王平3,
李隆1,,,
张轩1,
桂林国4,
黄建成4
1.中国农业大学资源与环境学院/植物与土壤相互作用教育部重点实验室 北京 100193
2.河南科技学院生命科技学院/现代生物育种河南省协同创新中心 新乡 453003
3.宁夏农林科学院作物研究所 银川 750105
4.宁夏农林科学院农业资源与环境研究所 银川 750002
基金项目: 国家重点研发计划项目2016YFD0300202
国家科技支撑计划项目2007BAD89B02
详细信息
作者简介:梅沛沛, 主要研究方向为生物多样性与资源高效利用。E-mail:meipeipei@126.com
通讯作者:李隆, 主要研究方向为生物多样性与资源高效利用。E-mail:lilong@cau.edu.cn
中图分类号:S344.2;S144.3计量
文章访问数:1003
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被引次数:0
出版历程
收稿日期:2017-07-07
录用日期:2017-09-28
刊出日期:2018-01-01
Construction of efficient nitrogen-fixing cropping pattern:Maize/faba bean intercrop with rhizobium inoculation in reclaimed low-fertility soils
MEI Peipei1, 2,,WANG Ping3,
LI Long1,,,
ZHANG Xuan1,
GUI Linguo4,
HUANG Jiancheng4
1. College of Resources and Environmental Sciences, China Agricultural University/Key Laboratory of Plant-Soil Interactions, Ministry of Education, Beijing 100193, China
2. College of Life Science and Technology, Henan Institute of Science and Technology/Henan Collaborative Innovation Centre of Modern Biological Breeding, Xinxiang 453003, China
3. Institute of Crop Science, Ningxia Academy of Agriculture and Forestry Sciences, Yinchuan 750105, China
4. Institute of Agricultural Resources and Environmental Science, Ningxia Academy of Agriculture and Forestry Sciences, Yinchuan 750002, China
Funds: the National Key Research and Development Program of China2016YFD0300202
the National Key Technology R & D Program of China2007BAD89B02
More Information
Corresponding author:Li Long, E-mail:lilong@cau.edu.cn
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摘要
摘要:为了在新开垦土壤上构建高效种植模式，本文采用温室盆栽和大田试验相结合的方法，选用4种根瘤菌接种方式（保水剂拌种、清水拌种、三叶期灌根和种子丸衣化）接种4种不同蚕豆根瘤菌（NM353、CCBAU、G254和QH258），分析接菌后新开垦土壤上玉米/蚕豆间作体系的生产潜力、地上部氮素吸收和结瘤特性以及生物固氮等方面的优势，拟为该体系筛选出高效的根瘤菌及其接种技术。结果表明：接种NM353后，玉米/蚕豆间作体系中蚕豆籽粒产量比单作平均增加152.84%，而玉米保持相对稳产；以保水剂拌种的方式接种NM353的间作蚕豆地上部氮素积累量最高，蚕豆结瘤数、瘤重、固氮比例和固氮量均高于本试验中其他3种方式接种的根瘤菌。在盛花期和盛花鼓粒期，接种NM353蚕豆的固氮比例比接种CCBAU的分别高19.1%和11.1%，在各个生育时期两者固氮量之间差异均达显著水平；接种NM353与接种其他菌种间固氮量和固氮比例差异更显著。因此，在新开垦土壤上，用保水剂拌种的方式对间作蚕豆接种NM353根瘤菌，构建玉米/蚕豆-根瘤菌高效固氮体系，为新开垦土壤合理开发利用的可持续发展模式。
Abstract:Lighe sierozem in the Ningxia Yellow River Irrigation area is characterized as newly reclaimed soil due to lower fertility and land consolidation. To rational use land resources with environmental friendly way of the soil, leguminous/non-leguminous crops intercropping plus nitrogen-fixing rhizobium is an important measure. In order to construct an efficient nitrogen-fixing cropping pattern in reclaimed lands, the efficiency of Rhizobium species and the inoculation methods were investigated aiming to the construction of a maize/faba bean-rhizobium intercropping systems through the greenhouse and field experiments with newly-reclaimed low-fertility soils. Four Rhizobium leguminosarum biovar viciae (NM353, CCBAU, G254 and QH258) and four inoculation methods (seed inoculation with water absorbent, seed inoculation with water, dip root at 3-leaf stage and seed pelleting & inoculation) were used and the relative advantages determined with respect to crop productivity, aboveground nitrogen acquisition, nodule characteristics, biological nitrogen fixation, etc. The greenhouse experiment results showed that the order of total biomass of intercropped