大豆-根瘤菌共生结瘤能力鉴定培养条件的筛选

doi:10.7668/hbnxb.20194991

摘要/Abstract

摘要：

为探究不同环境对大豆-根瘤菌共生结瘤能力的影响,筛选适宜大豆-根瘤菌共生结瘤的培养条件,设置4种环境因素共10种处理(培养温度:22,26,30 ℃;培养基质:蛭石,蛭石营养土;接种时期:0 d接种,10 d接种;摇菌终浓度:OD₆₀₀=0.5,OD₆₀₀=0.9,OD₆₀₀=1.3),组合出36种培养条件。测定36种培养条件下大豆-根瘤菌的5个结瘤相关表型,即单株根瘤数、单株根瘤干质量、根瘤大小、叶片生长10,24 d的SPAD值,运用主成分分析、模糊数学隶属函数法和聚类分析法对各条件下大豆-根瘤菌共生结瘤能力进行综合评价和分类,建立大豆-根瘤菌结瘤能力综合评价体系,并根据综合评价D值筛选最优培养条件。结果显示:培养温度是影响大豆-根瘤菌共生结瘤的主要环境因素,不同培养条件下的单株根瘤数、单株根瘤干质量、根瘤大小、叶片生长10,24 d的SPAD值的最大值对应的培养温度分别为30,26,26,30,26 ℃。在本研究设置的培养条件下,大豆-根瘤菌共生结瘤体系最优培养温度为26 ℃;通过基于综合评价D值的多元线性回归分析和灰色关联度分析得知,单株根瘤干质量和根瘤大小与大豆-根瘤菌共生结瘤能力的相关性较强,可作为结瘤能力鉴定的重要参考指标。该研究结果建立了一个大豆-根瘤菌共生结瘤能力的综合评价体系,为选择鉴定结瘤能力的表型和培养条件提供了理论依据。

关键词: 大豆, 根瘤菌, 共生培养, 结瘤能力, 综合评价

Abstract:

In order to investigate the impact of different environments on the symbiotic nodulation ability of soybean-rhizobia and to screen suitable culture conditions for symbiotic nodulation,this study established 10 treatments based on 4 environmental factors(culture temperatures:22,26,30 ℃;culture substrates:vermiculite,vermiculiteand nutrient soil;inoculation periods:0 day post-inoculation,inoculation at 10 days;final bacterial concentration:OD₆₀₀=0.5,OD₆₀₀=0.9,OD₆₀₀=1.3),resulting in 36 different culture conditions.Five nodulation-related phenotypes of soybean-rhizobia were measured under these conditions,including the number of nodules per plant,nodule dry weight per plant,nodule size,SPAD of leaf growth at 10 and 24 days post-inoculation(DPI).Principal component analysis,fuzzy mathematics membership function method,and cluster analysis were employed to comprehensively evaluate and classify the symbiotic nodulation ability of soybean-rhizobia under different conditions.A comprehensive evaluation system for the nodulation ability of soybean-rhizobia was established,and the optimal culture conditions were selected based on the comprehensive evaluation index D value.The results showed that culture temperature was the primary environmental factor influencing the symbiotic nodulation of soybean-rhizobia.The maximum values of the number of nodules per plant,nodule dry weight per plant,nodule size,SPAD of leaf growth at 10 and 24 days post-inocubion corresponded to culture temperatures of 30,26,26,30 and 26 ℃,respectively,under,different culture conditions.Under the culture conditions set,the optimal culture temperature for the symbiotic nodulation of soybean-rhizobia was 26 ℃.Multivariate linear regression analysis and grey relational analysis based on the comprehensive evaluation index D value indicated that nodule dry weight and nodule size had a strong correlation with the symbiotic nodulation ability of soybean-rhizobia,serving as important reference indicators for nodulation ability identification.The results of this study established a comprehensive evaluation system for the symbiotic nodulation ability of soybean-rhizobia,providing a theoretical basis for selecting phenotypes and culture conditions for nodulation ability identification.

