新疆农业科学 ›› 2023, Vol. 60 ›› Issue (9): 2248-2257.DOI: 10.6048/j.issn.1001-4330.2023.09.020
王丹丹1(), 李燕1, 张庆银1, 李世东2, 庞永超3, 马琨芝4, 马龙1, 牛瑞生1, 钟增明5, 齐连芬1(
), 师建华1(
)
收稿日期:
2022-11-20
出版日期:
2023-09-20
发布日期:
2023-09-19
通信作者:
齐连芬(1972-),女,河北任丘人,推广研究员,硕士,研究方向为设施蔬菜栽培与生理,(E-mail)nkyqlf@163.com;作者简介:
王丹丹(1991-),女,河北献县人,农艺师,硕士,研究方向为设施蔬菜栽培与生理,(E-mail)yuwenhanzhu@126.com
基金资助:
WANG Dandan1(), LI Yan1, ZHANG Qingyin1, Li Shidong2, PANG Yongchao3, MA Kunzhi4, MA Long1, NIU Ruisheng1, ZHONG Zengming5, QI Lianfen1(
), SHI Jianhua1(
)
Received:
2022-11-20
Published:
2023-09-20
Online:
2023-09-19
Supported by:
摘要:
【目的】评估不同微生物菌处理对番茄土壤微生物多样性的影响,为改善土壤微生物多样性、增加番茄产量提供理论依据与参考。【方法】以水果型番茄农博粉18109为试材,采用高通量测序技术,研究放线菌(T1)、枯草芽孢杆菌(T2)、哈茨木霉菌(T3)3种微生物菌处理,以不施用微生物菌对照(CK)进行番茄土壤微生物多样性及与环境相关性的影响。【结果】哈茨木霉(T3)处理的土壤中细菌含量最高,放线菌(T1)处理的土壤中真菌含量最高,T1、T3的细菌和真菌的比例均大于对照,土壤微生物呈现“细菌化”。Alpha多样性显示T1和T3的细菌种类多于CK,T1、T2、T3的真菌种类均少于CK;T2处理的细菌Shannon和Simpson指数均低于CK,T1和T3处理的细菌Shannon和Simpson指数均高于CK,3种微生物菌处理的真菌Shannon和Simpson指数均高于CK。对照、放线菌、枯草芽孢杆菌、哈茨木霉4个处理间存在差异,相同处理间存在较强的一致性。T1和T3微生物菌处理的细菌和真菌多样性较为相似。细菌中Unspecified_iii1_15、Unspecified_RB41、Unspecified_Gemm_5、Unspecified_Syntrophobacteraceae、Kaistobacter为优势细菌属。真菌中Fusarium(镰刀菌属)、Unspecified_Chaetomiaceae(未分类毛壳菌科)、Unspecified_Pezizales、Unspecified_Hypocreales_fam_Incertae_sedis、Mortierella(被孢霉属)为优势真菌属,镰刀菌属(Fusarium)在4个不同处理中相对丰度最高。细菌和真菌中叶宽与微生物群落分布间的相关性均最小,叶长、株高、茎粗和产量与微生物群落分布间的相关性高;放线菌和哈茨木霉对番茄土壤微生物的影响相对较大,叶长、叶宽、茎粗与放线菌的相关性最高,产量与哈茨木霉的相关性最高。【结论】不同微生物菌处理对番茄土壤微生物多样性影响不同,但放线菌和哈茨木霉对番茄土壤微生物的影响相对较大,叶长、叶宽、茎粗与放线菌的相关性最高,哈茨木霉与产量的相关性最高。
中图分类号:
王丹丹, 李燕, 张庆银, 李世东, 庞永超, 马琨芝, 马龙, 牛瑞生, 钟增明, 齐连芬, 师建华. 不同微生物菌处理对番茄土壤微生物多样性的影响[J]. 新疆农业科学, 2023, 60(9): 2248-2257.
WANG Dandan, LI Yan, ZHANG Qingyin, Li Shidong, PANG Yongchao, MA Kunzhi, MA Long, NIU Ruisheng, ZHONG Zengming, QI Lianfen, SHI Jianhua. Effects of different microbial treatments on tomato soil microbial diversity[J]. Xinjiang Agricultural Sciences, 2023, 60(9): 2248-2257.