maize and faba bean for different rhizobium was CCBAU > NM353 > G254 > QH258 > intercropping without rhizobium > sole crop without rhizobium, and for inoculation method was seed inoculation with water absorbent > seed pelleting & inoculation > seed inoculation with water > dip root at 3-leaf stage. The field experiment showed that inoculation with NM353 with seed inoculation via water absorbent method produced significant increase in yield of intercropped faba bean and maize by 79.7% and 42.8%. The intercropping system with G254 and NM353 rhizobium showed obvious yield advantage, the land equivalent ratio was >1.5. Also nitrogen content of faba bean and maize was highest among different treatments. In addition, the number of nodules, nodule biomass and nitrogen derived from air (Ndfa) and percent Ndfa (%Ndfa) were all greater in the newly-established system compared to non-inoculated faba beans. Particularly, %Ndfa of faba bean inoculated with NM353 was respectively 19.1% and 11.1% higher than that inoculated with CCBAU at peak flowering and pod-filling stages, and both had significant differences. The differences in Ndfa of faba bean were significant between rhizobium inoculation NM353 and CCBAU at each growth stage. Relatively, the difference between NM353 inoculation and the other strains was more significant for either%Ndfa or Ndfa. The inoculation with water absorbent method showed best inoculation effect with respect to nodules number, nodule biomass, %Ndfa and Ndfa. Thus an efficient cropping system was successfully set up for nitrogen fixation. The maize/faba bean-rhizobium intercropping system based on seed inoculated with Rhizobium NM353 was the most sustainable development model for rational development and utilization of the low-fertility soil.
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图1供试4种根瘤菌的OD值监测生长曲线
Figure1.OD value growth curves of the four tested rhizobia
下载: 全尺寸图片幻灯片
图2接种不同菌种对间作蚕豆、玉米地上部生物量和地上部氮积累量的影响(大田)
不同大写字母表示同一作物5个接菌处理间差异显著(P < 0.05); △, *, **和n.s.分别表示相同接菌方式下同一取样日期单、间作间差异在10%、5%和1%水平上显著, 在10%水平上差异不显著。
Figure2.Above-ground biomass and N acquisition of intercropped faba bean and maize affected by 4 different rhizobia (field experiment)
Different capital letters in the same column demonstrate significant differences among 5 treatments (4 rhizobia treatments and control with water) at 0.05 level; △, *, ** and n.s. refer to marginal significance at 10%, significance at 5%, 1% levels and not significant at 10% level, respectively, between sole and intercrop with the same rhizobium treatment at one growth stage.
下载: 全尺寸图片幻灯片
图3不同生育时期接种不同根瘤菌处理蚕豆的固氮比例(%Ndfa)和固氮量(Ndfa) (大田)
不同大写字母表示同一个生育时期5个接菌处理间差异显著(P < 0.05); △和*分别表示同一个生育时期同一接菌处理下单、间作间在10%和5%水平下差异显著。
Figure3.Percentage of nitrogen derived from air (%Ndfa) and nitrogen derived from air (Ndfa) of faba bean with different rhizobia treatments at different growth stages (field experiment)
Different capital letters demonstrate the significance of 5 treatments (4 rhizobia treatments and 1 control with water) at one growth stage at 0.05 level; △and * indicate the significant differences at 0.1 and 0.05 levels, respectively, of sole and intercrop with the same rhizobium treatment at one growth stage.
