Distribution and phylogenetic analysis of endophytic fungi in stems and leaves of Desmos chinensis based on histochemistry and rDNA ITS sequence
- Author:
Xiao-Xia GAO
1
Author Information
- Publication Type:Journal Article
- Keywords: Desmos chinensis; Endophytic fungi; Phylogenetic analysis; rDNA ITS
- From: Chinese Pharmaceutical Journal 2013;48(24):2102-2106
- CountryChina
- Language:Chinese
- Abstract: OBJECTIVE: To study the distribution and phylogenesis of endophytic fungi in the stems and leaves of Desmos chinensis. METHODS: Permanent paraffin-cut section, optical microscope photography, and histochemistry were used to observe the distribution of endophytic fungi in the stems and leaves of Desmos chinensis. Plate isolation method was used to separate and cultivate the endophytic fungi. The genomic DNA of endophytic fungi were extracted, then the rDNA ITS region was amplified using polymerase chain reaction (PCR). The PCR products were digested by Hae III and Hha I restriction enzymes to determine the species and genotypes, and sequenced according to the digestion genotyping results. Nucleotide sequences of the rDNA ITS of endophytic fungi were used for the identification and phylogenetic analysis. RESULTS: Endophytic fungi mainly existed in the parenchyma cells of the phloem in the stems, and in the parenchyma cells, palisade tissue or sponge tissue of the leaves. Fourteen strains of endophytic fungi were isolated from the stems and leaves. Four different genotypes and their sequences were obtained from RFLP analysis and sequencing. These strains were identified as Guignardia mangiferae, Colletotrichum gloeosporioides, and Phomopsis spp. The dominant group of those endophytic strains was Phomopsis, represented by 8 strains and accounting for 57.1% of all. CONCLUSION: The endophytic fungi distribute in both stems and leaves of Desmos chinensis without obvious tissue specificity. The result of the phylogenetic analyses with rDNA ITS sequences reveals the phylogenetic relationships of endophytic fungi in Desmos chinensis.
