黑曲霉ZJ-17配合有机肥和化肥施用对白芷生长、产量和品质的影响

刘思琴; 江美彦; 刘仁浪; 张宇柔; 侯凯; 冯冬菊; 徐东北; 郑全林; 吴卫

doi:10.11674/zwyf.2023115

黑曲霉ZJ-17配合有机肥和化肥施用对白芷生长、产量和品质的影响

1.
四川农业大学农学院，四川成都 611130
2.
四川全泰堂川白芷产业有限公司, 四川成都 629000

作者简介: 刘思琴 E-mail: 846547978@qq.com;

通讯作者: 吴卫, E-mail:ewuwei@sicau.edu.cn

基金项目: 四川省科技厅重点研发项目 (2022YFS0582)；四川省中医药重点学科建设项目(2021-16-4)；四川省科技厅重点研发项目 (2021YFYZ0012)；四川农业大学学科建设双支计划—学科创新团队项目 (P202108)。

Effects of Aspergillus niger ZJ-17 combined with organic and chemical fertilizers on the growth, yield and quality of Angelica dahurica var. formosana

1.
College of Agronomy, Sichuan Agricultural University, Chengdu, Sichuan 611130, China
2.
Sichuan Quantaitang Chuanbaizhi Industry Co., Ltd. Chengdu, Sichuan 629000, China

