摘要人工栽培半夏(Pinellia ternata)的土壤理化性质及微生物群落结构和功能类群发生变化,导致其产量低且质量不稳,而油樟(Cinnamomum longepaniculatum)加工剩余物中富含有机质和抗菌成分,因此本研究将油樟废渣应用于半夏的栽培中,以期提高半夏产量和品质,同时实现油樟资源的可持续利用。本研究设置全土壤(CK)、油樟废渣∶土壤=3∶1 (V/V)(T6)、全油樟废渣(T8) 3个处理组,于2022年3月15日开始种植,于4月15日和5月1日对半夏生长指标进行测定,5月15日对半夏生理指标进行测定,于11月15日实验结束,整个过程中对半夏病害情况进行统计,在半夏倒苗后采用宏基因组测序手段对半夏根际土壤微生物进行测序,对比分析其微生物群落结构和功能的差异,并测定半夏产量。结果显示,T6和T8均提高了半夏出苗数、苗高、叶长叶宽,增加了半夏蛋白质含量、丙二醛含量、过氧化物酶活性和脯氨酸含量,但降低了半夏SPAD (soil and plant analyzer development)值和超氧化物歧化酶活性;T6处理可缓解半夏病害,但T8处理病害更为严重;T6处理产量增加,但T8处理产量减少;对照组半夏根际土壤中细菌的占比极大(96.32%~99.57%),真菌占比极小(0.27%~0.82%),T6和T8组的细菌数量较CK分别增加了3.16%和3.25%,真菌数量较CK分别减少了0.41%和0.55%;添加油樟废渣后,T6和T8处理组均增加了有益菌变形菌门(Proteobacteria)、类固醇杆菌属(Steroidobacter)、Povalibacter、假单胞菌属(Pseudomonas)、壤霉菌属(Agromyces)和康奈斯氏杆菌属(Conexibacter)的相对丰度;降低了土壤有害菌酸杆菌门(Acidobacteria)的相对丰度;T6和T8组的直系同源蛋白簇(clusters of orthologous groups of proteins, COG)功能基因丰度增加,这与壤霉菌属(Agromyces)、康奈斯氏杆菌属(Conexibacter)和链霉菌属(Streptomyces)紧密相关。综上所述,油樟废渣能改良半夏的土壤条件,T6处理组效果更好。本研究为油樟废渣在半夏栽培中的利用提供理论依据。
Abstract:The soil physicochemical properties, microbial community structure, and functional groups of artificially cultivated Pinellia ternata have undergone changes, resulting in low yield and unstable quality. However, the processing residue of Cinnamomum longepaniculatum is rich in organic matter and antibacterial components. Therefore, this study applied the waste residue of C. longepaniculatum to the cultivation of P. ternata in order to improve the yield and quality of P. ternata and realize the sustainable utilization of C. longepaniculatum. In this study, 3 treatment groups were set: The whole soil (CK), waste residue∶soil=3∶1 (V/V)(T6), the whole waste residue (T8), which began on March 15, 2022 and ended on November 15, 2022. The growth indicators of P. ternata were measured on April 15 and May 1, and the physiological indicators of P. ternata were measured on May 15. The disease situation of P. ternata was statistically analyzed throughout the process. After P. ternata fell, metagenome platform was used to conduct sequencing studies on the differences of rhizospheric microbial community and function, and determined the yield of P. ternata. The results showed that T6 and T8 both increased the number of seedlings, seedling height, leaf length and width of P. ternata, increased its protein content, malondialdehyde content, peroxidase activity, and proline content, but decreased its SPAD (soil and plant analyzer development) value and superoxide dismutase activity; T6 treatment could alleviate the disease of P. ternata, but T8 treatment was more serious; The yield of T6 treatment increased, but the yield of T8 treatment decreased; The proportion of bacteria in P. ternata rhizosphere soil (CK) was very high (96.32%~99.57%), while the proportion of fungi was very low (0.27%~0.82%). The number of bacteria in the T6 and T8 groups increased by 3.16% and 3.25% compared to CK, respectively, while the number of fungi decreased by 0.41% and 0.55%, respectively; After adding waste residue of C. longepaniculatum, the relative abundance of beneficial bacteria Proteobacteria, Steroidobacter, Povalibacter, Steroidobacter, Agromyces and Conexibacter increased. The relative abundance of harmful bacteria Acidobacteria was decreased; The abundance of COG (clusters of homologous groups of proteins) functional genes in the T6 and T8 groups increased, which was closely related to the genera Agromyces, Conexibacter, and Streptomyces. In summary, waste residue of C. longepaniculatum could improve the soil condition of P. ternata, and the T6 treatment group had a better effect. This study provides theoretical basis for the utilization of waste residue of C. longepaniculatum in P. ternata cultivation.