Key words: Soybean, Rhizobia, Symbiotic culture, Nodulation ability, Comprehensive evaluation

赵圆, 刘智, 杨庆, 王宇, 王艳丽, 闫龙, 张锴, 史晓蕾, 刘晓燕. 大豆-根瘤菌共生结瘤能力鉴定培养条件的筛选[J]. 华北农学报, 2024, 39(S1): 125-134. doi: 10.7668/hbnxb.20194991.

ZHAO Yuan, LIU Zhi, YANG Qing, WANG Yu, WANG Yanli, YAN Long, ZHANG Kai, SHI Xiaolei, LIU Xiaoyan. Screening of Culture Conditions for the Identification of Nodulation Ability of Soybean-rhizobium Symbiosis[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(S1): 125-134. doi: 10.7668/hbnxb.20194991.

http://www.hbnxb.net/CN/Y2024/V39/IS1/125

图/表 11

参考文献 27

[1]	Hui K L, Xi B D, Tan W B, Song Q D. Long-term application of nitrogen fertilizer alters the properties of dissolved soil organic matter and increases the accumulation of polycyclic aromatic hydrocarbons[J]. Environmental Research, 2022, 215(Pt 2):114267.doi:10.1016/j.envres.2022.114267.
[2]	Yuan S L, Zhou S X, Feng Y, Zhang C J, Huang Y, Shan Z H, Chen S L, Guo W, Yang H L, Yang Z L, Qiu D Z, Chen H F, Zhou X N. Identification of the important genes of Bradyrhizobium diazoefficiens 113-2 involved in soybean nodule development and senescence[J]. Frontiers in Microbiology, 2021, 12:754837.doi:10.3389/fmicb.2021.754837.
[3]	陈文新, 汪恩涛, 陈文峰. 根瘤菌-豆科植物共生多样性与地理环境的关系[J]. 中国农业科学, 2004, 37(1):81—86.doi:10.3321/j.issn:0578-1752.2004.01.013.
	Chen W X, Wang E T, Chen W F. The relationship between the symbiotic promiscuity of rhizobia and legumes and their geographical environments[J]. Scientia Agricultura Sinica, 2004, 37(1):81-86.
[4]	Zhu Y G, Peng J J, Chen C, Xiong C, Li S L, Ge A H, Wang E T, Liesack W. Harnessing biological nitrogen fixation in plant leaves[J]. Trends in Plant Science, 2023, 28(12):1391-1405.doi:10.1016/j.tplants.2023.05.009.
[5]	Goyal R K, Mattoo A K, Schmidt M A. Rhizobial-host interactions and symbiotic nitrogen fixation in legume crops toward agriculture sustainability[J]. Frontiers in Microbiology, 2021, 12:669404.doi:10.3389/fmicb.2021.669404.
[6]	陆保福, 康文娟, 师尚礼, 关键, 南攀, 马瑞宏. 豆科植物-根瘤菌固氮系统及其碳氮互作[J]. 中国草地学报, 2023, 45(11):119-135,144.doi:10.16742/j.zgcdxb.20230023.
	Lu B F, Kang W J, Shi S L, Guan J, Nan P, Ma R H. Nitrogen fixation system of legume-rhizobia and its carbon-nitrogen interaction[J]. Chinese Journal of Grassland, 2023, 45(11):119-135,144.
