Antitumor Screening And Blological Characteristics Of Endophytic Fungus Isolated From Rumex Gmelini Turcz. In Vitro

Rumex gmelini Turcz. belongs to the family of Polygonaceae. Its root and rhizome have been used for antifungal, antitumor, antitussive and expectrant, antiasthma, antivirus and antioxidation. Endophytic living in a healthy plant is a variety of tissues and organs within the fungus infected plants do not show obvious external symptoms. We may proved their endogenous by separation from strict surface disinfectant plant tissue and direct amplification of DNA from plant tissues. The results are as follows.(1) Rumex gmelini endophyte strains in vitro screening of anti-tumor activityIn this experiment, sixteen strains of endophytic isolated from R. gmelini were tested on LOVO, SGC-7901, MCF-7, SMMC-7721, MGC-803, A549, SK-OV-3, HaLe eight strains of human tumor cells in vitro for antitumor. MTT was used in this experiment, and SPSS16.0 was used to analyze data. Four strains of endophytic were proved anti-tumor effect, RGT-S12 on MCF-7, LOVO, SK-OV-3 cells significantly inhibited; RGT-S31 on A549, MGC-803, HeLa cells significantly inhibited; RGT-S42 on SGC-7901, LOVO, SK-OV-3 cells significantly inhibited; RGT-S44 on SMMC-7721, MGC-803, A549, HeLa cells significantly inhibited.(2) Molecular identification of endophytic from R. gmeliniWe selected three strains which have antitumor effect, and identified them by sequencing their ITS, the results show that:the RGT-S31 strain is Fusarium Oxysporum; the RGT-S42 strain is Cercosporella; the RGT-S44 strain is Uncultured Fusarium.(3) Biological characteristics of Endophyte from R. gmeliniThe cultural characteristics of four selected strains of endophytic were studied by changing the different culture conditions. The results show that: RGT-S12 grow fastest in PDA medium; the most suitable PH value 6, the optimum temperature is 30℃; can be a good use of lactose and starch as carbon source, using KNO3 as nitrogen source. RGT-S31 could better growth medium in the four kinds of bacteria; optimum growth pH value 8, the optimum temperature is 30℃; can be a good use of maltose as carbon source, using KN03 as nitrogen source. RGT-S42 have a good growth in the basic medium while having a slow-growing in the investigation's; the optimum growth pH value 5, the optimal temperature was 25℃; can be a good use of xylose as the carbon source, use of peptone and NH4NO3 as nitrogen source. RGT-S44 can be good growth in the PDA medium while growth slowly in the investigation's; optimum growth pH value7, the optimum temperature is 30℃; can put to good use fructose as the carbon source, using NaNO3 as the nitrogen source.The experiments have studies the anti-tumor effect of endophytic in vitro and found out four strains finally, then we made the identification and studied the cultural characteristics of them. The results of these studies can offer the theoretic and the practical guide to develop the resources of the endophytic fungi isolated from Rumex gmelini Turcz.