王燕芝, 熊林, 张建, 王丽, 李娅武, 彦芳. 油樟废渣对半夏栽培和根际土壤微生物的影响[J]. 农业生物技术学报, 2024, 32(7): 1642-1659.
WANG Yan-Zhi, XIONG Lin, ZHANG Jian, WANG Li, LI Ya, WU Yan-Fang. Effects of Waste Residue of Cinnamomum longepaniculatum on the Cultivation of Pinellia ternata and Microorganisms of Rhizospheric Soil. 农业生物技术学报, 2024, 32(7): 1642-1659.
[1] 陈大超, 甘涛. 2008. 不同土壤类型对半夏产量和品质的影响[J]. 南方农业, 2(7): 17-18. (Chen D C, Gan T.2018. Effects of different soil types on yield and quality of Pinellia ternata[J]. South China Agriculture, 2(7): 17-18.) [2] 陈义光, 张晓蓉, 张丽, 等. 2008. 具抗菌活性海洋放线菌菌株JMC 06001的分离和鉴定[J]. 微生物学通报, 35(01): 40-44. (Chen Y G, Zhang X R, Zhang L, et al.2008. Isolation and identification of marine actinomycete JMC06001 exhibiting strong antibacterial activity[J]. Microbiology, 35(01): 40-44.) [3] 董艳, 鲁耀, 董坤, 等. 2010. 轮作模式对设施土壤微生物区系和酶活性的影响[J]. 土壤通报, (1): 53-55. (Dong Y, Lu Y, Dong K, et al. 2010. Effects of crop rotation patterns on soil microbial flora and enzyme activities in facilities[J]. Chinese Journal of Soil Science, (1): 53-55.) [4] 杜贵友, 方文贤. 2000. 有毒中药现代研究与合理应用[M]. 北京: 中国中医药出版社, pp. 417-420. (Du G Y, Fang W X.2000. Modern research and rational application of toxic traditional chinese medicine[M]. China Traditional Chinese Medicine Press, Beijing, China, pp. 417-420.) [5] 杜永华, 敖光辉, 魏琴, 等. 2014. 脱油油樟叶化学成分预试及其总黄酮含量测定[J]. 黑龙江农业科学, (10):120-123. (Du Y H, Ao G H, Wei Q, et al. 2014. Preliminary chemical analysis and total flavonoids assaying of de-oiled leaves from Cinnamomum longepaniculatum[J].Heilongjiang Agricultural Sciences, (10): 120-123.) [6] 高俊凤. 2006. 植物生理学实验指导[M]. 北京: 高等教育出版社, pp. 141-231. (Gao J F.2006. Experimental guidance of plant physiology[M]. Higher Education Press, Beijing, China, pp. 141-231.) [7] 郭魏. 2013. 不同耕作和培肥模式下豫中区麦田土壤微生物和腐殖质的研究[D]. 硕士学位论文, 河南师范大学, 导师: 李春喜, pp. 37-38. (Guo W.2013. Effects of different tillage and fertilization treatments on microbes and humus of wheat field soil in mid henan[D]. Thesis for M.S., Henan Normal University, Supervisor: Li X C, pp. 37-38.) [8] 杭烨, 罗夫来, 赵致, 等. 2018. 半夏间作不同作物对土壤微生物、养分及酶活性的影响研究[J]. 中药材, 41(07):1522-1528. (Hang Y, Luo F L, Zhao Z, et al.2018. Effect of Pinellia ternata with different intercropping crops on soil microorganism, nutrient and enzyme activity[J]. Journal of Chinese Medicinal Materials,41(07):1522-1528.) [9] 何志贵. 2019. 半夏连作障碍发生机制与轮作修复研究[D].博士学位论文, 西北农林科技大学, 导师: 董娟娥, pp. 38-49+59-88. (He Z G. 2019. Study on the mechanism of continuous cropping obstacle and rotation repair in Pinellia ternata[D]. Thesis for Ph.D., Northwest A&F University, Supervisor: Dong J E, pp. 38-49+59-88.) [10] 黄亚丽, 郑立伟, 黄媛媛, 等. 2020. 枯草芽孢杆菌菌剂不同施用方式对甜瓜土壤微生物多样性及生长的影响[J].生物工程学报, 36(12): 2644-2656. (Huang Y L, Zheng L W, Huang Y Y, et al.2020. Effects of different application methods of Bacillus subtilis agent on soil microbial diversity and growth of muskmelon[J]. Chinese Journal of Biotechnology, 36(12): 2644-2656.) [11] 黄之远, 赵桂茹, 刘涛, 等. 2023. 半夏玉米间作对根际微生物群落的影响[J/OL]. 分子植物育种: 1-13. http://kns.cnki.net/kcms/detail/46.1068.S.20220704.1333.007.html. (Huang Z Y, Zhao G R, Liu T, et al. 2023. The effect of intercropping Pinellia ternata and Zea mays L on the rhizosphere microbial community[J/OL]. Molecular Plant Breeding: 1-13.) [12] 李福生, 李志明, 于晓明, 等. 2021. 