[7]	Wang Y L, Liao J W, Wu J, Huang H M, Yuan Z X, Yang W, Wu X Y, Li X. Genome-wide identification and characterization of the soybean DEAD-box gene family and expression response to rhizobia[J]. International Journal of Molecular Sciences, 2022, 23(3):1120.doi:10.3390/ijms23031120.
[8]	Couchoud M, Salon C, Girodet S, Jeudy C, Vernoud V, Prudent M. Pea efficiency of post-drought recovery relies on the strategy to fine-tune nitrogen nutrition[J]. Frontiers in Plant Science, 2020, 11:204.doi:10.3389/fpls.2020.00204. pmid: 32174946
[9]	Gu J Y, Yu G Q, Zhu J B, Shen S J. The N-terminal domain of NifA determines the temperature sensitivity of NifA in Klebsiella pneumoniae and Enterobacter cloacae[J]. Science in China Series C:Life Sciences, 2000, 43(1):8-15.doi:10.1007/BF02881712.
[10]	吴月, 隋新华, 戴良香, 郑永美, 张智猛, 田云云, 于天一, 孙学武, 孙棋棋, 马登超, 吴正锋. 慢生根瘤菌及其与花生共生机制研究进展[J]. 中国农业科学, 2022, 55(8):1518-1528.doi:10.3864/j.issn.0578-1752.2022.08.004.
	Wu Y, Sui X H, Dai L X, Zheng Y M, Zhang Z M, Tian Y Y, Yu T Y, Sun X W, Sun Q Q, Ma D C, Wu Z F. Research advances of bradyrhizobia and its symbiotic mechanisms with peanut[J]. Scientia Agricultura Sinica, 2022, 55(8):1518-1528. doi: 10.3864/j.issn.0578-1752.2022.08.004
[11]	杨一晨, 杨习文, 周苏玫, 黄源, 徐利利, 付锦州, 郭芳芳, 贺德先. 不同培养条件下小麦幼苗与根系发育质量比较[J]. 西南农业学报, 2021, 34(12):2634-2639.doi:10.16213/j.cnki.scjas.2021.12.011.
	Yang Y C, Yang X W, Zhou S M, Huang Y, Xu L L, Fu J Z, Guo F F, He D X. Comparison of seedling quality and root development of wheat(Triticum aestivum L.) under different culture environments[J]. Southwest China Journal of Agricultural Sciences, 2021, 34(12):2634-2639.
[12]	Althabegoiti M J, López-García S L, Piccinetti C, Mongiardini E J, Pérez-Giménez J, Quelas J I, Perticari A, Lodeiro A R. Strain selection for improvement of Bradyrhizobium japonicum competitiveness for nodulation of soybean[J]. FEMS Microbiology Letters, 2008, 282(1):115—123.doi:10.1111/j.1574-6968.2008.01114.x. pmid: 18336548
[13]	李佳欢, 希娜, 漫静, 任成, 邓波. 苜蓿根瘤菌接种数量与方式对接种效果的影响[J]. 草地学报, 2022, 30(3):743-749.doi:10.11733/j.issn.1007-0435.2022.03.027.
	Li J H, Xi N, Man J, Ren C, Deng B. Effects of rhizobial population and inoculation method on the efficiency of Alfalfa rhizobium inoculation[J]. Acta Agrestia Sinica, 2022, 30(3):743-749.
[14]	吴树, 徐玥, 胥雅馨, 黄兴军, 吴全忠, 陈国栋, 翟云龙. 根瘤菌接种方式对复播大豆生长发育的影响[J]. 新疆农业科学, 2023, 60(1):52-60.doi:10.6048/j.issn.1001-4330.2023.01.007.
