The Community Diversity Of Endophytic Fungi In Camellia Oleifera And Isolation Of Endophytic Fungi Promoting Growth

Camellia oleifera Abel is an important woody plant in southern China.In order to improve the production of Camellia oleifera,farmers use a lot of fertilizers and pesticides,further aggravate the soil nutrient imbalance,and cause great pollution to the environment.Microbial fertilizer with pollution-free characteristics such as high output,low input,therefore,the development and utilization of microbial fertilizer for green agricultural sustainable development and barren hills of the efficient management to provide technical support,production is an urgent need to solve the problem.The species of endophytic fungi in plants are very rich that it must exist in almost all types of plant,and some of them can pomote the plant to growth.So for there is a lack of an overall view on endophytic fungi of Camellia oleifera.About screening and application of promoting strains on endophytic fungi of Camellia oleifera are seldom reported.So,in this study,we intend to isolate the endophytic fungu from different growth stages and different tissues of Camellia oleifera and screen dominant strains,and compare and analysis the community structure,distribution low and species composition of endophytic fungi in Camellia oleifera by high-throughput sequencing,we analyzsis the growth promoting effect of Endophytic Fungi by determine some relevant growth indexes of Camellia oleifera seedings after inoculating dominant strains.It lays a foundation for the exploration of endogenous fungal resources in Camellia oleifera and the development of microbial fertilizers for it.Following are main results from this study:1.The endophytic fungi isolated from root,stem and leaf of annual Camellia oleifera seedings and root,stem,leaf and fruit of mature trees add up to 88 strains by using tissue blocks cultivation method.Among,11 strains were isolate from root,13strains were isolated from stem and 10 strains were isolated from leaf of annual Camellia oleifera seedings;9 strains were isolated from root and 22 strains were isolated from stem,10 strains were isolated from leaf and 13 strains were isolated from fruit of mature Camellia oleifera trees.All these endophytes belong to 34 genera.12 dominant strains were screen out above them.These strains include Botryosphaeria,Ilyonectria,Phomopsis,Colletotrichum,Botryosphaeria,Penicillium genera.2、The number of effective sequences for annual Camellia oleifera’root,steam,leaf and adult tree’ root,steam,leaf and fruit was 749812 by high-throughput sequencing.The depth index of sequencing were higher than 90%.The diagram of Rarefaction curve and Shannon-wiener indicated that amount of sequence was enough.A total of 2364 OTUs were obtained by clustering analysis of OTUs under 97%similarity levels.Alpha diversity and Beta diversity analysis show that the endophytic fungal diversity of Le1 and St1 were similar,MLe、MSt、Sh and Se were similar,but they were not significant.Ro1 and MRo were very different from other examples.Ro1 was significant but MRo was not.After taxonomic analysis,we found the fungal strains from each sample of Camellia oleifera were classified into 8 phylums,31 classes,111orders,246 families and 602 genera.The species of fungal show that MRo>Le1>St1>MLe>MSt>Ro1>Sh>Se.The result of diversity analysis indicated that MRo’ diversity was higest than others,For annual Camellia oleifera’ root,stem and leaf,the dominant strain were Xylogone、Phomopsis and Strelitziana respectively.and the mature tree’root was Inocybe,stem and leaf were Pseudoidriella,shell and seed were Cladosporium.Above all,The species diversity of roots in annual Camellia oleifera was significantly lower than that of stems and leaves,and the species diversity of stems was significantly higher than that of leaves.The number of annual Camellia oleifera’ species were leaf>steam>root and mature tree were root>leaf>steam.The diversity of adult tree roots was significantly higher than that of stems and leaves,but there was no significant difference in the diversity between the two stems and leaves.This regularities were different from annual tree.3.Inoculated the selected 12 strains dominant bacteria(numberd A-L)into the Camellia oleifera seedings by the liquid of fungus.Determined the growth morphology and physiological indicators after 90 days.The results indicated that:1)Except treatment B,others treatments’ height growth rate were higher than CK,the D,L,C treatments’ height growth rate were significantly higher than CK(P<0.05),were 3.57 times,3.27times and 2.98 times than control group respectively.About ground diameter,except treatment A,B,H,others treatments’ ground diameter growth rate were higher than CK,the L treatments’ ground diameter growth rate significantly higher than control group(P<0.05),were 1.79 times than CK.Different treatments’ numbers of new buds indicate that H,D treatments’ average number of new buds were more than others obviously.2)Except treatment C,E,others treatments’ biomass were higher than CK,the H treatments’ biomass was significantly higher than CK(P<0.05).A,D,F,G,H,I,K treatments’ root and crown ratio were higher than control group but have no significant difference(P<0.05).3)The effect on Camellia oleifera seedings’ physiological indicators were different by inoculating different fungus.C,E,I,K,Ltreatments’chlorophyll content were significantly higher than CK(P<0.05).The promoting effect I>K>E>C>L.All treatments’ chlorophyll fluorescence parameters have no significant difference(P<0.05).The effects on Camellia oleifera seedings’ soluble suger content and soluble protein content are different by inoculating different fungus.L,G treatments’ soluble suger content were significantly higher than CK(P<0.05)and D,K treatments’ soluble suger content were significantly higher than CK(P<0.05).In summary,Endophytic fungi of Camellia oleifera were rich in diversity,and there are some fungal like D,L contributing to the growth of Camellia oleifera.The results of this study could provide theoretical basis for developing the biological fertilizer in Camellia oleifera,and it is meaningful to the economic,ecology and sociology in Camellia oleifera.