Corresponding author: WU Wei, E-mail:ewuwei@sicau.edu.cn

摘要: 【目的】 盲目施用化肥严重影响川白芷 (Angelica dahurica var. formosana)的产量和品质。研究白芷根际真菌配合有机无机肥施用的适宜用量，为白芷高产优质生产提供绿色施肥技术。 【方法】 田间试验在四川遂宁进行，供试菌剂为从白芷根际分离筛选出的黑曲霉ZJ-17 (Aspergillus niger ZJ-17) 制成的微生物菌剂 (ZJ-17)。共设8个处理：半肥 (T1)、全肥 (T2)、半肥+种施ZJ-17 (T3)、半肥+ZJ-17灌根 (T4)、全肥+ZJ-17种施 (T5)、全肥+ZJ-17灌根 (T6)，以不施肥 (T0)和全肥+种施市售黑曲霉 (有效活菌数≥100亿/g，T7)为对照。全肥量为油枯750 kg/hm²、尿素139.5 kg/hm²、过磷酸钙2193.8 kg/hm²、硫酸钾240 kg/hm²，半肥为上述各肥料施用量减半。菌液孢子含量3.0×10⁸ CFU/mL，施用量30 L/hm²，每个小区0.06 L，种施菌剂在播种时均匀浇在种子表面，灌根在次年3月进行。播种后每两个月取一次植株样品，共取5次，用于测定白芷农艺性状、植株养分含量。于收获期测定白芷根部品质和产量，并采用荧光定量法调查各处理土壤黑曲霉定植情况。 【结果】 种施菌剂有利于黑曲霉在土壤中定植，定植率显著优于菌剂灌根处理 (P<0.05)，市售黑曲霉也能定植于土壤。与仅全肥 (T2)处理比，种施菌剂 (T5)处理显著提高了收获期白芷株高、根长和根粗，分别显著增加11.57%、10.85%和16.70% (P<0.05)，根部产量和经济效益分别显著增加16.33%和19.94% (P<0.05)，根部氮、磷和钾积累量分别显著增加37.73%、21.18%和28.75%，香豆素类成分积累量也显著增加 (P<0.05)，且效果显著优于黑曲霉ZJ-17菌剂灌根 (T6)处理和市面购买黑曲霉 (T7)处理；半肥条件下，种施黑曲霉ZJ-17菌剂 (T3)使收获期白芷欧前胡素和异欧前胡素含量分别显著增加9.82%和22.45% (P<0.05)。 【结论】 常规有机肥和化肥用量下，种子表面浇施黑曲霉ZJ-17菌剂能显著促进白芷生长、养分吸收和香豆素类成分积累量，提高白芷产量和品质，提高经济效益。本实验室鉴定的黑曲霉ZJ-17菌株制备的菌剂效果显著好于市售菌剂，但采用生育中期 (3月)灌根的方法施用该菌剂的效果不明显。在半肥条件下，种施黑曲霉ZJ-17菌剂可显著提高白芷中的欧前胡素和异欧前胡素含量，但产量和经济效益低于全肥条件下种施黑曲霉ZJ-17菌剂处理。需进一步研究实际生产中保障产量且增加主要有效成分含量的肥料用量。
- 根际促生菌 /
- 白芷 /
- 产量 /
- 品质
Abstract: 【Objectives】 Unreasonable chemical fertilizer application impacts the yield and quality of Angelica dahurica var. formosana, a widely used Chinese medical herbs. We studied a green fertilization technology using the isolated rhizosphere fungi A. dahurica var. formosana with suitable chemical and organic fertilizers. 【Methods】 A field experiment was conducted in Suining, Sichuan Province. The tested microbial agent was prepared from Aspergillus niger ZJ-17 isolated from A. dahurica var. formosana rhizosphere. There were eight treatments in the experiment: half-fertilizer (T1), full-fertilizer (T2), half fertilizer+A. niger ZJ-17 seed application (T3), half fertilizer+A. niger ZJ-17 watering root (T4), full fertilizer+A. niger ZJ-17 seed application (T5), full fertilizer+A. niger ZJ-17 watering root (T6), no fertilizer (T0) and full fertilizer+commercial A. niger seed application (effective viable bacteria number≥10 billion/g, T7). Full fertilizer rate was composed of 750 kg/hm² oil cake, 139.5 kg/hm² urea, 2193.8 kg/hm² superphosphate, 240 kg/hm² potassium sulfate, and half fertilizer rate applied half amount of all the used fertilizers. The applying amount of the used microbial agent ( 3.0×10⁸ CFU/mL) was 30 L/hm², equaling to 0.06 L in each plot. Seed application was performed by pouring the microbial solution on the seed surface, and root irrigation by watering the microbial solution near the root soil in March of the next year. After seeding, plant samples were taken in frequency of every two months for determination of agronomic traits and plant nutrient content. At harvest, the quality and yield of A. dahurica var. formosana were analyzed, and the colonization of A. niger in soil was measured by fluorescence quantitative method. 【Results】 Seed application led to significantly higher colonization rate of A. niger than root application, commercial A. niger also colonized effectively in soil. Compared with T2, T5 treatment significantly increased plant height, root length and root diameter of A. dahurica var. formosana at harvest stage by 11.57%, 10.85% and 16.70% (P<0.05), the root yield and economic benefit by 16.33% and 19.94%, the accumulation of N, P and K in the underground part by 37.73%, 21.18% and 28.75%, increased the accumulation of total coumarins significantly. T5 also showed significantly higher promotion effect than T6 and T7 treatment. Under semi-fertilizer condition, T3 increased the contents of imperatorin and isoimperatorin ofA. dahurica var. formosana at harvest stage by 9.82% and 22.45% (P<0.05), T5 did not show significant effect on them. 【Conclusions】 Under conventional fertilizer application rate (full fertilizer), seed applying A. niger ZJ-17 can significantly improve the growth index, yield and benefit of A. dahurica var. formosana. Root application does not have the promotion effects. Under half fertilizer rate, seed applying A. niger ZJ-17 improved the imperatorin and isoimperatorin content, but decreased yield and benefit of A. dahurica var. formosanathan. Therefore, the suitable fertilizer rate needs further researching in practical production of A. dahurica var. formosana.
- rhizosphere growth-promoting bacteria /
- A. dahurica var. formosana /
- yield /
- quality

图 1 不同处理土壤中黑曲霉基因拷贝数

Figure 1. A. niger gene copies in different treatment soils

下载: 全尺寸图片幻灯片

图 2 收获期不同处理白芷养分积累量

Figure 2. Nutrient accumulation of A. dahurica var. formosana at harvest stage under different treatments

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图 3 不同处理白芷各时期香豆素类成分含量

Figure 3. Coumarin contents of A. dahurica var. formosana under different treatments in each period

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图 4 不同处理收获期白芷香豆素类成分积累量

Figure 4. Accumulation of coumarins in A. dahurica var. formosana at harvest stage

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表 1 实时荧光定量PCR反应体系

Table 1. Real-time fluorescence quantitative PCR

反应体系 Reaction system	体积 Volume (μL)
TB Green Premix Ex Taq II (Tli RNaseH Plus)	12.5
PCR正向引物 PCR forward primer	1
PCR反向引物 PCR reverse primer	1
DNA模板 DNA template (<100 ng)	2
灭菌水 Sterile water	8.5
总体积 Total volume	25

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表 2 不同处理各时期白芷株高 (cm)