	Wu S, Xu Y, Xu Y X, Huang X J, Wu Q Z, Chen G D, Zhai Y L. Effects of Rhizobium inoculation mode on growth and development of multiple soybean[J]. Xinjiang Agricultural Sciences, 2023, 60(1):52-60.
[15]	Hasan M M, Corpas F J, Fang X W. Light:a crucial factor for rhizobium-induced root nodulation[J]. Trends in Plant Science, 2022, 27(10):955-957.doi:10.1016/j.tplants.2022.07.002.
[16]	Verhoeven E, Barthel M, Yu L F, Celi L, Said-Pullicino D, Sleutel S, Lewicka-Szczebak D, Six J, Decock C. Early season N₂O emissions under variable water management in rice systems:source-partitioning emissions using isotope ratios along a depth profile[J]. Biogeosciences, 2019, 16(2):383-408.doi:10.5194/bg-16-383-2019.
[17]	侯慧云, 高峰, 高同国, 朱宝成. 施氮对大豆结瘤、固氮及产量影响的研究进展[J]. 江苏农业科学, 2022, 50(8):42-48.doi:10.15889/j.issn.1002-1302.2022.08.009.
	Hou H Y, Gao F, Gao T G, Zhu B C. Research progress on effects of nitrogen application on soybean nodulation,nitrogen fixation and yield[J]. Jiangsu Agricultural Sciences, 2022, 50(8):42-48.
[18]	Montiel J, García-Soto I, James E K, Reid D, Cárdenas L, Napsucialy-Mendivil S, Ferguson S, Dubrovsky J G, Stougaard J. Aromatic amino acid biosynthesis impacts root hair development and symbiotic associations in Lotus japonicus[J]. Plant Physiology, 2023, 193(2):1508-1526.doi:10.1093/plphys/kiad398. pmid: 37427869
[19]	杨涛, 黄雅婕, 李生梅, 任丹, 崔进鑫, 庞博, 于爽, 高文伟. 海岛棉种质资源表型性状的遗传多样性分析及综合评价[J]. 中国农业科学, 2021, 54(12):2499-2509.doi:10.3864/j.issn.0578-1752.2021.12.002.
	Yang T, Huang Y J, Li S M, Ren D, Cui J X, Pang B, Yu S, Gao W W. Genetic diversity and comprehensive evaluation of phenotypic traits in sea-island cotton germplasm resources[J]. Scientia Agricultura Sinica, 2021, 54(12):2499-2509. doi: 10.3864/j.issn.0578-1752.2021.12.002
[20]	王晓丽, 王敏, 岳爱琴, 郭数进, 王鹏, 王利祥, 杨婷婷, 张海生, 张永坡, 高春艳, 张武霞, 牛景萍, 杜维俊, 赵晋忠. 氮素营养和根瘤菌接种对大豆结瘤固氮和生长的影响[J]. 华北农学报, 2022, 37(1):95-102.doi:10.7668/hbnxb.20192484.
	Wang X L, Wang M, Yue A Q, Guo S J, Wang P, Wang L X, Yang T T, Zhang H S, Zhang Y P, Gao C Y, Zhang W X, Niu J P, Du W J, Zhao J Z. Effects of nitrogen nutrition and Rhizobium inoculation on nodulation and nitrogen fixation and growth of soybean[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(1):95-102.
