Differences of pH Values, Nutrients and Bacterial Diversities Between Canker Diseased and Healthy Carya cathayensis Trees
LIN Ma-Shui1*, ZHANG Mei1*, CHAI Ying-Fei1, WANG Bing-Xuan1, WANG Shi-Ying1, FAN Yan-Di1, WU Chou-Fei2, GUO Ming1, LIN Hai-Ping1**
1 Local and National Joint Engineering Laboratory of Biopesticide high-efficient Preparation, Zhejiang A&F University, Lin'an 311300, China;
2 College of Life Sciences, Huzhou University, Huzhou 313000, China
Abstract：The large outbreak of canker disease has posed a serious threat to the sustainable development of hickory (Carya cathayensis) industry. For the purpose of studying the effect of bacterial diversity and function of C. cathayensis on canker disease, this research adopted 16S rRNA gene high-throughput sequencing, compared the differences of pH values, nutrients and bacterial diversity in sick parts and healthy parts, bark and xylem, respectively of healthy hickory trees and diseased trees during diseased period of canker disease, and the correlation were also analyzed. The results indicated that the pH values of the bark and xylem of healthy hickory trees respectively were 7.03 and 7.63. The neutral pH was significantly higher than that of acidic hickory trees (P<0.05). The tannin and soluble sugar contents of healthy hickory bark were 0.317% and 3.348% respectively, while that of the xylem were 0.079% and 0.626% respectively, of which all the values were remarkably higher the values of diseased trees (P<0.05). The reducing sugar contents in tree bark and xylem of healthy hickory trees were 0.865% and 0.420%, respectively, of which both were lower than that of diseased hickory trees (P<0.05). The amount of bark bacterial operational taxonomic units (OTU) in healthy hickory trees was significantly higher than that in diseased trees, while the amount of xylem bacterial OTU was significantly lower than that in diseased trees. The amount of the main bacterial genus found in healthy hickory barks, sick parts and free-diseased parts of diseased hickory barks were 7, 8, and 8 respectively, while the amount of that in xylem respectively were 8, 4, and 8. The species and abundance of the main bacterial genus in the barks and xylem of healthy and diseased hickory trees presented remarkable differences (P<0.05). The dominant bacteria genus lived in healthy hickory bark were Bdellovibrio, Acidocella, and Sphingomonas, and their abundance were 0.13%, 0.01%, and 15.32%, respectively. While the dominant bacterial genus founded in xylem were Massilia and Sphingomonas, and their abundance were 1.2% and 4.41%, respectively, of which both were remarkably higher than that of diseased hickory trees (P<0.05). In the diseased hickory trees, the dominant bacterial genus of sick and free-diseased bark both were Byssovorax and Methylobacterium. In diseased parts, the abundance of these 2 genera were 7.56% and 6.46% respectively, both higher than that in free-diseased parts. Both of the abundance of these 2 bacteria genera in 2 parts, tree bark, of the diseased C. cathayensis trees were obviously higher than that of the healthy hickory trees (P<0.05). In diseased parts xylem and free-diseased parts xylem of diseased hickory trees, Methylobacterium, Luedemannella, and Byssovorax performed as the dominant genera, and their abundance were 2.36%, 0.35%, 2.35% and 1.76%, 1.81%, and 0.55% respectively. All these figures were significantly higher than that of healthy hickory trees (P<0.05). The index of Ace, Chao, Shannon, and Simpson in the bark and xylem of healthy hickory trees were 599, 603, 6.59, 0.98 and 440, 454, 6.35, 0.95. All values were significantly higher than that of diseased hickory trees (P<0.05). The analysis of redundancy analysis (RDA) and monte carlo testing indicated that the impact of pH value, soluble sugar, and reducing sugar on the dominant bacteria genera in the bark and xylem of C. cathayensis trees were remarkable and significant (P<0.05). This research result could be considered as a reference for a sustaining and healthy operation run by the hickory forest.
[1] 陈建勋, 王晓峰. 2015. 植物生理学实验指导[M]. 华南理工大学出版社, 广州. pp. 2-3.
(Chen J X, Wang X F.2015. Experimental Guidance of Plant Physiology[M]. South China University of Technology Press, Guangzhou. pp. 2-3.)
[2] 陈志雄, 应学兵, 谢飞军. 2011. 山核桃主要病害的发生与防治[J]. 现代农业科技, 19: 206-207.
