| Cinnamomum camphora(L.)Presl is the plant of Lauraceae and Cinnamomum. It's the main tree plant of evergreen and broad-leaved forest in Subtropics,and also one of city afforestation tree seeds in south of China. The chlorosis of camphor-tree is a common and serious disease in Shanghai city. It has the universal harm and seriously,lightly disease causes the plant growth to be blocked and the shape is abnormal. The heavy piece causes the adult plant death. Creates economic loss and ecology destruction which is unable torecall.The alkalinity physiological and biochemical characteristic of five camphor provenances and the ISSR molecular marker of Shanghai area's camphor provenance were studied. The chlorosis gene to perform through the molecular marker method,and to distinguish the discrimination of internal cause hereditary variation and the external factor soil environment lacks the iron which cause camphor chlorosis.Thus is better pointed out that take the measures to solve the camphor tree chlorosis problem,and provide certain scientific basis for the further research.The major results were as follows:1,Five provenances camphor seed was observed by the method of potted plant,and five rawness index of physiological and biochemical characteristic were determined,such as the highness,the content of proline,the content of chlorophy II,the SOD activity and the active iron content were mensurated, and the resistance performance of five provenance camphor was comprehensive evaluated. The results showed that the Shanghai provenance bears the alkalinity to be best.2,Based on the conventional SDS method, an efficient procedure was developed. The major improvement included elimination a lot of secondary metabolites such as polyphenols etc,and collection of nuclei before free DNA. The results from digestion with RNase, electrophoresis and PCR amplification suggested that isolated DNA was suitable for ISSR analysis.3,The optimal amplification procedure and the suitable PCR reaction system of camphor tree ISSR were screened and established. The optimal ISSR-PCR system in Camphor was established with L9(34) orthogonal diagram. In a total volume of 20μL ISSR-PCR system,it contains 1×PCR Buffer,0.25mmol/L dNTP,0.3μmol/L primer,2.0 mmol/L Mg2+,1 U Taq DNA polymerase and 50ng template DNA. The suitable PCR procedure is one cycle denaturing at 94℃for 4 min;42 cycles each involved denaturing at 94℃for 40 S,annealing at 58℃for 1 min,extending at 72℃for 2 min,one cycle extending at 72℃for 10 min,and then remaining at 4℃.4,20 primers were selected from 100 primers for ISSR analysis,which amplify a stable product,amplified with clear bands and specific. Carry on ISSR amplification to the Shanghai provenance camphor tree 18 normal leaf blades samples and 18 chlorosis leaf blades sample. The polymorphic loci ratio of 57.24% which 20 primers amplified all individuals. On average,each primer was amplified about 4.15 polymorphic loci .5,ISSR molecular marker analysis showed that Shanghai camphor provenance is mixed quite. There is a big difference between normal and chlorosis leaves amplification,and unable to determine whether the specificity with chlorosis gene have a chain. The cluster analysis indicated that camphor normal genetic material similar coefficient is big than chlorosis material,the difference between them is particularly small.
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