Table 2. Plant height of A. dahurica var. formosana under different treatments in each period

处理 Treatment	2021−11−30	2022−01−28	2022−03−27	2022−05−31	2022−07−21
T0	5.92±0.84 d	11.10±1.37 b	55.77±6.20 d	88.86±7.01 c	57.52±15.54 c
T1	6.77±1.15 bc	11.90±0.87 ab	57.36±5.92 d	96.04±10.55 b	76.52±11.01 b
T2	6.86±0.61 bc	12.21±1.54 ab	65.72±3.99 bc	100.48±6.30 b	78.42±10.66 b
T3	7.70±1.24 a	12.00±1.88 ab	64.13±3.58 c	101.36±6.63 b	77.94±13.67 b
T4	6.40±1.06 cd	11.69±2.87 b	63.64±4.61 c	100.64±8.69 b	80.91±13.05 ab
T5	7.34±1.18 ab	12.12±1.88 ab	68.79±6.45 ab	108.97±6.89 a	88.68±13.43 a
T6	6.83±0.97 bc	13.03±1.19 a	63.66±5.62 c	99.49±6.12 b	83.93±16.13 ab
T7	6.45±0.72 cd	12.13±1.38 ab	69.11±4.29 a	109.68±4.69 a	89.62±11.90 a
注：T0—不施肥；T1—半肥；T2—全肥；T3—半肥+种施ZJ-17；T4—半肥+ZJ-17灌根；T5—全肥+种施ZJ-17；T6—全肥+ZJ-17灌根；T7—全肥+种施市售黑曲霉。同列数据后不同小写字母表示处理间差异显著 (P<0.05)。 Note: T0—No fertilizer; T1—Half fertilizer rate; T2—Full fertilizer rate; T3—Half fertilizer rate+A. niger ZJ-17 seed application; T4—Half fertilizer rate+A. niger ZJ-17 watering root; T5—Full fertilizer rate+A. niger ZJ-17 seed application; T6—Full fertilizer rate+A. niger ZJ-17 watering root; T7—Full fertilizer rate+commercial A. niger seed application. Different lowercase letters after data in a column indicate significant difference among treatments (P<0.05).

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表 3 不同处理各时期白芷根长 (cm)

Table 3. Root length of A. dahurica var. formosana under different fertilizer treatments in each period

处理 Treatment	2021−11−30	2022−01−28	2022−03−27	2022−05−31	2022−07−21
T0	5.72±0.82 bc	7.68±1.33 d	13.97±1.82 d	17.49±2.04 c	18.18±2.18 c
T1	6.02±1.11 ab	8.29±1.51 cd	14.15±1.39 d	17.71±2.24 bc	18.41±2.56 c
T2	5.87±0.83 abc	8.91±1.03 bc	15.26±2.12 cd	18.82±1.46 ab	18.89±2.97 bc
T3	6.12±1.17 ab	9.13±1.61 ab	18.25±4.20 a	18.28±1.63 bc	20.43±2.35 ab
T4	5.43±0.77 bc	8.88±2.30 bc	15.12±1.28 cd	18.78±1.18 abc	18.64±2.72 bc
T5	6.50±1.02 a	10.09±1.46 a	17.34±1.94 a	18.78±1.84 abc	21.19±2.59 a
T6	5.62±1.18 bc	8.89±1.12 bc	15.88±1.66 bc	19.68±1.75 a	19.46±2.65 abc
T7	5.22±0.93 c	9.39±1.52 abc	17.19±1.60 ab	19.65±1.68 a	20.82±2.09 a
注：T0—不施肥；T1—半肥；T2—全肥；T3—半肥+种施ZJ-17；T4—半肥+ZJ-17灌根；T5—全肥+种施ZJ-17；T6—全肥+ZJ-17灌根；T7—全肥+种施市售黑曲霉。同列数据后不同小写字母表示处理间差异显著 (P<0.05)。 Note: T0—No fertilizer; T1—Half fertilizer rate; T2—Full fertilizer rate; T3—Half fertilizer rate+A. niger ZJ-17 seed application; T4—Half fertilizer rate+A. niger ZJ-17 watering root; T5—Full fertilizer rate+A. niger ZJ-17 seed application; T6—Full fertilizer rate+A. niger ZJ-17 watering root; T7—Full fertilizer rate+commercial A. niger seed application. Different lowercase letters after data in a column indicate significant difference among treatments (P<0.05).