下载: 全尺寸图片幻灯片表1播种前试验区耕层土壤(0~20 cm)基础肥力状况
Table1.Background properties of soil (0-20 cm depth) in the study area before sowing
| 年 Year | 有机质 Organic matter (g·kg-1) | 总氮 Total N (g·kg-1) | 无机氮 Mineral N (mg·kg-1) | 速效磷 Olsen P (mg·kg-1) | 速效钾 Quickly available K (mg·kg-1) | 缓效钾 Slowly available K (mg·kg-1) | pH |
| 2009 | 5.69 | 0.27 | 6.14 | 2.35 | 109.25 | 278.14 | 7.41 |
下载: 导出CSV表2不同接种方法接种4个根瘤菌种后间作蚕豆、玉米地上部生物量和地上部氮吸收(盆栽)
Table2.Above-ground biomass and N acquisition of intercropped faba bean and maize affected by 4 different rhizobia and 4 different inoculation methods (pot experiment)
| 接种方法 Inoculation method | 菌株 Rhizobium | 地上部生物量 Above-ground biomass (g·pot-1) | 地上部氮累积量 Above-ground N accumulation (mg×pot-1) | |||||
| 蚕豆?Faba bean | 玉米?Maize | 整盆?Whole pot | 蚕豆?Faba bean | 玉米?Maize | 整盆?Whole pot | |||
| 保水剂拌种 Seed inoculation with water absorbent | CCBAU | ?3.94±0.48a | 5.03±1.12a | 8.96±0.80a | 99.2±15.5a | 30.9±5.3a | ?130.1±13.6a | |
| G254 | ?3.03±0.22b | 6.05±0.45a | 9.08±0.43a | 80.1±6.6a | 36.1±2.5a | ?116.2±8.2a | ||
| NM353 | ?3.62±0.17ab | 6.04±0.45a | 9.66±0.52a | 93.2±4.9a | 38.2±1.7a | ?131.4±3.7a | ||
| QH258 | ?3.51±0.13ab | 6.63±0.51a | 10.14±0.41a | 91.0±3.8a | 37.8±2.1a | ?128.8±1.8a | ||
| 平均值?Mean | ?3.53±0.32AB | 5.93±0.71A | 9.46±0.58A | 90.9±9.0A | 35.8±3.4A | ?126.6±8.2A | ||
| 清水拌种 Seed inoculation with water | CCBAU | ?3.29±0.13a | 5.61±0.73a | 8.90±0.65a | 90.2±5.8a | 34.1±4.9ab | ?124.3±7.2a | |
| G254 | ?3.32±0.12a | 5.84±0.23a | 9.15±0.28a | 81.8±6.7a | 34.1±1.8ab | ?115.9±7.2a | ||
| NM353 | ?2.46±0.50b | 6.55±0.91a | 9.00±0.60a | 71.4±16.8a | 41.9±6.4a | ?113.3±14.5a | ||
| QH258 | ?3.32±0.12a | 5.27±0.05a | 8.58±0.13a | 84.6±11.7a | 30.5±1.0b | ?115.0±12.2a | ||
| 平均值?Mean | ?3.09±0.33B | 5.81±0.60AB | 8.91±0.44AB | 82.0±10.8A | 35.2±4.5A | ?117.1±9.9A | ||
| 三叶期灌根 Dip root at 3-leaf stage | CCBAU | ?3.37±0.40ab | 5.12±0.21a | 8.50±0.27a | 88.2±11.6ab | 30.4±1.3a | ?118.5±10.4ab | |
| G254 | ?3.56±0.37ab | 5.52±0.96a | 9.08±0.61a | 93.4±18.5ab | 31.9±5.3a | ?125.3±13.2ab | ||
| NM353 | ?4.30±0.26a | 4.34±0.39a | 8.64±0.43a | 116.1±7.4a | 28.2±2.5a | ?144.3±6.2a | ||
| QH258 | ?3.15±0.38b | 5.28±0.36a | 8.43±0.43a | 81.2±12.3b | 30.9±2.1a | ?112.1±11.3b | ||
| 平均值?Mean | ?3.59±0.41A | 5.07±0.57B | 8.66±0.43B | 94.7±14.1A | 30.3±2.9A | ?125.1±11.9A | ||
| 丸衣化 Seed pelleting & inoculation | CCBAU | ?3.61±0.58a | 5.55±0.87a | 9.16±0.75a | 90.7±17.5a | 34.8±6.1a | ?125.5±14.8a | |
| G254 | ?3.77±0.47a | 5.65±0.71a | 9.42±0.58a | 99.3±11.5a | 32.7±3.0a | ?131.9±9.0a | ||
| NM353 | ?3.39±0.51a | 5.51±0.77a | 8.89±0.33a | 89.8±19.7a | 33.9±5.0a | ?123.7±15.5a | ||
