Development of salt tolerant callus cultures by somatic hybridization between Oryza sativa and mangrove grass Myriostachya wightiana

M. Kiran Kumar, B.V. Sandeep, P. Sudhakar Rao
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引用次数: 7

Abstract

Rice is one of the agronomically and nutritionally important cereal crops. It provides food for more than 50% of the world's population. However, the yield of rice crop is decreased by the salinity stress. This can be overcome by the development of salt-resistant rice varieties. In this study salt resistant callus cultures were developed by the somatic hybridization between Oryza sativa and Myriostachya wightiana. To optimize the isolation and fusion of protoplasts, the experiments were carried out with three different enzyme mixtures and different incubation periods. To know the salt tolerant ability, growth and total antioxidant activity were measured in the calli of Oryza sativa, Myriostachya wightiana, and hybrid at the different NaCl concentrations. In Oryza sativa high viable protoplasts yield was obtained by Enzyme mixture B (6.1 ± 0.2 × 106) whereas, Myriostachya wightiana gives high viable protoplasts yield by Enzyme Mixture C (3.4 ± 0.2 × 106). Optimum digestion time for the Oryza sativa was 12 h, which gave the highest number that is, 6.36 ± 0.15 × 106 protoplasts/g FW and viability were 91.2 ± 1.05%. Optimum digestion time for the Myriostachya wightiana was 24 h, which gave the highest yield that is, 3.49 ± 0.08 × 106 protoplasts/g FW and viability were 60.3 ± 1.72%. The fusion was performed with 50% (w/v) PEG-MW4000. The optimal incubation time was found as 30 and 45 min, which gave the highest number of fused viable protoplasts. A maximum number of binary fusions was achieved at 45 min incubation. The use of liquid culture medium resulted in a range of 5–20 calli per Petri dish which continued their growth. The hybrid calli show better growth in all NaCl concentrations. Hence, it was concluded that the hybrids have a better tolerance to salt stress. These results are useful in the development of salt tolerant rice varieties.

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水稻与红木愈伤组织体细胞杂交培育耐盐愈伤组织
水稻是重要的农艺和营养谷类作物之一。它为世界上50%以上的人口提供食物。然而,盐胁迫使水稻产量下降。这可以通过开发耐盐水稻品种来克服。本研究通过水稻与密桃的体细胞杂交培养出耐盐愈伤组织。为了优化原生质体的分离和融合,采用三种不同的酶混合物和不同的孵育时间进行了实验。为了解耐盐能力,测定了不同NaCl浓度下水稻、杨桃和杂交种愈伤组织的生长和总抗氧化活性。在水稻中,混合酶B(6.1 ± 0.2 × 106)和混合酶C(3.4 ± 0.2 × 106)可获得较高的原生质体活产率。水稻的最佳消化时间为12 h,其原生质体数量最高,为6.36 ± 0.15 × 106个/g FW,活力为91.2 ± 1.05%。最佳消化时间为24 h,产率最高,为3.49 ± 0.08 × 106个原生质体/g FW,活力为60.3 ± 1.72%。采用50% (w/v) PEG-MW4000进行融合。最佳培养时间为30和45 min,原生质体融合数量最多。在45 min孵育时,实现了最大数量的二元融合。液体培养基的使用导致每个培养皿中有5-20个愈伤组织继续生长。杂种愈伤组织在各NaCl浓度下均表现出较好的生长。由此可见,杂交种具有较好的耐盐性。这些结果对耐盐水稻品种的选育具有指导意义。
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