History of Research on Nitrogen Fixation in Soybeans (1887-2018)

William Shurtleff, Akiko AoyagiISBN: 978-1-948436-00-7

Publication Date: 2018 Nov. 20

Number of References in Bibliography: 3002

Earliest Reference: 1024 BCE

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Brief Chronology/Timeline of Research on Nitrogen Fixation and Inoculation in Soybeans

The earliest entries here concern biological nitrogen fixation by legumes, but do not mention soybeans.

1000 BCE – The ancient Chinese character for soybeans and legumes is written on bronze vessels of around that date with a horizontal stroke in the middle and three dots below that which are widely interpreted to represent nodules on the roots (Hu Daojing 1963. May, p. 111-19; Ho Pi-ting 1969. Oct., p. 28-30, 35).

50-79 CE – Pliny, the Roman scholar, wrote: “The bean ranks first among the legumes. It fertilizes the ground in which it has been sown as well as any manure” (George T. Moore 1905, p. 12).

1888 May 17 – At a meeting of the Royal Society, J.B. Lawes, the founder of Rothamsted Experimental Station, and J.H. Gilbert, his colleague of many years, present a paper titled “On the present position of the question of the sources of nitrogen of vegetation.” They first describe in English the landmark work of H. Hellriegel and H. Wilfarth which had been reported a few months earlier at a meeting in Germany (Eaglesham 1989, p. 29).

1888 Nov. 16 – The first to propose that the organisms involved in nitrogen fixation are bacteria is Martinus W. Beyerinck (correctly spelled Beijerinck in Dutch).

He is also the first to successfully to name or cultivate these bacteria. He names them Bacillus Radicicola (Botanische Zeitung. Nov. 16, Nov. 23, Nov. 30).

1888 Nov. – Hermann Hellriegel and Hermann Wilfarth, German scientists, publish the key, classical paper on the acquisition of atmospheric nitrogen by legumes. The authors had performed seminal experiments at the agricultural station of Bernburg / Saale (later in eastern Germany) which proved that leguminous plants can utilize not only combined nitrogen in the soils but also atmospheric dinitrogen (N2) gas for growth. They showed that these plants cannot utilize dinitrogen gas directly, but are strictly dependent for this function on soil bacteria, which they harbour in nodules on their roots.

At first the observations of Hellriegel and Wilfarth were by no means universally accepted by botanists. Later, however, their results were fully substantiated by many other researchers, and the explanation of the long unsolved problem was made possible. Thus the discovery of the fact that the nodules of legumes enabled them to fix “free nitrogen” is usually ascribed to Hellriegel and Wilfarth.

1889 Oct. 25 – B. Frank in Germany changes the scientific name of the bacteria that inhabit the root nodules of legumes from Bacillus radicicola (Beijerinck, 1888) to Rhizobium leguminosarum.

Here the word “Rhizobium” first appears in connection with legume nodules.

1890 March – The earliest document seen concerning soybeans and atmospheric nitrogen fixation in the United States is by J.B. McBryde of the South Carolina Experiment Station (Bulletin No. 8, p. 79). He states, simply and without citation, that the soja bean probably obtains its “nitrogen from the air and subsoil...”

1891 Jan. – The word “tubercles” (or “tubercle,” so spelled) is first used in English in connection with soybeans to refer to (what was later called) the nodules on their roots (Goessmann p. 171-72).

1891 June – Herbert W. Conn, Professor of Biology at Wesleyan Univ. in Middletown, Connecticut, writes an excellent article titled “The nature of the root tubercles of leguminous plants – a review” in the Experiment Station Record (USDA, p. 686-93). He summarizes and cites important earlier work on the subject, only that of Ward being in English. He reviews the research of Woronin (1866), Erickson (1874), Brunchorst (1885), Tschirch (1887), Van Tieghem and Doulliot (1888), Hellriegel and Wilfarth (1886), Beyerinck (1888), Ward (1887), Prazmowski (1890), Frank (1888), and Laurent (1891. Soybeans are not mentioned.