| QH258 | ?2.77±0.12a | 5.88±0.31a | 8.65±0.22a | 69.9±3.1a | 32.5±1.0a | ?102.3±3.6a | ||
| 平均值?Mean | ?3.30±0.47AB | 5.64±0.63AB | 9.03±0.49AB | 87.4±14.4A | 33.5±3.9A | ?120.8±12.4A | ||
| 单作不接菌 CK-sole | 水?Water | ?3.56±0.19A | 7.12±0.33C | 86.0±4.9A | ?172.1±9.9A | |||
| 间作不接菌 CK-intercrop | 水Water | ?3.20±0.31C | 5.20±0.56B | 8.40±0.36BC | 72.9±11.8A | 29.40±2.2A | ?102.3±11.0A | |
| 表中所列数据为平均值±标准误差(n=4)。同列不同大写字母表示4个接种方式与两个对照共6个处理间差异显著(P < 0.05);同列不同小写字母表示同一接菌方式下不同根瘤菌间差异显著(P < 0.05)。Data are means ± S.E. (n= 4). Different capital letters in the same column demonstrate significant differences among 6 treatments (4 inoculation methods and 2 non-inoculation controls) at 0.05 level; different lowercase letters indicate significant differences among 4 rhizobia treatments with the same inoculation method at 0.05 level. | ||||||||
下载: 导出CSV表3根瘤菌接种处理对蚕豆/玉米间作作物籽粒产量及其间作优势的影响(大田)
Table3.Grain yields of faba bean and maize and land equivalent ratios (LER) of faba bean/maize intercropping system with 4 rhizobia inoculation (field experiment)
| 菌种 Rhizobium | 蚕豆?Faba bean | 玉米?Maize | 土地当量比 | ||||
| 单作?Sole | 间作?Intercrop | 单作?Sole | 间作?Intercrop | Land equivalent ratio | |||
| CCBAU | 2 975±533A | 5 578±1 295AB* | 7 413±315 | 8 536±832AB | 1.39±0.13AB | ||
| G254 | 2 488±638A | 5 438±1 685AB△ | 7 413±315 | 7 705±329AB | 1.53±0.74A | ||
| NM353 | 2 907±509A | 5 950±990A* | 7 413±315 | 9 029±268A** | 1.65±0.08AB | ||
| QH258 | 2 907±756A | 5 444±786AB* | 7 413±315 | 6 095±1066B | 1.28±0.33AB | ||
| Water | 1 765±293A | 3 311±905B△ | 7 413±315 | 6 322±817B | 1.22±0.44B | ||
| 增产率Over-yield ratio1) | 155.50% | 1.68% | |||||
| 表中数据为平均值±标准误差(n=3)。不同大写字母表示同一作物相同种植方式下5个接菌处理间差异显著(P < 0.05); △, *和**分别表示同一接菌处理下同一作物单、间作间在10%、5%和1%水平下差异显著。1)表示间作蚕豆(玉米)比单作蚕豆(玉米)的平均增产率。Data in the table are means ± S.E. (n = 3). Different capital letters demonstrate significant differences among 5 treatments (4 rhizobia treatments and 1 control with water) with the same crop and same cropping system at 0.05 level; △, * and ** indicate significant differences at 0.1, 0.05, and 0.01 levels, respectively, between sole and intercrop with the same rhizobium treatment.1) The over-yield ratio is average yield increasing rate of intercropped faba bean (maize) compared to that of sole faba bean (maize). | |||||||
下载: 导出CSV表4根瘤菌接种方法和根瘤菌菌种对间作蚕豆结瘤特性和生物固氮的影响(盆栽)
Table4.Nodule traits and biological N2 fixation of intercropped faba bean affected by different rhizobia with different inoculation methods (pot experiment)
| 接种方法 Inoculation method | 菌株 Rhizobium | 根瘤数 Nodule number (No×plant-1) | 根瘤干重 Nodule dry weight (g·plant-1) | 单瘤干重 Nodule size [mg(DW)×nodule-1] | 固氮比例 Percentage of nitrogen derived from air (%) | 固氮量 Nitrogen derived from air (mg·plant-1) |