中医药辨治江西地区2019新型冠状病毒肺炎验案3则[J]. 江西中医药大学学报, 33(05): 16-20. (Li F S, Li Z M, Yu X M, et al.2021. Three clinical cases of diagnoses and treatment of COVID-19 by traditional chinese medicine in jiangxi area[J]. Journal of Jiangxi University of Traditional Chinese Medicine, 33(5): 16-20.) [13] 刘诗蓉, 王红兰, 杨萍, 等. 2022. 不同土壤改良措施对连作半夏根际微环境的影响[J]. 成都中医药大学学报, 45(03): 1-6. (Liu S R,Wang H L,Yang P, et al.2022. Effects of different soil improvement measures on rhizosphere microenvironment of continuous cropping Pinellia ternata[J]. Journal of Chengdu University of Traditional Chinese Medicine, 45(03): 1-6.) [14] 刘建桥, 向更生, 刘西苑, 等. 2022. 不同基质配比对半夏生长和生理特性的影响[J]. 农业科学研究, 43(02): 40-43+54. (Liu J Q, Xiang G S, Liu X Y, et al. 2022. Effects of different substrate proportions on growth and physiological characteristics of Pinellia ternata[J]. Journal of Agricultural Sciences, 43(02): 40-43+54.) [15] 苗睿睿, 苏颖, 杨晨, 等. 2021. 生防芽孢杆菌微生态菌剂制备及在中药材GAP生产中的应用[J]. 山东化工, 50(05): 32-33. (Miao R R, Su Y, Yang C, et al.The preparation of microbial agents of Bacillus subtilis and application in GAP production of traditional chinese medicine[J]. Shandong Chemical Industry, 50(05): 32-33.) [16] 欧阳谅, 万淑婉, 涂国全, 等. 1993. 一株产生杀虫抗生素的链霉菌新种[J]. 江西农业大学学报, 1993(S4): 10-15. (Ou Y L, Wang S W, Tu G Q, et al.1993. A new species of Streptomyces producing insecticidal antibiotics[J]. Acta Agriculturae Universitis Jiangxiensis, 1993(S4): 10-15.) [17] 皮莉, 梁宗锁, 张跃进. 2007. 土壤含水量对半夏生长和抗氧化性的影响[J]. 西北农业学报,(3): 196-199+128. [18] 唐汉萌. 2019. 微生物菌剂和生物炭对半夏产量、品质及土壤微生态的影响[D]. 硕士学位论文, 华中农业大学, 导师: 朱端卫, pp. 9-81. (Tang H M.2019. Effects of microbial fertilizer and biochar applications on the yield and quality of Pinellia ternata and soil microecology[D].Thesis for M.S., Huazhong Agricultural University, Supervisor: Zhu D W, pp. 9-81.) [19] 王松, 叶雷, 罗运川, 等. 2017. 脱油油樟叶栽培茶树菇及品质分析[J]. 宜宾学院学报, 17(06): 115-119. (Wang S, Ye L, Luo Y C, et al.2017. Culture and quality analysis of Agrocybe aegerita using de-oiled leaves of C. longepaniculatum[J]. Journal of Yibin University, 17(06): 115-119.). [20] 万燕, 雨田, 郭晓恒, 等. 2017. 妇科千金片药渣发酵制备有机肥在半夏种植上的应用[J]. 成都大学学报(自然科学版), 36(01): 39-42. (Wan Y, Yu T, Guo X H, et al.2017. Application of organic fertilizer prepared by residue generated in production of gynecologic qianjin tablets through fermentation in Pinellia ternate cultivation[J]. Journal of Chengdu University (Natural Science Edition), 36(1): 39-42.) [21] 魏琴, 李群, 罗扬, 等. 2006. 油樟油对植物病原真菌活性的抑制作用[J]. 中国油料作物学报, 28(1): 63-66. (Wei Q, Li Q, Luo Y, et al.2006. Antifungal activity of leaf essential oil from Cinnamomum longepaniculatum (Gamble) N.Chao[J]. Chinese Journal of Oil Crop Sciences, 28(1): 63-66.) [22] 闫杨, 刘月静, 李晓静, 等. 2018. 生防芽孢杆菌对半夏根际土壤酶活性及产量的影响[J]. 聊城大学学报(自然科学版), 31(03): 99-105. (Yan Y, Liu Y J, Li X J,et al.2018.Effect of biocontrol bacillus strains on enzyme activities in rhizosphere soil and yield of Pinellia ternata[J]. Journal of Liaocheng University(Nat. Sci.), 31(3): 99-105.) [23] 游玲, 范玉琴, 叶雷, 等. 2014. 废弃油樟叶渣栽培平菇研究[J]. 江苏农业科学, 42(09): 205-207. (You L, Fan Y Q, Ye L,et al.2014. Cultivation of Pleurotus ostreat with waste camphor leaf residue[J]. Jiangsu Agricultural Sciences, 42(09): 205-207) [24] 张超, 魏琴, 杜永华, 等. 2011. 