[21]	刘函西, 吕浩, 郭广雨, 刘冬旭, 石岩, 孙志君, 张泽鑫, 张艳娇, 文莹楠, 王洁琦, 刘春燕, 陈庆山, 辛大伟, 王锦辉. 大豆根瘤菌HH103 rhcN突变对结瘤能力的影响[J]. 中国农业科学, 2021, 54(6):1104-1111.doi:10.3864/j.issn.0578-1752.2021.06.003.
	Liu H X, Lü H, Guo G Y, Liu D X, Shi Y, Sun Z J, Zhang Z X, Zhang Y J, Wen Y N, Wang J Q, Liu C Y, Chen Q S, Xin D W, Wang J H. Effect of rhcN gene mutation on nodulation ability of soybean Rhizobium HH103[J]. Scientia Agricultura Sinica, 2021, 54(6):1104-1111.
[22]	Hu Y A, Chen Y L, Yang X, Deng L S, Lu X. Enhancing soybean yield:the synergy of sulfur and rhizobia inoculation[J]. Plants, 2023, 12(22):3911.doi:10.3390/plants12223911.
[23]	姚佳睿, 吕金钊, 陈康, 李玉红. 不同油麦菜品种萌发期耐热指标筛选及耐热性评价[J]. 西北农业学报, 2024, 33(3):511-520.doi:10.7606/j.issn.1004-1389.2024.03.013.
	Yao J R, Lü J Z, Chen K, Li Y H. Screening of indexes for heat tolerance assessment during germination in various lettuce varieties[J]. Acta Agriculturae Boreali-occidentalis Sinica, 2024, 33(3):511-520.
[24]	代重阳, 孙继鑫, 王聪. 不同耐盐性菜用大豆与大豆根瘤菌的共生匹配性研究[J]. 北方农业学报, 2019, 47(6):90-95.doi:10.3969/j.issn.2096-1197.2019.06.16.
	Dai C Y, Sun J X, Wang C. Symbiotic matching between soybean rhizobium and vegetable soybean with different salt tolerance[J]. Journal of Northern Agriculture, 2019, 47(6):90-95. doi: 10.3969/j.issn.2096-1197.2019.06.16
[25]	朱丽丽, 张发玉, 安宁, 陈志国. 116份藜麦种质资源萌发期抗旱性综合评价[J]. 干旱地区农业研究, 2024, 42(1):23-31.doi:10.7606/j.issn.1000-7601.2024.01.03.
	Zhu L L, Zhang F Y, An N, Chen Z G. Comprehensive evaluation of drought resistance of 116 quinoa germplasm resources during germination[J]. Agricultural Research in the Arid Areas, 2024, 42(1):23-31.
[26]	薛贤滨, 贾琼, 陈峥峰, 黎瑞源, 陈庆富, 石桃雄. 基于主成分分析的苦荞麦重组自交系农艺性状综合评价[J]. 浙江农业学报, 2024, 36(4):748-759.doi:10.3969/j.issn.1004-1524.20230489.
	Xue X B, Jia Q, Chen Z F, Li R Y, Chen Q F, Shi T X. Comprehensive evaluation of agronomic characteristics of recombinant inbred lines of tartary buckwheat based on principal component analysis[J]. Acta Agriculturae Zhejiangensis, 2024, 36(4):748-759. doi: 10.3969/j.issn.1004-1524.20230489
[27]	杜普旋, 刘军, 陈荣华, 吴柔贤, 范呈根, 郭丹丹, 鲁清. 广东省花生种质资源收集与鉴定评价[J]. 植物遗传资源学报, 2023, 24(3):671-679.doi:10.13430/j.cnki.jpgr.20221010001.
	Du P X, Liu J, Chen R H, Wu R X, Fan C G, Guo D D, Lu Q. Systematic collection,identification and evaluation of peanut germplasm resources in Guangdong Province[J]. Journal of Plant Genetic Resources, 2023, 24(3):671-679.