1891 – An article by W.O. Atwater and C.D. Woods titled “The acquisition of atmospheric nitrogen by plants” appears in the Connecticut (Storrs) Agricultural Experiment Station, Annual Report for the year 1890 (p. 12-14).

This is the earliest document seen concerning inoculation of the soybean in the United States. It is also the earliest English-language document seen that uses the word “inoculated” (or any of its cognates) in connection with acquisition of atmospheric nitrogen by plants.

1893 Jan. – The word “bacilli” (or “bacillus”) and the term “tubercle bacilli” are first used in English in connection with nitrogen fixation in soybeans (Brooks, p. 152).

1893 March – The word “nodules” and the word “bacteria” are first used in English in connection with nitrogen fixation in soybeans (Sessions 1893).

1893. – William P. Brooks of the Massachusetts (Hatch) Agricultural Experiment Station conducts what is now considered a classic experiment. He placed never before cropped soil into pots and planted seed from three soybean cultivars originally from Japan. In one series of pots he added a pinch of dust collected from the floor where soybeans had been thrashed and the other series of pots were his control. The results were striking. In the pots receiving a pinch of dust, the plants were greener, more vigorous, and the seed yields much larger than the controls. In addition, the roots of the plants that received the pinch of dust were found to contain nodules. Soil from Brook's experiment was sent to New Jersey and Kansas stations and his results were confirmed (Brooks 1895).

1894 March 29 – In the prestigious British journal Nature, H. Marshall Ward writes (in English) a long, detailed, and carefully reasoned review and analysis of the various experiments concerning nitrogen fixation by legumes. He uses the words “fix” and “fixation” (referring to nitrogen) throughout (p. 511-14).

1896 Oct. 27 – J. Augustus Voelcker, in the Journal of the Royal Society of England, is the first to mention Nitragin. He begins with a detailed discussion of the development and commercialization (by Feb. 1896) of Nitragin, and the research of Messrs. Hellriegel, Wilfarth, and Prof. Nobbe of Tharland, Saxony.

1896 Dec. – Kalman Kerpely, writing in Hungarian in the journal Koztelek (Common Ground) is the second earliest person known to mention Nitragin.

1897 Jan. – Charles A. Goessman, writing in the Massachusetts (Hatch) Agricultural Experiment Station, Annual Report for the year 1896, is the first person in the United States to mention Nitragin. He also includes a translation (from German) of a publication on Nitragin by Nobbe and Hiltner.

Nitragin, a pure culture sold in a bottle which is sufficient for inoculation of 2.5 roods, is manufactured by Farbwerke vorm. Meister Lucius & Bruning, Hoechst on Main, Germany. A list of various kinds of Nitragin developed for 19 leguminous crops is given. Soy beans are not included in this list.

This article is also the first that uses the word “micro-organisms” (regardless of hyphenation) in connection with the acquisition of atmospheric nitrogen by plants.

1897 – David Fairchild writes to Japan and imports several pounds of soil from a soybean field (Fairchild 1948, p. 14).

1898 Dec. 6 – Nitragin is registered (by Nobbe and Hiltner) as a trademark in the United States.

1898 – M.W. Munson of the Maine Agricultural Experiment Station writes “The Acquisition of Atmospheric Nitrogen,” the longest and best review of the subject seen to date. Published in the Station’s Annual Report (p. 114-40), it contains 120 references and is 27 pages long.

In it, the words “inoculum” and “inoculation” are first used in connection with soybeans. The commercial inoculum is Nitragin, made in Germany (Munson, p. 114-40).

1900 June 1 – The scientific name Bacillus radicicola is first used to describe the organism producing nodules on the roots of soybeans (Los Angeles Times, p. I12).

1901 – The term “nitrogen fixation” and the terms “bacteriology” or “bacteriologists” (or any of its congeners except “bacteria”) is first used in connection with soybeans in English (Conn 1901, p. 136-37).