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表 4 各时期不同处理白芷的根粗 (mm)

Table 4. Root thickness of A. dahurica var. formosana under different fertilizer treatments in each period

处理 Treatment	2021−11−30	2022−01−28	2022−03−27	2022−05−31	2022−07−21
T0	1.98±0.37 c	3.24±0.95 e	12.51±1.68 f	27.04±2.14 e	37.10±3.35 d
T1	2.07±0.34 bc	3.81±0.55 d	12.96±1.87 ef	30.26±2.25 d	39.57±2.80 cd
T2	2.29±0.22 b	4.48±0.44 c	14.97±1.97 bc	32.35±3.41 cd	41.84±4.67 bc
T3	2.66±0.34 a	4.91±0.53 ab	14.45±1.56 cd	30.29±2.47 d	40.50±4.95 cd
T4	2.16±0.34 bc	3.62±0.42 de	14.11±1.51 cde	31.30±4.57 cd	40.58±5.15 cd
T5	2.74±0.36 a	5.14±0.48 a	16.39±1.79 a	35.20±3.03 ab	50.23±4.92 a
T6	2.12±0.22 bc	4.53±0.57 bc	13.75±1.66 de	33.52±3.31 bc	45.20±7.12 b
T7	2.00±0.34 c	4.52±0.43 bc	15.72±1.63 ab	36.08±3.24 a	44.86±4.33 b
注：T0—不施肥；T1—半肥；T2—全肥；T3—半肥+种施ZJ-17；T4—半肥+ZJ-17灌根；T5—全肥+种施ZJ-17；T6—全肥+ZJ-17灌根；T7—全肥+种施市售黑曲霉。同列数据后不同小写字母表示处理间差异显著 (P<0.05)。 Note: T0—No fertilizer; T1—Half fertilizer rate; T2—Full fertilizer rate; T3—Half fertilizer rate+A. niger ZJ-17 seed application; T4—Half fertilizer rate+A. niger ZJ-17 watering root; T5—Full fertilizer rate+A. niger ZJ-17 seed application; T6—Full fertilizer rate+A. niger ZJ-17 watering root; T7—Full fertilizer+commercial A. niger seed application. Different lowercase letters after data in a column indicate significant difference among treatments (P<0.05).

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表 5 各处理白芷产量和经济效益

Table 5. Yield and economic benefit of A. dahurica var. formosana under different fertilizer treatments

处理 Treatment	产量 Yield (kg/hm²)	投入成本 Input cost (yuan/hm²)			收入 Income (yuan/hm²)
处理 Treatment	产量 Yield (kg/hm²)	肥料 Fertilizer	除草 Weeding	菌剂 Microbial agent	总产值 Total output	经济效益 Benefit
T0	17857 d	0	33350		133929 d	100579 d
T1	20706 cd	2492	33350		155298 cd	119456 cd
T2	22197 bc	4984	33350		166481 bc	128147 bc
T3	22064 bc	2492	33350	566	165483 bc	129075 bc
T4	21803 bc	2492	33350	566	163524 bc	127116 bc
T5	26529 a	4984	33350	566	198966 a	160066 a
T6	23988 abc	4984	33350	566	179913 abc	141013 abc
T7	24819 ab	4984	33350	267	186146 ab	147545 ab
注：T0—不施肥；T1—半肥；T2—全肥；T3—半肥+种施ZJ-17；T4—半肥+ZJ-17灌根；T5—全肥+种施ZJ-17；T6—全肥+ZJ-17灌根；T7—全肥+种施市售黑曲霉。同列数据后不同小写字母表示处理间差异显著 (P<0.05)。 >Note: T0—No fertilizer; T1—Half fertilizer rate; T2—Full fertilizer rate; T3—Half fertilizer rate+A. niger ZJ-17 seed application; T4—Half fertilizer rate+A. niger ZJ-17 watering root; T5—Full fertilizer rate+A. niger ZJ-17 seed application; T6—Full fertilizer rate+A. niger ZJ-17 watering root; T7—Full fertilizer rate+commercial A. niger seed application. Different lowercase letters after data in a column indicate significant difference among treatments (P<0.05).