性状 Trait	平均值±标准差 Mean±s	最大值 Max	最小值 Min	变异系数/% CV
单株根瘤数Nodule number	18.27±12.19	57.75	4.92	66.72
单株根瘤干质量/g Nodule dry weight	0.02±0.01	0.03	7.50E-04	48.31
根瘤大小/g Nodule size	1.10E-03±7.50E-04	3.33E-03	1.47E-04	68.18
10 d叶绿素相对含量10 d SPAD	36.69±11.36	53.13	14.03	30.96
24 d叶绿素相对含量24 d SPAD	25.82±4.87	33.35	16.41	18.87

性状 Trait	平均值±标准差 Mean±s	最大值 Max	最小值 Min	变异系数/% CV
单株根瘤数Nodule number	18.27±12.19	57.75	4.92	66.72
单株根瘤干质量/g Nodule dry weight	0.02±0.01	0.03	7.50E-04	48.31
根瘤大小/g Nodule size	1.10E-03±7.50E-04	3.33E-03	1.47E-04	68.18
10 d叶绿素相对含量10 d SPAD	36.69±11.36	53.13	14.03	30.96
24 d叶绿素相对含量24 d SPAD	25.82±4.87	33.35	16.41	18.87

主成分Principal component	主成分1 PC1	主成分2 PC2
特征值Eigenvalues	2.26	1.97
贡献率Contribution	45.26	39.32
累计贡献率Cumulative contribution rate	45.26	84.58
单株根瘤数Nodule number per plant	0.74	-0.56
单株根瘤干质量Nodule dry weight per plant	0.84	0.47
根瘤大小Nodule size	0.26	0.93
10 d叶绿素相对含量10 d SPAD	0.87	0.12
24 d叶绿素相对含量24 d SPAD	-0.43	0.75

主成分Principal component	主成分1 PC1	主成分2 PC2
特征值Eigenvalues	2.26	1.97
贡献率Contribution	45.26	39.32
累计贡献率Cumulative contribution rate	45.26	84.58
单株根瘤数Nodule number per plant	0.74	-0.56
单株根瘤干质量Nodule dry weight per plant	0.84	0.47
根瘤大小Nodule size	0.26	0.93
10 d叶绿素相对含量10 d SPAD	0.87	0.12
24 d叶绿素相对含量24 d SPAD	-0.43	0.75

处理条件 Processing condition	综合指标(C) Comprehensive character indicators		隶属函数(μ) Membership function		综合得分(D) Composite scores	排名 Order
处理条件 Processing condition	X1	X2	μ1	μ2	综合得分(D) Composite scores	排名 Order
I₁₃	1.07	2.29	0.69	1.00	0.84	1
I₂₁	0.82	2.26	0.63	0.99	0.80	2
I₁₇	0.48	1.71	0.54	0.85	0.69	3
I₁₅	0.41	1.67	0.53	0.84	0.67	4
I₂₃	-0.18	1.55	0.38	0.81	0.58	5
I₃₄	2.26	-1.53	1.00	0.01	0.54	6
I₂₇	0.35	0.47	0.51	0.53	0.52	7
I₂₉	0.29	0.52	0.50	0.54	0.52	8
I₃₂	2.13	-1.58	0.97	0.00	0.52	9
I₁₆	-0.17	0.81	0.38	0.62	0.49	10
I₁₄	-0.26	0.86	0.35	0.63	0.48	11
I₃₆	1.54	-1.24	0.82	0.09	0.48	12
I₃₃	1.01	-0.67	0.68	0.23	0.47	13
I₁₉	-0.43	0.95	0.31	0.65	0.47	14
I₂₈	0.71	-0.37	0.60	0.31	0.47	15
I₂₅	0.40	-0.02	0.52	0.40	0.47	16
I₂₂	0.09	0.24	0.45	0.47	0.46	17
I₂₆	0.58	-0.34	0.57	0.32	0.45	18
I₂₄	-0.21	0.52	0.37	0.54	0.45	19
I₃₁	1.03	-1.02	0.69	0.14	0.43	20
I₁₈	-0.42	0.64	0.31	0.57	0.43	21
I₂₀	-0.09	0.12	0.40	0.44	0.42	22
I₂	0.07	-0.12	0.44	0.38	0.41	23
I₃₀	0.77	-0.93	0.62	0.17	0.41	24
I₃₅	0.38	-0.70	0.52	0.23	0.38	25
I₈	0.01	-0.41	0.43	0.30	0.37	26
I₄	-1.06	-0.52	0.15	0.27	0.21	27
I₇	-1.25	-0.41	0.10	0.30	0.19	28
I₁	-1.19	-0.49	0.12	0.28	0.19	29
I₁₁	-1.23	-0.49	0.11	0.28	0.19	30
I₉	-1.31	-0.51	0.09	0.28	0.18	31
I₆	-1.06	-0.79	0.15	0.20	0.17	32
I₃	-1.29	-0.54	0.09	0.27	0.17	33
I₅	-1.34	-0.53	0.08	0.27	0.17	34
I₁₀	-1.23	-0.66	0.11	0.24	0.17	35
I₁₂	-1.65	-0.74	0.00	0.21	0.10	36

选择文件类型/文献管理软件名称

选择包含的内容