(Chen Z X, Ying X B, Xie F J.2011. Occurrence and control of major Carya cathayensis canker disease[J]. Modern Agricultural Science and Technology, 19: 206-207.)
[3] 戴德江, 马海芹, 沈瑶, 等. 2015. 几种杀菌剂对山核桃干腐病的室内活性筛选与应用[J]. 农药, 54(3): 217-230.
(Dai D J, Ma H Q, Shen Y, et al.2015. Application and screening of fungicides for controlling Carya tree canker caused by Botryosphaeria dothidea[J]. Agrocheincals, 54(3): 217-230.)
[4] 戴胜利. 2015. 山核桃桃溃疡的发生及防治技术[J]. 安徽农学通报, 21(10): 97-103.
(Dai S L.2015. Occurrence and control measures of Botryosphaeria fusisporae[J]. Anhui Agricultural Science Bulletin, 21(10): 97-103.)
[5] 冯静, 施庆珊, 欧阳友生, 等. 2009. 醋酸菌多相分类研究进展[J]. 微生物学通报, 32(9): 1390-1396.
(Feng J, Shi Q S, Ou Yang Y S, et al.2009. Research progress on the polyphasic taxonomy of acetic acid bacteria[J]. Microbiology China, 32(9): 1390-1396.)
[6] 傅本重, 朱洁倩, 郭新梅, 等. 2017. 核桃内生菌XHE7对病原真菌的拮抗作用及种子发芽的影响[J]. 江西农业大学学报, 39(2): 272-277.
(Fu B Z, Zhu J Q, Guo X M, et al.2017. The antagonism of walnut endophyte XHE7 against plant pathogenic fungi and effect on crop germination[J]. Acta Agriculturae Universitis Jiangxiensis, 39(2): 272-277.)
[7] 傅锦婷. 2014. 山核桃干腐病的生物学特性与控制研究[D]. 硕士学位论文, 浙江农林大学, 导师:张传清. pp. 19-20.
(Zhang J T.2014. The biological property and molecular epidemiology of Carya cathayensis canker disease and its control[D]. Thesis for M.S., Zhejiang A & F University, Supervisor: Zhang C Q, pp. 19-20.)
[8] 侯天侦, 陈聚恒, 于凯. 1992. 新疆杨树干破腐问题的结构及生理特性的研究[J]. 林业科学, 28(5): 390-396.
(Hou T Z, Chen J H, Yu K.1992. Research on the structure and physiology of broken and rotten trunks of Populus bollena[J]. Scientia Silvae Sinicae, 28(5): 390-396.)
[9] 胡雪文. 2010. 山核桃病虫害的发生规律及防治方法[J]. 安徽林业, 2010(3): 70-71.
(Hu X W.2010. Occurrence regularity and control measures of diseases and insect pests of Carya cathayensis[J]. Journal of Anhui Forestry, 2010(3): 70-71.)
[10] 巨云为, 赵盼盼, 黄麟, 等. 2015. 薄壳山核桃主要病害发生规律及防控[J]. 南京林业大学学报, 39(4): 31-36.
(Ju Y W, Zhao P P, Huang L, et al.2015. Analysis of Carya illinoensis main diseases occurrence and control[J]. Journal of Nanjing Forestry University, 39(4): 31-36.)
[11] 孔利. 2015. 新疆林木腐烂病菌主要种Cytospora sacculus致病力分化机理研究[D]. 硕士学位论文, 石河子大学, 导师:赵思峰. pp.7.
(Kong L.2015. Study on pathogenicity differentiation of Cytospora sacculus of valsa canker mainly kinds isolated from woods in xinjing[D]. Thesis for M.S., Shihezi University, Supervisor: Zhao S F, pp. 7.)
[12] 李海燕, 刘惕若, 甄艳. 2006. 辣椒品种对疫病的抗性研究——氨酸、丙二醛与可溶性糖在抗病中的作用[J]. 中国农学通报, 22(11): 315-317.
(Li H Y, Liu X R, Zhen Y.2006. Study on the resistance to phytophthora blight of pepper and the effect of Pro, MAD and dissolubility sugar[J]. Chinese Agricultural Science Bulletin, 22(11): 315-317.)
[13] 李寒娥. 2000. 城市行道树pH值和电导率与交通环境[J]. 生态科学, 19(2): 80-83.