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中药白芷为伞形科植物白芷Angelica dahurica (Fisch. ex Hoffm) Benth. et Hook. f.或杭白芷Angelica dahurica (Fisch. ex Hoffm) Benth. et Hook. f. var. formosana (Boiss) Shan et Yuan的干燥根，始载于《神农本草经》，被列为中品，在我国有着悠久的药食两用的历史^[1−2]。白芷药材中的主要药效成分是香豆素^[3]，包括欧前胡素、异欧前胡素和氧化前胡素等^[4]。长期以来一直作为中药用于治疗感冒、发热引起的头痛和伤口引起的疼痛^[5]。在食品、保健品、香料和美容等方面也有广泛的用途^[6−7]。白芷适应性很强，喜温暖湿润气候，宜种植在土层深厚、疏松肥沃的砂质壤土地^[8]。现药用的白芷主要有川白芷、杭白芷、禹白芷和祁白芷等4大药材商品^[9]。川白芷主产于四川遂宁、安岳等地，其中遂宁白芷种植历史最久，其产量和品质都最佳，为道地药材^[10−11]。但目前川白芷生产中存在盲目施肥，严重影响其产量和品质等问题，采用适宜有效的绿色种植技术迫在眉睫。

我国农业生产中化肥过量施用问题较为普遍，肥料利用率不高、资源浪费、环境污染等问题时有发生。“十四五”期间，我国政府提出化肥减量增效目标为到2025年末全国农用化肥施用量实现稳中有降，三大粮食作物化肥利用率达到43%^[12]。随着人口的增加和耕地的减少，作物增产与生态环境保护的矛盾将日益尖锐，实现化肥减量增效是农业绿色可持续发展的必然要求^[13]。研究发现根际促生菌具有分解土壤内部营养物质^[14]、分泌生长素^[15]、调节植物生命周期某些或所有阶段植物体内促生菌含量^[16]、抑制有害菌生长^[17]等作用，从而可促进作物生长，提高作物产量^[18−21]和品质^[22]。但关于根际促生菌应用于白芷的研究报道很少。随着中药材市场的扩大与发展，中药材产业优质高效发展越来越关键。因此在白芷中应用根际促生菌并将其推广具有重要意义。本研究前期从白芷根际分离出一株具有固氮、溶磷和解钾活性的真菌黑曲霉ZJ-17，盆栽试验发现其对白芷具有促生作用，因此推测黑曲霉ZJ-17在田间对白芷生长、产量和品质也具有促进效果，本试验利用此黑曲霉ZJ-17制成微生物菌剂，与化肥和有机肥联合使用，探究其在田间的实际促生效果，以期为实现白芷高产、优质和绿色种植提供科学支撑。

1. 材料和方法

1.1. 试验地概况

试验点设在四川省遂宁市船山区唐家乡余建村白芷生产基地 (30°50′N，105°57′E)。试验地为平地，年平均气温为16.7℃～17.4℃，年平均降雨量是887.3～927.6 mm，前茬为白芷，土壤为灰潮土，土壤理化性质：pH7.8，有机质含量为10.41 g/kg，碱解氮含量为24.34 mg/kg，速效磷含量为12.66 mg/kg，速效钾含量为78.10 mg/kg。

1.2. 田间试验

田间试验始于2021年9月，为一年期试验，共设置8个处理，T0：不施肥；T1：半肥；T2：全肥；T3：半肥+种施黑曲霉ZJ-17；T4：半肥+黑曲霉ZJ-17灌根；T5：全肥+种施黑曲霉ZJ-17；T6：全肥+黑曲霉ZJ-17灌根；T7：全肥+种施市售黑曲霉，每个处理重复3次，共24个小区，小区面积20 m² (5 m×4 m)，随机区组排列。

黑曲霉ZJ-17真菌置于PDA培养基中，28℃恒温培养箱培养7 天后，收集黑曲霉孢子，加入无菌水，调整其浓度为3.0×10⁸ CFU/mL，按照30 L/hm²施用量，每个小区0.06 L，稀释100倍，将市面购买黑曲霉孢子粉的浓度调整一致。播前覆上油枯，施入磷酸钙和硫酸钾，开沟播种，按行距40 cm进行条播，最后将稀释后的菌液均匀浇在种子表面 (次年3月为灌根施用)。当白芷株高4～7 cm时，进行1～2次间苗，株距10 cm。全肥量为油枯750 kg/hm²、尿素139.5 kg/hm²、过磷酸钙2193.8 kg/hm²、硫酸钾240 kg/hm² ，半肥量为上述肥料施用量减半。尿素分3次施用，即苗肥 (12月)、冬肥 (次年2月)、春肥 (次年3月)，施用比例为2:5:3；过磷酸钙与硫酸钾分2次施用 (底肥、春肥)，施用比例为5:5。施用时将3种化肥混匀撒入土壤中。有机肥作为底肥施用。其余栽培措施与当地栽培措施保持一致。白芷品种为四川农业大学吴卫教授选育的‘川芷2号’白芷品种，原植物为杭白芷Angelica dahurica (Fisch. ex Hoffm) Benth. et Hook. f. var. formosana (Boiss) Shan et Yuan。供试菌株为课题组前期从‘川芷2号’根际土壤分离筛选的黑曲霉 (Aspergillus niger) ZJ-17菌株，具有较强固氮、溶磷和解钾活性；另市场购买黑曲霉孢子粉 (有效活菌数≥100亿/g，沂源康源生物科技有限公司)。

