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64 SEEDWORLD.COM SEPTEMBER 2015 RESEARCH SHOWCASE Welcome to Seed Worlds Research Showcase a new department within the magazine. This department is designed to bring more scientific information to readers as well as showcase the work being done by graduate students and their advisors. Potential for Application of Growth-Promoting Bacterial Endophytes in Turfgrasses Qiang Chen Marshall Bergen and James F. White Jr. Department of Plant Biology and Pathology Rutgers The State University of New Jersey New Brunswick NJ 08901 About The Author Qiang Chen is a doctoral student in Department of Plant Biology at Rutgers University. His research focuses on the bacterial endophytes from turf grasses. He intends to graduate in 2017 and hopes to work as a researcher studying bacterial endophytes and their func- tions either in the public or private sectors. Introduction The existence of non-pathogenic microbes in plants has long been known. These microbes are referred to as endophytes. Endophytes are microbes that enter into the tissues of plants without causing disease. Both fungal and bacterial endophytes are common in plants. How most endo- phytes interact with plant hosts is still unknown but several studies demonstrate that endophytes may benefit plants by increasing their competitive abilities and overall tolerance to stress. In turf grass fungal Epichlo endophytes have been well known to enhance plant growth and resistance to biotic and abiotic stresses. However the presence and effects of bacterial endo- phytes in turf grass are less well known. In our study we examined seed-transmitted bacterial endophytes in Chewings fescue creeping red fescue hard fescue Kentucky bluegrass peren- nial ryegrass sheep fescue slender creeping red fescue and tall fescue. Also we evaluated the effects of endophytic bacteria on the plants. Materials And Methods Bacterial Isolation and Identification Seeds of eight turfgrass species were obtained from breeding program selections available at the Rutgers Research Farm in Adelphia N.J. To remove surface bacteria grass seeds were surface disinfected in 4 percent sodium hypochlorite solution for 30 minutes and rinsed three times with water for 30 seconds. The surface disinfected seeds were put on Potato Dextrose Agar PDA Difco Laboratories Detroit Mich. and 10 percent Tripticase Soy Agar TSA Difco Laboratories Detroit Mich. for isolation of bacteria. For bacterial identification about 1450 base pairs of the 16S rDNA region were ampli- fied with primers 16S-27F 5- AGAGTTTGATCMTGGCTCAG-3 and 16S-1492R 5- AAGGAGGTGWTCCARCC-3 MAC WAT RAG. Polymerase chain reaction PCR amplification was conducted with the program of denaturing at 94o C for 60 seconds anneal- ing at 46o C for 30 seconds and extension at 72o C for 90 seconds 36 cycles. The PCR products were checked using agarose gel electrophoresis 1.2 percent and then sent for sequencing to Genewiz Inc Plainfield N.J.. The sequencing results were compared on NCBI for identification. Seed Germination Test Seeds of Kentucky bluegrass P. pratensis were disinfected in 4 percent sodium hypochlo- rite solution for 30 minutes and rinsed three times with sterile water for 30 seconds. Bacterial isolates Bacillus amyloliquefaciens and Bacillus pumilus from sheeps fescue and two Pantoea agglomerans isolates from tall fescue F. arundinacea and teosinte Zea mays Mexicana were cultured in Potato Dextrose Brouth. After 24 hours bacteria were concentrated by centrifuga- tion and then diluted in water to OD6000.8. Surface disinfected seeds of Kentucky bluegrass were inoculated with bacterial solutions by soaking with agitation for five minutes. Then seeds were placed on 1.5 percent water agar and 1.5 percent agar with 100mM NaCl. After a 7 10 14 and 17 day incubation period on the media the rates of germination were calculated for each treatment.