1903 Aug. 4 – The term to “fix nitrogen” is first used in connection with soybeans. The New York Times reports (p. 6): “The Secretary of Agriculture will ask Congress next Winter for a special appropriation for carrying on the work of soil inoculation. This is a new branch of work in the department, which has grown out of the discoveries made within the last few years in regard to the dependence of leguminous plants on bacteria which live on their roots. The discovery is described by scientists as one of the most important of those made as the results of modern agricultural experimentation.”

1903 Oct. 23 – The U.S. Department of Agriculture announces that it will provide free and effective nitrogen-fixing bacteria to farmers of soybeans and other legumes (Eureka Reporter (Utah), p. 6).

This same article is the first to use the word “tubercules” to refer to the nodules on the roots of leguminous plants.

1903 – George T. Moore of USDA’s Bureau of Plant Industry writes a long article titled “Bacteria and the nitrogen problem” in the Yearbook of the United States Department of Agriculture (for the year 1902, p. 333-42). He discusses the many failures that farmers have experienced trying to inoculate their legumes and says that the government has decided to start making legume inoculants and to distribute them free to charge to farmers. Photos show the roots of soybean plants with and without nodules.

In this article Moore is also the first to use the term “nitrogen fixing bacteria” in connection with soybeans.

1905 Jan. 23 – “Reports upon the Successful Use of Artificial Cultures by Practical Farmers,” by George T. Moore, lists the number of soy bean cultures sent free of charge by USDA to each of many U.S. states and foreign countries (USDA Bureau of Plant Industry 71. 72 p.).

1905 Sept. 18 – The legume inoculant “Nitro Cultures” is first mentioned in connection with soybeans; it is made by the National Nitro Culture Company of West Chester, Pennsylvania (Atlanta Constitution, p. 452).

1907 – The word “Rhizobium” is first used in English to refer to the genus to which soybean bacteria belong (Greig-Smith, p. 264-94). Note that this genus name had been coined in 1889 by B. Frank in Germany but was not use in connection with soybeans until 1907.

1908 April – The legume inoculant “Farmogerm” is first mentioned in connection with soybeans; it is made by Earp-Thomas Farmogerm Co. in Bloomfield, New Jersey (Country Gentleman, p. 452).

1910 – The word “bacteroids” is first used in connection with the root nodules of plants. Concerning bacteroids: “The first successful, artificial production of nodules by the aid of pure cultures was made by A. Prazmowski. This worker, in view of the absence of the sporogenic faculty in these organisms, changed the name of Bacillus radicicola, bestowed on them by Beyerinck, into Bacterium radicicola” (Lafar. 1910, p. 266-69).

1911 – Research on nitrogen fixation in soybeans is first reported in Latin America (Reports of the Botanic Station and Experiment Plots… Antigua, p. 16-29).

1911 – Research on nitrogen fixation in soybeans is first reported in Oceania (Agricultural Gazette of New South Wales, p. 592-94).

1914 April – The legume inoculant “Nitro-Germ” is first mentioned in connection with soybeans; it is made by Mulford Co. in Philadelphia, Pennsylvania (Northwestern Druggist, p. 82).

1915 May 20 – The term “cross inoculation” is first used in connection with nitrogen-fixing bacteria that will nodulate more than one group of legumes (W.D.R., p. 1021).

1923 July – Research on nitrogen fixation in soybeans is first reported in Africa (J. of the Department of Agriculture, Union of South Africa, p. 14-15).

1924 June – The word “nodulation” is first used in connection with nitrogen fixation in soybeans (Soil Science, p. 439-47).

1926 Nov. – The term “cross-inoculation group” and the term “cross-inoculation groups” are first used in connection with nitrogen-fixing bacteria that will nodulate more than one group of legumes (Fred et al. p. 40-43).

1926 – The scientific name Rhizobium japonicum is first used to describe the organism producing nodules on the roots of soybeans (Buchanan p. 81-90).

