酵母提取物蛋白胨共培养培养基促进热带玉米基因型瞬时GUS表达

J. Muli, C. Mweu, N. Budambula, S. Anami
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引用次数: 2

摘要

背景与目的:热带玉米是撒哈拉以南非洲种植最多的作物,是2.2亿多人的主食。研究了4个热带玉米(Zea mays L.)自交系在酵母提取液蛋白胨培养基上共培养的愈伤组织诱导和体细胞胚再生能力。材料与方法:采用瞬态GUS法评价不同基因型在分别添加2,4- d (yeah +2,4- d)、半胱氨酸(yeah +CYS)、脯氨酸(yeah +PRO)或混合添加(yeah +ALL)时农杆菌介导转化的能力。各参数数据采用多变量方差分析和SAS分析。结果:单独以叶青为基质的共培养培养基对愈伤组织诱导无影响,未成熟胚表现出前意识萌发。2,4-D浓度分别为1.5和3 mg LG1 2,4-D时,诱导胚性愈伤组织形成并开始再生。与农杆菌在yeah、yeah +PRO和yeah +ALL培养基上共培养的未成熟胚瞬时GUS表达量高于MS培养基,表明yeah促进了农杆菌介导的转基因在热带玉米中的整合。农杆菌浓度为0.07 (OD660)时瞬时GUS表达量最高(20.90%),浓度为0.8和0.2时瞬时GUS表达量较低(分别为9.17%和12.22%)。结论:在农杆菌介导的转化方案中整合YEP培养基可能有助于开发更有效的农杆菌介导的热带玉米基因型转化系统。
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Yeast Extract Peptone Based Co-cultivation Media Promotes Transient GUS Expression in Tropical Maize Genotypes
Background and Objective: Tropical maize is the most cultivated crop in sub-Saharan Africa and is a staple food to over 220 million people. This study, evaluated the competence of four tropical maize (Zea mays L.) inbred lines to callus induction and regeneration through somatic embryogenesis when co-cultivated on yeast extract peptone medium (YEP). Materials and Methods: Transient GUS assay was used to evaluate the competence of the genotypes to Agrobacterium-mediated transformation using YEP as co-cultivation media or when YEP was supplemented with 2,4-D (YEP+2,4-D), cysteine (YEP+CYS), proline (YEP+PRO) or in combination (YEP+ALL). Data on all parameters were analyzed using multivariate ANOVA and SAS. Results: Co-cultivation media based on YEP alone did not impact callus induction and immature embryos exhibited preconscious germination. When YEP was supplemented with 2,4-D at concentrations 1.5 and 3 mg LG1 2,4-D, the formation of embryogenic calli was induced and regeneration initiated. Immature embryos had high transient GUS expression when co-cultivated with Agrobacterium on YEP, YEP+PRO and YEP+ALL media than when co-cultivated on MS media, suggesting that YEP promotes Agrobacterium-mediated integration of transgenes in tropical maize. Agrobacterium tumefaciens at a concentration of 0.07 (OD660) gave the highest transient GUS expression (20.90%) while concentrations of 0.8 and 0.2 resulted in low transient GUS expression (9.17 and 12.22%, respectively). Conclusion: The integration of YEP media in the Agrobacterium-mediated transformation protocols is likely to contribute in the development of a more efficient Agrobacterium-mediated transformation system for tropical maize genotypes.
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