处理 Treatment | 细菌 Bacteria (107CFU/g) | 真菌 Fungi (105CFU/g) | 细菌/真菌 Bacteria/ Fungi |
---|---|---|---|
CK | 67.9±7.3b | 42.1±2.3ab | 161.1 |
T1 | 85.6±8.2ab | 48.6±5.4a | 176.3 |
T2 | 64.4±5.8b | 42.3±7.0ab | 152.4 |
T3 | 109.8±16.6a | 33.2±9.8b | 331.1 |
表1 土壤中的微生物数量
Tab.1 The number of microorganisms in the soil
处理 Treatment | 细菌 Bacteria (107CFU/g) | 真菌 Fungi (105CFU/g) | 细菌/真菌 Bacteria/ Fungi |
---|---|---|---|
CK | 67.9±7.3b | 42.1±2.3ab | 161.1 |
T1 | 85.6±8.2ab | 48.6±5.4a | 176.3 |
T2 | 64.4±5.8b | 42.3±7.0ab | 152.4 |
T3 | 109.8±16.6a | 33.2±9.8b | 331.1 |
[1] | 王丹丹. 基于实时生长量的日光温室袋培番茄灌溉模型研究[D]. 沈阳: 沈阳农业大学, 2017. |
WANG Dandan. Research on irrigation model of bag-grown tomatoes in solar greenhouse based on real-time growth[D]. Shenyang: Shenyang Agricultural University, 2017. | |
[2] | 张光星. 番茄无公害生产技术[M]. 北京: 中国农业出版社, 2003: 55-65. |
ZHANG Guangxing. Pollution-free tomato production technology[M]. Beijing: China Agriculture Press, 2003: 55-65. | |
[3] | 王丹丹, 张庆银, 李燕, 等. 日光温室袋培番茄氮、磷、钾、钙、镁营养吸收分配规律研究[J]. 河北农业大学学报, 2021, 44(1):33-40. |
WANG Dandan, Zhang Qingyin, Li Yan, et al. Regularity of absorption and distribution of nitrogen, phosphorus, potassium, calcium, and magnesium in growth bag tomatoes in solar greenhouse[J]. Jouranal of Hebei Agricultural University, 2021, 44(1):33-40. | |
[4] | 王丹丹, 田国英, 张庆银, 等. 不同腐熟剂处理后番茄秸秆和土壤配比对番茄产量的影响[J]. 河北农业大学学报, 2021, 44(4): 35-43. |
WANG Dandan, TIAN Guoying, ZHANG Qingyin, et al. Effects of the ratio of tomato stalks and soil on tomato yield after treatment with different composting agents[J]. Journal of Hebei Agricultural University, 2021, 44(4): 35-43. | |
[5] | 马云华, 魏珉, 王秀峰. 日光温室连作黄瓜根区微生物区系及酶活性的变化[J]. 应用生态学报, 2004,(6): 1005-1008. |
MA Yunhua, WEI Min, WANG Xiufeng. Changes of microbial flora and enzyme activity in continuous cropping cucumber root zone in solar greenhouse[J]. Chinese Journal of Applied Ecology, 2004,(6): 1005-1008.
