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missioners who, on 27 Feb. 1559–60, signed the contract with the English commissioners at Berwick, and his son Alexander was one of the pledges for the performance of the treaty (Cal. State Papers, For. Ser. 1559–60, No. 787). He also signed the band of 27 April 1560 in ‘defence of the liberty of the evangel,’ and for the expulsion of the French from Scotland (Knox, ii. 63).

In February 1563 Ruthven, at the instance of Maitland of Lethington, was chosen a privy councillor of Mary Queen of Scots. Referring to his election, Randolph affirmed that the appointment ‘misliked Moray’ on account of his sorcery; that ‘an unworthier there is not in Scotland than he,’ and that more might be spoken than he dared write (Cal. State Papers, For. Ser. 1563, No. 370). In a later letter he also mentions that the queen ‘cannot abide him,’ and that ‘all men hate him’ (ib. No. 839). The explanation of these rumours regarding Ruthven is partly supplied by Knox, who states that the queen in conversation referred to the ‘offering of a ring to her by Lord Ruthven,’ and declared that, though at Maitland's instance he had been made one of her privy council, she ‘could not love’ him, for she knew him ‘to use enchantment’ (Knox, Works, ii. 373).

Ruthven, notwithstanding his admission to the privy council, continued to be a staunch defender of protestantism; and at a meeting of the council, before which Knox was brought in 1563, he defended Knox's right to ‘make convocation of the queen's lieges’ (ib. p. 406). On 22 Sept. of this year Ruthven was appointed to expel the clan Gregor out of the bounds of Strathearn (Reg. P. C. Scotl. i. 249); and on 8 May 1564 the queen conceded to him the office of sheriff-clerk of Perthshire. On 1 Dec. 1564 he received a grant of a waste house adjoining Holyrood House (Reg. Mag. Sig. Scot. 1546–80, No. 1567), which he presumably fitted up for a residence, and in which he may have been living at the time of the murder of Rizzio, a fact which would sufficiently explain his appearance there from a sick-bed, and also the first thought of Mary's attendants, that he had escaped from his chamber while raving in a fever. On the same date on which he received a grant of the waste house, Ruthven also obtained a grant to him and his second wife, Janet Stewart, widow of Lord Methven, of the lands and lordship of Methven, Perthshire (ib. No. 1568).

The first wife of Ruthven having been a Douglas, and his children by her being cousins-german of Lord Darnley, Ruthven was naturally a supporter of the Darnley marriage. Randolph represents him as the ‘chief councillor’ of those who were bent on the marriage (Cal. State Papers, For. Ser. 1564–5, No. 1140); and Knox states that at Mary's council at this time were only the Earls of Atholl and Lennox and Lord Ruthven (Works, ii. 483). It was Ruthven and Atholl who, with three hundred horsemen, escorted the queen safely from Perth through Fife to Callendar House, when a plot was suspected to have been formed by Moray for her capture on the journey south. During the rebellion of Moray, after the queen's marriage to Darnley, Ruthven also joined the forces of the queen with a command in the rearguard of the battle (Reg. P. C. Scotl. i. 379).

The rise of Rizzio in the favour of the queen, accompanied as it was by the declining influence of Darnley and of the relatives and friends who had been the main supporters of the marriage, was observed by Ruthven with feelings of deep resentment. As early as 12 Oct. 1565 Randolph wrote that Morton and Ruthven ‘only spy their time, and make fair weather until it come to the pinch’ (Cal. State Papers, For. Ser. 1564–5, No. 1580). It was probably at the suggestion of Morton or Ruthven that George Douglas inspired Darnley to apply to Ruthven to aid him against the ‘villain David.’ Ruthven, although then so ill that he ‘was scarcely able to walk twice the length of his chamber’ (Ruthven, Relation), agreed to assist him to the utmost of his power, and formally made known the proposal to Morton. It was Ruthven and Morton who agreed to undertake the management of the arrangements for seizing Rizzio. Their names are the only ones known to have been attached to the band signed by Darnley, and probably they were attached as witnesses. Ruthven, in complete armour and pale and haggard from his long sickness, was the first of the conspirators to enter into the queen's supper chamber after Darnley had taken his seat beside the queen (9 March 1565–6). The first conjecture of the queen and her attendants was that he was ‘raving through the vehemency of a fever.’ In a stern voice Ruthven commanded Rizzio to come out from the presence of the queen, ‘as it was no place for him;’ and as he was about to seize Rizzio, who clung to the garments of the queen, the other conspirators broke in and hurried Rizzio to the outer chamber. When Atholl, Huntly, Bothwell, and other nobles then in attendance on the queen in the palace, alarmed at the uproar, appeared to be meditating a rescue, Ruthven went down, and explaining to them that