1927-1928 – According to the USDA, nitrogen-fixing bacteria specifically for soybeans are now available (apparently free of charge) from 15 state agricultural stations, one each in Georgia, Idaho, Maryland, Massachusetts, Michigan, Missouri, New York, North Carolina, Oregon, Pennsylvania, South Carolina, Virginia, Washington state, Wisconsin, and Wyoming.

For the for the year 1927/28 alone (according to a letter from Prof. Albrecht) the Missouri Agricultural Experiment Station (in Columbia) sent out enough bacteria for the inoculation of 38,500 bushels of soybean seed. This is equivalent to about 75% of the soybean area cultivated in that state (Prinz, 1929, p. 2).

1928 – I.L. Baldwin of the University of Wisconsin points out that there are various degrees of inoculation, ranging from well-inoculated / nodulated to poorly inoculated. The latter microorganisms are basically parasites, living at the expense of the plant.

1931 – The legume inoculant “Radicin” is first mentioned in connection with soybeans; it is made by the Radicin Institute, perhaps in Westerrade, Germany (Hospodarsky Ozor, p. 432).

1932 July – A landmark work by Edwin B. Fred and co-workers summarizes the published research in biological nitrogen fixation from approximately 1576 through 1930. A pioneering effort that brought together years of source material from English, German, French, Italian, Dutch and the Scandinavian languages, it discusses more than 975 publications on this subject.

The word “rhizobia” is also first used in connection with soybeans in this work.

1937 Dec. – In Wisconsin Agricultural Experiment Station Bulletin No. 439 we read: “Years ago it was thought that if legume plants had nodules of any kind on their roots, then they were performing satisfactorily their task of soil-building. This idea has now been exploded by the finding that a great many nodule forming strains of bacteria are not of much help, hence it is important to select good strains for inoculation purposes.”

1938 Sept. – The legume inoculant “Legume-Aid” is first mentioned in connection with soybeans; it is made by Agri Lab Inc. in Columbus, Ohio (Proceedings of the American Soybean Association, p. 69).

1939 Oct. – The word “serological” is first used to describe nitrogen fixing bacteria on soybeans (J. of Bacteriology, p. 401-10).

1941 Nov. – The legume inoculant “Vaccinograines” is first mentioned in connection with soybeans; it is made by Société des Produits Labotechniques in Paris, France (Revue Internationale du Soja, p. 215).

1964 Sept. – The word “serogroup” and the term “serogroup 123” are first used to describe nitrogen fixing bacteria on soybeans (Soybean Digest, p. 63-65).

1965 April – The genus name Bradyrhizobium is first used to refer to for nitrogen-fixing bacteria on soybeans (Agronomy Journal, p. 179-85).

1971 – The term “biological nitrogen fixation” is first used in connection with soybeans (Sundara Rao, p. 287-91).

1975 – The NifTAL Project is founded by USAID at the University of Hawaii’s College of Tropical Agriculture, Paia, Maui, Hawaii. NifTAL stands for Nitrogen Fixation by Tropical Agricultural Legumes. In 1978 it publishes a review of the literature titled The Legume-Rhizobium Symbiosis in Tropical Agriculture: a Selective Bibliography with Annotations, compiled by John Bose II, with 1181 references.

1977 – The words “promiscuous” and “promiscuously” are first used to refer to soybean varieties which are nodulated by indigenous/native bacteria (Johanna Döbereiner, p. 3-12).

By breeding such soybean varieties, with attention focused on the native soil bacteria, Brazilian farmers would be able to move into the huge cerrado area and African farmers would be able to develop a new soybean industry.

1982 Jan. – The scientific name Bradyrhizobium japonicum is first used for the slow-growing soybean root nodule bacterium so widespread in North America. The species had previously been named Rhizobium japonicum (International J. of Systematic Bacteriology, p. 136-39).

1982 – “The African Association for Biological Nitrogen Fixation (AABNF) was founded in 1982 with the support of the International Institute of Tropical Agriculture (IITA) at Ibadan, Nigeria, as part of the IITA/United Nations Development Programme (UNDP) project GLO/77/013 on 'Maximising nitrogen fixation by cowpeas and soybeans in farming systems in the humid tropics'. The AABNF is a multidisciplinary group, bringing together soil scientists, microbiologists, agronomists, climatologists, social scientists, breeders, plant physiologists, biotechnologists, nutritionists, policymakers and others interested in the promotion of biological nitrogen fixation systems in Africa.”