PMID |
|
[6] | 孙艳艳, 蒋桂英, 刘建国, 等. 加工番茄连作对农田土壤酶活性及微生物区系的影响[J]. 生态学报, 2010, 30(13): 3599-3607. |
SUN Yanyan, JIANG Guiying, LIU Jianguo, et al. Effects of continuous cropping of processed tomato on soil enzyme activities and microbial flora in farmland[J]. Acta Ecologica Sinica, 2010, 30(13): 3599-3607. | |
[7] | 高游慧, 郑泽慧, 张越, 等. 根际微生态防治作物土传真菌病害的机制研究进展[J]. 中国农业大学学报, 2021, 26(6): 100-113. |
GAO Youhui, ZHENG Zehui, ZHANG Yue, et al. Research progress on the mechanism of rhizosphere microecology in the prevention and control of soil-borne fungal diseases in crops[J]. Journal of China Agricultural University, 2021, 26(6): 100-113. | |
[8] | 唐小付, 刘岳飞, 张传进, 等. 设施甜瓜种植年限对土壤生物学特性和细菌多样性的影响[J]. 热带作物学报, 2018, 39(8): 1493-1500. |
TANG Xiaofu, LIU Yuefei, ZHANG Chuanjin, et al. Effects of planting years of muskmelon in facility on soil biological characteristics and bacterial diversity[J]. Chinese Journal of Tropical Crops, 2018, 39(8): 1493-1500. | |
[9] | 林先贵, 胡君利. 土壤微生物多样性的科学内涵及其生态服务功能[J]. 土壤学报, 2008, 45(5): 892-900. |
LIN Xiangui, HU Junli. The scientific connotation of soil microbial diversity and its ecological service function[J]. Acta Pedologica Sinica, 2008, 45(5): 892-900. | |
[10] | 张凡. 日光温室黄瓜连作土壤和基质微生物多样性与有机质组分关系研究[D]. 沈阳: 沈阳农业大学, 2017. |
ZHANG Fan. Study on the relationship between soil and substrate microbial diversity and organic matter components in continuous cucumber cropping in solar greenhouse[D]. Shenyang: Shenyang Agricultural University, 2017. | |
[11] | 韩雪, 潘凯, 吴凤芝. 不同抗性黄瓜品种根系分泌物对枯萎病病原菌的影响[J]. 中国蔬菜, 2006,(5): 13-15. |
HAN Xue, PAN Kai, WU Fengzhi. The effect of root exudates of different resistant cucumber varieties on the pathogen of Fusarium wilt[J]. Chinese Vegetables, 2006, (5): 13-15. | |
[12] | 庞师婵, 郭霜, 任奎瑜, 等. 番茄/茄子嫁接对其根际土壤生物学性状及细菌群落结构的影响[J]. 园艺学报, 2020, 47(2): 253-263. |
PANG Shichan, GUO Shuang, REN Kuiyu, et al. Effects of tomato/eggplant grafting on the biological properties of rhizosphere soil and bacterial community structure[J]. Acta Horticulturae Sinica, 2020, 47(2): 253-263. | |
[13] |
Hong S H, Lee E Y. Phytostabilization of salt accumulated soil using plant and biofertilizers: Field application[J]. Int Biodeterior Biodegrad, 2017, 124: 188-195.
DOI URL |
[14] | Bhattacharyya P N, Jha D K. Plant growth-promoting rhizobacteria (PGPR): emergence in agriculture[J]. World Jouornal of Microbial Biotechnology, 2012, 28(4): 1327-1350. |
[15] | 王光飞, 高晓东, 马艳, 等. 生物有机类复合调理剂在设施叶菜障碍土壤上的应用效果[J]. 中国土壤与肥料, 2020,(2): 56-65. |
WANG Guangfei, GAO Xiaodong, MA Yan, et al. Application effects of bio-organic compound conditioners on the soil with leafy vegetable barriers[J]. China Soil and Fertilizer, 2020,(2): 56-65. | |
[16] |
赖宝春, 姚锦爱, 戴瑞卿, 等. 2株拮抗放线菌复合防治番茄青枯病的研究[J]. 中国生物防治学报, 2021, 37(5):1035-1040.
DOI |
LAI Baochun, YAO Jinai, DAI Ruiqing, et al. Study on the compound control of tomato bacterial wilt with two antagonistic actinomycetes[J]. Chinese Journal of Biological Control, 2021, 37(5):1035-1040.
DOI |
|
[17] | 王迪, 徐慧春, 杜志强, 等. 生防放线菌Act12对棚室连作甜瓜根域微生物的调整效应[J]. 黑龙江农业科学, 2015,(11): 55-59. |
WANG Di, XU Huichun, DU Zhiqiang, et al. Regulation effect of biocontrol actinomycetes Act12 on the microorganisms in the roots of continuous cropping melon in the greenhouse[J]. Heilongjiang Agricultural Sciences, 2015,(11): 55-59. | |
[18] |
宁楚涵, 李文彬, 张晨, 等. 丛枝菌根真菌与放线菌对辣椒和茄子的促生防病效应[J]. 应用生态学报, 2019, 30(9): 3195-3202.
DOI |
NING Chuhan, LI Wenbin, ZHANG Chen, et al. Effects of arbuscular mycorrhizal fungi and actinomycetes on growth and disease prevention of pepper and eggplant[J]. Chinese Journal of Applied Ecology, 2019, 30(9): 3195-3202.