1.3. 测定项目及方法

2021年9月播种后连续取样至次年收获期，每两月进行1次采样，分别为2021年11月及2022年1月、3月、5月和7月收获期。每次每个小区取5株有代表性、长势一致、健康无病虫害的白芷植物样品，测定株高、根长和根粗。于收获期测产^[23]。采用高效液相色谱法^[24]测定白芷欧前胡素、异欧前胡素、氧化前胡素、白当归素、佛手柑内酯、白当归脑和水合氧化前胡素含量，参照《土壤农化分析》测定氮、磷、钾含量^[25]，荧光定量法测定收获期白芷土壤中黑曲霉ZJ-17定植率。根据黑曲霉ZJ-17基因序列，参考Rinttilä等^[26]的方法进行实时荧光定量PCR反应引物序列设计，荧光定量PCR反应体系见表1。

表 1 实时荧光定量PCR反应体系

Table 1. Real-time fluorescence quantitative PCR

反应体系 Reaction system	体积 Volume (μL)
TB Green Premix Ex Taq II (Tli RNaseH Plus)	12.5
PCR正向引物 PCR forward primer	1
PCR反向引物 PCR reverse primer	1
DNA模板 DNA template (<100 ng)	2
灭菌水 Sterile water	8.5
总体积 Total volume	25

1.4. 数据处理

采用Excel 2016、SPSS 23.0和GraphPad Prism 8等软件进行统计分析和作图。

3. 讨论

3.1. 白芷的生长指标和产量

研究表明植物根际促生真菌具有促进植物生长、提高植物抗逆性，增加作物产量和提高质量的作用^[27]。在目前化肥农药大量施用破坏生态环境的背景下，促生菌作为生物制剂，在农业、林业和环境修复等方面具有良好应用前景^[28]。

课题组前期通过筛选白芷根际促生菌发现，所筛促生综合潜力最强的真菌为黑曲霉ZJ-17，其具有较强的固氮、溶磷、溶钾活性，与不施菌对照处理相比，接种菌株ZJ-17后，遂宁土壤白芷的株高、根粗、地上鲜重和根鲜重均显著增长，崇州土壤白芷的株高和根鲜重均显著增加^[29]。本试验结果进一步证实，在大田生产条件下，黑曲霉ZJ-17与有机肥和化肥联合使用确实可明显促进白芷生长，其植株株高、根长和根粗均明显高于不施肥处理以及全肥处理，白芷单位面积产量也明显增加。在全肥条件下，播种期施用菌剂处理的产量明显高于未施用菌剂处理，增产率16.33%，产量最大值为26529 kg/hm²，与市面售卖黑曲霉处理间差异不显著；半肥条件下施用黑曲霉ZJ-17菌剂能保证其不减产，且与仅全肥处理差异不明显，说明黑曲霉ZJ-17菌剂确实能替代部分化肥用量。究其原因，可能是施入土壤中的根际促生菌株能改善植物根际营养环境，增加了对作物营养的供应，对植物生长和发育起到促进作用^[30]。王丹等^[31]发现巨大芽孢杆菌在正常氮磷供应条件下改善了油菜生长，促进了油菜对P、K、Zn和B 等营养元素的吸收。申佳丽等^[32]将胶质芽孢杆菌菌液对小黄瓜进行灌根，结果发现小黄瓜根中钾、磷，茎中钾、氮，小黄瓜叶中磷、钾受到显著影响。本试验的进一步测定分析也发现，全肥条件下，播期施加黑曲霉ZJ-17菌剂后白芷地下部分全氮、全磷和全钾的积累量均显著增加。半肥条件下，播期施加黑曲霉ZJ-17菌剂白芷地下部分全氮和全钾的积累量也显著增加。据此认为，施加黑曲霉ZJ-17能增强白芷获得养分的能力，进而促进其生长和产量提升。

