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saddest bass line ever?

Orion by Metallica (just consider that Cliff Burton, the bassist here, would tragically die later that year aged only 24)

Atmosphere by Joy Division (also consider that Ian Curtis, the singer, committed suicide aged 23)

New Dawn Fades by Joy Division (very sombre)

Day of the Lords by Joy Division (Peter Hook sure does have a knack for writing some truly depressing basslines)

With or Without You by U2 (just...damn...)

Don't Forget Me by the RHCP (simple, sad and effective)

Otherside by the RHCP (written about their deceased former guitarist and friend. It's bound to be full of emotion)

Black by Pearl Jam (this is just about the most depressing song ever written. The bassline must be on this list)

High Hopes by Pink Floyd (this song just gets me every time)


That should just about cover it!
 
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It's difficult for me to say that a particular bass line is sad strictly on it's own, when the actual lyrics and other instrumentation of the song may be melancholy or bluesy as well.

To my ears, the upright bass is the easiest of basses (solo/on its own) to be able to convey the feeling that the OP is seeking.

To be able to fully master the "Growl" and "slides" and then not overuse them is pure musical heaven, as well as being able to get gradually louder and slower (and vice-versa) while interpreting an ascending or descending figures.

The closest that an electric bass can come to that would be a fretless, for me.

Getting back to the OP's question, to me, some of the standard bass lines in "Brown Ballad", Track 9 @ 19:21 help convey the feel of the song:

 
Both David Gilmour's fretless playing on Pink Floyd's "Hey You" (1979) and Pino Palladino's fretless playing on Don Henley's "New York Minute" (1989) are both standouts in my book. Of course, both songs are rather melancholy to begin with, but the amazing bass work really makes the tunes what they are.
 
There's been a couple of Hooky bass lines listed already...i'd go for
All day Long - New Order:


or,
Elegia - New Order


Something about that Chorus sound that instantly sounds melancholic IMO, Simon Gallup has that same vibe