DOI |
|
[19] | 乔俊卿, 张心宁, 梁雪杰, 等. 枯草芽孢杆菌PTS-394诱导番茄对灰霉病的系统抗性[J]. 中国生物防治学报, 2017, 33(2): 219-225. |
QIAO Junqing, ZHANG Xinning, LIANG Xuejie, et al. Bacillus subtilis PTS-394 induces tomato systemic resistance to gray mold disease[J]. Chinese Journal of Biological Control, 2017, 33(2): 219-225. | |
[20] | 张晓云, 李宝庆, 郭庆港, 等. 枯草芽孢杆菌CAB-1抑菌蛋白对黄瓜白粉病的防治作用[J]. 中国生物防治学报, 2012, 28(3): 375-380. |
ZHANG Xiaoyun, LI Baoqing, GUO Qinggang, et al. The effect of Bacillus subtilis CAB-1 bacteriostatic protein on cucumber powdery mildew[J]. Chinese Journal of Biological Control, 2012, 28(3): 375-380. | |
[21] | 何嘉, 马婷慧, 白小军, 等. 微生物菌剂对枸杞生长发育、产量品质及土壤养分的影响[J]. 江苏农业科学, 2021, 49(14): 149-154. |
HE Jia, MA Tinghui, BAI Xiaojun, et al. Effects of microbial agents on the growth and development, yield and quality of Lycium barbarum and soil nutrients[J]. Jiangsu Agricultural Sciences, 2021, 49(14): 149-154. | |
[22] | 马慧媛, 黄媛媛, 刘胜尧, 等. 微生物菌剂施用对设施茄子根际土壤养分和细菌群落多样性的影响[J]. 微生物学通报, 2020, 47(1): 140-150. |
MA Huiyuan, HUANG Yuanyuan, LIU Shengyao, et al. Effects of microbial inoculants application on soil nutrients and bacterial community diversity in the rhizosphere of eggplant in greenhouses[J]. Microbiology Bulletin, 2020, 47(1): 140-150. | |
[23] | 赵远征, 王东, 徐利敏, 等. 不同微生物菌剂对马铃薯黑痣病的田间防效比较[J]. 中国植保导刊, 2020, 40(9): 90-92. |
ZHAO Yuanzheng, WANG Dong, XU Limin, et al. Comparison of field control effects of different microbial agents on potato mole disease[J]. China Plant Protection Guide, 2020, 40(9): 90-92. | |
[24] | 周芳, 马新远, 成金豪, 等. 不同微生物菌剂对苹果幼树白绢病的影响研究[J]. 上海蔬菜, 2021,(1): 76-78. |
ZHOU Fang, MA Xinyuan, CHENG Jinhao, et al. Effects of different microbial agents on sclerotium sclerotium of young apple trees[J]. Shanghai Vegetables, 2021,(1): 76-78. | |
[25] | 周晨, 张绍武, 陈伟. 微生物分类单元聚类算法比较研究[J]. 北京生物医学工程, 2014, 33(6): 591-597. |
ZHOU Chen, ZHANG Shaowu, CHEN Wei. Comparative study of clustering algorithms for microbial taxonomies[J]. Beijing Biomedical Engineering, 2014, 33(6): 591-597. | |
[26] | Aliasgharzad N, Pourmirzaei Z, Dehnad A R, et al. Streptomyces species favour spore germination and hyphal growth of arbuscular mycorrhizal fungus[J]. International Journal of Agriculture Research &Review, 2012, 68: 765-773. |
[27] |
Kumar G P, Kishore N, Amalraj E LD, et al. Evaluation of fluorescent Pseudomonas spp.with single and multiple PGPR traits for plant growth promotion of sorghum in combination with AM fungi[J]. Plant Growth Regulation, 2012, 67: 133-140.
DOI URL |
[28] | Desai S, Kumar G P, Amalraj L D, et al. Exploiting PGPR and AMF Biodiversity for Plant Health Management[J]. Springer India, 2016. |
[29] |
Thatoi H, Behera B C, Mishra R R, et al. Biodiversity and biotechnological potential of microorganisms from mangrove ecosystems: a review[J]. Ann Microbiol, 2013, 63(1): 1-19.