| 保水剂拌种 Seed inoculation with water absorbent | CCBAU | 96.0±23.9a | 0.273±0.027a | 3.88±1.96a | 51.64±5.72b | 49.63±4.38ab |
| G254 | 69.5±18.8b | 0.218±0.022a | 3.75±1.05a | 51.62±2.17b | 41.16±2.54b | |
| NM353 | 102.0±14.9a | 0.280±0.036a | 2.78±0.30ab | 65.21±2.33a | 60.82±3.56a | |
| QH258 | 105.5±17.0a | 0.210±0.021a | 2.12±0.39b | 55.70±3.45ab | 51.02±4.82ab | |
| 平均值?Mean | 93.3±18.9A | 0.245±0.036A | 3.13±1.10BC | 56.04±3.42AB | 50.66±3.83BC | |
| 清水拌种 Seed inoculation with water | CCBAU | 85.0±11.3a | 0.230±0.007a | 2.85±0.52ab | 65.14±8.35a | 57.49±4.16a |
| G254 | 94.8±25.6a | 0.213±0.018a | 2.52±0.55b | 68.10±6.66a | 56.19±8.55a | |
| NM353 | 71.5±21.8a | 0.228±0.029a | 3.85±0.99a | 67.85±2.60a | 63.33±4.71a | |
| QH258 | 95.8±7.7a | 0.223±0.026a | 2.35±0.33b | 64.22±1.32a | 54.30±6.73a | |
| 平均值 ?Mean | 86.8±17.2A | 0.223±0.040AB | 2.89±0.67BC | 66.33±4.73A | 57.83±6.04A | |
| 三叶期灌根 Dip root at 3-leaf stage | CCBAU | 55.3±8.0a | 0.210±0.031a | 3.82±0.50b | 64.38±1.42a | 56.78±6.80a |
| G254 | 31.5±7.1b | 0.180±0.053a | 5.26±1.10a | 58.25±3.85a | 53.45±8.75a | |
| NM353 | 50.3±7.6a | 0.238±0.021a | 4.81±0.27ab | 58.85±1.74a | 61.04±2.45a | |
| QH258 | 52.8±4.0a | 0.190±0.019a | 3.65±0.46b | 58.43±4.55a | 46.17±3.16a | |
| 平均值 ?Mean | 47.4±8.3BC | 0.204±0.043B | 4.38±0.71AB | 59.98±2.89A | 54.36±5.29B | |
| 丸衣化 Seed pelleting & inoculation | CCBAU | 40.5±14.0ab | 0.200±0.046a | 6.65±2.98a | 47.97±0.53a | 38.73±3.03b |
| G254 | 64.8±19.6ab | 0.245±0.039a | 4.04±0.38b | 54.37±6.93a | 54.86±10.67a | |
| NM353 | 69.5±10.0a | 0.240±0.041a | 3.71±0.94b | 50.85±1.64a | 46.00±9.26b | |
| QH258 | 32.3±5.3b | 0.160±0.018b | 5.43±1.49a | 59.31±2.45a | 41.53±2.82b | |
| 平均值 ?Mean | 51.8±15.2B | 0.211±0.026AB | 4.96±1.71A | 53.13±2.89B | 45.28±6.45C | |
| 单作不接菌 CK-sole | 水Water | 26.8±2.4C | 0.125±0.017C | 2.34±0.27C | 31.07±0.59C | 26.86±4.04E |
| 间作不接菌 CK-intercrop | 水 ?Water | 38.5±14.6BC | 0.090±0.031C | 2.89±0.86BC | 42.11±1.37C | 30.55±2.32D |
| 表中所列数据为平均值±标准误差(n=4)。同列不同大写字母表示4个接种方式与两个对照共6个处理间差异显著(P < 0.05);同列不同小写字母表示同一接菌方式下不同根瘤菌间差异显著(P < 0.05)。Data are means ± S.E. (n = 4). Different capital letters in the same column demonstrate significant differences among 6 treatments (4 inoculation methods and 2 non-inoculation controls) at 0.05 level; different lowercase letters indicate significant differences among 4 rhizobia treatments with the same inoculation method at 0.05 level. | ||||||
下载: 导出CSV表5接种根瘤菌对间作蚕豆结瘤特性的影响(大田)
Table5.Dynamic characteristics of intercropped faba bean nodule traits affected by different rhizobia inoculation (field experiment)
