生物炭与甲壳素配施对连作平邑甜茶幼苗及土壤环境的影响

doi:10.3864/j.issn.0578-1752.2017.04.011

摘要/Abstract

摘要： 【目的】研究生物炭与甲壳素配施对连作条件下平邑甜茶幼苗生物量、根系呼吸速率、根系保护酶和土壤环境的影响，为防治苹果连作障碍提供依据。【方法】盆栽条件下，以苹果常用砧木-平邑甜茶为试材，试验设苹果连作土壤作为对照（CK），用1‰甲壳素（w/w，T1）、2%生物炭（w/w，T2）、1‰甲壳素+2%生物炭（T3）以及溴甲烷灭菌（T4）处理苹果连作土壤，共5个处理。采用常规方法测定不同处理对平邑甜茶幼苗生物量、根系呼吸速率及根系抗氧化酶活性的影响，同时用末端限制性片段长度多态性（T-RFLP）和实时荧光定量PCR测定不同处理土壤中真菌群落结构和尖孢镰刀菌数量。【结果】T3和T4可显著增加平邑甜茶幼苗株高、地茎和干鲜重，T3使平邑甜茶幼苗的株高、地径、鲜重、干重分别增加了44.6%、33.0%、76.8%和77.1%，T4各指标分别增加了73.1%、76.9%、117.0%和123.7%；与对照相比，不同处理均使根系呼吸速率明显提高，T1、T2、T3和T4处理的幼苗根系呼吸速率分别是对照的1.37、1.70、1.87和2.02倍；T4处理超氧化物歧化酶（SOD）活性最高，其次为T3处理，T2和T1处理也增加了SOD活性，所有处理都与对照达到了显著差异；过氧化物酶（POD）和过氧化氢酶（CAT）活性变化与SOD变化趋势一致，T4处理根系POD和CAT活性分别为对照的2.68和2.64倍，其次为T3，3种酶活分别为对照的2.52和2.18倍；与对照相比，T2、T3和T4均可显著降低根系丙二醛（MDA）的含量，分别为对照的82.0%、61.9%和43.1%。T2和T3明显提高细菌数量，细菌/真菌比值分别是对照的1.7和2.2倍，溴甲烷灭菌（T4）则使土壤中的细菌、真菌显著降低，但是真菌降低的数量大于细菌的，因此，细菌/真菌比值高于对照；T4具有最高的多样性、丰富度和均匀度指数，分别比对照增加了24.4%、41.3%和13.6%，其次是T3处理；溴甲烷灭菌、生物炭配施甲壳素处理土壤中尖孢镰刀菌基因拷贝数均显著低于连作土，说明连作土壤中尖孢镰刀菌为主的有害真菌数量明显减少；与对照相比，T3土壤中根皮苷、根皮素、肉桂酸、对羟基苯甲酸和间苯三酚含量分别降低了28.1%、30.6%、50.8%、33.6%和46.2%。【结论】生物炭与甲壳素配施相较于单施生物炭或者甲壳素，能更好的提高连作条件下平邑甜茶幼苗的生物量、根系呼吸速率和根系保护酶活性。并且二者配合施用优化了连作土壤的真菌群落结构，增加了土壤细菌/真菌比值，降低了土壤中尖孢镰刀菌基因拷贝数，减少了酚酸类物质含量。因此，生物炭配施甲壳素能更好地缓解苹果连作障碍。

关键词: 苹果连作障碍, 生物炭, 甲壳素, 平邑甜茶幼苗

Abstract: 【Objective】 The experiment was conducted to explore the effects of biochar and chitin combined application on the plant biomass, root respiration rate, root protective enzymes and soil environment under replanting conditions, and to provide a basis for prevention of apple replanting disease (ARD). 【Method】 The Malus hupehensis Rehd. seedlings were planted in pots with apple replanting soil. The experiment included 5 treatments, which were the control (CK), 1‰ chitin (T1), 2% biochar (T2), 1‰ chitin + 2% biochar (T3) and methyl bromide sterilization (T4). The biomass, root respiration rate and protective enzymes activities of M. hupehensis seedlings were measured by conventional method. The number of microorganism, fungal group structure and the Fusarium oxysporum numbers were detected by terminal restriction fragment length polymorphism (T-ＲFLP) and a real-time quantitative PCR detection system. 【Result】 The results showed that compared with the control, T3 and T4 significantly enhanced the plant height, ground diameter, fresh and dry weight, and that in T3 increased by 44.6%, 33.0%, 76.8% and 77.1%, and in T4 increased by 73.1%, 76.9%, 117.0% and 123.7%, respectively. T1, T2, T3 and T4 treatments made the root respiration rate increased significantly, which were 1.37, 1.70, 1.87 and 2.02 times more than the control. The four different treatments also increased the root activity of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT), and the increase reached a significant difference. The SOD, POD and CAT activities of T4 treatments were 1.34, 2.68 and 2.64 times compared with the control, and T3 were 1.21, 2.52 and 2.18 times, respectively. T2 and T3 improved the numbers of bacteria, enhanced the ratio of bacteria and fungi and made the soil become “bacterial soil”. The ratios of T3 and T4 were 1.7 and 2.2 times as large as the control, methyl bromide sterilization made the numbers of bacteria and fungi decreased significantly, and the fungi numbers decreased more than bacteria’s, so the bacteria/fungi ratio was higher than the control. T4 had the highest Shannon diversity index, evenness index and richness index, increased by 24.4%, 41.3% and 13.6% compared with CK, and the second was the T3 treatment. Compared with the control, T3 and T4 reduced the gene copie’s number of F. oxysporum, which explained that the F. oxysporum harmful fungi number decreased significantly in replanting soils. Compared with CK, the concentration of phlorizin, phloretin, cinnamic acid, p-hydroxybenzoic acid and phloroglucinol reduced by 28.1%, 30.6%, 50.8%, 33.6% and 46.2% in T3, respectively.【Conclusion】The biochar and chitin combined application improved the condition in apple replanting soil better than solely biochar or chitin addition. The combined application improved the M. hupehensis. seedlings biomass, root respiration rate and root activity of protective enzymes. And the combined application optimized the soil fungi community structure, increased the soil bacteria/fungus ratio, and reduced the F. oxysporum gene copy numbers and contents of phenolic acids. Therefore, biochar and chitin combined application can better relieve ARD.

Key words: apple replant disease, biochar, chitin, Malus hupehensis Rehd. seedlings

王艳芳，相立，徐少卓，王森，王晓伟，陈学森，毛志泉，张民. 生物炭与甲壳素配施对连作平邑甜茶幼苗及土壤环境的影响[J]. 中国农业科学, 2017, 50(4): 711-719.

WANG YanFang, XIANG Li, XU ShaoZhuo, WANG Sen, WANG XiaoWei, CHEN XueSen, MAO ZhiQuan, ZHANG Min. Effects of Biochar and Chitin Combined Application on Malus hupehensis Rehd. Seedlings and Soil Environment Under Replanting Conditions[J]. Scientia Agricultura Sinica, 2017, 50(4): 711-719.

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