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《Acta Biologiae Experimentalis Sinica》 1986-03
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IN VITRO SELECTION OF NaCl-TOLERANT VARIANT OF TOBACCO CALLUS AND DIFFERENTIATION OF REGENERATED PLANT

Zhou Rong-ren Yang Shi-rong Yu Shu-wen (ShanghaiI nstitute of Plant Physiology,Academia Sinica,)  
Tobacco leaf segments as explants weredirectly cultured on modified MS medium con-taining 0.5% NaCl to induce NaCl-tolerantcallus.Then the concentration of NaCl inmedium was increased step by step.Cell linestolerated to 0.5,1.0,1.5,and 2.0% NaClwere established separately.The cell line tole-rated to 1.5% NaCl maintained its salt-tole-rance after subculture 9-11 passages in NaCl-free medium.The regenerated plants from various NaCl-tolerant cell lines were obtained.The cell line 04-9,tolerant to 2.0% NaCl,produced 15 regenerated plants with narrowand indented leaves and more pedicel branches.The style stretched out of coralla and thefilament of stamens shortened.Most of thepollen grains appeared abnormal.A smallamount of seeds were obtained by means ofartificial pollination.The regenerated plantof 04-9 variant grown in Hogland's solutionwith 1—2% NaCl for 85 days were stillalive. The cells of original type were irregularellipse,while the cells of NaCl-tolerant celllines spherical.The higher the concentrationof salt to which the screened callus tolerated,the more the cell size decreased.The chloro-phyll content of tolerant cell lines increasedwith the increasing level of NaCl to whichthey can tolerated.The osmotic potential oftolerated cell lines decreased with the increa-sing concentration of NaCl.Free proline con-tent in 2.0% NaCl-tolerant cell line 04-9was 40.7 times higher than that in the ori-ginal type cells.Regenerated plants of 04-9grown in non-salted soil also contained morefree proline than original one.When theirleaf discs treated with NaCl solution for 24hours,leaves of former plants showed a lar-ger increase in proline content than leaves of ??latter.Isozymes of peroxidase in adult andyoung leaves of 04-9 regenerated plants wassomewhat different from that of original type.All these results showed that cell line 04-9and its regenerated plant is a NaCl-tolerantvariant.
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