(Li H E.2000. The pH value and electric conductivity of urban roadside tree's bark and traffic environment[J]. Ecological Science, 19(2): 80-83.)
[14] 李雪莹, 王文杰, 武永刚. 2005. 植物单宁的生理作用及经济价值[J]. 西部林业科学, 34(1): 66-69.
(Li X Y, Wang W J, Wu Y G.2005. Physiological function and economic value of plant tannin[J]. Journal of West China Forestry Science, 34(1): 66-69.)
[15] 欧倩. 2006. 丝氨酸生产菌MB200的筛选鉴定及丝氨酸羧甲基转移酶基因的研究[D]. 硕士学位论文, 广西大学, 导师:武波, pp. 6.
(Ou Q.2006. Isolation and identification of L-serine prdoucer strain MB200 and ITS serine hydroxymethyltransferase gene[D]. Thesis for M.S., Guangxi University, Supervisor: Wu B, pp. 6.)
[16] 秦彦杰. 2005. 黄檗主要药用成分的分布规律研究[D]. 硕士学位论文, 东北林业大学. 导师: 阎秀峰, pp. 21.
(Qin Y J.2005. Study on the distribution of principle medicinal compositions in Amur corktree[D]. Thesis for M.S., Northeast Forestry University, Supervisor: Yan X F, pp. 21.)
[17] 孙丽. 2012. 水稻蔗糖转化酶GIF1调控抗病性的机制研究[D]. 博士学位论文, 浙江大学, 导师: 何祖华, pp. 20.
(Sun L.2012. Function and mechanism of the cell-wall invertase GIF1 in regulating disease resistance in rice (Oryza sativa L.)[D]. Thesis for Ph.D, Zhejiang University, Supervisor: He Z H, pp. 20.)
[18] 吴平. 2006. 几种植物对室内污染气体甲醛的净化能力研究[D]. 硕士学位论文, 南京林业大学. 导师: 芦建国, pp. 17-43.
(Wu P.2006. Study on the purification ability of several plants to indoor polluted gas formaldehyde[D]. Thesis for M.S., Nanjing Forestry University, Supervisor: Lu J G, pp. 17-43.)
[19] 王月霞. 2016. 浙江省主要亚热带森林群落类型的物种和谱系α和β多样性研究[D]. 硕士学位论文, 浙江大学, 导师: 于明坚, pp. 27-36.
(Wang Y X.2016. The study of the taxonomic and phylogenetic α and β diversities of the main subtropical forest types in Zhejiang[D]. Thesis for M.S., Zhejiang University, Supervisor: Yu M J, pp. 27-36.)
[20] 杨淑贞, 丁立忠, 楼君芳, 等. 2009. 山核桃干腐病发生发展规律及防治技术[J]. 浙江林学院学报, 26(2): 228-232.
(Yang S Z, Ding L Z, Lou J F, et al.2009. Occurrence regularity of Carya cathayensis canker disease and its control[J]. Journal of Zhejiang Forestry College, 26(2): 228-232.)
[21] 张璐璐. 2012. 山核桃干腐病流行规律及田间防治的研究[D]. 硕士学位论文, 浙江农林大学. 导师:马良进, pp. 8.
(Zhang L L.2012. Epidemic regularity of cathayensis canker disease and its control[D]. Thesis for M.S., Zhejiang A & F University, Supervisor: Shu Q L, pp. 8.)
[22] 张璐璐, 贾桂民, 叶建丰, 等. 2013. 浙江临安山核桃干腐病发生发展规律[J]. 浙江农林大学学报, 30(1): 148-152.
(Zhang L L, Jia G M, Ye J F, et al.2013. Frequency of Carya cathayensis canker disease in Lin' an city, zhejiang province[J]. Journal of Zhejiang Agriculture and Forestry University, 30(1): 148-152.)
[23] 张耀辉. 1998. 生物多样性及生态平衡原理的探讨[J]. 农业环境科学学报, 17(5): 235-236.
(Zhang Y H.1998. Discussion on the principle of biodiversity and ecological balance[J]. Journal of Agro-Environment Science, 17(5): 235-236.)
[24] 张治安, 陈展宇. 2008. 植物生理学实验技术[M]. 吉林大学出版社, 吉林. pp. 100-133.
