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Regeneration of soapnut tree through somatic embryogenesis and assessment of genetic fidelity through ISSR and RAPD markers

机译:通过体细胞胚发生再生皂树并通过ISSR和RAPD标记评估遗传保真度

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摘要

Somatic embryogenic system was developed in Sapindus mukorossi Gaertn. using rachis as explants from a mature tree. Explants showed callus initiation on Murashige and Skoog medium supplemented with TDZ (1-Phenyl-3-(1, 2, 3-thiadiazol-5-yl) urea), zeatin or 6-benzylaminopurine. Induction of somatic embryogenesis was achieved on both MS basal medium and MS medium supplemented with 8.88 µM 6-benzylaminopurine. Hundred percent embryogenesis was observed on MS medium supplemented with 8.88 µM 6-benzylaminopurine with maximum intensity of embryogenesis (51.92 ± 0.40 a). Maximum maturation of somatic embryos (92.86 ± 0.34 a) was observed on induction medium supplemented with 0.0378 µM abscisic and treated for 21 days. Germination of somatic embryos was maximum (77.33 ± 0.58 a) on MS medium supplemented with 8.88 µM 6-benzylaminopurine. In vitro raised plantlets were hardened, acclimatized and transferred to the field. Survival frequency of plantlets was 80 % in field conditions. The genetic fidelity of in vitro regenerated plants was also evaluated and compared with mother plant using random amplified polymorphic DNA and inter simple sequence repeat. Both markers showed similarity in molecular profile of mother plant and in vitro regenerated plants.
机译:在Sapindus mukorossi Gaertn开发了体细胞胚发生系统。使用rachis作为成熟树的外植体。外植体在补充了TDZ(1-Phenyl-3-(1,2,3-thiadiazol-5-yl)尿素),玉米蛋白或6-苄基氨基嘌呤的Murashige和Skoog培养基上显示出愈伤组织起始。在MS基础培养基和补充8.88 µM 6-苄基氨基嘌呤的MS培养基上均实现了体细胞胚发生的诱导。在添加8.88μM6-苄基氨基嘌呤的MS培养基上观察到百分百的胚胎发生,最大的胚胎发生强度(51.92±0.40 a)。在补充了0.0378μM脱落酸并处理21天的诱导培养基上观察到体细胞胚的最大成熟度(92.86±0.34 a)。在补充了8.88μM的6-苄基氨基嘌呤的MS培养基上,体细胞胚的萌发最大(77.33±0.58a)。体外培育的小苗被硬化,适应环境并转移到田间。在田间条件下,小苗的成活率为80%。还评估了体外再生植物的遗传保真度,并使用随机扩增的多态性DNA和内部简单序列重复与母本植物进行了比较。两种标记在母本植物和体外再生植物的分子谱上均显示相似性。

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