| In order to grasp Aconitum (Aconitum carmichaeli Debx). entophytic bacteria community composition and its seasonal variation, recognize the synergistic relation between Aconitum carmichaeli Debx. and its host provide a reference for the scientific management and further development and utilization of this medicinal plant resource. In this study, the different growth stages of potted Aconitum are studied as samples, with the Terminal Restriction Fragment Length Polymorphism (T-RFLP) to measure Aconitum carmichaeli Debx. and fungi community composition to calculate the Patrick richness index (S), Shannon-Wiener diversity index (H) and Pielou index (J) so that it can study the dominant species composition and its changes. Besides, it will utilize the measurement of PCA to explore similar relationships among Aconitum carmichaeli Debx. community within various growth stages. The results are indicated as followed:1. There are 1067 T-RFs fragments produced through consolidating three restriction enzymes (Hha Ⅰ, Hae Ⅲ and Msp Ⅰ) that digested 16S rDNA; the overall trends of 16S rDNA Patrick richness index (S) showed decrease, which can be divided into two different high and low stages, they are January to May and June to October. In the further step (Duncan, α= 0.05) it can be found that January to May Aconitum root entophytic bacteria richness was significantly higher than that from June to October (P<0.05). There is significant fluctuations between January and May (P<0.05), while less significant variations from June to October (P>0.05). The general trends of Shannon-Wiener diversity index (H) are similar to Patrick richness index (S). However, the difference between 1-5 and 6-10 of each month were not significant (P>0.05). In entire growth process of Aconitum, the overall fluctuation of entophytic bacteria diversity index is displayed that richness index (CVS= 0.1725)> evenness index (CVJ= 0.0935)> Shannon-Weiner index (CVH= 0.0896). According to digestion results of Hha Ⅰ, Hae Ⅲ and Msp Ⅰ that combined with MiCA PAT+ online test, the advanced bacterial population of Aconitum root has been selected initially, they are:Microbacterium, Bacillus, Clostridium, Rhodococcus, Stenotrophomonas, Streptomyces, Clostridiacea, Deinococcus, Rhizobium and Prevotella and so on. Based on different endonuclease, the dominant groups showed changes in the composition according to the month:For Hha Ⅲ, MspⅠ and Hha Ⅰ,10 months a total of only one kind of groups, respectively, and its restriction fragment size ranges of 72.5-74.5bp,98.5-100.5bp and 464.5-466.5bp; but in terms of each month, each has its different composition groups.2. There are 1130 T-RFs fragments produced through consolidating three restriction enzymes (Hha Ⅰ, Hae Ⅲ and Msp Ⅰ) that digested ITS; the overall trends of ITS Patrick richness index (S) showed decrease, which can be divided into three different high and low stages, they are January to April, May to June and July to October. In the further step (Duncan, α= 0.05) it can be found that from January to April Aconitum root entophytic bacteria richness shows fluctuation (P<0.05), which has more significant difference than May and June (P<0.05). While the richness index of July decline to the lowest level (P<0.05), and the difference between 7 and 10 is not significant in each month (P>0.05). The general trends of Shannon-Wiener diversity index (H) are similar to Patrick richness index (S), but increasing in June. In entire growth process of Aconitum, the overall fluctuation of entophytic bacteria diversity index is displayed that richness index (CVS= 0.1953)> evenness index (CVJ= 0.1316)> Shannon-Weiner index (CVH= 0.0643).3. During Aconitum growth process, the richness index of entophytic bacteria is significantly correlated with (P<0.01) entophytic fungi, and shows significantly correlated with Shannon-Wiener index of entophytic fungi (P<0.05). The Shannon-Wiener index of entophytic bacteria is significantly correlated with (P<0.01) richness index of entophytic fungi, and shows significantly correlated with Shannon-Wiener index of entophytic fungi (P<0.05).4. Cluster analysis and PCA analysis showed that within the growth process of Aconitum, as the seasons change, entophytic bacteria and fungi community structure are taking changes accordingly. It shows obviously structural change of entophytic bacteria community from January to May, which has great differences on the composition. Entophytic bacteria community from June to October has high similarity, and there is no significant change. The entophytic fungi community structure changes more obviously between January and May, and there are great differences on the composition. Especially in the second principal component that represents variable composition shows significantly different. There is high similarity from June to October of entophytic fungi community structure and no significant change. Overall, entophytic bacteria community structure changes more obviously from January to May as well as and entophytic fungi, but there are large differences in composition. The after 5 months (6-10) have high similarity of both entophytic bacteria and entophytic fungi community structure, but no obvious changes.In conclusion, it can be seen that entophytic bacteria community of Aconitum root has significant spatial and temporal variation during its growth process from January to October. It mainly indicates that the composition of root entophytic bacteria community (such as dominant groups and special groups) between first and second half year; community characteristics parameters (such as richness, diversity, and volatility) and correlation (such as bacteria and fungi internal and mutual) and so on have different performance and change. Under the condition of environmental variation, these changes are associated with stages of Aconitum growth, which reflects the monthly internal physiological state changes of Aconitum, and illustrates the systematical changed relationship between entophytic bacteria and Aconitum. |
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