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首页> 外文学位 >SOMATIC HYBRIDIZATION OF CYTOPLASMIC MALE STERILE BRASSICA OLERACEA AND ATRAZINE RESISTANT B. CAMPESTRIS (PROTOPLAST).
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SOMATIC HYBRIDIZATION OF CYTOPLASMIC MALE STERILE BRASSICA OLERACEA AND ATRAZINE RESISTANT B. CAMPESTRIS (PROTOPLAST).

机译：胞质雄性不育油菜和抗阿特拉津的B. Campestris（原生质体）的体细胞杂交。

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Techniques for plant regeneration in Brassica oleracea L. var. italica cv. Green Comet broccoli were developed. Using media developed by Pelletier et al. (Mol. Gen. Genet, 191: 244-252), high frequency regeneration from leaf protoplasts was obtained when plants regenerated from hypocotyl explants (GCR) were used as a protoplast source. Over 70% of protoplast-derived cells divided and up to 77% of the calli produced shoots. Some of the 178 plants grown to maturity had variant phenotypes. Cotyledon protoplasts from selfed progeny of GCR showed higher totipotency than those from seed-grown plants.; Brassica lines carrying the ogu cytoplasmic male sterility (cms) derive from Raphanus show temperature sensitive chlorosis due to nuclear-plastid incompatibility. Protoplast fusion was used to replace Raphanus chloroplasts in this cytoplasm with atrazine resistant Brassica chloroplasts. An assay using nitro-blue tetrazolium was developed to verify the presence of atrazine resistant chloroplasts. After 30 min light, protoplasts from atrazine sensitize B. oleracea remained green in photosynthesis medium with 50 uM atrazine and 0.01% nitro blue tetrazolium; protoplasts from atrazine resistant B. campestris stained blue black.; Protoplasts from cms Green Comet and atrazine resistant B. campestris were fused using polyethylene glycol. The facts that B. campestris does not regenerate on the media used and that atrazine inhibits the growth of light-grown Green Comet calli were used to select for hybrid tissue. One plant, regenerated on medium containing 50 uM atrazine, contained nuclear genomes from both parents, as determined from rDNA hybridization, chromosome counts and morphology. This plant contained atrazine resistant chloroplasts and was male fertile. Mitochondrial DNA showed homology with both parents; novel fragments were also present. On medium lacking atrazine a plant regenerated that contained the Green Comet nucleus and atrazine sensitive chloroplasts. Mitochondrial DNA showed homology with cms mitochondria but also contained novel fragments. The original goal, cms plants with Brassica chloroplasts, was not accomplished; however the regenerated plants may be of value in studying mitochondrial genetics.
机译：甘蓝型油菜植物再生技术italica简历开发了绿色彗星西兰花。使用Pelletier等开发的媒体。 （Mol.Gen.Genet，191：244-252），当从下胚轴外植体（GCR）再生的植物用作原生质体来源时，从叶片原生质体获得了高频再生。超过70％的原生质体衍生细胞分裂，多达77％的愈伤组织产生芽。在178种成熟植物中，有些具有不同的表型。 GCR自交后代的子叶原生质体比种子生长的植物具有更高的全能。携带来自萝卜的ogu胞质雄性不育（cms）的芸苔属品系由于核质体不相容性而表现出对温度敏感的萎黄病。原生质体融合用于用耐阿特拉津的芸苔属叶绿体代替该细胞质中的萝卜叶绿体。开发了使用硝基蓝四唑鎓的测定法以验证耐阿特拉津的叶绿体的存在。光照30分钟后，来自阿特拉津的原生质体致敏油菜芽胞杆菌在含有50 uM阿特拉津和0.01％硝基蓝四唑的光合作用培养基中保持绿色。来自耐阿特拉津的野营双歧杆菌的原生质体染成蓝黑色。使用聚乙二醇融合来自cms Green Comet的原生质体和耐阿特拉津的弯曲杆菌。营地芽孢杆菌不能在所用的培养基上再生并且阿特拉津抑制了浅生的绿色彗星愈伤组织的生长的事实被用于选择杂交组织。一株植物在含有50 uM阿特拉津的培养基上再生，包含来自两个亲本的核基因组，这是根据rDNA杂交，染色体数和形态确定的。该植物含有抗阿特拉津的叶绿体，雄性可育。线粒体DNA与父母双方都具有同源性。还存在新的片段。在缺乏阿特拉津的培养基上，再生出的植物含有绿色彗星核和对阿特拉津敏感的叶绿体。线粒体DNA显示出与cms线粒体的同源性，但也含有新的片段。最初的目标是使用芸苔属叶绿体的cms植物没有实现；然而，再生植物可能在研究线粒体遗传学方面具有价值。

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