effects of genotypes on plant regeneration were analyzed.【Result】The best results were obtained by co-culture with MS+0.030 mg/L NAA+4.5 mg/L6-BA+6 mg/L AgNO_3 as culture basis.The callus induction rate,explant induction rate and adventitious bud induction rate were the highest of 100.0%,90.0%and 430.0%respectively.After 3weeks,the differentiation buds grew robust and leaves were green.The callus induction rate of 20 droot age explants was the highest of 96.0%,and the callus bud points were more with less browning around bud points.The explant induction rate and adventitious bud induction rate were the highest of 84.0% and420.0%,respectively.The DH plant genotype was a major factor determining the effect of root regeneration plant.In the 3 tested genotypes,the root segment regeneration of DH15-2 Bwas the best.The callus induction rate,explant induction rate and adventitious bud induction rate were 83.3%,76.7%and 430.0%,respectively.【Conclusion】The root segment of DH15-2 Bgenotype cabbage at age of 20 dcultured on MS+0.03 mg/L NAA+4.5 mg/L 6-BA+6 mg/L AgNO_3 was the most beneficial to the formation of root regeneration plant,and the plant regeneration rate was as high as 100%.
【Objective】This study investigated the technology of root regeneration plants in cabbage to create conditions for rapid propagation of DH plants.【Method】The root of Brassica variety DH15-1 Awas used as explant.Two culture methods including pre-culture combined with co-culture and direct co-culture.The co-culture medium was MS+4.5 mg/L 6-BA+6 mg/L AgNO_3,and then NAA was added with different mass concentrations(0.045,0.03,0.025,0.018 and 0.015 mg/L)to screen suitable culture mode and NAA mass concentration.The root segments at the age of 15,20 and 25 days were selected for culture in suitable medium,and the induction effect of root age was compared.The fibrous roots of aseptic seedlings of DH15-1 A,DH15-2 Band DH15-3 Cwere used as explants,and the effects of genotypes on plant regeneration were analyzed.【Result】The best results were obtained by co-culture with MS+0.030 mg/L NAA+4.5 mg/L6-BA+6 mg/L AgNO_3 as culture basis.The callus induction rate,explant induction rate and adventitious bud induction rate were the highest of 100.0%,90.0%and 430.0%respectively.After 3weeks,the differentiation buds grew robust and leaves were green.The callus induction rate of 20 droot age explants was the highest of 96.0%,and the callus bud points were more with less browning around bud points.The explant induction rate and adventitious bud induction rate were the highest of 84.0% and420.0%,respectively.The DH plant genotype was a major factor determining the effect of root regeneration plant.In the 3 tested genotypes,the root segment regeneration of DH15-2 Bwas the best.The callus induction rate,explant induction rate and adventitious bud induction rate were 83.3%,76.7%and 430.0%,respectively.【Conclusion】The root segment of DH15-2 Bgenotype cabbage at age of 20 dcultured on MS+0.03 mg/L NAA+4.5 mg/L 6-BA+6 mg/L AgNO_3 was the most beneficial to the formation of root regeneration plant,and the plant regeneration rate was as high as 100%.
引文
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