At first, in 1857, a proposal via Siegelsdorf, Windsheim, Marktbreit and Ochsenfurt had been favoured, which led to an alternative route being put forward from the Neustadt and Kitzingen area, which ultimately prevailed.

On 1 July 1886, a heavy head-on collision of two trains took place between Würzburg and Rottendorf, causing the death of 16 people.

On 8 December 1889, the Bavarian State Parliament approved the doubling of the single-track line between Fürth and Rottendorf,[4] which was completed by 1891.

On 10 June 1928 at 02:30, the locomotive of the night train D 47, a class S3/6, derailed while crossing the Zenngrund bridge shortly before the entry to Siegelsdorf station.

In the Nazi period, the first preliminary work began to connect the line from Mainbernheim via a wye with the Würzburg–Treuchtlingen railway (near Marktbreit).

[5] The electrification of the line began on 8 August 1952 with the installation of the first catenary mast in Neustadt (Aisch) station and it was formally opened on 2 October 1954 with a special train.

[5] A pilot installation of train radio, which had been approved in 1959, was introduced in 1962; this culminated in the nationwide introduction of the ZBF 70 system in 1974.

As a result, the hand-operated block and junction signal boxes at Kohlenhof, Rothenburger Straße and Neusünderbühl on the Nuremberg–Fürth section were replaced.

The 19th century line had a minimum railway curve radius of 2,000 Bavarian feet (about 600 m) causing numerous speed limits of 100 km/h (62 mph).

The first Federal Transport Infrastructure Plan (Bundesverkehrswegeplan, 1973) identified the line from Würzburg to Nuremberg and continuing to Augsburg as one of eight railway development projects.

[11] Further development was foreseen as part of the ABS/NBS Würzburg–Nürnberg–München, which was included as one of 13 new projects that were identified as urgent needs in the 1985 Federal Transport Infrastructure Plan.

[2] Step-free platform access was created at the stations of Dettelbach, Kitzingen, Iphofen, Markt Bibart and Emskirchen.

[2] A number of measures were envisaged within the scope of the further upgrades envisaged in the Federal Transport Infrastructure Plan of 1985: in addition to the provision of alignment improvements and the shortening of blocks, short overtaking loops, level crossings and slow points (especially in Würzburg Hauptbahnhof) were to be eliminated.

The Linienzugbeeinflussung cab signalling and train protection system would be installed and a freight detour route (Würzburg–Ansbach–Nuremberg) would be developed.

[16] As a result of line improvements, the speed in the section between Neustadt (Aisch) and Iphofen was raised by 40 to 60 km/h to 200 km/h by May 1999; this also required the installation of the Linienzugbeeinflussung system.

[7] Away from the high-speed section, smaller increases in speed were realised, for example, by raising the cant up to 160 mm, minor changes to transition curves and shifts of tracks of up to 60 cm.

[7] As part of the upgrade, Markt Bibart station was equipped with two external platforms and a crossover and a turnback track were built for commuter trains.

[7] Due to a lack of funding, plans for further upgrading of the line estimated to cost €140 million were abandoned at the end of 2004.

[22] The list of "urgent needs" identified in the Federal Transport Plan of 1992 included a new line between Iphofen and Rottendorf as part of proposed upgrades on the Hanau–Nantenbach/Würzburg–Iphofen route, which were estimated to cost DM 1.495 billion.

Near Markt Einersheim the new line would connect with the section towards Neustadt (Aisch) / Nuremberg, which is designed for 200 km/h operations.

Also, the estimated cost of DM 850 million and the ecological sensitivity of the areas crossed by the proposed line were emphasised.

LINT sets run between Nuremberg and Siegelsdorf, continuing over the Zenn Valley Railway to Markt Erlbach.

A total of eight pairs of Regionalbahn services operate from Monday to Friday during peak hours between Kitzingen and Würzburg.

With the Bayerische Eisenbahngesellschaft mbH (Bavarian Railway Company, BEG) putting operations on this line to tender, there has been some increase of services in the off-peak (especially in the evening and at the weekend).

The number of passengers on regional services on the line increased by 220 percent between 2003 and 2012 according to the Bayerische Eisenbahngesellschaft.

However, the portion working concept was rejected and instead additional Alstom Coradia Continental services were commissioned.

A DB Regio subsidiary won the tender and started operations in December 2009 with new electric railcars.

When the Verkehrsverbund Großraum Nürnberg (Greater Nuremberg Transport Association, VGN) was established on 27 September 1987, the line between Nuremberg and Neustadt an der Aisch was in its tariff area, it was extended to Markt Bibart on 31 May 1992, to Iphofen on 10 December 2006, to Kitzingen on 9 December 2007 and lastly to Dettelbach Bahnhof on 1 September 2016.

Between Iphofen and Dettelbach the line lay in the tariff area of Kitzinger Nahverkehrs-Gemeinschaft (Kitzing local transport association, KiNG), until the district of Kitzingen joined the Verkehrsunternehmens-Verbund Mainfranken (Main-Franconia transport association, VVM) on 1 February 2009 after years of coordination problems.