DOI URL |
[30] | 胡亚杰, 韦建玉, 卢健, 等. 枯草芽孢杆菌在农作物生产上的应用研究进展[J]. 作物研究, 2019, 33(2): 167-172. |
HU Yajie, WEI Jianyu, LU Jian, et al. Research progress of Bacillus subtilis application in crops production[J]. Crop Research, 2019, 33(2): 167-172. | |
[31] | 邓开英, 凌宁, 张鹏, 等. 专用生物有机肥对营养钵西瓜苗生长和根际微生物区系的影响[J]. 南京农业大学学报, 2013, 36(2): 103-109. |
DENG Kaiying, LING Ning, ZHANG Peng, et al. Effects of bio-organic fertilizers on the growth and rhizosphere microflora of watermelon plants in nursery pots[J]. Journal of Nanjing Agricultural University, 2013, 36(2): 103-109. | |
[32] | Arroyo P, Demiera L E S, Ansola G. Influence of environmental variables on the structure and composition of soil bacterial communities in natural and constructed wetlands[J]. Sci Total Environ, 2015, 506-507,380-390. |
[33] |
Vanelsas J D, Chiurazzi M, Mallon C A, et al. Microbial diversity determines the invasion of soil by a bacterial pathogen[J]. Proceedings of the National Academy of Sciences of the United States of America, 2012, 109(4):1159-1164.
DOI PMID |
[34] | 李凤霞, 赵营. 氮肥减量配施微生物菌剂对灌淤土花椰菜产量及土壤微生物的影响[J]. 水土保持研究, 2017, 24(2): 94-100. |
LI Fengxia, ZHAO Ying. Effect of nitrogen fertilizer reduction with microbial inoculants on broccoli production and the influence of the soil microbial characteristics[J]. Research of Soil and Water Conservation, 2017, 24(2): 94-100. | |
[35] |
任天志. 持续农业中的土壤生物指标研究[J]. 中国农业科学, 2000, 33(1): 68-75.
DOI |
REN Tianzhi. Soil bioindicators in sustainable agriculture[J]. Scientia Agricultura Sinica, 2000, 33(1): 68-75. | |
[36] | 辛冰牧, 刘亚, 朱德斌, 等. 应用 Illumina MiSeq高通量测序技术分析活性银离子处理饮用水微生物多样性[J]. 载人航天, 2017, 23(6): 824-828. |
XIN Bingmu, LIU Ya, ZHU Debin, et al. Microbiota diversity analysis of potable water treated with silver ion using Illumina Miseq sequencing technology[J]. Manned Spaceflight, 2017, 23(6): 824-828. | |
[37] | 张斌, 杨晓云, 陈志谊. 番茄枯萎病致病镰刀菌种类鉴定及优势种群的研究[J]. 植物病理学报, 2016, 46(4): 561-565. |
ZHANG Bin, YANG Xiaoyun, CHEN Zhiyi. Identification of fusarium species and dominant populations of tomato fusarium wilt[J]. Chinese Journal of Phytopathology, 2016, 46(4): 561-565. | |
[38] |
王芳, 王晓倩, 王莹, 等. 6种杀菌剂对黄瓜尖孢镰刀菌的毒力测定试验[J]. 中国农学通报, 2019, 35(9): 143-148.
