| Endophytes can affect the metabolic process of the host plant and then influence the biochemical quality of the host plant.The endophytes that inhabit the flesh and seeds of wine grapes are closely related to the quality and characteristics of wine grapes.Few researches have concerned the endophytes in grape flesh and seeds to date.In this study,high-throughput sequencing technology and culture methods were used to analyze the endophytic communities in young and mature fruit flesh and the seeds of three grape varieties（Rose honey（Vitis.Vinifera L.× V.labrusca L.）,Crystal grape（V.Vinifera L.×V.labrusca L.）and Qiuzi grape（V.davidii L.））from the same vineyard.At the same time,the biochemical effects of some isolated endophytic strains on grape cells by means of dual culture method were evaluated and main results as follows:1.Results of the fungal endophytes communities in grape flesh and seeds based on highthroughput sequencing analysis.From all of the samples,a total of 394 endophytic fungi features were detected,which belong to 134 species,109 genera and 5 phyla.And a total of 288 endophytic fungi features were detected in grape flesh,and these fungi belong to 120 species102 genus,and 5 phyla.Inseeds,178 endophytic fungi features were detected,belonging to 65 species,55 genus,and 3 phyla.The most 3 dominant distributed endophy-tic fungi genus in the flesh tissue were Nakaseomyces（46.51%）、Kabatiella（32.74%）、and Didymella（13.16%）,occupied 92.41% of he otal dominance.While 3 most dominant distributed endophytic fungi genus in seeds were Ascomycota＿unclassified（77.06%）、Nakaseomyces（6.90%）、Malassezia（5.21%）,occupied 89.17% of the total dominance.The similarity coefficient of endophytic fungi community structure in different varieties and developmental stage of grape flesh varied from 0.05 to 0.36,the diversity index varied from 0.14 to 1.49.The similarity coefficient of endophytic fungi community structural in different grape cultivars and developmental stage of seed varied from 0.05 to 0.49,and the diversity index varied from 1.01 to 2.03.As to the differrent varities and developmental stages,the endophytic fungi communities in flesh are not obvious difference between young and mature fruits,while quite different between seeds from young or from mature fruits.2.The diversity distribution of bacterial endophytes in grape flesh and seeds based on high-throughput sequencing analysis.From all of the samples,a total of 208 endophytic bacteria of feature were detected,which belong to 158 species,116 genus and 9 phyla.Endophytic bacteria were not detected from fruit.3 most dominant distributed endophytic bacteria genus in seeds are Sphingomonas（47.04%）、Acinetobacter（8.87%）、Enhydrobacter（5.50%）、Ralstonia（4.70%）、Bradyrhizobium（4.26%）,occupied 70.37% of the total dominance.In different grape cultivars and developmental stage,the similarity coefficients of endophytic bacteria varied from 0.22 to 0.36 in flesh,whereas varied from 2.23 to 2.88 in the seeds.3.Results of culturable endophytes from grape flesh and seeds based.From all samples,a total of 1458 endophytic fungal strains were isolated,which belong to 60 species and 45 genera.The separation rate of endophytic fungi in flesh from different varieties varied from 0.31 to129.90 CFU/g,and the diversity index varied from 1.12 to 2.25.While in see-ds,the isolation rate of endophytic fungi of different cultivars varied from 2.78 to 37.78 CFU/g,and the diversity index between 0.80 and 2.03.The most dominant genera of fungal endophytes in flesh are Clonostachys、Cylindrocladium、Cladosporium、Plenodomus、Physisporinus,occupied 96.91%of the relative abundance.While the most dominant genera in the seeds were Malassezia、Cladosporium、Vanderbylia、Psathyrella,and Stagonosporopsis,ccupied 71.83% of the relative abundance.Seven endophytic fungi genera were co-detected in seeds and flesh.14 genera of endophytic fungi were specifically detected only in seeds.4.Nine endophytic fungal strains from different genera which isolated from grape flesh and seeds were sellected to evaluate their physio-chemical effects on grape cells by dual culture method.The results showed that the physiological effects of each strain on grape cells were different.Dual cultured with flesh endophyte CMF-17 significantly increased the SOD activity of grape cells,and dual cultivation with flesh endophyte RYF-3 obviously increased the PAL activity of grape cell.The treatment of seed endophyte RYS-8 promoted the content of grape cell flavonoids and total phenols.And the seed endophytes QYS-3 enhanced the total protein content of grape cells,while CMS-2 can increase the soluble sugar content of grape cells.The paper invesgated the distribution characteristics of endophytes communities in flesh and seeds of different varieties and development stages of grape fruits,which harvested from the same vineyard,And explored some grape flesh and seed endophytic resources for further utilization and researches. |
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