脱油油樟叶提取物的体外抑菌活性研究[J]. 广西植物, 31(05): 690-694. (Zhang C, Wei Q, Du Y H, et al.2011. The antibacterial study on extracts from de-oiled leaves of Cinnamomum longepaniculatum against three of the pathogenic bacteria[J].Guihaia, 31(5): 690-694.) [25] 张皓, 何腾兵, 林昌虎, 等. 2015. 不同轮作方式对黔产半夏土壤机械组成与养分含量的影响[J]. 作物杂志, (02): 101-106. (Zhang H, He T B, Lin C H, et al. 2015. Effects of different rotation methods on soil mechanical composition and nutrient content of Pinellia ternata in guizhou province[J]. Crop Journal, (02): 101-106.) [26] 张丽霞. 2015. 泥炭对半夏生长的促进作用[J]. 农业与技术,35(8): 22-22. (Zhang L X.2015. Promoting effect of peat on Pinellia ternata growth[J]. Agriculture and Technology, 35(8): 22-22.) [27] 展丽然, 张克诚, 冉隆贤, 等. 2008. 苹果轮纹病菌拮抗放线菌的筛选与鉴定[J]. 中国农学通报, (04): 341-344. (Zhan L R, Zhang K C, Ran L X, et al. 2008. Selection and identification of the antagonistic actinomycetes against apple ring rot disease[J]. Chinese Agricultural Science Bulletin, (04): 341-344.) [28] 周成强, 刘斌, 郑仁红, 等. 2021. 油樟枝叶加工剩余物处理与资源化利用途径探讨[J]. 再生资源与循环经济, 14(10): 21-25. (Zhou C Q, Liu B, Zheng R H, et al.2021. Discussion on treatment and resource utilization of processing residues of Cinnamomum longipaniculatum branches and leaves[J]. Recyclable Resources and Circular Economy, 14(10): 21-25.) [29] 朱振兴, 包婉玉, 江林波, 等. 2021. 间作大豆对半夏生长环境及产量与药材质量的影响[J]. 湖北农业科学, 60(10): 85-90. (Zhu Z X, Bao W Y, Jiang L B, et al.2021. (Effects of intercropping soybean on growth environment, yield and quality of Pinellia ternata (Thunb.) Breit[J]. Hubei Agricultural Sciences, 60(10): 85-90.) [30] Bodenhausen N, Horton M W, Bergelson J.2013.Bacterial communities associated with the leaves and the roots of Arabidopsis thaliana[J]. PLOS ONE, 8(2): e56329. [31] Chen X F, Wang J C, You Y M, et al.2021. When nanoparticle and microbes meet: The effect of multi-walled carbon nanotubes on microbial community and nutrient cycling in hyperaccumulator system[J]. Journal of Hazardous Materials, 423(Pt A): 126947-126947. [32] Guo J, Liu W, Zhu C, et al.2018. Bacterial rather than fungal community composition is associated with microbial activities and nutrient-use efficiencies in a paddy soil with short-term organic amendments[J]. Plant and Soil, 424:335-349. [33] Liu M, Sui X, Hu Y,et al.2019. Microbial community structure and the relationship with soil carbon and nitrogen in an original Korean pine forest of changbai mountain[J]. BMC Microbiol, 19: 218. [34] Rangjaroen C, Rerkasem B, Teaumroong N, et al.2014. Comparative study of endophytic and endophytic diazotrophic bacterial communities across rice landraces grown in the highlands of northern thailand[J]. Archives of Microbiology, 196: 35-49. [35] Shi Y W, Lou K, Li C, et al.2015. Illumina-based analysis of bacterial diversity related to halophytes Salicornia europaea and Sueada aralocaspica[J]. Journal of Microbiology, 53: 678-685. [36] Wai W C N, San P S, Fung T J W, et al.2022. Intercropping of Pinellia ternata (herbal plant) with Sedum alfredii (Cd-hyperaccumulator) to reduce soil cadmium (Cd) absorption and improve yield[J]. Environmental Pollution (Barking, Essex: 1987).