The first AABNF meeting was held in July 1984 at Nairobi, Kenya.

1984 Oct. – The scientific name Rhizobium fredii is first used for the fast-growing soybean root nodule bacteria with generation times of less than 6 hours (International J. of Systematic Bacteriology, p. 484-86).

Note that this bacterium was first reported by Keyser et al. in March 1982.

1990 Nov. – Burias and Planchon of France ask the important question: What would happen if, instead of breeding for primarily yield, soybean breeders worked to increase soybean productivity through selection for nitrogen fixation? (Agronomy Journal, Nov/Dec., p. 1031-34).

1990 – Assessment of the Soil Nutrient Depletion in sub-Saharan Africa, 1983-2000. Volume I: Main report, by Stoorvogel and Smaling is published. It is the earliest known study of nutrient balances on smallholder farming systems in Africa. Soil fertility in Africa is being rapidly depleted. An important part of the solution is nitrogen fixation by legumes, especially cowpeas and soybeans.

1991Nitrogen Fixation in Tropical Cropping Systems, by Ken E. Giller and Kate J. Wilson is published by CAB International (xi, 313 p.). It contains several excellent sections on soybeans.

One notes (p. 130): “In the centre of diversity of soyabean and in countries of South-east Asia, where the crop has been grown for centuries, soyabean usually nodulates without inoculation. Strains of Bradyrhizobium isolated from nodules of other legume hosts, such as cowpea, grown in the same soils, can also nodulate the local soyabean genotypes effectively. When the nodulation of soyabean genotypes from North America and from Asia was compared in the field in Tanzania, many of the Asian varieties nodulated effectively with indigenous strains of Bradyrhizobium whilst nodulation of the North American varieties was poor (Chowdhury, 1977). Asian varieties also nodulated well in the field in Nigeria whilst American varieties formed very few nodules (Nangju, 1980).” The reasons for this important difference are explained.

Revised and expanded editions of this book are published in 2001 and 2011. The new frontier of nitrogen-fixation research is in the tropics.

1991 – In an article titled “Effect of deep placement of controlled release nitrogen fertilizer (coated urea) on growth, yield and nitrogen fixation of soybean plants,” by Y. Takahashi et al., Japanese researchers in Niigata start a completely new line of research on how to prevent soybean plants from reducing nitrogen fixation when grown in soil amply fertilized with nitrogen fertilizer (Soil Science and Plant Nutrition 37(2):223-31).

This new idea contributed to a stable increase in soybean yield. But is it practical and economically feasible?

1994 – In this key paper. Japanese researchers continue to explore the “Effect of deep placement of coated urea slow release nitrogen fertilizer…” (Japanese J. of Soil Science and Plant Nutrition 65:41-47).

2000 March – Mpepereki et al. write “Soyabeans and sustainable agriculture: 'promiscuous' soyabeans in southern Africa” – a superb, long article (Field Crops Research p. 137-49).

2005 – M. Hungria and co-workers of Brazil write a brilliant 30+ page article titled “Inoculant preparation, production and application.” Basic principles must be fundamentally revised when soybeans are grown in hot, acidic, tropical soils.

2006 Sept. – Anne Willems writes “The taxonomy of rhizobia: An overview,” in Plant and Soil (287(1/2):3-14. This extremely interesting article also contains an excellent history of biological nitrogen fixation.

2014 – International Institute of Tropical Agricola (IITA) in Ibadan, Nigeria starts to make NoduMax commercial legume inoculant suited for Nigerian conditions. (IITA Annual Report 2014, 25-26).

2018 – Although biological nitrogen fixation has been understood for more than 130 years, scientists today are still working to solve many unanswered puzzles. They are increasingly working on the level of molecules and genes.

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