| 菌株 Rhizobium | 初花期?Initial flowering stage | 盛花期?Peak flowering stage | 盛花鼓粒期?Pod-filling stage | 成熟期?Maturity | |||||||
| 单作 Sole | 间作 Intercrop | 单作 Sole | 间作 Intercrop | 单作 Sole | 间作 Intercrop | 单作 Sole | 间作 Intercrop | ||||
| 根瘤数Nodule number (No·plant -1) | |||||||||||
| CCBAU | 147.3±42.9A | 136.8±33.0A | 145.6±16.0AB | 188.0±51.4A | 143.7±46.5A | 156.1±38.2A | 159.8±33.2A | 119.6±41.8A | |||
| G254 | 181.1±49.4A | 140.3±11.0A | 162.4±36.6AB | 156.9±15.1A | 125.7±29.7A | 127.2±24.0A | 158.1±16.7A | 164.7±72.9A | |||
| NM353 | 185.7±18.7A | 123.6±17.2A△ | 231.8±35.9A | 193.7±7.6A | 191.1±15.7A | 147.8±22.0A | 115.2±14.1A | 158.2±22.6A | |||
| QH258 | 194.0±16.2A | 181.2±30.5A | 172.0±6.3AB | 168.2±34.1A | 142.7±11.6A | 149.2±16.1A | 185.3±71.7A | 165.3±44.8A | |||
| Water | 124.7±48.1A | 165.7±41.5A | 112.4±29.5B | 115.1±22.5A | 124.1±26.0A | 190.2±33.2A | 155.0±5.2A | 134.8±18.9A | |||
| 根瘤干重Nodule biomass (g·plant -1) | |||||||||||
| CCBAU | 0.251±0.128A | 0.264±0.054B | 0.944±0.040AB | 1.367±0.107A* | 1.300±0.268A | 1.674±0.435AB | 1.733±0.564A | 1.371±0.427A | |||
| G254 | 0.402±0.021A | 0.298±0.084AB | 0.867±0.120B | 0.944±0.022A | 1.318±0.233A | 1.032±0.183B | 1.650±0.196A | 2.464±0.712A | |||
| NM353 | 0.457±0.055A | 0.448±0.014A | 1.267±0.058A | 1.386±0.062A | 1.931±0.360A | 2.054±0.185A | 1.557±0.183A | 1.459±0.177A | |||
| QH258 | 0.362±0.122A | 0.415±0.064AB | 0.978±0.239AB | 1.200±0.366A | 1.399±0.272A | 1.773±0.023AB | 2.113±0.680A | 1.548±0.174A | |||
| Water | 0.229±0.055A | 0.328±0.040AB | 0.978±0.044AB | 0.978±0.308A | 1.342±0.467A | 1.884±0.349AB | 1.308±0.200A | 1.533±0.094A | |||
| 单瘤干重Nodule size [mg(DW)·nodule -1] | |||||||||||
| CCBAU | 1.3±0.7A | 2.0±0.1B | 6.7±1.0A | 8.5±2.3A | 10.8±2.8A | 10.7±0.2A | 10.5±2.9A | 12.7±2.0A | |||
| G254 | 2.5±0.5A | 2.1±0.5B | 6.6±2.9A | 6.1±0.5A | 11.0±1.4A | 8.2±0.1A△ | 10.7±1.7A | 17.0±2.9A | |||
| NM353 | 2.5±0.3A△ | 3.8±0.6A | 5.7±0.7A | 7.0±0.3A | 9.9±1.2A | 14.8±3.3A△ | 9.9±3.8A | 14.4±2.7A | |||
| QH258 | 1.9±0.6A | 2.3±0.4B | 5.6±1.2A | 7.7±2.7A | 9.6±1.2A | 12.1±1.1A | 12.2±3.0A | 11.1±3.4A | |||
| Water | 2.4±0.8A | 2.2±0.5B | 9.8±2.2A | 8.0±1.2A | 10.0±2.2A | 11.0±3.3A | 8.4±1.2A | 11.8±1.5A | |||
| 同列不同大写字母表示同一作物相同种植方式下5个接菌处理间差异显著(P < 0.05); △和*分别表示同一个生育时期同一接菌处理下单、间作间在10%和5%水平下差异显著。Different capital letters demonstrate significant differences among 5 treatments (4 rhizobia treatments and 1 control with water) with the same crop and same cropping system at 0.05 level; △ and * indicate significant differences at 0.1 and 0.05 levels, respectively, between sole and intercrop with the same rhizobium treatment. | |||||||||||
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