Influence of nitrogen and phosphorus on induction embryogenic callus of sorghum
Influence of nitrogen and phosphorus on induction embryogenic callus of sorghum
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DOI：
10.1023/a:1006472418218
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发表时间：
2000-01-01
影响因子：
3
通讯作者：
Pakhomova, NV
中科院分区：
文献类型：
--
作者：
Elkonin, LA;Pakhomova, NV
以不同品种高粱幼穗和幼胚为外植体，在添加L-天冬酰胺（6.7 mM）、L-脯氨酸（17.4 mM）和不同浓度的NO3-、NH 4+和PO 43-的改良MS和N6培养基上进行培养。在不同氮源条件下，以穗源培养物为材料，研究了胚源培养物和穗源培养物中PO 43-水平的影响。在含氨基酸的培养基中，NO3-和NH 4+浓度的增加显著提高了胚性愈伤组织的诱导和生长及其再生能力。一种新的M2培养基，其中含有62.5 mM NH 4+和72.4 mM NO3-，超过了以前有效获得高粱EC的其他培养基。添加L-天冬酰胺和L-脯氨酸的培养基中NO3-水平和NO3-/NH 4+比值是高粱脆弱EC形成的关键因素。高浓度NH ~（4+）和低浓度NO ~（3-）有利于EC的形成，而在MS或N_6培养基中，NO ~（3-）浓度的增加（39.9- 82.4mM）有利于某些基因型EC的增殖。紧凑的EC在NH 4+丰富的介质中形成伴随着强烈的色素沉着的介质，这是从来没有观察到在文化与易碎EC。结果表明，提高PO_（43）~-浓度（8.8- 14.2mM），体细胞胚发生频率提高3-4倍，EC质量提高1.5-2.0倍，其再生能力也提高。增加氮或磷的培养基可用于高粱EC的4-5代维持;为了更长时间的维持，应降低培养基中的氮或磷水平。
Fragments of young panicles and immature embryos of different cultivars of grain sorghum were cultured on modified MS and N6 media supplemented with L-asparagine (6.7 mM), L-proline (17.4 mM) and different concentrations of NO3-, NH4+ and PO43-. The panicle-derived cultures were used in experiments with different nitrogen sources; the influence of PO43- level was studied in embryo-derived and panicle-derived cultures. An increase of the NO3- and NH4+ levels in the media with amino acids significantly increased induction and growth of embryogenic callus (EC) and its regeneration ability. A new M2 medium, which contained 62.5 mM NH4+ and 72.4 mM NO3-, exceeded other media which were previously effective for obtaining EC of sorghum. The level of NO3- and NO3-/NH4+ ratio in the media supplemented with L-asparagine and L-proline were established to be the critical factors of friable EC formation in sorghum. High level of NH4+ with low level of NO3- resulted in formation of compact EC, while increase of NO3- concentration (39.9-82.4 mM) in MS or N6 media favoured proliferation of friable EC in some genotypes. Formation of compact EC in the NH4+-rich media was accompanied by strong pigmentation of the medium, which was never observed in cultures with friable EC. An increase of the PO43- level (up to 8.8-14.2 mM) was shown to increase the frequency of somatic embryogenesis by 3-4 times, the EC mass by 1.5-2.0 times and its regeneration ability. Media with increased nitrogen or phosphorus could be used for maintenance of sorghum EC for 4-5 passages; for a more prolonged maintenance, their level in the medium should be decreased.