DOI |
WANG Fang, WANG Xiaoqian, WANG Ying, et al. Toxicity test of six fungicides to Fusarium oxysporum in cucumber[J]. Chinese Agricultural Science Bulletin, 2019, 35(9): 143-148. | |
[39] | 王泽楠. 尖孢镰刀菌对不同紫花苜蓿品种生长和营养品质的影响[D]. 兰州: 兰州大学, 2021. |
WANG Zenan. The effect of Fusarium oxysporum on the growth and nutritional quality of different alfalfa varieties[D]. Lanzhou: Lanzhou University, 2021. | |
[40] | 刘新月, 李凡, 陈海如, 等. 致病性尖孢镰刀菌生物防治研究进展[J]. 云南大学学报(自然科学版), 2008, 30(S1):89-93. |
LIU Xinyue, LI Fan, CHEN Hairu, et al. Advances in biocontrol of pathogenic Fusarium oxysporum[J]. Journal of Yunnan University, 2008, 30 (S1): 89-93. | |
[41] | 李伟, 王金亭. 枯草芽孢杆菌与解磷细菌对苹果园土壤特性及果实品质的影响[J]. 江苏农业科学, 2018, 46(3): 140-144. |
LI Wei, WANG Jinting. Effects of Bacillus subtilis and phosphobacteria on soil properties and fruit quality in apple orchards[J]. Jiangsu Agricultural Sciences, 2018, 46(3): 140-144. |
[1] | 徐毛毛, 高杰, 李君明, 李鑫, 刘磊, 潘峰. 20个番茄商业品种群体的多样性分析[J]. 新疆农业科学, 2024, 61(9): 2191-2196. |
[2] | 田海燕, 张占琴, 颉建辉, 王建江, 杨相昆. 加工番茄果实番茄红素与主要品质性状的关系[J]. 新疆农业科学, 2024, 61(9): 2197-2202. |
[3] | 田超, 李玉姗, 马越, 宋羽. 不同浓度苦豆子浸提液对连作番茄生长及土壤肥力的影响[J]. 新疆农业科学, 2024, 61(9): 2203-2210. |
[4] | 张庭军, 李字辉, 崔豫疆, 孙孝贵, 陈芳. 微生物菌剂对棉花生长及土壤理化性质的影响[J]. 新疆农业科学, 2024, 61(9): 2269-2276. |
[5] | 陈芳, 李字辉, 孙孝贵, 张庭军. 不同剂量的微生物菌剂对加工番茄产量及品质的影响[J]. 新疆农业科学, 2024, 61(9): 2285-2289. |
[6] | 帕孜丽耶·艾合麦提, 王新勇, 周燕, 宋彬, 玉苏甫·阿不力提甫. 微生物菌剂对核桃叶片生理及光合特性的影响[J]. 新疆农业科学, 2024, 61(9): 2299-2306. |
[7] | 陈芳, 李字辉, 王兵跃, 孙孝贵, 张庭军. 微生物菌剂对冬小麦生长发育及产量的影响[J]. 新疆农业科学, 2024, 61(8): 1853-1860. |
[8] | 奚瑞, 陈怡佳, 李宁, 余庆辉, 王强, 秦勇. 外源2, 4-表芸苔素内酯对盐胁迫下不同盐敏感型番茄种子萌发的影响[J]. 新疆农业科学, 2024, 61(8): 1983-1992. |
[9] | 张彩虹, 王国强, 姜鲁艳, 刘涛, 德贤明. 低能耗组装式深冬生产型日光温室环境因子变化及番茄性状分析[J]. 新疆农业科学, 2024, 61(8): 2043-2053. |
[10] | 张福林, 李宁, 刘宇翔, 陈怡佳, 余庆辉, 闫会转. 外源2,4-表油菜素内酯及褪黑素对樱桃番茄果实品质和果皮形态结构的影响[J]. 新疆农业科学, 2024, 61(7): 1738-1747. |
[11] | 贾东海, 宋贤明, 顾元国, 李强, 曾幼玲, 苗昊翠, 郭美丽, 侯献飞. 化肥减量配施微生物菌肥对膜下滴灌红花生长发育及产量的影响[J]. 新疆农业科学, 2024, 61(4): 781-790. |
[12] | 阮向阳, 蒲敏, 肖乐乐, 罗林毅, 陈瑞杰, 李然, 陈国永, 冶军. 镁肥施用策略对加工番茄产量和品质的影响[J]. 新疆农业科学, 2024, 61(4): 916-925. |
[13] | 王贺亚, 罗静静, 王康, 王瑞楠, 王旭, 高光瑞, 房艳. 不同微生物菌剂对甜菜根腐病的防效及产量影响[J]. 新疆农业科学, 2024, 61(2): 448-454. |
[14] | 李春雨, 谭占明, 程云霞, 高源, 马全会, 李志国, 马兴. 水肥耦合对沙培番茄叶绿素含量以及光合特性日变化的影响[J]. 新疆农业科学, 2024, 61(12): 3006-3013. |
[15] | 李亚莉, 哈丽哈什·依巴提, 唐亚莉, 段婧婧, 李青军. 氮磷减施与钾协同共效对加工番茄产量和养分吸收的影响[J]. 新疆农业科学, 2024, 61(12): 3014-3019. |
阅读次数 | ||||||||||||||||||||||||||||||||||||||||||||||||||
全文 92
|
|
|||||||||||||||||||||||||||||||||||||||||||||||||
摘要 209
|
|
|||||||||||||||||||||||||||||||||||||||||||||||||