3.2. 白芷的品质

本研究中供试黑曲霉ZJ-17对白芷品质也有明显促进作用。至于全肥条件下施加黑曲霉ZJ-17菌剂处理白芷的生长性状及产量高于半肥条件下施加菌剂者，但半肥条件下施加菌剂处理白芷的香豆素含量明显升高的原因，推测与全肥条件下施加黑曲霉ZJ-17菌剂处理的白芷根生长较快，根比较粗大，韧皮部在根中所占比例较小，其分泌组织分布较少，香豆素含量相对较低有关。曾有学者研究发现白芷香豆素相对含量与直径间呈非正相关关系^[33]。马逾英等^[34]测定了两个产地不同商品等级 (注：主要根据根大小划分等级)川白芷样品，也发现三等品总香豆素含量均高于一、二等品。雷雨恬等^[35]研究也表明白芷6种香豆素成分含量的总和以三等品最高，一等品最低。本试验还发现，全肥条件下施用黑曲霉ZJ-17，尽管白芷香豆素含量未提升，但其香豆素成分积累量均明显增加，这与江美彦等^[29]的研究结果一致，进一步表明黑曲霉ZJ-17对白芷品质确实具有明显影响，但如何将黑曲霉ZJ-17与化肥和有机肥合理配施，在提高产量的同时也增加主要有效成分含量值得进一步探讨。本试验中，施用黑曲霉ZJ-17条件下白芷香豆素累积量显著高于市售黑曲霉处理，充分说明从白芷根际分离的促生菌黑曲霉ZJ-17更适合用于白芷专用微生物肥料的开发利用。

3.3. 黑曲霉ZJ-17施用时间分析

本试验还探讨了黑曲霉ZJ-17菌剂的最佳施用时间。研究表明，微生物肥料不是速效肥，所以，要在作物的营养临界期和大量吸收期前7~10天施用效果最佳。同时对不同作物或同一作物的不同时期，要选用不同的施用方法。最适施用时间是清晨和傍晚或无雨阴天，避免阳光中紫外线将肥料中的微生物杀死^[36]。种肥15 kg配施不同量微生物菌剂，玉米根茎性状及产量均明显优于微生物肥料和高于常规施肥^[37]。本研究中，半肥条件下，播期施用该菌剂使苗期白芷株高和根粗明显增加；全肥条件下播期施用该菌剂使大部分生长阶段白芷的株高、根长和根粗均明显增加。至于次年3月施用黑曲霉ZJ-17菌剂对白芷生长无明显促进作用，可能与菌剂施入时间短，菌的定植率相对低有关。综合考虑，建议实际生产中于播期施入该菌剂。

4. 结论

常规有机肥和化肥用量下，播期种子表面浇施黑曲霉ZJ-17菌剂能显著促进白芷生长、养分吸收和香豆素类成分积累量，促进白芷生长，提高白芷产量和品质，提高了经济效益。本实验室鉴定的黑曲霉ZJ-17菌株制备的菌剂效果显著好于市售菌剂，但采用生育中期 (3月)灌根的方法施用该菌剂的效果不明显。在半肥条件下，种施黑曲霉ZJ-17菌剂可显著提高白芷根中的欧前胡素和异欧前胡素含量，但产量和经济效益不及全肥条件下种施黑曲霉ZJ-17菌剂处理。需进一步研究实际生产中，保障产量且增加主要有效成分含量的肥料用量。

参考文献 (37)

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黑曲霉ZJ-17配合有机肥和化肥施用对白芷生长、产量和品质的影响

作者简介: 刘思琴 E-mail: 846547978@qq.com;

通讯作者: 吴卫, E-mail:ewuwei@sicau.edu.cn

Effects of Aspergillus niger ZJ-17 combined with organic and chemical fertilizers on the growth, yield and quality of Angelica dahurica var. formosana

Corresponding author: WU Wei, E-mail:ewuwei@sicau.edu.cn

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黑曲霉ZJ-17配合有机肥和化肥施用对白芷生长、产量和品质的影响

作者简介:刘思琴 E-mail: 846547978@qq.com

通讯作者: 吴卫, ewuwei@sicau.edu.cn

English Abstract