Biological Properties Of Endophytes/Calama Grostis Symbiota

Epichloae endophytes are obligate stable symbiosis with their host through co-evolution. Until now,12 species of Epichloe and 23 species and 5 varieties of Neotyphodium had been documented. E. amarillans, E. baconii and N. stromatolongum isolated from Calamagrostis spp. had been reported.We collected 29 Calamagrostis sp. samples without stromata from Huangshan, Anhui. All samples were infected by endophyte. The fungal hyphae grown in the intercellular space,2μm width, parallel the prolate axis of plant cell, rarely branched and uncrossed each other. We isolated 9 strains from the samples. On PDA plates, obverse of.colonies was round, white, cottony and reverse of colonies was brown. Diameter of fungus colonies expand to 15.4-16.8 mm after 21 days’ incubation on PDA. Conidium single, ellipses mostly,3.6-3.7×2.3～2.6μm. Length and base of the phialides were 17.1～19.7μm and 2.1～2.6μm respectively. Morphological of isolates is distinct with N. stromatolongum. Maximum parsimony (MP) trees based on tubB and actG genes illustrated that the isolates clustered into one clade with N. stromatolongum, but the tefA gene had one substitution difference and formed one small clade. These indicated that the endophyte collected in Huangshan is distinguishable from our previously reported N. stromatolongum, a symptomatic asexual Neotyophodium species in Nanjing.Endophytes can affect the morphological and physiological charateristics of host plants and improve agronomic performance. Through measure the width and length of leaf, diameter of culm and height of host, we discuss the effects of endophyte on Calamagrostis spp.. When comparing HEI and NIEF plants, width of the lower leaves was small, length of fresh leaves was longer, culm was slender and height was taller. Comparison between NEI with NIEF plants indicated that width of the leaves was small, length of fresh leaves was bigger and width was small, culm was slender and height was shorter. Precipitation was richness during the experimental process. We speculate the genotype of host also cause the different growth status besides the effects of endophyte.The physiological effects of fungal endophytes on Calamagrostis spp. include photosynthesis and the soluble carbohydrates were investigated in this research. Stomatal conductance, CO2 concentration of intercellular space, transpiration rate and net photosynthesis rate were surveyed in before and after inflorescence. Stomatal conductance, CO2 concentration of intercellular space, transpiration rate and net photosynthesis rate were similar of NEI with NIEF plants in before and after inflorescence. In addition to transpiration rate of HEI plants was significant greater than NIEF plants in after inflorescence. Photosynthesis of HEI and NIEF plants was not significantly difference. We can conclude that the endophyte and the formation of stromata were not influence the photosynthesis of the host.Soluble carbohydrate content of HEI was significant less than that of NIEF in before inflorescence and overturn in inflorescence but was not significant difference, no significant difference in after inflorescence. The endophyte can influence the soluble carbohydrate content of HEI in before inflorescence. The endophyte did not influence the soluble carbohydrate content of NEI from the analysis.N. stromatolongum can form stromata on culms of some host tillers, while isolates in this study could not. PCR-based mating-type was used for detecting and analyzing 29 strains isolated from 6 genus hosts.21/25 isolates of Neotyphodium spp. were detected content MAT1-1-1 gene and 22/25 isolates of Neotyphodium spp. were detected content MAT1-2-1 gene,4 isolates of Epichloe sp. possessed both MAT1-1-1 and MAT1-2-1 genes. Presence or absence of MAT genes detected in this study could not clearly determine the mating type of the strains and effectively distinguish the Epichloe and Neotyphodium species. Maximum parsimony (MP) trees based on MAT1-1-1 and MAT1-2-1 illustrate that partial of epichloae species were cluster into one clade. It is a complex question of mating relationship among epichloae endophytes. MAT1-2-1 could be a certria to distinct strains and N. stromatolongum.Fungal endophytes were isolated from Calamagrostis sp. obtained from Huangshan, Anhui and is distinguishable from our previously reported N. stromatolongum, a symptomatic asexual Neotyophodium species in Nanjing. The morphological difference among the HEI, NEI and NIEF was effect by endophytes and the genomic difference of the hosts. There is no significant effect of endophyte on photosynthesis of Calamagrostis spp.. Endophytes can influence the soluble carbohydrate content of HEI in inflorescence and after inflorescence, but was no significant effect on NEI during the experiment. Presence or absence of MAT genes detected in this study could not clearly determine the mating type of the Epichloe species and effectively distinguish the Epichloe and Neotyphodium species. Some different species of epichloae endophytes are not distinct each other in MAT1-1-1 and MAT1-2-1 phylograms. MAT1-2-1 could affect fungal even symbiotic properties.