(Zhang Z A, Chen Z Y.2008. Experimental Techniques of Plant Physiology[M]. Jilin University Press, Jilin, pp. 100-133.)
[25] 赵联正, 谢占玲, 赵朋. 2015. 一种新的镰刀菌Q7-31木聚糖酶Xyn9的分离纯化鉴定及酶学特性[J]. 江苏农业科学, 43(5): 42-45.
(Zhao L Z, Xie Z L, Zhao P.2015. To separate and characterize a new Fusarium Q7-31 xylanase Xyn9[J]. Jiangsu Agricultural Sciences, 43(5): 42-45.)
[26] 赵莹. 2017. 根肿菌、寄主及根部内生微生物组交互作用研究[D]. 博士学位论文, 华中农业大学, 导师:姜道宏. pp. 109-111.
(Zhao Y.2017. Interaction among plasimodiophora brassicae, host and endo-microbiome of root[D]. Thesis for PH.D., Huazhong Agricultural University, Supervisor: Jang D H, pp. 109-111.)
[27] 郑宏兵. 2004. 山核桃抗溃疡病的机理及相关因素的研究[D]. 硕士学位论文, 安徽农业大学. 导师: 束庆龙, pp. 41.
(Zheng H B.2004. Studies on the resistance mechanism of Dothiorella canker of Carya tree and some factors[D]. Thesis for M.S., Anhui Agricultural University, Supervisor: Shu Q L, pp. 41.)
[28] 郑万钧. 1985. 中国树木志(2)[M]. 中国林业出版社, 北京. pp. 38.
(Zheng W J.1985. Sylva Sinica (2)[M]. China Forestry Press, Beijing. pp. 38.)
[29] 周军, 丁立忠, 张慧. 2016. 山核桃干腐病综合防治技术[J]. 浙江林业, 2016(4): 30-31.
(Zhou J, Ding L Z, Zhang H.2016. Integrated control measures of Botryosphaeria dothidea[J]. Journal of Zhejiang Forestry , 2016(4): 30-31.)
[30] 朱斌. 2003. 粘细菌NUST03的生物学特性和吸附重金属性质及其胞外活性物质的研究[D]. 硕士学位论文, 南京理工大学. 导师: 林吉文, pp. 1-5.
(Zhu B.2003. Study on the biological characteristics of NUST03 and the properties of heavy metals and their extracellular active substances[D]. Thesis for M.S., Nanjing University of Science and Technology, Supervisor: Lin J W. pp. 1-5.)
[31] Adhikari T B, Joseph C M, Yang G, et al.2001. Evaluation of bacteria isolated from rice for plant growth promotion and biological control of seedling disease of rice[J]. Canadian Journal of Microbiology, 47(10): 916-924.
[32] Ara I, Kudo T.2007. Luedemannella gen. nov., a new member of the family Micromonosporaceae and description of Luedemannella helvata sp. nov. and Luedemannella flava sp. nov[J]. Journal of General & Applied Microbiology, 53(1): 39-51.
[33] Caporaso J G, Kuczynski J, Stombaugh J, et al.2010. QIIME allows analysis of high-throughput community sequencing data[J]. Nature Methods, 7(5): 335-336.
[34] Edgar R C.2013. Uparse: Highly accurate OTU sequences from microbial amplicon reads[J]. Nature Methods, 10(10): 996-998.
[35] Gan H Y, Gan H M, Tarasco A M, et al.2014. Whole-genome sequences of five oligotrophic bacteria isolated from deep within lechuguilla cave, New Mexico[J]. Genome Announcements, 2(6): 2-14.
[36] Jurkevitch E.2006. The Genus Bdellovibrio[M] //: In Dworkin M, Falkow S, Rosenberg E,et al. The Prokaryotes. Springer, New York. pp. 12-30.
[37] Lepš J, Šmilauer P.2003. Multivariate Analysis of Ecological Data Using Canoco[M]. Cambridge University Press, New York,, pp. 60-75.
[38] Magoč Tanja, Steven L. Salzberg.2011. Flash: Fast length adjustment of short reads to improve genome assemblies[J]. Bioinformatics, 27(21): 2957-2963.
[39] Yu Y, Lee C, Kim J, et al.2005. Group-specific primer and probe sets to detect methanogenic communities using quantitative real-time polymerase chain reaction[J]. Biotechnology & Bioengineering, 89(6): 670-679.
